SUSTAINABLE

HORTICULTURE
Volume 2:
Food, Health, and Nutrition
Innovations in Horticultural Science

SUSTAINABLE
HORTICULTURE
Volume 2:
Food, Health, and Nutrition

Edited by
Debashis Mandal, PhD
Amritesh C. Shukla, DPhil, DSc
Mohammed Wasim Siddiqui, PhD
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Library and Archives Canada Cataloguing in Publication

Sustainable horticulture / edited by Debashis Mandal, PhD, Amritesh C. Shukla,

DPhil, DSc, Mohammed Wasim Siddiqui, PhD.
(Innovations in horticultural science)
Includes bibliographical references and indexes.
Contents: Volume 2. Food, health, and nutrition.
Issued in print and electronic formats.
ISBN 978-1-77188-647-5 (v. 2 : hardcover).--ISBN 978-1-315-14799-4 (v. 2 : PDF)
1. Sustainable horticulture. I. Mandal, Debashis, editor II. Shukla, Amritesh
C., editor III. Siddiqui, Mohammed Wasim, editor IV. Series: Innovations in
horticultural science
SB319.95.S87 2018 635.028'6 C2018-903327-4 C2018-903328-2

CIP data on file with US Library of C

​ ​ongress

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CONTENTS

Innovations in Horticultural Science Book Series....................................... ix

Books in the Series....................................................................................... xi
About the Editors....................................................................................... xiii
List of Contributors................................................................................... xvii
List of Abbreviations.................................................................................. xxi
Preface.......................................................................................................xxv

PART I: POSTHARVEST MANAGEMENT AND

PROCESSED FOOD................................................................................ 1
1. Emerging Fruit Juice Processing Technologies:
Quality Improvement.................................................................................. 3
Chandran Somasundram, Zuliana Razali, and
Vicknesha Santhirasegaram
2. Development of Postharvest Processing Technology for
Ginger, Turmeric, and Chilli in Mizoram............................................... 13
Pradip K. Chatterjee, and George Srzednicki
3. Waste Products of Horticultural Crops: An Alternative
for Developing Entrepreneurship............................................................ 27
Amritesh C. Shukla and D. Mandal
4. Osmotic Dehydration of Mango Varieties Alphonso and
Totapuri...................................................................................................... 39
J. Lenka and R. B. Tiwari
5. Breadnut: Innovative Products for the Agro Food Sector..................... 55
S. Subramaniam, N. Ramburn, and R. Chacoory
6. Storage Study of Jamun-Aonla Blended Ready-to-Serve
Beverages........................................................................................................ 69
Preeti Singh, Neelima Garg, and Sanjay Kumar
vi Contents

7. Development of Blended Aonla Squash.................................................. 79

Neelima Garg, Sanjay Kumar, Preeti Singh, and Kaushlesh K. Yadav
8. Saccharomyces cerevisiae Postharvest Dip Treatment for
Improving Quality and Storability of Mango (Mangifera indica
cv. Dashehari)............................................................................................ 87
Bharati Killadi, Neelima Garg, Rekha Chaurasia,
Kaushlesh K. Yadav, and D. K. Shukla
9. Postharvest Quality Evaluation of Winter Annuals............................. 103
Sellam Perinban, Babita Singh, Jayoti Majumder, and Puja Rai
10. Optimizing the Shelf Life of Whole and Fresh Cut Breadfruit
in Mauritius............................................................................................. 117
Roop Soodha Munbodh
11. Standardization of Dehydration Techniques of Some
Ornamental Foliages............................................................................... 139
Moumita Malakar, Sukanta Biswas, and Pinaki Acharyya
12. Dependence on Non-Timber Forest Products from a Community
Forest as a Safety Net for Livelihood Security Among the
Villagers of Mamit District, Mizoram................................................... 151
K. Lalhmingsangi and U. K. Sahoo
13. Effects of Black Rot on the Antioxidant Properties of Morris
and Sarawak Pineapple (Ananas comosus)........................................... 175
Hasvinder Kaur Baldev Singh, Vicknesha Santhirasegaram,
Zuliana Razali, and Chandran Somasundram

PART II: PROTECTION OF HORTICULTURE CROPS..................... 187

14. Essential Oils as Green Pesticides for Plant Protection
in Horticulture......................................................................................... 189
Murray B. Isman and Rita Seffrin
15. Integrated Health Management in Horticultural Crops..................... 217
Ajanta Birah and C. Chattopadhyay
16. Technology for Diagnosis of Plant Viruses in Horticultural
Crops in India.......................................................................................... 229
Bikash Mandal
Contents vii

17. Acaricidal Activity of Petroleum Ether Extract from Seed

of Custard Apple, Annona squamosa L. (Annonaceae) Against
Red Spider Mite, Oligonychus coffeae (Nietner) Infesting Tea........... 235
Biplab Tudu and Sibdas Baskey
18. Evaluation of Some Plant Extracts Against Early Blight of
Tomato Under Net House Condition..................................................... 251
Manisha Dubey, T. S. Thind, S. K. Jindal, and R. K. Dubey
19. Bio-Efficacy of Eco-Friendly Pesticides on the Management
of Spodoptera litura (Fab.) on Cabbage................................................. 261
R. Mandi and A. Pramanik
20. Foliose Lichen Species: A Potential Source for Bio-Control Agent
Against Colletotrichum capsici............................................................... 279
M. Chinlampianga, A. C. Shukla, A. R. Logesh, and D. K. Upreti
21. Fungal Diseases of Medicinal and Aromatic Plants and Their
Biological Management.......................................................................... 291
Ramesh S. Yadav, D. Mandal, and Amritesh C. Shukla

PART III: HORTICULTURE FOR HEALTH AND NUTRITION...... 313

22. Broad Spectrum of Indian Peppermint Oil Against
Disease-Causing Human Bacterial Pathogens...................................... 315
Awadhesh Kumar, Amritesh C. Shukla, and Anupam Dikshit
23. Underexploited Vegetables in North Eastern India:
A Gateway to Food Security................................................................... 335
B. Lalramhlimi, Akoijam Ranjita Devi, and Khaidem Nirja
24. Essential Oil of Thymus saturejoides Coss. in the High Atlas
of Morocco: From Traditional Medicine to Community Natural
Product Development.............................................................................. 353
Bernadette Montanari and Amritesh C. Shukla
25. Potent Nutrimental and Ethnomedicinal Horticultural Flora
from North Central Terai Forests of U.P. India................................... 369
S. C. Tripathi and T. P. Mall
viii Sustainable Horticulture Volume 2: Food, Health, and Nutrition

26. Elemental Determination of Two Medicinal Plants of Mizoram

Using EDXRF.......................................................................................... 379
R. Lawmzuali, K. Birla Singh, and N. Mohondas Singh
Index.......................................................................................................... 387
INNOVATIONS IN
HORTICULTURAL SCIENCE

Editor-in-Chief:
Dr. Mohammed Wasim Siddiqui Assistant Professor-cum- Scientist
Bihar Agricultural University | www.bausabour.ac.in
Department of Food Science and Post-Harvest Technology
Sabour | Bhagalpur | Bihar | P. O. Box 813210 | INDIA
Contacts: (91) 9835502897
Email: [email protected] | [email protected]

The horticulture sector is considered as the most dynamic and sustainable seg-
ment of agriculture all over the world. It covers pre- and postharvest management
of a wide spectrum of crops, including fruits and nuts, vegetables (including pota-
toes), flowering and aromatic plants, tuber crops, mushrooms, spices, plantation
crops, edible bamboos etc. Shifting food pattern in wake of increasing income
and health awareness of the populace has transformed horticulture into a vibrant
commercial venture for the farming community all over the world.

It is a well-established fact that horticulture is one of the best options for improv-
ing the productivity of land, ensuring nutritional security for mankind and for
sustaining the livelihood of the farming community worldwide. The world’s pop-
ulace is projected to be 9 billion by the year 2030, and the largest increase will be
confined to the developing countries, where chronic food shortages and malnutri-
tion already persist. This projected increase of population will certainly reduce
the per capita availability of natural resources and may hinder the equilibrium and
sustainability of agricultural systems due to overexploitation of natural resources,
which will ultimately lead to more poverty, starvation, malnutrition, and higher
food prices. The judicious utilization of natural resources is thus needed and must
be addressed immediately.

Climate change is emerging as a major threat to the agriculture throughout the

world as well. Surface temperatures of the earth have risen significantly over the
past century, and the impact is most significant on agriculture. The rise in temper-
ature enhances the rate of respiration, reduces cropping periods, advances ripen-
ing, and hastens crop maturity, which adversely affects crop productivity. Several
climatic extremes such as droughts, floods, tropical cyclones, heavy precipitation
events, hot extremes, and heat waves cause a negative impact on agriculture and
are mainly caused and triggered by climate change.
x Innovations in Horticultural Science

In order to optimize the use of resources, hi-tech interventions like precision

farming, which comprises temporal and spatial management of resources in hor-
ticulture, is essentially required. Infusion of technology for an efficient utilization
of resources is intended for deriving higher crop productivity per unit of inputs.
This would be possible only through deployment of modern hi-tech applications
and precision farming methods. For improvement in crop production and returns
to farmers, these technologies have to be widely spread and adopted. Considering
the above-mentioned challenges of horticulturist and their expected role in ensur-
ing food and nutritional security to mankind, a compilation of hi-tech cultivation
techniques and postharvest management of horticultural crops is needed.

This book series, Innovations in Horticultural Science, is designed to address the

need for advance knowledge for horticulture researchers and students. Moreover,
the major advancements and developments in this subject area to be covered in
this series would be beneficial to mankind.
BOOKS IN THE SERIES

• Spices: Agrotechniques for Quality Produce

Amit Baran Sharangi, PhD, S. Datta, PhD, and Prahlad Deb, PhD

• Sustainable Horticulture, Volume 1: Diversity, Production, and

Crop Improvement
Editors: Debashis Mandal, PhD, Amritesh C. Shukla, PhD, and
Mohammed Wasim Siddiqui, PhD

• Sustainable Horticulture, Volume 2: Food, Health, and Nutrition

Editors: Debashis Mandal, PhD, Amritesh C. Shukla, PhD, and
Mohammed Wasim Siddiqui, PhD

• Underexploited Spice Crops: Present Status, Agrotechnology, and

Future Research Directions
Amit Baran Sharangi, PhD, Pemba H. Bhutia,
Akkabathula Chandini Raj, and Majjiga Sreenivas
ABOUT THE EDITORS

Debashis Mandal, PhD

Assistant Professor, Department of Horticulture, Aromatic and Medicinal
Plants, Mizoram University, Aizawl, India

Debashis Mandal, PhD, is an Assistant Professor in the Department of

Horticulture, Aromatic and Medicinal Plants at Mizoram University in
Aizawl, India. Dr. Mandal started his academic career as Assistant Profes-
sor at Sikkim University, India. He had postdoctoral research experience
as a project scientist at the Precision Farming Development Centre, Indian
Institute of Technology, Kharagpur, India. He has done research on sus-
tainable hill farming for the past seven years and has published 25 research
papers in reputed journals and three book chapters and two books. He is a
working member of the global working group on Lychee and Other Sap-
indaceae Crops, International Society for Horticultural Science (ISHS),
Belgium, and as Editor-in-Lead (Hort.) for the International Journal of
Bio-resource & Stress Management. In addition, he is a consultant horti-
culturist to the Department of Horticulture and Agriculture (Research and
Extension), Government of Mizoram, India, where he works on various
research projects. He has visited many countries for his work, including
Thailand, China, Nepal, Bhutan, Vietnam, South Africa, and South Korea.

Amritesh C. Shukla, DPhil, DSc

Professor, Department of Botany, University of Lucknow, India

Amritesh C. Shukla, DPhil, DSc, is currently a Professor in the Department

of Botany, University of Lucknow, India. He was formerly a Professor in
the Department of Horticulture, Aromatic and Medicinal Plants, Mizoram
University, Aizawl, India. He began his academic career as Associate
Professor at the same university. He has more than 20 years of experi-
ence on natural products and drug development and standardized various
scientific methods, viz., MSGIT, MDKT, MS-97, NCCLS (2002–2003).
xiv About the Editors

He has developed six commercial herbal formulations and holds four pat-
ents. He has published 75 research papers and 15 book chapters. He has
also authored six books and has handled many externally funded research
projects. He is a Fellow of five national and international scientific societ-
ies and also a Visiting Professor at the University of British Columbia,
Canada, and the University of Mauritius. In addition, Dr. Shukla is work-
ing as an Editor, Associate Editor, and editorial board member of many
internationally reputed journals, including the American Journal of Food
Technology, Journal of Pharmacology & Toxicology, Research Journal of
Medicinal Plants, etc. He has also been a visiting scientist at universities
in Australia, Germany, China, and USA.

Mohammed Wasim Siddiqui, PhD

Assistant Professor and Scientist in the Department of Food Science and
Post-Harvest Technology, Bihar Agricultural University, Sabour, India

Mohammed Wasim Siddiqui, PhD, is an Assistant Professor and Scientist

in the Department of Food Science and Post-Harvest Technology, Bihar
Agricultural University, Sabour, India, and author or co-author of 31
peer-reviewed research articles, 26 book chapters, two manuals, and 18
conference papers. He has 11 edited and an authored books to his credit,
published by Elsevier, CRC Press, Springer, and Apple Academic Press.
Dr. Siddiqui has established the international peer-reviewed Journal of
Postharvest Technology. He is Editor-in-Chief of two book series (Post-
harvest Biology and Technology and Innovations in Horticultural Sci-
ence), published by Apple Academic Press. Dr. Siddiqui is also a Senior
Acquisitions Editor in Apple Academic Press, New Jersey, USA, for Hor-
ticultural Science. He has been serving as an editorial board member and
active reviewer of several international journals.
Dr. Siddiqui has received several grants and respected awards for his
research work by a number of organizations. He is an active member of the
organizing committees of several national and international seminars, con-
ferences, and summits. He is one of key members in establishing theWorld
Food Preservation Center (WFPC), LLC, USA, and is currently an active
associate and supporter.
About the Editors xv

Dr. Siddiqui acquired a BSc (Agriculture) degree from Jawaharlal

Nehru Krishi Vishwa Vidyalaya, Jabalpur, India. He received his MSc
(Horticulture) and PhD(Horticulture) degrees from Bidhan Chandra Kri-
shi Viswavidyalaya, Mohanpur, Nadia, India, with specialization in Post-
harvest Technology. He is a member of Core Research Group at the Bihar
Agricultural University (BAU), which provides appropriate direction and
assistance in prioritizing research.
LIST OF CONTRIBUTORS

Pinaki Acharyya
Department of Horticulture, University of Calcutta, 51/2, Ballygunge Circular Road,
Kolkata–700019, India

Sibdas Baskey
Regional Research Station (Hill Zone), North Bengal Agriculture University, Kalimpong,
Darjeeling, West Bengal – 734301, India

Ajanta Birah
ICAR-National Research Centre for Integrated Pest Management, New Delhi–110012, India

Sukanta Biswas
Department of Horticulture, University of Calcutta, 51/2, Ballygunge Circular Road,
Kolkata–700019, India

R. Chacoory
Crop Department, Food and Agricultural Research and Extension Institute, Reduit, Mauritius

Pradip K. Chatterjee
Thermal Engineering Division, CSIR Central Mechanical Engineering Research Institute,
Ministry of Science and Technology, Government of India, M. G. Avenue, Durgapur – 713209, India
C. Chattopadhyay
Uttar Banga Krishi Viswavidyalaya, Pundibari, Coochbehar–736165, West Bengal, India

Rekha Chaurasia
Division of Post Harvest Management, ICAR–CISH Lucknow 226106, India
M. Chinlampianga
Department of Horticulture, Aromatic and Medicinal Plants, Mizoram University,
Aizwal – 796004, India

Akoijam Ranjita Devi

Department of Spices and Plantation Crops, Faculty of Horticulture, Bidhan Chandra Krishi
Viswavidyalaya, Mohanpur–741252, Nadia, West Bengal, India

Anupam Dikshit
Biological Product Lab, Department of Botany, University of Allahabad, Allahabad–211004, India

Manisha Dubey
Department of Plant Pathology, Punjab Agricultural University, Ludhiana–141004, India

R. K. Dubey
Department of Floriculture and Landscape, Punjab Agricultural University, Ludhiana–141004, India

Neelima Garg
Central Institute for Subtropical Horticulture, Rehmankhera, P.O. Kakori, Lucknow–226101, India
xviii List of Contributors

Murray B. Isman
Faculty of Land and Food Systems, University of British Columbia, Vancouver V6T1Z4, Canada

S. K. Jindal
Department of Vegetable Science, Punjab Agricultural University, Ludhiana–141004, India
Bharati Killadi
Division of Post Harvest Management, ICAR–CISH Lucknow 226106, India

Awadhesh Kumar
Department of Horticulture, Aromatic and Medicinal Plants, Mizoram University, Aizawl–796004,
India

Sanjay Kumar
Central Institute for Subtropical Horticulture, Rehmankhera, P.O. Kakori, Lucknow–226101, India

K. Lalhmingsangi
Department of Forestry, School of Earth Sciences and Natural Resource Management,
Mizoram University, Aizawl–796004, India
B. Lalramhlimi
Department of Vegetable Crops, Faculty of Agriculture, Bidhan Chandra Krishi Viswavidyalaya,
Mohanpur–741252, Nadia, West Bengal, India

R. Lawmzuali
Department of Chemistry, School of Physical Sciences, Mizoram University, Aizawl–796004, India

J. Lenka
Division of Post Harvest Management, ICAR–CISH, Rehmankhera, P.O. Kakori,
Lucknow–226 101, India
A. R. Logesh
Department of Horticulture, Aromatic and Medicinal Plants, Mizoram University, Aizwal – 796004,
India

Jayoti Majumder
Bidhan Chandra Krishi Viswavidyalaya (BCKV), Kalyani, West Bengal, India

Moumita Malakar
Department of Horticulture, University of Calcutta, 51/2, Ballygunge Circular Road,
Kolkata–700019, India
T. P. Mall
Postgraduate Department of Botany, Kisan PG College, Bahraich, U.P., India

Bikash Mandal
Advanced Centre of Plant Virology, Division of Plant Pathology, Indian Agricultural Research
Institute, New Delhi–110012, India
D. Mandal
Department of Horticulture, Aromatic & Medicinal Plants, Mizoram University,
Aizawl-796001, India.
R. Mandi
Department of Agricultural Entomology, Bidhan Chandra Krishi Viswavidyalaya, Mohanpur,
Nadia, 741252, West Bengal, India
List of Contributors xix

Bernadette Montanari
Department of Geography, University of Urbana Champaign, USA [Social Dimensions of
Environmental Policy (SDEP)]

Roop Soodha Munbodh

Food and Agricultural Research and Extension Institute (FAREI), Reduit, Mauritius
Khaidem Nirja
Department of Agricultural Extension, Faculty of Agriculture,
Bidhan Chandra Krishi Viswavidyalaya, Mohanpur–741252, Nadia, West Bengal, India

Sellam Perinban
Directorate of Floricultural Research, Indian Agricultural Research Institute, Pusa Campus,
New Delhi–110012, India

A. Pramanik
Department of Agricultural Entomology, Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, Nadia,
741252, West Bengal, India

Puja Rai
Bidhan Chandra Krishi Viswavidyalaya (BCKV), Kalyani, West Bengal, India

N. Ramburn
Crop Department, Food and Agricultural Research and Extension Institute, Reduit, Mauritius

Zuliana Razali
Institute of Biological Sciences, Faculty of Science and the Centre for Research in Biotechnology for
Agriculture (CEBAR), University of Malaya, 50603, Kuala Lumpur, Malaysia
U. K. Sahoo
Department of Forestry, School of Earth Sciences and Natural Resource Management,
Mizoram University, Aizawl–796004, India

Vicknesha Santhirasegaram
Institute of Biological Sciences, Faculty of Science and the Centre for Research in Biotechnology for
Agriculture (CEBAR), University of Malaya, 50603, Kuala Lumpur, Malaysia

Rita Seffrin
Faculty of Land and Food Systems, University of British Columbia, Vancouver V6T1Z4, Canada

Amritesh C. Shukla
Department of Botany, University of Lucknow, Lucknow–226007, India

D. K. Shukla
Division of Post Harvest Management, ICAR–CISH Lucknow 226106, India

Babita Singh
Directorate of Floricultural Research, Indian Agricultural Research Institute, Pusa Campus,
New Delhi–110012, India

N. Mohondas Singh
Department of Chemistry, School of Physical Sciences, Mizoram University, Aizawl–796004, India

Preeti Singh
Central Institute for Subtropical Horticulture, Rehmankhera, P.O. Kakori, Lucknow–226101, India
xx List of Contributors

Hasvinder Kaur Baldev Singh

Institute of Biological Sciences, Faculty of Science and the Centre for Research in Biotechnology for
Agriculture (CEBAR), University of Malaya, 50603, Kuala Lumpur, Malaysia

K. Birla Singh
Department of Zoology, Pachhunga University College, Aizawl–796001, India
Chandran Somasundram
Institute of Biological Sciences and Centre for Research in Biotechnology for Agriculture (CEBAR),
Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia

George Srzednicki
School of Chemical Engineering, Food Science and Technology, University of New South Wales,
Sydney 2052, Australia

S. Subramaniam
Crop Department, Food and Agricultural Research and Extension Institute, Reduit, Mauritius

T. S. Thind
Department of Plant Pathology, Punjab Agricultural University, Ludhiana–141004, India

R. B. Tiwari
Division Post Harvest Technology, IIHR, Bangalore–560089, India

S. C. Tripathi
Postgraduate Department of Botany, Kisan PG College, Bahraich, U.P., India

Biplab Tudu
Regional Research Station (Hill Zone), North Bengal Agriculture University, Kalimpong,
Darjeeling, West Bengal – 734301, India
D. K. Upreti
Lichenology Laboratory, Plant Biodiversity, Systematics and Herbarium Division,
CSIR-National Botanical Research Institute, Lucknow – 226001, Uttar Pradesh, India

Kaushlesh K. Yadav
Central Institute for Subtropical Horticulture, Rehmankhera, P.O. Kakori, Lucknow–227107, India

Ramesh S. Yadav
Department of Plant Pathology, Sardar Vallabhbhai Patel University of Agriculture & Technology,
Meerut–250110, India
LIST OF ABBREVIATIONS

AAE ascorbic acid equivalent

ACPV Advanced Center of Plant Virology
ANS average number of spores
AOA aminooxy acetic acid
APSA All Purpose Suspension Agent
ASA acetyl salicylic acid
BA benzyl adenine
CASE custard apple seed
CBCS Central Biological Control Stations
CD critical difference
CE catechin equivalent
CMV cucumber mosaic virus
CP coat protein
CPPS Central Plant Protection Stations
CRBD completely randomized block design
CSS Central Surveillance Stations
DAT days after treatment
DMRT Duncan’s multiple range test
DMSO dimethyl sulfoxide solvent
DPPH diphenyl-1-picrylhydrazyl
EIL economic injury level
ELISA enzyme-linked immunosorbent assay
EOs essential oils
EPA entry point activities
ETL economic threshold
FAO Food and Agriculture Organization
FFSs Farmers’ Field Schools
FGI fungal growth inhibition
GAE gallic acid equivalent
GAP good agricultural practices
GBNV groundnut bud necrosis virus
xxii List of Abbreviations

GC gas chromatography
GoI Government of India
HACCP hazard analysis critical control point
HPLC high performance liquid chromatography
HSD honestly significant difference
HTST high temperature short time
IA intensive agriculture
IARI Indian Agricultural Research Institute
ICAR Indian Council of Medical Research
ICT Information Communication Technology
IDS International Drying Symposium
IDSS integrated decision support system
IFW initial fresh weight
IgG immunoglobulin G
IPM integrated pest management
ISR induced systemic resistance
IVM initial volume of medium
IVPD in vitro multienzyme protein digestibility
JFM Joint Forest Management
K potassium
LFA lateral flow assay
LPS lipopolysaccharides
MAb monoclonal antibody
MAPs medicinal and aromatic plants
MARDI Malaysian Agricultural Research and Development
Institute
MBC minimum bactericidal concentration
MCCs minimum cidal concentration
MEC minimum effective concentration
MIC minimum inhibitory concentration
MKT minimum killing time
MPL maximum permissible limit
MPTs multipurpose tree species
MR moisture ratio
MRLs maximum residue limit
MS mass spectrometry
List of Abbreviations xxiii

MSGIT modified spore germination inhibition technique

MT metric tons
NBRI National Botanical Research Institute
NCIPM National Research Centre for Integrated Pest
Management
NE north-east
NGO Non Governmental Organization
NPV nucleopolyhedrovirus
NSKE neem seed kernel extract
NTFPs non timber forest products
OPD out patient department
PAb polyclonal antibody
PCR polymerase chain reaction
PDI percentage disease index
PEF pulsed electric field
PR pathogenesis related
PRA participatory rural appraisal
PRSV papaya ringspot virus
PT persistent toxicity
RBD randomized block design
RDA recommended dietary allowance
RFW relative fresh weight
RSM red spider mite
scF single chain variable fragment
SD standard deviation
SDA sabourad dextrose agar medium
SEM standard error of mean
SG solid gain
SHGs self help groups
TA titratable acidity
TLC thin layer chromatography
TSS total soluble solids
UV-C ultraviolet-c
VFDC Village Forest Development Committee
VMF volume of microscopic field
VOCs volatile organic compounds
xxiv List of Abbreviations

VSUR vase solution uptake rate

WHO World Health Organization
WL water loss
WR weight reduction
YMA Young Mizo Association
YPDA yeast potato dextrose agar
PREFACE

Global food demand is expected to be doubled by 2050, while the pro-

duction environment and natural resources are continuously shrinking and
deteriorating. Horticulture, a major sector of agriculture, needs to take
part in enhancing crop production and productivity in parity with agri-
cultural crops to meet the emerging food demand. There are projections
that demand for food grains would increase from 192 million tonnes in
2000 to 345 million tonnes in 2030. Hence, in the next 15 years, produc-
tion of food grains needs to be increased at the rate of 5.5 million tonnes
annually. The demand for high-value commodities (such as horticulture,
dairy, livestock and fish) is increasing faster than food grains—for most of
the high-value food commodities demand is expected to increase by more
than 100% from 2000 to 2030. These commodities are all perishable and
require, different infrastructure for handling, value-addition, processing
and marketing.
Asia, the major crop-producing continent, thought to be the global super
power because of its increasing skilled and energetic human resources and
faster developing technological and economic growth, has to play a major
role to meet the projected global food demand. However, the developing
Asiatic countries are more or less facing the common problem of diminu-
tion of cultivable land with massive rapid urbanization. Apart from that,
the contemporary production limitations—depleting land fertility, unequal
cross-subsidy and, more predominantly, vagaries of climate change—put
forth a tough task to perform.
India, the second most important Asiatic food grain producer, is fac-
ing a more grim production situation. The average size of land holding
declined to 1.32 ha in 2000–01, from 2.30 ha in 1970–71, and absolute
number of operational holdings increased from about 70 million to 121
million. If this trend continues, the average size of holding in India would
be mere 0.68 ha in 2020, and would be further reduced to a low of 0.32 ha
in 2030. This is a very complex and serious problem, when the share of
agriculture, including horticulture, in gross domestic product is declining,
xxvi Preface

average size of landholding is contracting and fragmenting, and numbers

of operational holdings are increasing. In addition, annually, India is los-
ing nearly 0.8 million tonnes of nitrogen, 1.8 million tonnes of phosphorus
and 26.3 million tonnes of potassium—the deteriorating quality and health
of soil is something to be checked. Problems are further aggravated by
imbalanced application of nutrients (especially nitrogen, phosphorus and
potash) and excessive mining of micronutrients, leading to deficiency of
macro- and micro-nutrients in the soils. Similarly, the water-table is lower-
ing steeply in most of the irrigated areas, and water quality is also deterio-
rating, due to leaching of salts and other pollutants.
Amidst this situation, fulfilling the target with a sustainable model of
crop production is really an enormous endeavor. There is an earnest need
to develop promising technologies and management options to raise pro-
ductivity to meet the growing food demand in this situation of deteriorat-
ing production environment at the lowest cost; and to develop appropriate
technologies, to create required infrastructures, and to evolve institutional
arrangements for production, postharvest and marketing of high-value and
perishable commodities and their value-added products. Improved agro-
techniques, quality planting materials, improved varieties, climate resilient
production models, involvement of information technology and biotech-
nology, improved postharvest handling-storage, and marketing are the key
issues on which to focus on to bring about the desired metamorphosis
in global horticulture. However, to achieve the goal with a sustainable
production environment (i.e., sustainability in terms of economy-ecology-
society), is the greatest challenge to meet.
The International Symposium on Sustainable Horticulture, orga-
nized by the Department of Horticulture, Aromatic and Medicinal Plants,
Mizoram University, Aizawl, India, from 14–16th March, 2016, provided
a platform for the exchange of ideas and research experience and dis-
cussed several facets of sustainable horticulture with the thematic areas
such as management of genetic resources and biodiversity conservation;
production technology of horticultural crops; crop improvement and bio-
technology; plant protection in horticultural crops; postharvest technology
and value addition; trade, marketing, entrepreneurship development and
extension; and horticulture for food, health and nutrition.
Preface xxvii

In this regard, this research compendium has been categorically divided

to form two volumes; Sustainable Horticulture, Volume 1: Diversity, Pro-
duction, and Crop Improvement, and Sustainable Horticulture, Volume 2:
Food, Health, and Nutrition. The first volume outlines the contemporary
trends in sustainable horticulture research, in particular: crop diversity,
species variability and conservation strategies, production technology, tree
architecture management, plant propagation, and nutrition management,
organic farming, new dynamics in breeding, and marketing of horticul-
ture crops. The second volume depicts the research trends in sustainable
horticulture comprising postharvest management and processed food pro-
duction from horticulture crops, crop protection and plant health manage-
ment, and horticulture for human health and nutrition.
We extend our sincere thanks to the contributors, reviewers, and Apple
Academic Press for their efforts and contributions. It is hoped that these
book volumes will be useful for students, teachers, scientists, extension
workers, and researchers in horticulture and allied disciplines.
—Debashis Mandal
Amritesh C. Shukla
Wasim Siddiqui
 PART I

POSTHARVEST MANAGEMENT
AND PROCESSED FOOD
CHAPTER 1

EMERGING FRUIT JUICE

PROCESSING TECHNOLOGIES:
QUALITY IMPROVEMENT
CHANDRAN SOMASUNDRAM, ZULIANA RAZALI,
and VICKNESHA SANTHIRASEGARAM
Institute of Biological Sciences and Centre for Research in
Biotechnology for Agriculture (CEBAR), Faculty of Science,
University of Malaya, 50603 Kuala Lumpur, Malaysia,
Tel: +60379674423; Fax: +60379674178,
E-mail: [email protected]

CONTENTS

Abstract...................................................................................................... 3
1.1 Introduction....................................................................................... 4
1.2 Fruit Juice Processing....................................................................... 5
1.3 Conclusion........................................................................................ 9
Keywords................................................................................................... 9
References................................................................................................ 10

ABSTRACT

Fruit juice has the highest acceptability among other beverages, generally
due to its natural taste as well as its nutritional value. The presence of vari-
ous phytochemicals in fruit juice is related to various health-promoting
properties such as protection against several chronic human diseases such
4 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

as cancer, cardiovascular diseases, and diabetes. However, the number of

outbreaks and cases of illness caused by consumption of contaminated
juices, especially unpasteurized juices, has increased over the last decade.
Currently, conventional thermal treatment is the preferred technology to
inactivate microorganisms and enzymes causing spoilage, thus prolonging
the shelf-life of juice. Because of the relatively high temperatures generally
needed to inactivate food poisoning- and spoilage-causing microorgan-
isms, thermal treatment can adversely affect the quality of food products,
by reducing their nutritional value and altering sensory attributes such
as color and flavor. The growing interest for fresh-like products has pro-
moted the effort for the development of innovative nonthermal food pres-
ervation methods. Nonthermal processing techniques have been explored
for their efficacy in extending shelf-life and enhancing the safety of fresh
juice while preserving organoleptic and nutritional qualities. As consum-
ers continue to seek food products with improved nutritional value and
functionality, juice producers have the opportunity of improving product
marketability through the use of these novel technologies that provide bet-
ter retention of phytonutrients. Therefore, there is a need for nonthermal
processing techniques to be tested on a pilot scale, so that these methods
can be developed as an alternative to thermal pasteurization.

1.1 INTRODUCTION

According to Business Insights (2010), the global market for juices val-
ued about US$ 93 billion in 2014. The key driver for the growth of fruit
juice market is the increase in awareness among consumers on preventive
healthcare and wellness benefits. Natural fruit juices are susceptible to
spoilage, mainly due to their intrinsic properties such as pH, water activity,
redox potential, and nutrients (Odumeru, 2012).
Fruit juice deterioration is mostly caused by enzymatic, chemical, and
microbial reactions. Enzymes in fruit juices such as polyphenol oxidase
and peroxidase may react with oxygen, thus contributing to juice browning
and off-flavor (Bharate and Bharate, 2014). The causal agents of microbial
spoilage of fruit juices are bacteria, yeast, and molds. Yeast and molds are
the main spoilage agents due to the low pH of fruit juices. According to the
Emerging Fruit Juice Processing Technologies: Quality Improvement 5

Centre for Disease Control and Prevention (1996), one of the current food-
borne disease outbreaks has been linked to pathogens such as Escherichia
coli O157:H7, where the emphasis was on unpasteurized juices.
Spoilage of fruit juice and related products as a result of microbial
growth may contribute to physical and chemical changes in food products.
These alterations include unacceptable flavor and odor, changes in color
and turbidity, gas production, and formation of slime. Usually, growth of
microorganisms to high numbers is necessary before spoilage becomes
noticeable. Hence, it is important to control the growth of spoilage organ-
isms in order to inhibit microbial spoilage (Odumeru, 2012).
The increase in outbreaks and cases of illness related to consumption of
unpasteurized juices have urged the development of a more effective food
safety control program, known as the hazard analysis and critical control
point (HACCP) program. HACCP is a systematic approach to identify,
assess, and control microbiological, chemical, and physical hazards of
public health concern (Odumeru, 2012). Currently, there are a number of
fruit juice preservation technologies for controlling microbial growth and
survival. These preservation methods must be evaluated to avoid signifi-
cant organoleptic changes in food products (Bates et al., 2001).
This paper will provide an overview of emerging thermal and non-
thermal processing methods. This information is necessary to improve the
progress of positive implementation of novel processing methods in the
fruit juice industry.

1.2 FRUIT JUICE PROCESSING

The main objective of fruit juice processing is to prevent microbiologi-

cal spoilage while assuring safety and maintaining quality characteristics.
Fruit juice processing technologies can be divided into two groups, namely
thermal and nonthermal processing.

1.2.1 THERMAL PROCESSING

Conventional thermal treatment is the preferred technology to inactivate

microorganisms and enzymes causing spoilage, thus prolonging the shelf-
6 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

life of juice. Traditional thermal processing depends on the generation of

heat outside the product to be heated and its transfer into the product via
convection and conduction mechanisms (Pereira and Vicente, 2010). Pas-
teurization is an example of thermal treatment that is commonly practiced
in the food industry. The most common pasteurization method for fruit
juice is high temperature short time (HTST) or also known as flash pas-
teurization (David et al., 1996).
According to Nagy et al. (1993), HTST treatment for fruit juices range
from 90°C to 95°C for 15 to 60 seconds to assure at least 5-log reduction
in microbial count. The time and temperature variables for pasteurization
of juice depend on the type of juice, initial microbial count, pH, water
activity, and thermal inactivation kinetics of microorganisms present in
juice. Hence, pasteurization conditions should be selected appropriately
to avoid overprocessing. However, underprocessing may not completely
inactivate microbial growth, thus resulting in juice spoilage (Rawson et
al., 2011).
Because of the relatively high temperatures generally needed to inacti-
vate food poisoning- and spoilage-causing microorganisms, conventional
pasteurization can adversely affect the quality of food products by reduc-
ing their nutritional value or altering sensory attributes such as color and
flavor (Rawson et al., 2011). Some studies on thermally treated fruit juices
such as orange (Cortes et al., 2008) and strawberry (Aguilo-Aguayo et
al., 2009) reported significant loss of quality and degradation of bioac-
tive compounds such as ascorbic acid. In addition, Rattanathanalerk et al.
(2005) reported significant color degradation in thermal-treated pineapple
juice (at 85°C and 95°C for 60 seconds).

1.2.2 NON-THERMAL PROCESSING

The growing interest for fresh-like products has promoted the effort for
developing innovative nonthermal food preservation methods. Nonther-
mal processing techniques have been explored for their efficacy to extend
shelf-life and enhance the safety of fresh juice while preserving organo-
leptic and nutritional qualities. In addition, these preservation methods
Emerging Fruit Juice Processing Technologies: Quality Improvement 7

are considered to be more energy efficient and provide better retention of

quality when compared to conventional thermal processing. Some of the
nonthermal processing methods extensively studied for juice preservation
include ultrasound, ultraviolet light irradiation, and pulsed electric field
(Morris et al., 2007).

1.2.2.1 Ultrasound

Power ultrasound (10–1000 W/cm2) is used to alter food properties, either

physically or chemically, such as by disrupting cells and inactivating
enzymes. Ultrasonic processing equipment includes ultrasonic bath and
probe system (Carcel et al., 2012). The mechanism of action for ultrasonic
processing or sonication is explained in three different approaches, which
include cavitation, localized heating, and formation of free radicals. When
high power ultrasound at low frequencies (20–100 kHz) propagates in a liq-
uid, cavitation (formation and collapse of bubbles) occurs. As a result, there
is elevation of localized pressure (up to 500 MPa) and temperature (up to
5000°C). These “tiny hotspots” provide the energy to alter the properties of
food product either physically or chemically. Accordingly, these cavitation
bubbles induce microstreaming and shear stress, resulting in the disinte-
gration of the microbial cells. Besides that, cavitation causes intracellular
micromechanical shock that disrupts the functional components of the cell,
thus inactivating enzymes (O’Donnell et al., 2010; Abid et al., 2013).
Sonication is a potential technology to achieve the US FDA condition
of a 5-log reduction of foodborne pathogens in fruit juices. Several stud-
ies using ultrasonic processing on fruit juice reported minimal effect on
the degradation of quality parameters and improved functionalities, such
as in kasturi lime (Bhat et al., 2011a), apple (Abid et al., 2013), and car-
rot juice (Jabbar et al., 2014). Besides that, Rawson et al. (2011) reported
that sonication provides better retention of bioactive compounds. In addi-
tion, Tiwari et al. (2009) reported that ultrasonic processing (25 kHz for 2
min) improves the cloud value and stability of orange juice during storage.
However, significant color degradation was observed in orange juice sub-
jected to sonication (Tiwari et al., 2009).
8 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

1.2.2.2 Ultraviolet-C (UV-C) Light

Ultraviolet-C (UV-C) light is a part of the electromagnetic spectrum with

wavelengths between 200 to 280 nm and exhibits germicidal properties as
it inactivates bacterial and viral microorganisms. Although UV-C radia-
tion technology is considered as an effective method for food preserva-
tion, consumers’ misconception about this process have delayed many of
its potential applications in the food industry. Actually, UV-C radiation is
a physical treatment that does not result in chemical residues. Hence, the
consumption of UV-C-treated food products is not harmful to humans.
One of the limitations of the application of UV in juice processing is
associated with the high absorbance coefficients of juice. According to
Koutchma et al. (2004), UV-C penetration largely depends on the presence
of dissolved organic solutes (suspended solids) and colored compounds
that act as a barrier, thus exhibiting UV-C attenuation effects. Hence, an
appropriate UV reactor should be designed to reduce the interference of
UV absorbance and improve microbial inactivation efficiency. The reac-
tor design should include a narrow laminar flow or conditions with high
turbulence, where juices are mixed resulting in all parts of the juice being
exposed to the UV light source (Koutchma et al., 2004).
The US FDA criterion of a 5-log reduction of the chosen pathogen in fruit
juices can be achieved by UV-C radiation. Several studies using short-wave
UV-C light treatment on fruit juices reported better retention of nutritional
and quality attributes, such as in starfruit (Bhat et al., 2011b) and orange
juice (Pala and Toklucu, 2013). Besides that, Bhat et al. (2011b) reported
that UV-C processing (for 30 and 60 minutes) induces a significant increase
in polyphenol and flavonoid content of starfruit juice. However, there is an
increasing trend in browning degree and color changes corresponding to
increased UV-C treatment time, as previously reported by Bhat et al. (2011b).

1.2.2.3 Pulsed Electric Field (PEF)

Pulsed electric field (PEF) treatment applies short pulses (1 to 100 microsec-
onds) with high voltage (10 to 50 kV/cm) to liquid products in a continuous
system. A simple PEF system consists of a high voltage power supply, a pulse
generator, treatment chamber, and a switch to discharge energy to electrodes.
Emerging Fruit Juice Processing Technologies: Quality Improvement 9

In addition, there is a cooling system to balance moderate temperature rise

during treatment (Morris et al., 2007; Lopez-Gomez et al., 2009). The effec-
tiveness of PEF processing is dependent on variables such as pulse width,
electric field strength, flow rate, treatment temperature, and time of exposure.
Some studies reported that juices treated with lower intensity pulse
fields and shorter pulse width exhibited higher retention of ascorbic acid,
such as observed in orange (Elez-Martinez and Martin-Belloso, 2007). In
addition, an enhancement of antioxidant capacity was observed in PEF-
processed juices due to increased extraction yield of secondary metabo-
lites and generation of free radicals (Rawson et al., 2011).

1.3 CONCLUSION

The value for fruit juices has been increasing in the global market due to
their health benefits. Conventional thermal pasteurization is the preferred
technology used to achieve microbial inactivation and extend the shelf-
life of juices. Lately however, consumers’ demand for a new preserva-
tion technology that retains freshness and at the same time ensures food
safety has resulted in growing interest for nonthermal processing methods.
Therefore, there is a need for other nonthermal processing techniques to
be tested on a pilot scale, so that these methods can be developed as an
alternative to thermal pasteurization. In future, the combination of non-
thermal processing methods as a hurdle concept would be a new trend of
preservation of fruit juices that improves the microbiological quality and
safety with minimal impact on the quality of the food product.

KEYWORDS

•• fruit juice
•• non-thermal processing
•• quality
•• safety
•• thermal processing
10 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

REFERENCES

Abid, M., Jabbar, S., Wu, T., Hashim, M. M., Hu, B., Lei, S., Zhang, X., & Zeng, X.,
(2013). Effect of ultrasound on different quality parameters of apple juice. Ultrason-
ics Sonochemistry, 20, 1182–1187.
Aguilo-Aguayo, I., Oms-Oliu, G., Soliva-Fortuny, R., & Martin-Belloso, O., (2009).
Changes in quality attributes throughout storage of strawberry juice processed by
high-intensity pulsed electric fields or heat treatments. LWT- Food Science and Tech-
nology, 42, 813–818.
Bates, R. P., Morris, J. R., & Crandall, P. G., (2001). Principles and practices of small
and medium scale fruit juice processing: Food and Agriculture Organization (FAO),
Agricultural Services Bulletin Issue, 146, FAO, Rome.
Bharate, S. S., & Bharate, S. B., (2014). Non-enzymatic browning in citrus juice: Chemical
markers, their detection and ways to improve product quality. Journal of Food Sci-
ence and Technology, 51(10):2271-2288. DOI: 10.1007/s13197–012–0718–8.
Business Insights (2010). Innovations in fruit and vegetable juices: Emerging opportunities
in premiumization, sustainability and positive health. Available online: <http://www.
scribd. com/doc/50026681/Innovations-in-Fruit-and-Vegetable-Juices>.
Carcel, J. A., Garcia-Perez, J. V., Benedito, J., & Mulet, A., (2012). Food process innova-
tion through new technologies: Use of ultrasound. Journal of Food Engineering, 110,
200–207.
Centers for Disease Control and Prevention (CDC), (1996). Outbreak of Escherichia coli
O157:H7 infections associated with drinking unpasteurised commercial apple juice:
British Columbia, California, Colorado, and Washington. Morbidity and Mortality
Weekly Report, 45, 975.
David, J. R. D., (1996). Principles of thermal processing and optimization. In: David, J.
R. D., Graves, R. H., & Carlson, V. R., (eds.), Aseptic Processing and Packaging of
Food: A Food Industry Perspective. CRC Press LLC. Florida, pp. 14.
Elez-Martinez, P., & Martin-Belloso, O., (2007). Effects of high intensity pulsed electric
field processing conditions on vitamin C and antioxidant capacity of orange juice and
gazpacho, a cold vegetable soup. Food Chemistry, 102, 201–209.
Jabbar, S., Abid, M., Hu, B., Wu, T., Hashim, M. M., Lei, S., Zhu, X., & Zeng, X., (2014).
Quality of carrot juice as influenced by blanching and sonication treatments. LWT-
Food Science and Technology, 55(1), 16–21.
Koutchma, T., Keller, S., Chirtel, S., & Parisi, B., (2004). Ultraviolet disinfection of juice
products in laminar and turbulent flow reactors. Innovative Food Science and Emerg-
ing Technologies, 5, 179–189.
Lopez-Gomez, A., Fernandez, P. S., Palop, A., Periago, P. M., Martinez-Lopez, A., Marin-
Iniesta, F., & Barbosa-Canovas, G. V., (2009). Food safety engineering: An emergent
perspective. Food Engineering Reviews, 1, 84–104.
Morris, C., Brody, A. L., & Wicker, L., (2007). Non-thermal food processing/preservation
technologies: A review with packaging implications. Packaging Technology and Sci-
ence, 20, 275–286.
Nagy, S., Chen, C. S., & Shaw, P. E., (1993). Fruit Juice Processing Technology. Ag. Sci-
ence, Florida.
Emerging Fruit Juice Processing Technologies: Quality Improvement 11

O’Donnell, C. P., Tiwari, B. K., Bourke, P., & Cullen, P. J., (2010). Effect of ultrasonic
processing on food enzymes of industrial importance. Trends in Food Science and
Technology, 21, 358–367.
Odumeru, J. A., (2012). Microbial safety of food and food products. In: Simpson, B. K.,
Nollet, L. M. L., Toldra, F., Benjakul, S., Paliyath, G., & Hui, Y. H., (eds.), Food Bio-
chemistry and Food Processing, 2nd edition. John Wiley & Sons, UK, pp. 787–797.
Pala, C. U., & Toklucu, A. K., (2013). Microbial, physicochemical and sensory properties
of UV-C processed orange juice and its microbial stability during refrigerated stor-
age. LWT - Food Science and Technology, 50, 426–431.
Pereira, R. N., & Vicente, A. A., (2010). Environmental impact of novel thermal and non-
thermal technologies in food processing. Food Research International, 43, 1936–
1943.
Rattanathanalerk, M., Chiewchan, N., & Srichumpoung, W., (2005). Effect of thermal
processing on the quality loss of pineapple juice. Journal of Food Engineering, 66,
259–265.
Rawson, A., Patras, A., Tiwari, B. K., Noci, F., Koutchma, T., & Brunton, N., (2011).
Effect of thermal and non-thermal processing technologies on the bioactive content
of exotic fruits and their products: Review of recent advances. Food Research Inter-
national, 44, 1875–1887.
Tiwari, B. K. O.,’Donnell, C. P., Muthukumarappan, K., & Cullen, P. J., (2009). Ascorbic
acid degradation kinetics of sonicated orange juice during storage and comparison
with thermally pasteurised juice. LWT-Food Science and Technology, 42, 700–704.
CHAPTER 2

DEVELOPMENT OF POSTHARVEST
PROCESSING TECHNOLOGY FOR
GINGER, TURMERIC, AND CHILLI IN
MIZORAM
PRADIP K. CHATTERJEE1 and GEORGE SRZEDNICKI2
1
Thermal Engineering Division, CSIR Central Mechanical
Engineering Research Institute, Ministry of Science and Technology,
Government of India, M. G. Avenue, Durgapur – 713209, India
2
School of Chemical Engineering, Food Science and Technology,
University of New South Wales, Sydney 2052, Australia,
E-mail: [email protected]

CONTENTS

Abstract.................................................................................................... 14
2.1 Introduction..................................................................................... 14
2.2 Equipment Development................................................................ 16
2.3 Studies Conducted in the Experimental Dryer................................ 21
2.4 Project Achievements...................................................................... 23
2.5 Conclusions..................................................................................... 24
Keywords................................................................................................. 25
References................................................................................................ 25
14 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

ABSTRACT

Ginger (Zingiber officinale Roscoe) and turmeric (Curcuma longa) belong

to Zingiberaceae family. They are known for their pungent and aromatic
flavor and for their medicinal properties. Both crops are often grown by
smallholders in mountain areas on rich former forest soils without the
need for fertilizers and pesticides. They can be consumed fresh or dried.
Both of them are major cash crops in Mizoram. Given the lack of proper
postharvest processing and storage, a significant amount of the crop per-
ishes, and the remainder is sold in the local market at very low price. The
traditional sun drying is not effective due to insufficient availability of
sunshine. Therefore, a complete postharvest processing package consist-
ing of a rotary drum washer, a slicing unit, a dryer, and a grinder was
developed and implemented by Central Mechanical Engineering Research
Institute (CMERI) in collaboration with a local NGO Community Devel-
opment Action and Reflection (CDAR). Freshly harvested ginger/turmeric
is first washed, sliced, and then dried in a cabinet dryer. Proper washing
of ginger/turmeric is essential for the quality of the dried product as fresh
rhizomes coming from the field are covered with mud. The mud is difficult
to remove by hand wash due to the critical shape of the rhizomes. Dried
product is ground into a fine powder and stored.
The project resulted in the following achievements:
• Establishment of two centers for postharvest processing and research.
• Processing technology for ginger and turmeric transferred (two pat-
ents and two copyrights registered).
• Training of rural youths on ginger/turmeric processing.
• Employment of local youths.
• Enhancement of farmers’ economy.

2.1 INTRODUCTION

Ginger (Zingiber officinale Roscoe) and turmeric (Curcuma longa), like

cardamom and galangal, are members of Zingiberaceae family. They
are known for their pungent and aromatic flavor and for their medicinal
Development of Postharvest Processing Technology 15

properties. Chili is the fruit of plants from the genus Capsicum, members
of the nightshade family Solanaceae. In Mizoram, ginger, turmeric, and
chili are generally grown by smallholders in mountain areas on rich for-
mer forest soils without the need for fertilizers and pesticides.
Various bioactive compounds have been identified in their rhizomes,
and their content affects the price of the dried product. The bioactive
compounds in ginger rhizome (Z. officinale Roscoe) are gingerols (poly-
phenolic compounds including 10-gingerol, 8-gingerol, 6-gingerol, and
its derivatives and also shogaols that are produced by the dehydration
reaction of gingerols under high temperature). They are present in the
rhizomes of ginger, and their extracts have been proved to have high
antioxidant activity (Stoilova et al., 2007; Sakulnarmrat et al., 2015) and
anti-inflammatory effect (Dugasani et al., 2010). New phenolics have also
been identified, which have antioxidant and anti-inflammatory properties.
Turmeric (C. longa) is known for its bright yellow color and pharmaco-
logical properties due to curcumin, a phenolic compound. Curcumin is
the component of turmeric responsible for its color and all its medicinal
properties. Its structure has been identified as diferuloylmethane. Cur-
cumin and its two related demethoxy compounds, namely demethoxyc-
urcumin and bisdemethoxycurcumin, are known as curcuminoids. These
components have been identified as antioxidants. Cyclocurcumin is a
newly identified curcuminoid isolated from the fraction of turmeric found
to be active as a nematicide. In chili, the substances that give chili pep-
pers their intensity when ingested or applied topically are capsaicin and
several related chemicals, collectively called capsaicinoids that belong
to the group of amides. Although all the three crops can be consumed
fresh, the distance from the major markets or industrial clients makes
it essential for them to be preserved, mostly by dehydration. Given the
economic importance of the bioactive compounds used in pharmaceutical
and food industry, it is important to maximize the retention of bioactive
compounds in the dried products.
Ginger and turmeric are among the major cash crops in northeastern
India and particularly in Mizoram. They are generally harvested in Octo-
ber–November. So far, there has been no proper postharvest processing
for ginger and turmeric rhizomes. Open sun drying is a widespread post-
harvest practice, but given that the sunshine is erratic, this process is not
16 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

very effective in this part of India. The relative humidity is generally high
during the nighttime in the mountain areas, and the drying process is slow,
often leading to fungal contamination. Moreover, the dried product also
gets contaminated by dust and insects (Loha et al., 2008).
Therefore, CSIR-Central Mechanical Engineering Research Institute
(CMERI) in collaboration with a local NGO, Community Development
Action and Reflection (CDAR), set up a project to establish and imple-
ment rural R&D centers at Mizoram, which will continuously cater to the
technology need of the people of Mizoram in the area of postharvest pro-
cessing and enhance the economy of the local farmers. It was expected
that as a result of the project, Mizoram farmers would get a better access
to the market, finance, and education about their crops. The initial project
(RSP-0011), started in 2007, received Rs. 3 crore (INR 30 million) fund-
ing for 5 years from the Indian Government. The primary goal was to
develop a rapid, safe, and controllable drying system. The forced convec-
tion hot air drying is an efficient method to produce a uniform, hygienic,
and attractively colored product. Therefore, a forced convection cabinet
dryer was developed to address such issues (Loha et al., 2012). The project
is currently focusing on processing of ginger and turmeric but will con-
sider expanding the technology to process other cash crops such as chili
at a later stage.

2.2 EQUIPMENT DEVELOPMENT

A complete postharvest processing package consisting of a rotary drum

washer, a slicing unit, a dryer, and a grinder was developed and imple-
mented as shown in Figure 2.1.

2.2.1 ROTARY DRUM WASHER

Freshly harvested ginger/turmeric rhizomes from the field are first washed
and cut into slices before being subjected to the drying process. Proper
washing of ginger/turmeric is very important to maintain the quality of the
dried product because the ginger/turmeric rhizomes coming from the field
Development of Postharvest Processing Technology 17

FIGURE 2.1 Complete postharvest processing system for ginger/turmeric.

contain a lot of mud on it. It is very difficult to remove the mud by hand
washing due to the particular shape of the rhizomes and also because hand
wash is not practicable for large-scale application. Water consumption for
washing is another important aspect due to the scarcity of water in the
eastern Indian states. Therefore, a continuous rotary drum washing unit of
500 kg/h capacity was developed as shown in Figure 2.2, where the waste
water is filtered and re-circulated.

2.2.2 SLICING UNIT

After washing, the material is cut into slices before being introduced into
the dryer. The material is sliced to achieve faster and uniform drying by
providing a larger surface area for heat and mass transfer. Therefore, a 50
kg/h capacity slicing unit (SU-50 KPH) was developed as shown in Figure
2.3. The slicing unit consists of three blades, which can be adjusted to get
different slice thickness according to the requirement.
18 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 2.2 Rotary drum washer with filtration and recirculation of water.

FIGURE 2.3 Slicing unit.

2.2.3 CABINET DRYER

Ginger/turmeric slices should be dried from an initial moisture content

of 85% to 90% to a final moisture content of less than 10% to preserve
Development of Postharvest Processing Technology 19

them for long time. Therefore, a 50 kg/batch cabinet dryer was developed
as shown in Figure 2.4. The dryer has four chambers, and each chamber
houses six perforated stainless steel trays. Air is heated by an electrical
heater placed at the bottom of the drying chamber. Air is forced to move
in a round-about path inside the chamber to increase the heat and mass
transfer rate before being expelled by an exhaust fan at the top. Figure 2.5
shows an experimental 10 kg/batch dryer used for studies of drying kinet-
ics and design optimization. To evaluate the thin layer drying characteris-
tics of ginger slices, the 10 kg/batch dryer was slightly modified. Top four
trays were removed and a perforated sample tray was suspended between
two trays and attached to an electronic balance placed at the top of the dry-
ing chamber in order to monitor the weight loss during drying.

2.2.4 GRINDER

The dried material should be grinded into powder for the end use. There-
fore, a grinder of 50 kg/h capacity was developed as shown in Figure 2.6.

FIGURE 2.4 Cabinet dryer (50 kg/batch capacity).

20 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 2.5 Cabinet dryer (10 kg/batch capacity).

FIGURE 2.6 Grinder for dry ginger/turmeric chips.

Development of Postharvest Processing Technology 21

2.3 STUDIES CONDUCTED IN THE EXPERIMENTAL DRYER

A series of experiments were conducted in the experimental dryer (10 kg/

batch capacity).
The first set of experiments was related to the effects of drying air
temperature on the drying rates and total drying time required to reach the
equilibrium moisture content. The second set of experiments dealt with the
effects of moisture content of the slices on thermal conductivity.

2.3.1 EFFECT OF DRYING AIR TEMPERATURE ON THE

DRYING RATES

Experiments were conducted at 45°C, 50°C, 55°C, and 60°C drying air
temperature and a constant air velocity of 1.3 m/s. Moisture contents of
ginger slices were monitored online using an electronic balance until a
constant weight of the product was reached. The moisture content was
expressed as moisture ratio (MR) (see Eq. (1)).

( w t − w e ) = a exp
MR = ( −kt ) (1)
wi − we

where Wi is the initial moisture content, We the equilibrium moisture con-

tent, Wt the moisture content at time t, and a and k are temperature-depen-
dent constants.
The variation in MR is plotted against the drying time for four different
drying air temperatures of 45°C, 50°C, 55°C, and 60°C as shown in Figure
2.7. It is evident from the figure that the MR decreases exponentially with
the drying time. With an increase in drying air temperature, the drying
time decreases considerably. The drying time required to reach equilib-
rium moisture content starting with an initial moisture content of around
88–87% (w.b.) to a final moisture content of around 6–7% (w.b.) are 8.5,
7.5, 6, and 4.5 h with drying air temperatures of 45°C, 50°C, 55°C, and
60°C, respectively.
The experimental data were fitted to various mathematical models.
The model developed by Midilli et al. (2002) gave the highest R2 value
22 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 2.7 MR of ginger slices at different air temperatures and air velocity of 1.3 m/s.

and lowest RMSE value for all the temperatures. R2 values and root mean
square error (RMSE) values for the Midilli et al. (2002) model varied from
0.9993 to 0.9996 and 0.007072 to 0.009637, respectively.

2.3.2 EFFECT OF MOISTURE CONTENT ON THERMAL

CONDUCTIVITY

The experimental thermal conductivity variation with moisture content (%

w.b.) is shown in Figure 2.8. The thermal conductivity decreases with the
reduction in moisture content. It varies from 0.571 to 0.358 W/m.K with
the reduction in moisture content from 80% to 40% (w.b.) at room tem-
perature of 24°C. The moisture content has the most significant effect on
thermal conductivity, whereas the effect of temperature is negligible.
As the moisture content has the most significant effect on thermal con-
ductivity, a correlation was developed to calculate the thermal conductiv-
ity of sliced ginger (see Eq. (2)).

κ = 3.098 ×10−006 ( M 3 ) − 0.0004412 ( M 2 ) + 0.02294 ( M ) − 0.02775 (2)

The correlation predicts the value within 1.5% accuracy for a moisture
range of 80% to 40% (w.b.) and a temperature of 24°C.
Development of Postharvest Processing Technology 23

FIGURE 2.8 Variation in thermal conductivity of sliced ginger with moisture content at
24oC.

2.4 PROJECT ACHIEVEMENTS

The project Development of Postharvest Processing Technology for Gin-

ger, Turmeric, Chili, and its Implementation for augmenting Regional
Economy of Mizoram (RSP-0011) produced the following achieve-
ments:
• Establishment of two centers for postharvest processing and re-
search.
• Cabinet dryer for ginger and turmeric (Reference No. 0090NF2009)
patented and copyrighted (Reference No: IPMG/Copyright/2009).
• Rotary drum washer for ginger and turmeric (Reference No.
0151NF2009) patented and copyrighted (Reference No: IPMG/
Copyright/2008/05).
• The know-how of the technology of (1) cabinet dryer, (2) rotary
drum washer, and (3) slicing unit was transferred on a nonexclusive
basis with one-time license fee and royalty on the sold items using
the technologies.
• Processing technology for ginger & turmeric transferred.
• Training of rural youths on ginger/turmeric processing.
24 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

• Employment of local youths.

• Enhancement of farmers’ economy.
Moreover, after the establishment of the postharvest processing sys-
tem and market linkages, more than 6000 farmer families (30,000 peo-
ple approx.) in Mizoram benefitted from the technology by getting good
value of their organic products. About 14,110 hectares of land (see Table
2.1) and a number of self-help groups (SHGs) in Mizoram have been
brought under the project. The technology has also led to employment
generation in the states in the form of workers engaged in the processing
units; 35–40 tribal youths are earning their daily livelihood in these cen-
ters. More than 10000 man-days have been generated in Mizoram. The
use of technology has reduced the drudgery of the farmers in processing
their produce.

2.5 CONCLUSIONS

The project RSP-0011, carried out by the CSIR-CMERI in collaboration

with a local NGO, CDAR, has introduced the modern processing technol-
ogy for ginger and turmeric in Mizoram. The project team developed a
complete system including a rotary washing unit, a slicing unit, a cabinet
dryer, and a grinder. The project has not only developed the processing
technology (including two patents and copyrights) but also transferred this
technology to the users. A total of 68 villages were included and more than
6000 farmers were trained and are now capable of adding value to their
cash crops and to significantly improve their revenue.

TABLE 2.1 Statistics Regarding the RSP-0011 Project in Mizoram (Aizawl District)
Block Name No. of Villages No. of Farmers Area (hectares)
1. Tlangnuam 18 1452 3212
2. Thingsulthliah 17 1713 3430
3. Aibawk 15 1172 2731
4. Phullen 7 653 1749
5. Darlawn 11 1288 2988
Total 68 6278 14110
Development of Postharvest Processing Technology 25

KEYWORDS

•• ginger
•• grinding
•• income generation
•• slicing
•• training
•• turmeric

REFERENCES

Dugasani, S., Pichika, M. R., Nadarajah, V. D., Balijepalli, M. K., Tandra, S., & Kor-
lakunta, J. N., (2010). Comparative antioxidant and anti-inflammatory effects of
[6]-gingerol, [8]-gingerol, [10]-gingerol and [6]-shogaol. Journal of Ethnopharma-
cology, 127(2), 515–520.
Loha, C., Choudhury, B., & Chatterjee, P. K., (2008). Development of tray dryer for pro-
cessing ginger for the rural people in Mizoram. Proceedings of the 16th International
Drying Symposium (IDS 2008), Hyderabad, 1358–1362.
Loha, C., Das, R., Choudhury, B., & Chatterjee, P. K., (2012). Evaluation of air drying
characteristics of sliced ginger (Zingiber officinale) in a forced convective cabinet
dryer and thermal conductivity measurement. J., Food Process Technol. 3, 130.
doi:10.4172/2157–7110.1000160.
Midilli, A., Kucuk, H., & Yapar, Z., (2002). A new model for single-layer drying. Drying
Technology, 20, 1503–1513.
Sakulnarmrat, K., Srzednicki, G., & Konczak, I., (2015). Antioxidant, enzyme inhibitory
and antiproliferative activity of polyphenolic-rich fraction of commercial dry ginger
powder. International Journal of Food Science & Technology (in print). doi:10.1111/
ijfs.12889.
Stoilova, I., Krastanov, A., Stoyanova, A., Denev, P., & Gargova, S., (2007). Antioxidant
activity of a ginger extract (Zingiber officinale). Food Chemistry, 102(3), 764–770.
CHAPTER 3

WASTE PRODUCTS OF HORTICULTURAL

CROPS: AN ALTERNATIVE FOR DEVELOPING
ENTREPRENEURSHIP

AMRITESH C. SHUKLA1 and D. MANDAL2

1
Department of Botany, University of Lucknow, Lucknow–226007,
India, E-mail: [email protected]
Department of Horticulture, Aromatic & Medicinal Plants,
2

Mizoram University, Aizawl-796001, India.

CONTENTS
Abstract.................................................................................................... 27
3.1 Introduction..................................................................................... 28
3.2 Materials and Methods.................................................................... 29
3.3 Results............................................................................................. 34
3.4 Conclusion...................................................................................... 36
Acknowledgments.................................................................................... 36
Keywords................................................................................................. 36
References................................................................................................ 37

ABSTRACT

Antimicrobial activity of the secondary metabolites extracted from the

leafy wastes of some species of Curcuma, viz., Curcuma angustifolia, C.
aromatica, C. domestica, and C. zedoaria, was screened against three com-
mon dermatophytic fungi causing ringworm infection in human beings.
The volatile oil of C. domestica Valet (Family Zingiberaceae) was found
28 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

to be the strongest toxicant against the test fungi. The minimum inhibi-
tory concentration (MIC) of the oil was 1.6 μL/mL against Epidermophyton
floccosum and 1.4 μL/mL against Microsporum gypseum and Trichophy-
ton rubrum; however, it was fungicidal at 1.6 μL/mL against M. gypseum
and T. rubrum and 2.0 μL/mL against E. floccosum. The efficacy contains
heavy doses of inoculums (25 discs of 5 mm each.). The minimum kill-
ing time (MKT) of the oil was 30 s against E. floccosum and M. gypseum
and 20 s against T. rubrum, while, its minimum fungicidal concentration
(MFCs) required 6.30 h against E. floccosum and M. gypseum and 5.30 h
against T. rubrum. The oil efficacy was thermostable up to 100°C and for
36 months of storage, with the maximum unit taken into consideration.
Moreover, the oil of C. domestica did not exhibit any adverse effect on
mammalian skin up to 5% conc. The clinical trial of the oil in the form of
ointment (at 1% v/v conc) to topical testing on patients attending outpatient
department (OPD) of mLN Medical College, Allahabad, is still in progress.

3.1 INTRODUCTION

The World Health Organization (WHO) estimated that 80% of the popu-
lation of developing countries rely on traditional medicine, mostly plant
drugs, for their primary healthcare needs. Medicinal plants being natural,
non-narcotic, having no side effects, safe, and cost-effective offer preven-
tive and curative therapies that could be useful in achieving the goal of
“Health for all” in a cost-effective manner. Demand for medicinal plants is
increasing in both developing and developed countries, but 90% malarial
la harvested from wild sources without applying scientific management
hence many species are under threat to become extinct.
In fact, traditional herbal remedies have led the scientists to the devel-
opment of numerous modern drugs. At this point, the discovery of reser-
pine from Rauvolfia serpentine can be cited as an example of how a plant
utilized by the indigenous people eventually becomes the source of one of
the most important pharmaceuticals of the world.
Keeping these views in mind, in the present investigations, a scientific
attempt has been made to explore the possibilities of Curcuma spp., as a
protecting measure against ringworm infection in human beings.
Waste Products of Horticultural Crops 29

3.2 MATERIALS AND METHODS

3.2.1 IN VITRO INVESTIGATION

3.2.1.1 Extraction and Isolation of Essential Oil

The essential oil(s) were extracted separately from the wasted leaves of
Curcuma angustifolia, C. aromatica, C. domestica, and C. zedoaria (Fam-
ily- Zingiberaceae) by hydro distillation using Clevenger’s apparatus
(Clevenger, 1928). A clear light yellow colored oily layer was obtained on
the top of the aqueous distillate, which was then separated and dried over
anhydrous sodium sulfate. The oils thus obtained were subjected to vari-
ous antimicrobial investigations.

3.2.1.2 In vitro Antimicrobial Investigations of the Essential Oil(s)

The minimum effective concentration (MEC) of the oil against some com-
mon human pathogenic fungi, namely, Epidermophyton floccosum Hartz,
Microsporum gypseum (Bodin) Guiart et Grigorakis, and Trichophyton
rubrum Castellani, was determined using the technique of Shahi et al.
(2001), with a slight modification. Two sets were maintained: one for the
treatment set and another for the control. The treatment set at different
concentrations of the oil was prepared by mixing the required quantity of
the oil samples in acetone (2% of the total quantity of the medium) and
then added in pre-sterilized Sabouraud dextrose agar (SDA) medium. In
the control set, sterilized water (in place of the oil) and acetone were used
in the medium in appropriate amounts. The fungistatic/fungicidal (MSC/
MCC) action of the oil was tested by aseptically reinoculating the fungi
in culture tubes containing Sabouraud dextrose broth (Tables 3.1–3.3).
The data recorded were the mean of triplicates, repeated twice. The
percentage of fungal growth inhibition (FGI) was calculated according to
the formula:
Dc-Dt
FGI(%)= ×100
Dc
30 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

TABLE 3.1 Minimum Effective Concentration of Oil of Four Different spp. of Curcuma
against Some Common Human Pathogenic Fungi
Curcuma spp. Human Pathogenic Fungi
Epidermophyton Microsporum Trichophyton
floccosum gypsum rubrum
Curcuma angustifolia 2.6 μL/mL 2.2 μL/mL 2.4 μL/mL
C. aromatica 1.8 μL/mL 1.6 μL/mL 1.8 μL/mL
C. domestica 1.6 μL/mL 1.4 μL/mL 1.4 μL/mL
C. zedoaria 2.2 μL/mL 1.8 μL/mL 2.0 μL/mL

TABLE 3.2 Minimum Effective Concentration of the Oil Extracted from the Waste
Leaves of Curcuma Domestica Against Test Fungi
Concentra- Human Pathogenic Fungi
tion (μL/ Epidermophyton floc- Microsporum gypsum Trichophyton ru-
mL) cosum brum
2.0 100C 100C 100C
1.8 100 s
100 C
100C
1.6 100s 100s 100C
1.4 88 60 100s
1.2 60 — 80
1.0 — — 60
c
indicates cidal and indicates static.
s

where Dc indicates colony diameter in the control set and Dt indicates

colony diameter in the treatment sets.

3.2.1.3 Effect of Inoculum Density

The effect of inoculum density on the minimum cidal concentration

(MCC) of the oil against the test fungi was determined using the method
of Shukla et al. (2001). Mycelial discs of 5 mm diameter of 7-day oil
cultures were inoculated in culture tubes containing oil at their respective
MCCs. In the control set, sterilized water was used in place of the oil and
Waste Products of Horticultural Crops 31

TABLE 3.3 Detailed in-Vitro Investigations of Curcuma Domestica Against the Test
Fungi
Properties studied Observations
Epidermophy- Microsporum gyp- Trichophyton
ton floccosum sum rubrum
Minimum Inhibitory Concentration
MEC (μL/mL) 1.6 μL/mL 1.4 μL/mL 1.4 μL/mL
MFC (μL/mL) 2.0 μL/mL 1.6 μL/mL 1.6 μL/mL
Minimum Killing Time
Pure oil 30 sec 30 sec 20 sec
MFC 6.30 hrs 6.30 hrs 5.30 hrs
Inoculum Density (25 No Growth No Growth No Growth
disc, 5 mm diam)
Thermostability (up to No Growth No Growth No Growth
100ºC)
Effect of Storage (36 No Growth No Growth No Growth
months)
*MEC indicates minimum effective concentration.; MFC indicates minimum fungicidal concentra-
tion.

run simultaneously. The numbers of mycelial discs in the treatment as well

as control sets were increased progressively up to 25 discs, in multiply of
five. Observations were recorded up to the 7th day of incubation. Absence
of mycelial growth in the treatment sets up to 7th day exhibited the oil
potential against heavy doses of inoculums (Table 3.3).

3.2.1.4 Effect of Some Physical Factors

Effect of some physical factors, viz., temperature (40°C, 60°C, and 80°C)
and autoclaving (up to 15 lb/sq inch pressure for 30 min) on the efficacy
of the oil at MCC was also determined following the method of Shukla
et al. (2001) and Shahi et al. (2001). Samples of oil in small vials, each
containing 1 mL, were exposed to 40°C, 60°C, and 80°C temperatures in a
hot water bath. Further, the oil’s efficacy was tested against the test fungi
at their respective MCCs (Table 3.3).
32 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

3.2.1.5 Minimum Killing Time

The minimum killing time (MKT) of the pure oil and their respective
MCCs of C. domestica against the test fungi was determined using the
method of Shahi et al. (1999) (Table 3.4).

3.2.1.6 Fungitoxic Spectrum

The fungitoxic spectrum of the oil at lethal and hyperlethal concentra-

tion (i.e., 2 and 4 μL/mL) was determined against some common human
pathogenic fungi, viz., Microsporum auddouinii Gruby, M. canis Bodin,

TABLE 3.4 Minimum Killing Time of the Oil Extracted from the Waste Leaves of
Curcuma Domestica Against Test Fungi
Mycelial Growth Inhibition (%)
Minimum Killing Epidermophy- Microsporum gyp- Trichophyton
Time (MKT) ton floccosum seum rubrum
P.O. MFC P.O. MFC P.O. M.F.C.
7.0 100 100 100 100 100 100
6.30 100 100 100 100 100 100
6.0 100 60 100 80 100 100
5.30 100 — 100 — 100 100
5.0 100 100 100 80
2.30 100 100 100 —
2.0 100 100 100
1.30 100 100 100
1.00 100 100 100
30 min 100 100 100
15 min 100 100 100
5 min 100 100 100
60 sec 100 100 100
30 sec 100 100 100
20 sec 90 80 100
10 sec 60 — 70 — 88 —
*P.O. indicates pure oil; MFC indicates minimum fungicidal concentration.
Waste Products of Horticultural Crops 33

M. nanum Fuentes, Trichophyton mentagrophytes (Robin) Blanchard, T.

tonsurans Malmstem, and T. violaceum Bodin. This was done using the
method of Shahi et al. (2001) (Table 3.5).
The oil’s efficacy was also tested against some plant pathogenic fungi,
viz., Aspergillus parasiticus Speare, Cladosporium cladosporioides (Fre-
senius) de Vries, Curvularia lunata (Wakker) Boedijin, Colletotrichum
capsici (Syd.) Butler and Bisby, C. falcatum Went, Fusarium oxyspo-
rum Schlecht, F. udum de vries, Helminthosporium maydis Nisikado &
Miyakel, H. oryzae Breda de Haan, Penicillium implicatum Biourge, and
P. minioluteum Dierckx by using the technique of Shukla et al. (2001)
(Table 3.5).

TABLE 3.5 Fungitoxic Spectrum of the Oil of Curcuma Domestica Against Some
Common Pathogenic Fungi
Fungi Tested Lethal Concentra- Hyper Lethal Concentra-
tion (2.0 μL/mL) tion (4.0 μL/mL)
Human Pathogens
Microsporum auddouinii 100s 100c
M. canis 100 s
100c
M. nanum 100c 100c
Trichophyton mentagrophytes 100 c
100c
T. tonsurans 100c 100c
T. violaceum 100 c
100c
Plant Pathogens
Aspergillus parasiticus 100s 100c
Cladosporium cladosporioides 100 c
100c
Curvularia lunata 100c 100c
Colletotrichum capsici 100 c
100c
C. falcatum 100c 100c
Fusarium oxysporum 100 c
100c
F. udum 100c 100c
Helminthosporium maydis 100c 100c
H. oryzae 100 c
100c
Penicillium implicatum 100c 100c
P. minio-luteum 100 c
100c
s
indicates static; c indicates cidal in nature.
34 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

3.2.1.7 Comparison with Some Synthetic Fungicides:

The comparative efficacy of oil of C. domestica with some synthetic anti-

fungal drugs was carried out by comparing MECs. This was done using
the method of Shahi et al. (1999) (Tables 3.6 and 3.7).
All the experiments were repeated twice, and each contained three rep-
licates; the data presented in the tables are the mean values.

3.3 RESULTS

On comparing the MECs of the oils of Curcuma angustifolia, C. aromat-

ica, C. domestica, and C. zedoaria against the test fungi, the MEC of the
oil of C. domestica was found most effective (Table 3.1).
The MEC of C. domestica oil was 1.4 μL/mL against M. gypseum and
T. rubrum and 1.6 μL/mL against E. floccosum; however, it was fungicidal
at 1.6 μL/mL against M. gypseum and T. rubrum and at 2.0 μL/mL against
E. floccosum (Table 3.2).
The oil’s efficacy contains heavy doses of inoculums (i.e. up to 25
discs, each of 5 mm), thermostable up to 80°C and also persisted after
autoclaving at 15 lb/ sq inch pressure for 30 min (Table 3.3).
The pure oil kills the test fungi within 30 s; however, its MCC ranges
from 5.30 to 6.30 h to kill all the fungi (Table 3.4).
Fungitoxic spectrum of the oil at lethal and hyperlethal concentration
(i.e., 2 μL/mL and 4 μL/mL), against some common pathogenic fungi
reveals that the oil has a broad fungicidal spectrum (Table 3.5).

TABLE 3.6 Comparative MECs of the Oil Extracted From the Waste Leaves of
Curcuma domestica With Some Synthetic Antifungal Drugs
Oil & Trade Active Ingredi- Minimum Effective Concentration (μL/mL)
Name of Anti- ents Epidermophy- Microsporum Trichophyton-
fungal Drugs ton floccosum gypseum rubrum
Curcuma do- Essential oil 1.6 1.4 1.4
mestica
Dactrine Miconazole 6.0 6.0 6.0
nitrate
Nizaral Ketoconazole 6.0 0.5 5.0
Tenaderm Tolnaftate 2.0 1.5 0.8
TABLE 3.7 Comparative Efficacy of the Oil Extracted from the Waste Leaves of Curcuma Domestica With Some Synthetic Antifungal
Drugs
Antimycotic Drugs Drugs % Cost (Rs.) Adverse Effects Expiry Duration Environmental
Ointment/g Lotion/mL (months) impact
C. domestica 1%v/v 0.90 0.70 No adverse effects 24-36 Renewable, biode-
gradable, non-
residual toxicity.
Dactrine 2%w/w 2.80 —- Occasionally produced 35 Non-renewable,
gastrointestinal side non-biodegradable
Waste Products of Horticultural Crops

effects viz., nausea, and residual toxic-

vomiting, diarrhea ity
Nizaral 2%w/w 3.75 3.17 Adverse reaction 24 ——do——
observed were mainly
burning, irritation.
Drug may block tes-
tosterone synthesis
Tenaderm 1%w/v 1.06 1.30 Adverse effects were 24 ——do——
fever, nausea, vomit-
ing, diarrhoea & skin
rash, rarely produced
irritation
Batrafine 1%w/v 1.50 1.60 —-do—- 24 —-do—-
35
36 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Furthermore, comparison of the MECs of the oil with those of some

synthetic antifungal drugs revealed that the MECs of the oil was higher
than those of Dactrine, Nizaral, and Tenaderm (Tables 3.6 and 3.7).

3.4 CONCLUSION

The preliminary in vitro investigations of the extracted oil from the wastes
of Curcuma spp. against some common human pathogenic fungi E. flocco-
sum, M. gypseum, and T. rubrum revealed that on the basis of the detailed
in vivo and multicenter clinical trials, Curcuma domestica can not only be
an effective antimicrobial agent against dermatophytes but can also be an
alternative for developing entrepreneurship in pharmaceutical industries.

ACKNOWLEDGMENTS

Authors are thankful to Prof. Anupam Dikshit, Biological Product Labora-

tory, Department of Botany, University of Allahabad, for providing some
research facilities during this research work. Authors are also thankful
to Prof. A. K. Bajaj, Former Head of Department of Dermatology, MLN
Medical College, Allahabad, and to Dr. Uma Banerjee, Division of Micro-
biology All India Institute of Medical Sciences, New Delhi, for providing
the fungal cultures and their identification. Author are also highly thankful
to the authorities of the Mizoram University, Aizawl, as well as University
of Lucknow, for providing various kinds of support as and when required
during compilation of the work.

KEYWORDS

•• antimicrobial activity
•• dermatophtes
•• herbal drug
•• medicinal plants
•• minimum inhibitory concentration
Waste Products of Horticultural Crops 37

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CHAPTER 4

OSMOTIC DEHYDRATION OF
MANGO VARIETIES ALPHONSO
AND TOTAPURI
J. LENKA1 and R. B. TIWARI2
1
Division of Post Harvest Management, ICAR–CISH,
Rehmankhera, P.O. Kakori, Lucknow–226 101, India
2
Division Post Harvest Technology, IIHR, Bangalore–560089, India

CONTENTS

Abstract.................................................................................................... 39
4.1 Materials and Methods.................................................................... 42
4.2 Results and Discussion................................................................... 44
4.3 Conclusion...................................................................................... 52
Keywords................................................................................................. 52
References................................................................................................ 53

ABSTRACT

Osmotic dehydration is one of the new preservation methods for value

addition to fruits. It is preferred over others due to better retention prop-
erty of vitamin and minerals, color, flavor, and taste. It is less energy
intensive than air or vacuum drying process because partial dehydration
takes place at low or ambient temperature. The study on osmotic dehy-
dration of mango varieties Alphonso and Totapuri was done. The treat-
ments were Alphonso slices treated with 55 °Brix (T1) and 70 °Brix (T2)
and Totapuri slices 40 °Brix (T3), 40 °Brix (Used) (T4), 45 °Brix (Used)
40 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

(T5), 50 °Brix (T6), and 60 °Brix (T7) for 24 h. The conditions used in the
dehydration process were syrup/fruit ratio of 2:1 (v/w) in room tempera-
ture. Higher weight loss and solid gain (SG) were observed in Alphonso
slices (T2) for 24 h than in Totapuri slices. Solid gain, water loss (WL),
and weight reduction (WR) ranged from 6.60% to 7.70%, 34.50% to
44.40%, and 27.96% to 36.70% in Alphonso and from 7.55% to 13.00%,
27.15% to 48.89%, and 19.60% to 36.10% in Totapuri, respectively.
It was also observed that an increase in duration of osmosis and syrup
concentration increased weight loss, moisture loss, and SG in slices of
both the varieties. Final dried yield ranged from 25.34% to 28.52% in
Totapuri and was highest in Alphonso slices (T2) at 29.46%. The sensory
acceptance of osmo-dehydrated fruits was determined for the attributes
of aroma, flavor, texture, and overall acceptance using a hedonic scale.
Highest sensory score (84.08) was found in Alphonso slices made using
70 °Brix (T2). Osmotic dehydration process also significantly affected
other parameters and values were moisture content (10.07 to 15.45%), aw
(0.631 to 0.684) and Total solids (83.97 to 86.77%). Osmo-dehydrated
slices were found to be microbially safe for direct consumption after 2
months of storage at room temperature. The process of osmotic dehydra-
tion could be applied in rural areas for value addition to fruits by entre-
preneurs and at home scale, as it is a simple process and can contribute
to sustainable horticulture.
Mango (Mangifera indica L.) is one of the most important commer-
cial crops worldwide in terms of production, marketing, and consump-
tion. India, China, and Mexico are the main producers of mango. Unripe
mangoes are rich in vitamin C; the ripe fruits are rich in provitamin A and
contain moderate levels of vitamin C and are an excellent source of fiber.
All mango varieties represent a potential source of natural antioxidants.
A vast diversity of products can be prepared from fresh mango. However,
mangoes are extremely perishable like other farm produce, especially fruits
and vegetables. Besides the fresh fruit, the range of processed products
includes juices, nectars, concentrates, jam, fruits bars, flakes, chutney, and
dried fruit (Schieber et al., 2000; Berardini et al., 2005). Dried mango is
the commonly preserved form of the fruit in Asia, and it has also become
increasingly popular in Europe (Tedjo et al., 2002). However, convention-
ally dried mango fruit has undesirable tough texture, poor color, and cooked
Osmotic Dehydration of Mango Varieties Alphonso and Totapuri 41

flavor with a loss of nutritive value, which reduce its economic importance
(Durance et al., 1999).
India is the second largest producer of fruits after China. In India, the
area of mango cultivation is 2516,000 ha and productivity is 7.3 MT/ha
(NHB, 2014). Fruit and vegetable losses in the developing countries are
considerably high. In India, postharvest losses of fruits and vegetables are
estimated to be more than 25%. Osmotic dehydration is an important pre-
treatment that involves the immersion of the fruit in concentrated solutions
where both partial dehydration of water from the fruit and solid uptake are
obtained. Mass transfer rates during osmotic dehydration are influenced by
several factors including temperature, concentration of osmotic medium,
size and geometry of the samples, sample to solution ratio, and the level of
agitation of the solution (Torreggiani, 1993; Raoult-Wack, 1994). Osmotic
dehydration technique is an effective method for the preservation of fruits
that has gained more attention due to its potential application in the food
processing industry. The main advantages of osmotic dehydration include
better color, texture, flavor, nutrient retention, and prevention of micro-
bial spoilage. The product obtained by osmotic dehydration is more stable
during storage due to low water activity imparted by solute gain and WL
(Tiwari, 2005).
Osmotic dehydration has been reported to be useful prior to drying
to produce a variety of shelf-stable dried products with improved quality
attributes such as color, texture, and aroma (Heng et al., 1990). Osmotic
dehydration is a complementary treatment in the processing of dehydrated
foods, as it presents some advantages such as minimizing heat damage to
the color and flavor, inhibiting enzymatic browning, and reducing energy
costs (Torres et al., 2006, 2007). The technique aims to dehydrate food
products by immersing them in a hypertonic solution. Water is removed
due to the difference of osmotic potential between the food and the
osmotic solution, thus reducing the water activity of the food and conse-
quently the water availability for chemical and biological deterioration.
Osmotic concentration is the process of water removal from fruits
because the cell membranes are semi-permeable and allow water to pass
through them more rapidly than sugar. During osmosis, small quantity
of fruit acid is removed along with water. It is a dynamic process, in
which water and acid are removed at first and then move slowly, while
42 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

sugar penetration is very slight at first but increases with the time. The
characteristics of the product can be varied by controlling temperature,
sugar syrup concentration, concentration of osmosis solution, time of
osmosis, etc., to make the osmotic concentration process faster. The
present study on osmotic dehydration of mango varieties Alphonso and
Totapuri has the following objectives: to assess the effect of osmotic
pre-treatment, viz., concentration of sugar syrup, on WL, SG, WR,
yield, and physiochemical and quality parameters and to evaluate sen-
sory quality of osmo-dehydrated Alphonso and Totapuri mango slices
(Figure 4.1).

4.1 MATERIALS AND METHODS

The materials used in present investigation on osmotic dehydration

of mango are Alphonso and Totapuri. The experiment was conducted
at the Processing Laboratory of Division of Post Harvest Technology,
Indian Institute of Horticultural Research (IIHR), Bengaluru. Fruits
of Alphonso and Totapuri varieties were harvested from IIHR farms.
Fruits were allowed to ripe at room temperature, and hard ripe fruits

FIGURE 4.1 Effect of sensory quality on osmo-dehydrated mango slices.

Osmotic Dehydration of Mango Varieties Alphonso and Totapuri 43

were selected for the experiment. Fresh fruits with uniform shape and
size free from insect damage and diseases were selected for osmo-dehy-
drated products. The selected mango fruits were weighed and peeled.
The edible fruit portion was cut into slices. Prepared slices were again
weighed to record the yield recovery of fresh slices for osmotic dehy-
dration. Prepared Alphonso mango slices of 1 kg each were dipped in
55 and 70 °Brix fresh sugar syrup solution and Totapuri slices were
treated with 40 °Brix fresh and used syrup, 45 °Brix used syrup, and 50
and 60 °Brix fresh sugar syrup in the ratio of 1:2 of fruit to syrup and
allowed to continue osmosis for 24 h at room temperature (20–30°C).
Changes in syrup Brix was observed periodically. During the process of
osmosis, water flows out of the fruit pieces to the syrup and a fraction
of solute moves into the fruit slices. At the end of the treatment, the
fruit slices were taken out of the osmotic solution, and water adhering
to the surface of the slices was removed. These osmosed mango slices
were weighed to know the extent of water removal from the slices by
osmosis.

4.1.1 PRE-TREATMENTS DETAILS

• T1: Alphonso Slices 55 °Brix for 24 h

• T2: Alphonso Slices 70 °Brix for 24 h
• T3: Totapuri Slices 40 °Brix for 24 h
• T4: Totapuri Slices 40 °Brix (Used) for 24 h
• T5: Totapuri Slices 45 °Brix (Used) for 24 h
• T6: Totapuri Slices 50 °Brix for 24 h
• T7: Totapuri Slices 60 °Brix for 24 h
The following characteristics of fresh and osmo-dehydrated mango
slices were recorded:

initial weight − weight at time

Weight loss (% ) = × 100
initial weight

Solid gain (%) = Moisture loss (%) − Weight loss (%)

44 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Weight of prepared fruit pieces

Yield (% ) = ×100
Weight of fresh fruuit

Moisture content of fresh pulp, osmosed slice, and osmotically dehy-

drated samples was determined on percentage basis. Five grams of sam-
ple was taken in a pre-weighed China dish and kept in a hot air oven for
overnight, and the weight was then recorded using an electronic balance.
Moisture content was determined on fresh weight basis. Total solids were
calculated by subtracting moisture content from 100.

Moisture loss
Moisture content ( % ) = ×100
Sample weight

4.2 RESULTS AND DISCUSSION

Data given in Table 4.1 indicate that various osmotic treatments signifi-
cantly affected the different physical parameters of mango slices in both
the varieties.

4.2.1 MOISTURE CONTENT

Moisture content of osmo-dehydrated mango slices ranged from 10.07%

to 16.03% . The maximum moisture content (16.03%) of osmo-dehydrated
products was recorded in the treatment of Totapuri slices at 40 °Brix used
syrup for 24 h (T4), followed by 15.45% in Alphonso mango slices at 55
°brix for 24 h (T1). Treatments T2, T7, and T5 were found to be statistically
at par with each other (Table 4.1).

4.2.2 TOTAL SOLID CONTENT

Total soluble solid (TSS °brix) in fresh mango slices was 16 to 17 °brix in
Alphonso and 13 to 13.5 °brix in Totapuri. Total solid of osmo-dehydrated
mango slices ranged from 83.97% to 89.93%. In Alphonso, total highest
TABLE 4.1 Effect of Different Osmotic Treatments on Physical Composition, Yield and Mass Transfer Kinetics in Osmotically
Dehydrated Alphonso and Totapuri Mango Slices
Treatments Osmosed-mango slices Osmo-dehydrated samples
Moisture (%) Total Solid Yield (%) Water Activ- Water Loss Solid gain (%) Weight Reduc-
(%) ity (aw) (%) tion (%)
T1 Alphonso slices at 15.45 84.55 27.81 0.643 34.50 6.60 27.96
55⁰Brix for 24 h
T2 Alphonso slices at 14.93 85.07 29.46 0.642 44.40 7.70 36.70
70⁰ Brix for 24 h
T3 Totapuri slices at 13.23 86.77 27.98 0.64 32.93 13.00 19.93
40⁰B for 24 h
T4 Totapuri slices at 16.03 83.97 27.62 0.655 34.55 12.68 21.87
40⁰Brix (Used) for
24 h
T5 Totapuri slices at 14.18 85.82 27.03 0.631 40.71 11.11 29.60
45⁰B (Used) for
24 h
T6 Totapuri slices at 10.07 89.93 25.34 0.684 27.15 7.55 19.60
50⁰B for 24 h
Osmotic Dehydration of Mango Varieties Alphonso and Totapuri

T7 Totapuri slices at 14.80 85.20 28.52 0.648 48.89 12.79 36.10

60⁰B for 24 h
Sem ± 0.02 0.02 0.22 0.008 0.15 0.01 0.27
CD at 5% 0.43 0.43 1.49 0.001 1.05 0.09 1.81
45
46 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

solid (85.07%) was recorded in Alphonso at 70 °brix (T2) and lowest in

Alphonso at 55 °brix (T1) In Totapuri slices, maximum total solid was
recorded in Totapuri slices treated at 50 °brix (T6) and lowest in Totapuri
slices at 40 °brix used syrup (T4) and the values ranged from 83.97% to
86.77%.

4.2.3 YIELD OF OSMOTICALLY DEHYDRATED MANGO

SLICE

There was a statistically significant difference among the various

osmotic treatments of final dried yield of mango slices, and it ranged
from 25.34% to 29.46%. Highest yield (29.46%) was recorded in
Alphonso at 70 °brix (T2) because of maximum SG followed by Tota-
puri slices (28.52%) at 60 °brix (T7). Treatments T3, T1, and T5 were
found to be statistically at par with each other and lowest in Totapuri
slices at 50 °brix (T6) due to minimum SG among the Totapuri mango
slices. WL in osmosed mango slices ranged from 27.15 to 48.89. It also
showed that the drying rate was better in concentrated syrup due to
the increased osmotic pressure in the sugar syrup at higher concentra-
tions, which increased the driving force available for water transport.
These results are in conformity with the findings of Thippanna (2005)
for banana.

4.2.4 WATER LOSS AND SOLID GAIN

Maximum WL of 48.89% was recorded in Totapuri slices at 60 °brix for

24 h (T7) followed by Alphonso (44.4%) slices treated with 70 °brix for
24 h (T2), while minimum WL of 27.15% was found in Totapuri slices at
50 °brix (T6) . Highest SG of 13.00% was recorded in Totapuri slices at 40
°brix for 24 h (T3), followed by 12.79% in Totapuri slices at 60 °brix for
24 h (T7). These findings are also in conformity with observations made
by other workers in case of mango (Varany-Anond et al., 2000), pineapple
(Rahaman and Lamb, 1990). Sharma et al. (2003) reported that during
osmotic dehydration, WL is always favored over solid uptake that leads to
mass loss of pear fruit.
Osmotic Dehydration of Mango Varieties Alphonso and Totapuri 47

4.2.5 WEIGHT REDUCTION DURING OSMOSIS

Weight reduction in osmosed mango slices ranged from 19.60% to 36.70%.

Highest WR (36.70%) was recorded in Alphonso slices treated with 70
°brix for 24 h (T2), followed by 36.10% in Totapuri slices at 60 °brix for
24 h (T7), while minimum WR (19.60%) was found in Totapuri slices at
50 °brix for 24 h (T6). Further, it was also observed that increase in syrup
concentration led to higher WR in osmosed mango slices. Variation in
weight loss during osmotic dehydration among the varieties of apricot was
observed by Sharma et al. (2004). Lombard et al. (2008) studied the effect
of osmotic dehydration on mass fluxes (WL, SG, and WR).

4.2.6 WATER ACTIVITY (AW) IN DEHYDRATED SAMPLE

Water activity in osmo-dehydrated mango slices ranged from 0.640 to

0.684. Minimum water activity was recorded in Totapuri slices at 40 °brix
for 24 h (T3) followed by Alphonso at 70 °brix for 24 h (T2), while maxi-
mum water activity was found in Totapuri slices at 50 °brix for 24 h (T6)
treatment. Similar results were reported by Pointing (1973). The product
obtained by osmotic dehydration is more stable during storage due to low
water activity imparted by solutes gain and WL (Tiwari, 2005). They also
found that all these parameters depend on the concentration of syrup and
syrup to fruit ratio.

4.2.7 EFFECT OF VARIOUS OSMOTIC TREATMENTS ON

ACIDITY, ASCORBIC ACID, CAROTENOIDS, AND NON-
ENZYMATIC BROWNING IN OSMO-DEHYDRATED MANGO
SLICES

Data given in Table 4.2 indicate that various osmotic treatments signifi-
cantly affected acidity, ascorbic acid, carotenoids and non-enzymatic
browning (NEB) in osmo-dehydrated mango slices. Acidity content
in fresh Alphonso and Totapuri slices was 1.45% and 0.90%, respec-
tively. Acidity content in osmo-dried mango slices ranged from 0.60 to
1.79%. Highest acidity was recorded in Totapuri slices at 45 °brix used
 48

TABLE 4.2 Effect of Different Osmotic Treatments on Chemical and Sensory Quality Parameters of Osmotically Dehydrated Alphonso
and Totapuri Mango Slices
Treatments Total Acidity Ascorbic Acid Carotenoids NEB
(%) (mg/100 g) (mg/100 g) (OD 440 nm)
T1 Alphonso slices at 55⁰Brix for 24 h 0.77 123.03 13.60 0.167
T2 Alphonso slices at 70⁰ Brix for 24 h 1.54 96.43 20.25 0.196
T3 Totapuri slices at 40⁰B for 24 h 0.90 36.93 2.35 0.110
T4 Totapuri slices at 40⁰Brix (Used) for 24 h 1.54 24.90 3.67 0.100
T5 Totapuri slices at 45⁰B (Used) for 24 h 1.79 53.95 4.82 0.090
T6 Totapuri slices at 50⁰B for 24 h 0.90 41.50 6.22 0.129
T7 Totapuri slices at 60⁰B for 24 h 0.60 52.29 5.67 0.100
Sem ± 0.01 0.24 0.02 0.001
CD at 5% 0.08 1.59 0.17 0.008
Sustainable Horticulture Volume 2: Food, Health, and Nutrition
Osmotic Dehydration of Mango Varieties Alphonso and Totapuri 49

for 24 h (T5), followed by Totapuri slices at 40 °brix used for 24 h (T3)

and Alphonso at 70 °brix for 24 h (T2). Lowest acidity was recorded in
Totapuri slices at 60 °brix fresh for 24 h (T7). Ascorbic acid content in
osmo-air dried samples ranged from 24.90 to 123 mg per 100 g. High-
est ascorbic acid was recorded in Alphonso slices treated with 55 °brix
for 24 h (T1) followed by Alphonso at 70 °brix for 24 h (T2) while lowest
was recorded in Totapuri slices at 40 °brix used for 24 h (T4). Carotenoid
content in osmo-dried mango slices ranged from 2.35 to 20.25 mg/100 g.
Highest carotenoid content (20.25 mg/100 g) was recorded in Alphonso
slices treated with 70 °brix sugar syrup for 24 h (T2) followed by Alphonso
at 55 °brix for 24 h (T1), while lowest carotene content (2.35 mg/100 g)
was recorded in Totapuri slices at 40 °brix fresh for 24 h (T3) (Figure 4.2).
Non-enzymatic browning (NEB OD at 440 nm) in osmo-air dried mango
slices ranged from 0.090 to 0.196. Lowest NEB value (0.100 OD at 440
nm) was observed in Totapuri slices treatment with 45 °brix used for 24
h (T5) followed by Totapuri slices at 60 °brix fresh for 24 h (T7), while
maximum NEB value (0.196 OD at 440 nm) was recorded in the treatment
Alphonso at 70 °brix for 24 h (T2). Torregianni et al. (1986) reported on
sugar content, color, acidity, vitamin C, pH, and organoleptic distinctive-
ness on osmo-dehydrated cherry.

FIGURE 4.2 Effect of carotenoid contents on osmo-dehydrated mango slices.

50 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

4.2.8 COLOR, TEXTURE, FLAVOR OF OSMO-DRIED

MANGO SLICES

Significant differences were recorded due to different treatments for color

score in osmo-dehydrated mango slices. Sensory score for color in osmo-
dehydrated mango slices ranged from 24 to 27.08, which was in accept-
able range (Table 4.3, Plate 1). The highest score recorded in Alphonso
at 70 °brix for 24 h (T2) followed by Alphonso at 55 °brix for 24 h (T1)
and Totapuri slices at 50 °brix for 24 h (T6), and lowest score was found
in Totapuri slices at 40 °brix used for 24 h (T4). Sensory score for texture
in osmo-dehydrated mango slices ranged from 22.50 to 27.00. The high-
est score recorded was in Alphonso at 70 °brix for 24 h (T2), followed by

TABLE 4.3 Effect of Different Osmotic Treatments on Sensory Quality Parameters of

Osmotically Dehydrated Alphonso and Totapuri Mango Slices
Treatments Sensory Score
Colour (30) Texture (30) Flavour (40) Total (100)
T1 Alphonso slices 26.00 25.00 27.08 78.08
at 55⁰Brix for
24 h
T2 Alphonso slices 27.08 27.00 30.00 84.08
at 70⁰ Brix for
24 h
T3 Totapuri slices at 24.50 22.50 24.91 71.91
40⁰B for 24 h
T4 Totapuri slices at 24.00 23.20 24.80 72.00
40⁰Brix Used for
24 h
T5 Totapuri slices 25.00 23.00 25.41 73.41
at 45⁰B Used for
24 h
T6 Totapuri slices at 26.00 25.00 27.00 78.00
50⁰B for 24 h
T7 Totapuri slices at 24.80 22.65 24.30 71.75
60⁰B for 24 h
Sem ± 0.31 0.33 0.27 0.78
CD at 5% 2.11 2.23 1.82 5.26
Osmotic Dehydration of Mango Varieties Alphonso and Totapuri 51

PLATE 1 (See color insert.) Osmotic dehydration of mango slices.

Alphonso at 55 °brix for 24 h (T1) and Totapuri slices at 50 °brix for 24

h (T6) while lowest score was found in Totapuri slices at 40 °brix for 24
h (T3). Sensory score for flavor in osmo-dehydrated mango slices ranged
from 24.30 to 30.00 (Figure 4.1). The highest score was recorded in
Alphonso at 70 °brix for 24 h (T2), followed by Alphonso at 55 °brix for
24 h (T1) and Totapuri slices at 50 °brix for 24 h (T6), while lowest score
was found in Totapuri slices at 60 °brix fresh for 24 h (T7). Overall, sen-
sory score in osmo-dehydrated mango slices ranged from 71.75 to 84.08.
The highest score 84.08 was recorded in Alphonso slices treated with 70
°brix sugar syrup for 24 h (T2). Treatments T1 and T6 were statistically at
par with each other, while lowest score was found in Totapuri slices at
60 °brix for 24 h (T7). These results are in conformity with observations
made by other workers Heng et al. (1990); Torres et al. (2006, 2007, 2008)
improved quality attributes such as color, texture and aroma by inhibit-
ing enzymatic browning by osmotic dehydration. Ramanuja and Jayara-
man (1980) prepared intermediate moisture banana with better flavor and
appearance with good storability.
52 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

4.2.9 MICROBIAL QUALITY IN OSMO-DRIED MANGO

SLICES

Microbial quality in the samples was evaluated after 2 months of storage at

room temperature in the Microbiology Laboratory of post-harvest technol-
ogy (PHT) Division at IIHR. Different media were used for yeast, mold,
lactic acid bacteria, E. coli, and total count. Total plate count was recorded
in 13 × 10², 4 × 104, and Nil × 106 dilution in Totapuri slices at 60 °brix
fresh for 24 h (T7), and all other treatments were recorded nil in different
dilutions. E. coli, yeast, and mold counts were negligible in osmo-dehy-
drated mango slices, and they were found to be microbially safe. There-
fore, these slices were safe for direct consumption. Similar results were
reported by Ramanuja and Jayaraman (1980) and Kandekar et al. (2005).

4.3 CONCLUSION

It was concluded that various osmotic treatments had significant effect on

product quality. Osmotic dehydration improved acceptability in mango sam-
ples. Variation in syrup concentration significantly affected WL, SG, WR,
and final osmotically dehydrated yield. Varietal variation was also noticed.
The overall sensory score in osmo-dehydrated mango slices ranged from
71.75 to 84.08. The highest score of 84.08 was noted in Alphonso slices
treated with 70 °brix sugar syrup for 24 h (T2), which was nonsignificantly
followed by Totapuri slices treated with 50 °brix sugar syrup for 24 h (T6).

KEYWORDS

•• mango
•• moisture loss
•• organoleptic quality
•• osmotic dehydration
•• preservation
Osmotic Dehydration of Mango Varieties Alphonso and Totapuri 53

REFERENCES

Berardini, N., Knodler, M., Schieber, A., & Carle, R., (2005). Utilization of mango peels as
a source of pectin and polyphenolics. Innovative Food Science and Emerging Tech-
nologies, 6, 442–452.
Durance, T. D., Wang, J. H., & Meyer, R. S., (1999). Processing for Drying Mango and
Pineapples. US Patent No. 5962057.
Heng, W., Guilbert, S., & Cug, J. L., (1990). Osmotic dehydration of papaya: influence of
process variables on the quality. Science Des. Aliments, 10, 831–848.
Khandekar, S. V., Chavan, U. D., & Chavan, J. K., (2005). Preservation of pulp and prepa-
ration of toffee from fig fruit. Beverage and Food World, 32, 55–56.
Lombard, G. E., Oliveira, J. C., Fito, P., & Andres, A., (2008). Osmotic dehydration of
pineapple as a pre-treatment for further drying. J. Food Eng., 85, 277–284.
National Horticulture Board Database (2015). National Horticulture Board, Government
of India, Gurugram, Haryana. (http://www.nhb.gov.in).
Ponting, J. D., (1973). Osmotic dehydration of fruits-recent modifications and applica-
tions. Process Biochem., 8, 18–20.
Rahman, M. S., & Lamb, J. (1990). Osmotic dehydration of pineapple. J. Food Sci. Tech-
nol. (India), 27(3), 150–152.
Ramarjuna, M. N., & Jayaraman, K. S., (1980). Studies on the preparation and storage
stability of intermediate banana. J. Food Sci. Technol, 17, 183.
Raoult-Wack, A. L., (1994). Recent advances in the osmotic dehydration of fruits. Trends
in Food Sci. Technol., 5, 255–260.
Sharma, H. K., Pandey, H., & Kumar, P., (2003). Osmotic dehydration of sliced pears. J.
Agric. Eng., 40(1), 65–68.
Sharma, K. D., Kunen, R., & Kaushal, B. B. L., (2004). Mass transfers characteristics of
yield and quality of five varieties of osmotically dehydrated apricot. J. Food Sci.
Technol., 41, 264–275.
Tedjo, W., Taiwo, K. A., Eshtiaghi, M. N., & Knorr, D., (2002). Comparison of pretreat-
ment methods on water and solid diffusion kinetics of osmotically dehydrated man-
gos. Journal of Food Engineering, 53, 133–142.
Thippanna, K. S., (2005). Studies on osmotic dehydration of banana (Musa spp.) fruits. M.,
Sc. (Hort.) Thesis, University of Agricultural Sciences, Bangalore.
Tiwari, R. B., (2005). Application of osmo–air dehydration for processing of tropical fruits
in rural areas. Indian Food Industry, 24, 62–69.
Torreggiani, D., (1993). Osmotic dehydration in fruit and vegetable processing. Food Res.
Intl., 26, 59–68.
Torreggiani, D., Giagiacamo, R., Bertolo, G., & Abbo, E., (1986). Research on the
osmotic dehydration of fruits-I. Suitability of cherry varieties. Ind. Conserv., 61(2),
101–107.
Torres, J. D., Castello, M. L., Escriche, I., & Chiralt, A., (2008). Quality characteristics,
respiration rates, and microbial stability of osmotically treated mango tissue (Man-
gifera indica L.) with or without calcium lactate. Food Sci. Technol. Int., 14(4),
355–365.
54 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Torres, J. D., Talens, P., Carot, J. M., Chiralt, A., & Escriche, I., (2007). Volatile profile of
mango (Mangifera indica L.), as affected by osmotic dehydration. Food Chem., 101,
219–228.
Torres, J. D., Talens, P., Escriche, I., & Chiralt, A., (2006). Influence of process condi-
tions on mechanical properties of osmotically dehydrated mango. J. Food Engg., 74,
240–246.
Varney, W., Wongkrajang, K., Warunee, V. A., & Wongkrajan, K., (2000). Effects of some
parameters on the osmotic dehydration of mango Cv. Kaew. Thai. J. Agri. Sci., 33,
123–135.
CHAPTER 5

BREADNUT: INNOVATIVE
PRODUCTS FOR THE AGRO FOOD
SECTOR
S. SUBRAMANIAM, N. RAMBURN, and R. CHACOORY
Crop Department, Food and Agricultural Research and Extension
Institute, Reduit, Mauritius, Tel.: +230 6708249,
E-mail: [email protected], [email protected]

CONTENTS

5.1 Introduction..................................................................................... 56
5.2 Material and Methods..................................................................... 57
5.3 Results and Discussion................................................................... 60
5.4 Conclusion...................................................................................... 67
Keywords................................................................................................. 67
References................................................................................................ 67

Breadnut (Artocarpus camansi) is an untapped crop in Mauritius. Bread-

nut grows with minimum inputs and presents high potential for food
security. Breadnut seeds are considered to be nutritious, being a good
source of protein, namely essential amino acids, minerals, and carbohy-
drates. This study aimed at evaluating the potential of breadnut as (i) a
cooked nut; and (ii) processed products in view of providing new healthy
innovative products for the local population and for the export market
targeting celiac patients in particular. The work confirmed that locally
available immature and mature green breadnut fruits can be consumed as
a vegetable, while the mature seeds from fully ripened fruits can be used
56 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

as a staple, a vegetable, or a snack with high acceptability. Proximate

analysis showed that breadnut is low in fat and is gluten free. Breadnut
seeds can be preserved in brine or can be frozen for subsequent use as a
vegetable. A protocol was also developed for the production of breadnut
flour. A 100-g portion of breadnut flour contains 74 g carbohydrates, 9.74
g protein, 2.27 g ash, 1.65 g fat, and minerals like potassium (390 mg),
phosphorous (260 mg), calcium (110 mg), and magnesium (70 mg). The
breadnut flour was found to be suitable for enrichment of wheat flour
and can be used in different forms for the preparation of various nutri-
tious dishes like porridge, stews, and milk-based drink with high sensory
value, thereby indicating the potential of an unexploited crop for value
addition.

5.1 INTRODUCTION

Breadnut (Artocarpus camansis Blanco, Family: Moraceae), also known as

chataigne, castana, kamansi, and rima, is a tropical tree nut distinct from
breadfruit with its spiny texture and numerous seeds embedded in the fruit
pulp. It is an underutilized crop with significant potential as a nutritious food
source (Ragone, 2006). Immature fruits are bright green, firm with white
pulp and soft immature seeds. Mature fruits are light green, firm with a light
yellow pulp and mature seeds enclosed within a brown thick, hard endocarp
and an inner thin brown testa. Most fruits are harvested at this stage as the
seeds are at their best eating quality. Ripe fruits are yellow brown with a soft
pulp and mature seeds; ripe fruits should be immediately harvested as any
delay leads to declination of seed eating quality due to its high perishable
nature (Robrts-Nkrumah, 2015). Breadnut seeds are considered to be nutri-
tious: a 100-g portion of dried seeds contain 13.3–20 g protein, 6.2−12.8 g
fat, 76.2 g carbohydrates, 2.5−3.9 g fiber, and essential amino acids (Qui-
jano et al., 1979; Negro de Bravo et al., 1983); they are lower in fat content
compared to almond, brazil nut, cashew, and macademia (Moreira et al.,
1998).
Immature breadnut fruits are used in Trinidad and Tobago and Guyana
as a curried dish. Seeds from mature fruits are consumed after boiling, fry-
Breadnut: Innovative Products for the Agro Food Sector 57

ing, or roasting and used for flour production in some Caribbean and Latin
American countries (Williams and Badrie, 2005; Ordonez, 2011; Roberts-
Nkrumah, 2015).
Breadnut is an untapped crop in Mauritius; it is not widely distrib-
uted, and it is mainly used for the production of seedlings that are used as
rootstock for the propagation of breadfruit locally. Breadnut trees, which
require minimum inputs, can be introduced in agro-forestry in Mauritius
and can have high potential as a food and nutrition security crop. It can
also be a new source of produce and products for celiac patients particu-
larly as breadnut seeds are gluten free.
This study aimed at evaluating the potential of breadnut as (i) a cooked
nut and (ii) processed products in view of providing healthy innovative prod-
ucts for the local population and for the export market through low-cost pro-
cessing methods and creating new commercial opportunities in Mauritius.

5.2 MATERIAL AND METHODS

Mature ripe breadnut fruits were harvested or collected at St. Aubin and
Pamplemousses over a period of 4 years during the production season
that ranged between October–February (summer) and April–June (early
winter) in Mauritius. Immature fruits were harvested for evaluation as a
vegetable.
Fully mature firm fruits that contained mature seeds and ripe fruits
were harvested or collected for evaluation of the mature breadnut seeds as
a staple, vegetable, or healthy snack and for processing.
The breadnut seeds were extracted from the ripe pulp, the seeds were
thoroughly washed, and the surface microbial load was reduced by soak-
ing the seeds for 5 minutes in chlorinated water (25 mL of 3.25% active
chlorine/5 liters of water); subsequently, they were (i) characterized, (ii)
evaluated as fresh produce or processed immediately, or (iii) stored for 1
day at ambient prior to processing into selective products. The fruits were
characterized after harvest for weight, circumference, length, number of
seeds, total seed weight; the seeds were also characterized for the same
parameters.
58 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

5.2.1 EVALUATION OF BREADNUT AS A VEGETABLE,

STAPLE, AND NUT

For the evaluation as a vegetable, both immature and mature green fruits
were harvested. The immature fruits were thoroughly washed, peeled
and pulped, and seeds cut into small pieces prior to cooking. Seeds were
extracted from ripe fruits, washed and cooked after removal of the endo-
carp and testa (peeling) or boiled, peeled, and cooked.

5.2.2 EVALUATION OF BREADNUT FOR SELECTED

PROCESSED PRODUCTS

For processing, different treatments and methods were evaluated, and the
processing methods were reviewed based on outcomes of each trial until

FIGURE 5.1 Preparation of breadnut seeds for cooking or processing.

Breadnut: Innovative Products for the Agro Food Sector 59

the optimum protocol was developed for each processed product. Sensory
evaluation was carried out, and shelf-life determined for each selected
product. The breadnut seeds were prepared and pretreated prior to pro-
cessing as shown in Figure 5.1. For use as a vegetable, snack, or frozen
nuts, the breadnut seeds were precooked in a pressure cooker until fully
cooked but firm (15 minutes), while for processing, the seeds were pres-
sure cooked (10 minutes) until 80% cooked to maintain firmness. Boiling
the breadnut seeds and removal of the testa prior to processing was found
to be the most effective processing steps as it eliminated the “raw” and
unappealing flavor associated with raw breadnut seeds.

5.2.2.1 Breadnut Seeds in Brine

The breadnut was boiled in pressure cooker in salted water (1.5% w/v)
until cooked but firm; two treatments were evaluated: fully peeled boiled
breadnut seeds and boiled breadnut seeds with testa. The breadnut seeds
were canned in hot 3.0% brine (salinity of 3.1–3.2) to which citric acid
(0.075% w/v, pH 4.5) had been added, pasteurized for 10 minutes at 85°C,
cooled, and stored at ambient.

5.2.2.2 Frozen Breadnut Seeds

Peeled raw and pre-boiled (0.5% salt water) peeled breadnut were sub-
jected to individual quick-freezing technique, packed in polyethene vac-
uum bags, and stored at −15°C up to 15 months; they were evaluated at
every 3 months interval to check product quality.

5.2.2.3 Breadnut Seeds Flour

Different pretreatments (eight methods) were investigated for flour pro-

duction with a view to reduce the labor-intensive procedure of cleaning
the nuts and to yield dried breadnut/flour of good acceptability and quality.
Mature breadnut seeds were peeled (removal of endocarp, with
and without testa), sliced (1 mm) or shredded (2 × 2 mm) and dried or
60 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

blanched, prior to drying or boiled, before drying at 50 and 55°C. The pre-
treated breadnut seeds were dried until brittle. The dried breadnut shreds
generated from the different combination treatments were grinded into
flour (0.5–0.8 µm) or granules (0.75 mm, 1.0–1.5 mm diameter; similar
to semolina and broken wheat) using a grinding mill and stored in high
density polyethylene (HDPE) packs.
The production method that yielded promising results in terms of prod-
uct’s taste, flavor, color, and texture and that was less labor intensive was
selected. Based on the recommended method, breadnut seed flour, and
breadnut seed granules were produced for further evaluation. The bread-
nut flour of different grades (coarse granules, small granules, and flour)
was assessed into a range of products (sweet and savory). The nutritional
value and shelf life of the flour was determined.

5.3 RESULTS AND DISCUSSION

5.3.1 FRUIT AND SEED CHARACTERISTICS

Fruits are oval to round in shape and texture varied according to maturity
of fruits.
Immature fruits were green with sharp pointed firm spines and hold
immature seeds; these seeds are susceptible to oxidative browning after
extraction and when cut exudates a latex. The immature seeds have a very
low acceptability. Mature and ripe fruits were light green to greenish brown
in color with soft spines and pulp and contain the mature seeds. The seeds
are embedded within the soft pulp; the edible part is enclosed within a brown
thick pattern veined endocarp and wrapped in a thin layer membrane (testa).

TABLE 5.1 Breadnut Fruit Characteristics

Fruit circumference 32–43 cm
Fruit length 13–22 cm
Fruit weight 500–1170 g
Fibrous core 59–68 g
Number of seeds/fruit 13–61
Seed weight 160–680 g
Breadnut: Innovative Products for the Agro Food Sector 61

Fruits’ characteristics are shown in Table 5.1. Characterization indi-

cated that a fruit weighed between 500 and 1170 g with around 13–61
seeds. The seed recovery rate was between 30% and 45%.
Each seed weighed between 5.25 and 10.50 g with a diameter of 2.20–
3.73 cm and length of 2.7–3.5 cm; the seeds formed 22–45% of the total
fruit weight. The endocarp constituted an average of 21% of fruit weight,
while the testa weight can vary between 10 and 14% of fruit weight. The
physical characteristics are within the range reported by Ragone (2006)
and Roberts-Nkrumah (2005).
The peeled breadnut seeds that are of main commercial interest are off
white in color, firm with light trace of latex. The recovery rate of peeled
seeds varied between 25% and 30% of fruit weight.

5.3.2 USE OF BREADNUT

5.3.2.1 Immature Fruits as a Vegetable

The cooked dish was slightly bitter and was rated as moderate by consum-
ers, thereby indicating its limited use locally.

5.3.2.2 Mature Seeds as Vegetable, Staple, and Snack

Mature breadnut seeds were cooked into dish similar to local potato-based
dishes. Raw breadnut seeds and boiled breadnut seeds were evaluated. The
curry from raw breadnut had a longer cooking time, required more water,
and maintained a very firm and rigid texture. Boiling of the seeds with
salted water (1.5–2.0%) in a pressure cooker and removal of the seed peel
prior to cooking/preparation of different dishes improved the taste with
the product having a sweet cooked flavor similar to chestnut and edoes,
and soft and creamy texture. These observations are in line with that of
Mathews et al. (2001) wherein cooked breadnut steamed at atmospheric
pressure and above atmospheric pressure for 10–15 minutes were most
preferred. Boiled seeds consumed as a snack was rated with high accept-
ability as indicated by a consumer perception study. The findings are in
line with those reported by Roberts-Nkrumah (2015), Ragone (2006), and
Williams and Badrie (2005).
62 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

The seeds may also be roasted in pan or oven at 200°C for 15–20 min-
utes or steam cooked yielding cooked nuts of very good acceptability.
Seeds from over ripe fruits that were collected (brown pulp and strong
fermentation flavor and seeds were enclosed in a dark brown endocarp)
had a slight bitter taste and astringent off taste after boiling, indicating
declining quality of the seeds due to physiological changes.

5.3.3 NUTRITIONAL VALUE OF BOILED BREADNUT SEEDS

Table 5.2 shows the proximate composition of boiled breadnut seeds and
compares it with the nutritional value of that reported by Williams and
Badrie (2005) in West Indies.

TABLE 5.2 Proximate Composition of Boiled Breadnut Seeds for Mauritius and
Comparison with Values for West Indies
Composition Breadnuts seeds (Mauritius) Breadnuts seeds (West Indies)
Moisture (g) 62.46 61.59
Protein (g) 4.08 6.89
Fat (g) 1.87 4.20
Carbohydrate (g) 28.36 23.90
Ash (g) 1.14 3.42
Crude fiber (g) 2.09 N/A
Gluten (g) NIL N/A
Reducing sugar (g) 1.53 N/A
Calcium (mg) 47 9.62
Magnesium (mg) 36 44.78
Phosphorus (mg) 120 4.28
Sodium (mg) 43 204.61
Potassium (mg) 420 734.62
Iron (mg) 1.25 1.18
Zinc (mg) 2.14 0.69
Copper (mg) 0.29 0.34
Manganese (mg) 0.14 0.36
Energy Kcal 150 -
Source: FAREI (2014); Williams & Badrie (2005).
Breadnut: Innovative Products for the Agro Food Sector 63

The boiled breadnut is rich in carbohydrates and a good source of min-

erals like calcium, phosphorous, and potassium.
The fat content of the breadnut seeds collected in Mauritius is lower
than that reported for West Indies, and calcium, phosphorous and iron con-
centration is slightly higher for nuts under the current study.
The high carbohydrate and protein content and presence of minerals
confirms that breadnut seeds has a good nutritional profile and can be con-
sidered and included as a food and nutrition security crop in the Mauritian
diet as a vegetable, staple, and healthy snack.

5.3.4 BREADNUT IN BRINE

An average product yield of 65% was recorded (seed weight basis).

Sensory evaluation after 1 week yielded a product of moderate accept-
ability. Breadnut preserved without the testa appeared to disintegrate
slightly after pasteurization and during storage as observed by the
slightly turbid brine and tiny bits of cooked seeds. The brine solution
for breadnut seeds with the testa or thin peel was clear, and these bread-
nuts had a better sensory value; it is therefore recommended to remove
only the endocarp. These observations are in line with that of Mathews
et al. (2001) whereby precooked, thin peel covered breadnuts canned in
2.5% brine were most preferred. Salinity and pH were stable, brine was
clear, and a shelf life of 12 months is recommended for the breadnut
seeds with the thin membrane or testa that should be removed prior to
consumption.

5.3.5 FROZEN BREADNUT

Raw frozen seeds suffered from chilling injury with water accumula-
tion, the seeds became spongy, and when cooked (unthawed and thawed)
yielded a product with low acceptance. However, precooked or pre-
boiled peeled breadnut was stable when frozen to −15°C and keep up to
15 months.
The boiled frozen breadnut is not affected by chilling injury and after
thawing; the breadnut can be used for cooking, as a snack, or for process-
64 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

ing into flour with very good results. Assessment of the product indicated
that it was suitable for preparation of curries with its taste close to fresh
ones. A yield of 62–72% (seed weight basis) or 30% (fruit weight basis)
was recorded.

5.3.6 BREADNUT FLOUR

5.3.6.1 Flour Production

Flour from raw breadnut seeds had a slight raw, astringent, and off fla-
vor, and the same was detected in the prepared products. However, flour
and prepared dishes from boiled breadnut seeds had a high sensory value
indicating that boiling improves the flour quality by inactivation some
anti-nutritional factors. This is in line with the findings of Fagbemi et al.
(2005) who showed that breadnut seeds contain some level of phytic acid,
trypsin inhibitors, and tannins; boiling was the most effective pretreat-
ment technique to reduce the tannin and trypsin inhibitor activity by 37%
and 20%, respectively, thereby improving the digestibility of the bread-
nut seed flour by at least 71% when compared to raw dried flour. Dry-
ing the seeds with the testa yielded flour that was lightly brown in color
and slightly bitter while drying fully peeled breadnut shreds resulted into
off white colored flour with a higher sensory appeal and flavor although
manual peeling is highly intensive. The following protocol is recom-
mended for flour production:
The mature seeds are cooked in a pressure cooker (up to 3–4 whistles
are recommended). The boiled seeds are dried for 30 minutes at 50°C to
facilitate removal of the endocarp (brittle and easy to crack); the seeds are
peeled to remove the testa, reduced into shreds of 2 × 2 mm, dried at 50°C
until brittle, and grinded coarsely into granules or finely into flour.
Yield of dried breadnut seeds was estimated at 23–29% (seed weight
basis) at a moisture content of 8–10%. It was found that the dried bread-
nut when grinded could yield three categories of flour: granules similar to
broken wheat, coarse powder like semolina, and flour. The three types of
products with moisture content of 10–12% and packed in moisture proof
packages can keep above 12 months.
Breadnut: Innovative Products for the Agro Food Sector 65

5.3.6.2 Uses of Flour

The breadnut flour of different grades (coarse granules, similar to broken

wheat; small granules; and fine particles) were assessed into a range of
products (sweet and savory). For best results, it was found that rehydrating
the coarse and small granules in hot water for 1 h prior to cooking reduced
cooking time and improved taste. The recovery rate of the soaked granules
was three times more than the dried granules. The soaked granules and the
breadnut coarse granules were found to be highly suitable for the prepa-
ration of porridge (sweet or savory), and the dish was similar to oatmeal
or rice meal or “couscous.” The coarse breadnut granule was found to
be highly suitable for thickening soups and stews and preparation of dal
(Indian pulse-based dish) with high sensory value.
The coarse breadnut was also added to breadfruit-based burgers with a
view to improve as coating in the form of roasted granules.
With a view to improve protein content of breadfruit burger, breadfruit
burgers were coated with the (roasted) breadnut granules and breadfruit
mash was mixed with pre-soaked breadnut granules at 5%.
The smaller granules were found to be suitable for the preparation of
semolina-based dish and dessert cream. Both breadnut smaller granules
and flour after presoaking in hot water and blending were found to be
highly suitable for the preparation of a delicious milk-based beverage and
therefore constitute a good nutritional supplement.
The breadnut seeds flour can be used up to 50% as a supplement with
wheat flour for a wide range of prepared dishes like chapatti, tortillas,
pizza, cookies, and pastries with high acceptability.

5.3.6.3 Nutritional Value of Breadnut Flour

The proximate composition of breadnut flour is shown in Table 5.3 and

indicates that dried breadnut seeds are highly nutritious. These results
are comparable with those of previous studies (Ragone, 2003; Malamo
et al., 2011). The high carbohydrate (74.07 g) and energy value (350
kcal) confirm breadnut as a high-energy food. The protein content of
66 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

TABLE 5.3 Proximate Composition of Dried Breadnut Seeds (100 g)

Composition Breadnut seeds flour
Moisture (g) 11.25
Protein (g) 9.74
Fat (g) 1.65
Saturated fat (g) 38
Carbohydrate (g) 74.07
Ash (g) 2.27
Crude fiber (g) 1.02
Gluten (g) NIL
Reducing sugar (g) 1.90
Calcium (mg) 110
Magnesium (mg) 70
Phosphorus (mg) 260
Sodium (mg) 10
Potassium (mg) 390
Iron (mg) 1.42
Zinc (mg) 1.54
Copper (mg) 0.48
Manganese (mg) 0.28
Energy Kcal 350
Sample is analyzed in 2014.

breadnut seeds flour for Mauritius is much higher (9.74 g) than the one
reported by Malamo et al. (2011), which could be due to differences in
the production method and agro-climatic factors. Breadnut flour con-
tains a significant amount of minerals, particularly potassium (390 mg),
phosphorous (260 mg), calcium (110 mg), and magnesium (70 mg), and
low fat (1.65 g/100 g). Breadnut being gluten free can be a good healthy
and nutritious source of food for celiac patients. Breadnut flour can be
used as a nutritional supplement or for the preparation of food products
and as a supplemental flour with wheat and other flour for a wide num-
ber of uses.
Breadnut: Innovative Products for the Agro Food Sector 67

5.4 CONCLUSION

This study showed that breadnut seeds can be used as a vegetable, staple,
and healthy snack in the fresh form and has wide application in the pro-
cessed form either as processed products or as nutritional supplements.
Being gluten free, it is also highly suitable for use by celiac patients. Its
commercial cultivation in agro-forestry systems and exploitation will be
highly beneficial in terms of sustainable development and food nutrition
security.

KEYWORDS

•• breadnut
•• nutritional value
•• processing
•• uses

REFERENCES

Fagbemi, T. N., Oshodi, A. A., & Ipimmoroti, K. O., (2005). Processing effects on some
anti-nutritional factors and In vitro multienzyme protein digestibility (IVPD) of three
tropical seeds: Breadnut (Artocarpus altilis, cashewnut (Anacardium occidentale)
and Fluted Pumpkin (Telfairia occidentalis). Pakistan Journal of Nutrition, 4(4),
250–256.
Malomo, S. A., Eleyinmi, A. F., & Fashakin, J. B., (2011). Chemical composition, rheo-
logical properties and bread making potential of composite flours from breadfruit,
breadnut and wheat. African Journal of Food Science, 5(7), 400–410.
Mathews, R., Commissiong, E., Baccus-Taylor, G., & Badrie, N., (2001). Effect of peeling
methods on breadnut (Artocarpus altilis) seeds and acceptability of canned seeds in
brine. Journal of Food Science and Technology, 38(4), 402–404.
Negron de Bravo, E., Graham, H. D., & Padovani, M., (1983). Composition of breadnut
(seeded breadfruit), Caribb. J., Sci., 19(3–4), 27–32.
Quijano, J., & Arango, G. J., (1979). The breadfruit from Colombia - A detailed chemical
analysis, Econ. Bot., 33(2), 199–202.
Ragone, D., (2006). Species Profiles for Pacific Island Agroforestry. www. traditional tree.
org (accessed 15 March 2012).
68 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Roberts-Nkrumah, L. B., (2005). Fruit and seed yields in chataigne (Artocarpus camansi
Blanco) in Trinidad and Tobago. Fruits, 60(6), 387–393.
Roberts-Nkrumah, L., (2015). Breadfruit and Breadnut Orchard Establishment and Man-
agement (A manual of commercial production). FAO.
William, K., & Badrie, N., (2005). Nutritional composition and sensory acceptance of
boiled breadnut (Artocarpus camansi Blanco) seeds. Journal of Food Technology,
3(4), 546–557.
Woodroof, J. G., (1979). Tree Nuts, 2nd edn., AVI Publ. Co Inc.: Westport, Connecticut,
USA.
CHAPTER 6

STORAGE STUDY OF
JAMUN-AONLA BLENDED
READY-TO-SERVE BEVERAGES
PREETI SINGH, NEELIMA GARG, and SANJAY KUMAR
Central Institute for Subtropical Horticulture, Rehmankhera P.O.,
Kakori, Lucknow–226101, India, E-mail:
[email protected]

CONTENTS

Abstract.................................................................................................... 69
6.1 Introduction..................................................................................... 70
6.2 Materials and Methods.................................................................... 71
6.3 Results and Discussion................................................................... 72
6.4 Conclusion...................................................................................... 76
Acknowledgment..................................................................................... 76
Keywords................................................................................................. 76
References................................................................................................ 76

ABSTRACT

Jamun (Syzygium cumini) and aonla (Emblica officinalis) are two impor-
tant medicinal plants of India, and their respective juices are used for
curing number of health ailments. Juice blending is one of the methods
to improve the nutritional quality of the product. Ready-to-serve (RTS)
beverages were prepared by blending juices of jamun and aonla in three
70 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

ratios (9:1, 8:2, and 7:3). Pure aonla and jamun juice were kept as control.
Microbiological, biochemical, and sensory evaluations of RTS beverages
were carried out at 0 day and after 3 and 6 months of storage at 12 ± 2°C.
No microbial growth could be detected in any of the samples during the
storage period. Among the treatments, RTS beverage prepared from jamun
and aonla juices in 9:1 ratio was found to be the best on sensory score
(8.72 out of 9), followed by blend in the ratio of 8:2 (8.3 out of 9). Increas-
ing the aonla juice further reduced the acceptability of the RTS beverage.
After 6 months of storage, the RTS beverage blend of 9:1 ratio had 15.8
°B total soluble solids (TSS), 0.27% titratable acidity, 9.18 mg/100 mL
ascorbic acid, 6.17% reducing sugar, 23.15 mM/mL antioxidants (fluores-
cence recovery after photobleaching (FRAP) value), and 1.96 mg/100 mL
anthocyanin content compared to pure jamun juice (15.8 °B TSS, 0.32%
titratable acidity, 11.9 mg/100 mL ascorbic acid, 5.73% reducing sugar,
19.39 mM/mL antioxidants, and 2.29 mg/100 mL anthocyanins. The study
reflected that jamun–aonla blend in the ratio of 9:1 could be stored up to
6 months with higher antioxidant level and better sensory acceptability.

6.1 INTRODUCTION

Jamun (Syzygium cumini) is a minor fruit crop belonging to family Myrta-

ceae. It is considered to be indigenous to India and West Indies, being
cultivated in Philippines, West Indies, and Africa (Shrivastava and Kumar,
2009). It is gaining popularity among the rural as well as urban masses due
to its high nutraceutical values. The ripe berries are rich sources of iron and
pectin with a fair amount of ascorbic acid. It is used as an effective thera-
peutic medicine against diabetes, heart, and liver trouble (Garande and
Joshi, 1995). However, jamun fruit is highly perishable; the short shelf-life
of fruit makes it available only for a short period, which makes its popu-
larity unrealized. Jamun is very popular as a dessert fruit, because of its
slight astringent but sweet-sour taste and excellent color. In jamun grow-
ing regions, a glut of surplus fruit is available that needs to be processed
and converted into value-added products. As the fruit is a rich source of
anthocyanin, it imparts antioxidant properties too. Aonla (Emblica offici-
nalis Gaertn.), also known as Indian gooseberry, is famous for medicinal
Storage Study of Jamun-Aonla Blended Ready-to-Serve Beverages 71

values including antisorbutic, diuretic, laxative, and antibiotic properties.

The fruits also possess pronounced expectorant, antiviral, cardiotonic, and
hypoglycemic activity (Mehta and Tomar, 1979). Fresh aonla is too sour
to consume and hence is preferred in the form of preserves, dried aonla,
trifala, jam, juice, pickle and chavyanprash, toffees, and fruit bar (Singh
and Kumar, 1995). Although aonla juice and beverages have poor con-
sumer acceptance, it could be utilized for vitamin C enrichment of other
fruit juice-based beverages. These vitamin C and antioxidant-rich natural
drinks if given due publicity can replace synthetic drinks and overcome
the vitamin and antioxidant deficiency in people as well.

6.2 MATERIALS AND METHODS

The mature fully ripened jamun and aonla fruits procured from the experi-
mental farm of Central Institute for Subtropical Horticulture, Lucknow,
were collected in clean polythene bags and brought to laboratory. The
fruits were washed separately under running tap water, crushed in fruit
mill, and the juice was extracted by applying pressure of 1500–2000 kg/
square inch with a hydraulic press. Ready to serve beverages were pre-
pared by blending juices of jamun and aonla in ratios, viz., control jamun,
9:1 (T1), 8:2 (T2), 7:3 (T3), and control aonla. The treatments as well as
controls were adjusted with requisite proportion of water, sugar, and cit-
ric acid in order to maintain 10% juice, 14 °B total soluble solids (TSS),
and 0.24% acidity and pasteurized at 90°C for 1 min before packing in
sterilized glass bottles. The beverages were stored for 6 months at 12 ±
2°C. Biochemical analysis of jamun–aonla RTS beverage was carried out
for TSS by using hand refractometer (Erma, Japan), acidity, and ascor-
bic acid, according to the methods of Ranganna (2000). The antioxidant
property of juice in terms of FRAP values was determined according to
Benzie and Strain (1999). The amount of reducing sugars was determined
by the spectrophotometric method according to Folin and Wu (1920).
Microbiological quality of jamun–aonla RTS beverage was carried out as
described by Speck (1985). For judging the sensory attributes of the RTS
drink, sensory evaluation was conducted by a panel of seven semi-skilled
judges. The attributes considered in the scoring were color, clarity, aroma,
72 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

taste, tannin, astringency, freedom from acetic acid, sugar, and impression
evaluated (Amerine et al., 1965). The overall final rating was obtained by
calculating the average of scores.

6.3 RESULTS AND DISCUSSION

The microbial quality assessment of the jamun-aonla RTS beverage

revealed that all the treatments as well as controls were microbiologi-
cally safe during the storage period. No significant increase in TSS was
observed during storage (Table 6.1).
Slight increase in total acidity was observed at the end of 6 months of
storage (Table 6.1). Priyadevi et al. (2002) reported that pectic substances
are responsible for increasing the acidity of fruits. Hence, in the present
study, degradation of pectic substances into soluble solids might have
contributed toward increase in the acidity of jamun beverages. Ascorbic
acid content of all the treatments decreased continuously during the entire
period of storage (Table 6.1). Highest ascorbic acid (19.74 mg/100 mL)
was observed in pure aonla juice and lowest (4.23 mg/100 mL) in T3
(Table 6.1). This reduction might be due to oxidation of ascorbic acid into
dehydroascorbic acid by oxygen (Sethi et al., 1980). The reducing sugar
content slightly increased during storage of jamun–aonla RTS beverages
(Table 6.1). The increase is attributable to the hydrolysis of sucrose in
glucose and fructose by the acid present in the beverages (Lotha, 1992) or
gradual inversion of nonreducing sugars into reducing sugars in the acidic
medium (Malav et al., 2014). Anthocyanins and antioxidants decreased
in all treatments during storage. Among the beverages containing jamun,
the lowest anthocyanin content (1.54 mg/100 mL) was observed in T3
treatment (Figure 6.1), while the highest content (2.16 mg/100 mL) was
observed in pure jamun juice followed by T1 (1.90 mg/100 mL) after 6
months of storage. Total antioxidants content in the samples ranged from
19.39 to 55.49 mM/mL of jamun-aonla RTS at 0 day, being highest in pure
aonla juice RTS and lowest in pure jamun juice RTS (Figure 6.2).
Antioxidants gradually decreased after 6 months of storage. Raj et al. (2011)
have reported gradual decline in polyphenol contents from 332 to 305 mg %
in sand pear and pear-apple juice beverage during storage period of 6 months.
TABLE 6.1 Biochemical Analysis of Jamun–Aonla Blended Ready-to-Serve Beverage after Six Months of Storage
Parameters Storage period (Months) Treatments
Control jamun T1 T2 T3 Control aonla
0
T.S.S ( B) 0 15.8 15.8 15.8 15.8 15.6
2 15.8 15.8 15.8 15.4 15.6
4 15.8 15.8 15.8 15.4 15.8
6 16.0 16.0 15.8 15.6 15.8
Acidity (%) 0 0.24 0.24 0.24 0.24 0.23
2 0.24 0.29 0.31 0.27 0.27
4 0.26 0.30 0.32 0.32 0.30
6 0.29 0.30 0.33 0.38 0.34
Ascorbic acid 0 11.9 9.52 4.76 4.76 21.4
(mg/100ml) 2 11.22 9.18 4.59 4.59 20.4
4 11.01 8.89 4.44 4.37 19.88
6 10.34 8.46 4.37 4.23 19.74
Reducing sugar 0 5.73 6.01 5.61 5.73 5.53
(%) 2 5.77 6.17 5.67 5.80 5.71
Storage Study of Jamun-Aonla Blended Ready-to-Serve Beverages

4 5.85 6.21 7.71 5.83 5.76

6 5.96 6.23 5.77 5.88 5.81
73
74 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 6.1 (See color insert.) Changes in anthocyanin content during storage of
blended jamun-aonla ready-to-serve beverages.

FIGURE 6.2 (See color insert.) Changes in phenolics in blended jamun–aonla ready
to-serve beverages during storage.

Organoleptic analysis indicated treatment T1, RTS prepared from jamun

and aonla juices in 9:1 ratio, scored as best (8.72 out of 9) during storage
(Figure 6.3 and Plate 1), followed by blend in the ratio of 8:2 (8.3 out of 9).
Storage Study of Jamun-Aonla Blended Ready-to-Serve Beverages 75

FIGURE 6.3 (See color insert.) Sensory evaluation of blended jamun–aonla ready-to-
serve beverages during storage.

PLATE 1 Blended jamun–aonla ready-to-serve beverages.

76 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Increasing the aonla juice further reduced the acceptability of the RTS
beverage.

6.4 CONCLUSION

The study reflected that jamun–aonla blend in the ratio of 9:1 could be
stored up to 6 months at room temperature with higher antioxidant level
and better sensory acceptability.

ACKNOWLEDGMENT

The authors are thankful to Director, Central Institute for Subtropical

Horticulture, Lucknow, for his keen interest in the work and constant
support. The research was conducted under the Women Scientist program
funded by Department of Science and Technology, New Delhi.

KEYWORDS

•• anthocyanins
•• antioxidants
•• beverage
•• jamun
•• juice blends
•• ready-to-serve
•• syzygium cumini

REFERENCES

Amerine, M. A., Pangborn, R. M., & Roessler, E. B., (1965). Principles of Sensory Evalu-
ation of Food. New York, Academic Press.
Benzie, F. F., & Strain, J. J., (1999). Ferric reducing/antioxidant power assay: direct mea-
sure of total antioxidant activity of biological fluids and modified version for simul-
Storage Study of Jamun-Aonla Blended Ready-to-Serve Beverages 77

taneous measurement of total antioxidant power and ascorbic acid concentration.

Methods in Enzymology, 299, 15.
Folin, O., & Wu, H., (1920). Estimation of blood sugar by alkaline copper reducing method.
J., Biol. Chem., 41, 367.
Garande, V. K., & Joshi, G. D., (1995). Storage of jamun fruit products. Asian Food J.,
10, 54–56.
Lotha, R. E., (1992). Studies on processing and storage of kinnow mandarin juice. PhD,
Thesis, IARI, New Delhi.
Malav, M., Gupta, R., & Nagar, T., (2014). Studies on bio-chemical composition of orange
based blended ready-to-serve (RTS) beverages. Biosci. Biotech. Res. Comm., 7(1),
78–83.
Mehta, G. L., & Tomar, M. C., (1979). Studies on simplification of preserve making II.
Amla (Phyllanthus emblica L.). Indian Food Packer, 33 (5), 27–30.
Priyadevi, S., Thangam, M., & Desai, A. R., (2002). Studies on variability in physico-
chemical characters of different jamun (Syzygium cuminii). Indian J., Hort., 59,
153–156.
Raj, D., Sharma, P. C., & Vaidya, D., (2011). Effect of blending and storage on quality
characteristics of blended sand pear-apple juice beverage. J., Food Sci. Technol.,
48(1), 102–105.
Ranganna, S., (2000). Handbook of Analysis and Quality Control for Fruit and Vegetable
Products. IInd edn. Tata Mc Graw Hill Publication Co Ltd, New Delhi, pp. 1112.
Sethi, V., Anand, J. C., & Saxena, S. K., (1980). Kinnow orange in juice and beverage mak-
ing. Indian Hort., 25, 13.
Shrivastava, R. P., & Kumar, S., (2009). Fruit and Vegetable Preservation. Principles and
practices, IBDC, New Delhi.
Singh, I. S., & Kumar, S., (1995). Studies on processing of aonla fruits II. Aonla products.
Prog Horticult., 27(1/2), 39–47.
Speck, M., (1985). Compendium of methods for the microbiological examination of foods.
Second edition, American Public Health Association Inc., pp. 644–649.
CHAPTER 7

DEVELOPMENT OF BLENDED AONLA

SQUASH
NEELIMA GARG, SANJAY KUMAR, PREETI SINGH,
and KAUSHLESH K. YADAV
Central Institute for Subtropical Horticulture Rehmankhera,
P.O. Kakori, Lucknow–227107, India,
E-mail: [email protected]

CONTENTS

Abstract.................................................................................................... 79
7.1 Introduction..................................................................................... 80
7.2 Materials and Methods.................................................................... 81
7.3 Results and Discussion................................................................... 82
Acknowledgment..................................................................................... 84
Keywords................................................................................................. 85
References................................................................................................ 85

ABSTRACT

Aonla (Emblica officinalis), despite being a highly nutritious and thera-

peutically important fruit, is highly acidic. Beverages prepared from aonla
have low acceptability owing to their high acidity. Blending appears to
be one of the tools for the preparation of acceptable beverage from aonla.
Litchi possesses pleasant aroma, while grape contains good flavor in addi-
tion to attractive color in purple variety. In the current study, blended
squash from aonla was prepared with litchi and grape (purple variety)
80 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

separately in 3:1 and 1:1 ratio each. The ascorbic acid and total pheno-
lic content was highest in aonla–litchi squash with 3:1 ratio (152 mg/100
g and 652 mg/100 g, respectively). In general, the ascorbic acid content
decreased while phenolics increased after 6 months of storage at 12 ± 2°C.
The anthocyanin content of aonla–grape squash was 3.7 and 7.7 mg/100g
in 3:1 and 1:1 blends, respectively, which declined to 1.2 and 2.9 mg/100
g, respectively, after 6 months of storage. Initial sensory evaluation of the
product revealed that aonla–litchi (1:1) blend was liked most, scoring 8.4
out of 9, followed by aonla–grape (1:1) with 8.1 score. Sensory scoring
of squashes after 3 and 6 months of storage also indicated higher prefer-
ences for 1:1 blend than for 3:1 blend in both aonla–litchi and aonla–grape
squashes. It may be inferred that 50% litchi or grape juice could be mixed
with aonla juice to obtain acceptable quality of squash.

7.1 INTRODUCTION

Aonla (Emblica Officinalis, Gaertn), commonly known as Indian goose-

berry, belongs to family Euphorbiaceae. India produces a huge quantum
of aonla as the tree is well adapted to various kinds of soils and climatic
conditions. The glistening, translucent, pale green fruits are highly fibrous
in nature. The astringent flesh contains fairly rich amount of vitamin C
and polyphenols and has high antioxidant property. The ascorbic acid in
aonla is considered highly stable, apparently protected by tannins, which
retards oxidation (Morton, 1987). The juice extracted from aonla fruit,
despite having unparallel nutritional and medicinal qualities, is not suit-
able for fresh consumption due to lack of any flavor, high acidity, and
astringent taste. Even beverages prepared from aonla juice are not very
appealing due to poor sensory properties. Acceptability of aonla drink,
however, could be drastically increased by addition of juices of some other
fruits possessing attractive color, pleasant aroma, and taste. Fruits like
litchi (Litchi chinensis) and grape (Vitis vinifera) may find good suitability
in this context. Both the fruits are easily available in the market. Litchi
bears a good blend of taste and aroma. Similarly, purple varieties of grape
possess attractive color due to the presence of anthocyanin pigments. It
Development of Blended Aonla Squash 81

also has soothing taste and flavor. The beverages from these two fruits are
already well established in the market and have good public demand.
In the present investigation, an attempt has been made to enhance the
sensory qualities of aonla squash through blending of litchi or grape juice
and evaluate the biochemical and sensory qualities of products during
storage.

7.2 MATERIALS AND METHODS

Aonla, litchi, and grape fruits were brought from Central Institute for
Subtropical Horticulture experimental farm located at Rehmankhera, Luc-
know. Healthy fruits were selected and washed thoroughly with tap water.
Whole aonla fruits were subjected to fruit mill, and coarse pulp contain-
ing ground seeds was obtained. The pulp was wrapped in thick cloth and
pressed in a hydraulic press to extract juice. The filtered juice was pasteur-
ized at 90°C and preserved with 500 ppm SO2 in the form of potassium
metabisulfite. Litchi fruits were peeled, stones separated manually, and
the pulp was homogenized in a blender. It was then squeezed through a
muslin cloth and the juice was collected. The juice was pasteurized and
preserved like aonla juice. Healthy grape berries were wrapped in thick
cloth and pressed in a hydraulic press to obtain juice. Grape juice was also
pasteurized in the similar manner and preserved with 500 ppm sodium
benzoate. Aonla juice was blended with litchi and grapes juice separately
in 3:1 and 1:1 ratios, and a squash was prepared from each of the blends
using 40% juice, 50% sugar, and 0.9% acidity. The final combinations
were as follows:
1. Aonla-litchi (3:1): Aonla-Litchi I
2. Aonla-litchi (1:1): Aonla-Litchi II
3. Aonla-grape (3:1): Aonla-Grape I
4. Aonla-grape (1:1): Aonla-Grape II
The squashes were filled hot in glass bottles, sealed, and stored in dry
and cool place. They were analyzed for various biochemical and sensory
parameters at 0, 3, and 6 months of storage at 12 ± 2°C.
82 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

The total soluble solids (TSS) of squashes were recorded using a

hand refractometer (Erma, Japan). Titratable acidity, ascorbic acid, and
total phenolics were determined according to the methods described by
Ranganna (2000). Ascorbic acid content of beverage was measured by
titrating samples against dye (2, 6-dichloro phenol indophenol, sodium
salt) solution, while total phenolics were estimated spectrophotometri-
cally using Folin–Ciocalteu’s reagent. The microbial examination of fer-
mented beverage was carried out according to the method detailed by
Speck (1985). The organoleptic evaluation of the beverage was carried
out on the basis of color, flavor, and taste by a panel of semi-skilled
judges by using a 9-point hedonic scale as reported by Amerine et al.
(1965).

7.3 RESULTS AND DISCUSSION

The microbial examination carried out at 0, 3, and 6 months of storage

revealed no microbial growth at any stage in any sample. The TSS of
squash samples ranged from 50 to 55 °B, while titratable acidity ranged
from 0.92% to 0.95% at 0 day. Minor changes in these values were
observed during storage. The ascorbic acid content was found to be the
highest in 3:1 aonla–litchi blend (152 mg/100 mL), while lowest was in
1:1 aonla-grape blend (92 mg/100 mL). The amount of ascorbic acid in
the samples was directly proportional to the volume of aonla juice added,
as it was the main source of ascorbic acid. Decrease in the ascorbic acid
content to an extent 33–35% and 24–29% was observed in aonla–litchi
and aonla–grape blends, respectively, after 6 months of storage at 12 ±
2°C (Figure 7.1).
Decrease in ascorbic acid content of aonla juice was also reported by
Bhattacherjee et al. (2013). Loss in ascorbic acid is attributed to oxida-
tion of ascorbic acid molecules during storage. The total phenolic content
in the samples ranged from 443 to 652 mg per 100 mL of squash at 0
day, being highest in 3:1 aonla–litchi blend and lowest in 1:1 aonla–litchi
blend. It increased to a range of 613 to 875 mg per 100 mL after 6 months
of storage (Figure 7.2).
The increase in phenolic content might be due to gradual solubiliza-
tion of phenolics or transformation of molecules into some other forms
Development of Blended Aonla Squash 83

FIGURE 7.1 (See color insert.) Changes in ascorbic acid content of blended aonla
squash during storage.

during storage. The total anthocyanin content of aonla–grape squash

was 3.7 and 7.7 mg/100g in 3:1 and 1:1 blends, respectively, which
declined to 1.2 and 2.9 mg/100 g, respectively, after 6 months of storage
(Figure 7.3).

FIGURE 7.2 (See color insert.) Changes in total phenolic content of blended aonla
squash during storage.
84 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 7.3 (See color insert.) Changes in anthocyanin content of aonla–grape squash
during storage.

Decrease in anthocyanins was also observed by Yadav et al. (2014) in

mulberry juice during storage. The sensory evaluation of squash, carried
out at 0 day by semi-skilled judges on the basis of color, flavor, and taste
revealed that aonla–litchi (1:1) blend was liked most, scoring 8.4 out of 9,
followed by aonla–grape (1:1) with 8.1 score (Figure 7.4).
Similar trends were observed during sensory scoring of squashes after
3 and 6 months of storage, indicating higher preferences for 1:1 blend than
for 3:1 blend in both aonla–litchi and aonla–grape squashes. This indi-
cated that the addition of higher quantity of litchi or grape juice resulted
in better quality of product due to enhanced color, flavor, and taste. It may
therefore be concluded from the study that 50% litchi or grape juice could
be mixed with aonla juice to obtain an acceptable quality of squash with
improved color, flavor, and taste.

ACKNOWLEDGMENT

The authors are thankful to Director, Central Institute for Subtropical Hor-
ticulture, Lucknow, for providing infrastructure facility for carrying out
the research work.
Development of Blended Aonla Squash 85

FIGURE 7.4 (See color insert.) Changes in sensory scores of blended aonla squash
during storage.

KEYWORDS

•• aonla
•• beverages
•• grape
•• litchi
•• sensory qualities

REFERENCES

Amerine, M. A., Pangborn, R. M., & Roessler, E. B., (1965). Principles of Sensory Evalu-
ation of Food. Academic Press. New York and London.
Bhattacherjee, A. K., Dikshit, A., Kumar, S., Shukla, D. K., & Tandon, D. K., (2013).
Quality evaluation in storage of aonla (Emblica officinalis Gaertn.) juice extracted
from fruits preserved by steeping in water. Internatl. Food Res. J., 20(4), 1861–1865.
Morton, J., (1987). Emblic., In: ‘Fruits of Warm Climate’, by Julia, F., Morton, Miami,
FL., pp. 213.
Ranganna, S., (2000). Handbook of Analysis and Quality Control for Fruit and Vegetable
Products. IInd edn. Tata Mc Graw Hill Publication Co Ltd, New Delhi, pp. 1112.
Speck, M. L., (1984). Compendium of Methods for the Microbiological Examination of
Foods. American Public Health Association, Washington DC.
Yadav, P., Garg, N., & Kumar, S., (2014). Improved shelf stability of mulberry juice by
combination of preservatives. Indian J., Natural Products Resources, 5(1), 62–66.
CHAPTER 8

SACCHAROMYCES CEREVISIAE
POSTHARVEST DIP TREATMENT
FOR IMPROVING QUALITY AND
STORABILITY OF MANGO cv.
DASHEHARI
BHARATI KILLADI, NEELIMA GARG, REKHA CHAURASIA,
KAUSHLESH K. YADAV, and D. K. SHUKLA
Division of Post Harvest Management,
ICAR–CISH Lucknow 226106, India,
E-mail: [email protected]

CONTENTS

Abstract.................................................................................................... 87
8.1 Introduction..................................................................................... 88
8.2 Materials and Methods.................................................................... 89
8.3 Results and Discussion................................................................... 91
8.4 Conclusion...................................................................................... 99
Keywords................................................................................................. 99
References.............................................................................................. 100

ABSTRACT

Bio-agents are natural antagonists capable of inhibiting and keeping the

target pathogens at low level, besides being nontoxic and environmen-
tally safe. Anthracnose is one of the major postharvest diseases of mango.
88 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

The use of chemical pesticide to treat this disease is avoided from food
safety point of view. The objective of the present study is to understand
the use of Saccharomyces cerevisiae as a bio-agent for controlling spoil-
age and improving the quality of mangoes. Mature green fruits of Dashe-
hari were treated with three strains of S. cerevisiae as (T1), (T2), and (T3)
@ 108 cells/mL for 10 minutes and control (T4) dip treated with water and
stored under ambient conditions (34 ± 2°C and 85% to 90% RH). Fruits
were assessed for physico-chemical parameters at regular intervals of 0,
4, 6, 8, and 10 days. The cumulative physiological loss in weight (CPLW)
was highest in T4 (12.87%) followed by T2 (12.44%), T1 (12.41%), and
T3 (11.57%) on the 10th day of storage. The total soluble solids (TSS)
increased and titratable acidity (TA) and firmness of the fruits decreased
during the course of fruit storage. The total carotenoid content was high-
est in T2 (7.22 mg/100 g), followed by T3 6.88 (mg/100 g), T1 (5.43 mg/100
g) and T4 (4.88 mg/100 g) on the 10th day of storage. The antioxidant
content estimated by fluorescence recovery after photobleaching (FRAP)
was maximum in T1 (4602.22 µmolar trolox equivalent/g), followed by
T2 (3353.02 µmolar trolox equivalent/g) and T3 (2975. 24 µmolar trolox
equivalent/g) on the 10th day of storage. Percent inhibition of antioxidants
estimated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) was maximum in T3
(76.47%), followed by T1 (61.52%) and T2 (58.36%) on the 10th day of
storage. The spoilage of the fruit was minimum in T3 (7.33%), followed
by T2 (11.33%), T1 (11.66%), and T4 (15.33%) on the 10th day of storage.
In vitro studies on the efficacy of S. cerevisiae for controlling Colletotri-
chum gloeosporioides (pathogen for anthracnose) confirmed the efficacy
of the bio-agent. Thus, S. cerevisiae may be used as a potential bio-agent
for improving the postharvest quality of mango.

8.1 INTRODUCTION

Mango (Mangifera indica L.) is one of the major and important fruits of
the tropics and subtropical areas. India is the largest producer of mango,
with the production of about 12 million tons of fruits. About 21 million
tons of vegetables are spoiled each year, with a total estimated value of
240 billion rupees. India loses about 35–40% of the produce estimated
Saccharomyces cerevisiae Postharvest DIP Treatment 89

at Rs. 40,000 crores per year due to improper postharvest management.

Postharvest pathogen developed may be considered as a minor problem
for local markets with short time period of harvest and selling; however,
when the fruit is exported to foreign countries, prolonged storage requires
control of the postharvest pathogens. Mango cv. Dashehari is harvested
in the month of June, with the onset of monsoon accompanied with high
temperature and relative humidity.
Postharvest pathogens are the causes of loss during storage of fruits.
The most important and major postharvest pathogen of mango is anthrac-
nose. The primary means to control postharvest disease of fruits is the use
of synthetic fungicides. Yeasts have been explored as one of the promising
alternatives for the control of postharvest pathogen (Rosa et al., 2010; Liu
et al., 2013). Biocontrol of postharvest spoilage in mango by antagonistic
microorganisms seems promising in reducing the use of synthetic fungi-
cides (Lima et al., 1999; Janisiewicz and Korsten, 2002). Incidence and
severity of postharvest disease significantly reduced in mango (Kefalew
and Ayalew, 2008). Several yeasts have been reported as bio-control
agents in sweet cherry (Wang and Tian, 2008); apple (Yu et al., 2008);
jujube fruit (Cao et al., 2012), pear fruit (Yu et al., 2012); mandarins (Guo
et al., 2014), and mango (Bautista-Rosales et al., 2014). S. cerevisiae has
been used for postharvest management of postharvest diseases of grape
(Suzzi et al., 1995) and apple (Scherma et al., 2003). Antifungal activ-
ity of S. cerevisiae and S. pombe against Botrytis cineraria on grape was
reported for the first time (Nally et al., 2012). In the present investigation,
S. cerevisiae was used as pretreatment for improving the quality and stor-
ability of mango cv. Dashehari.

8.2 MATERIALS AND METHODS

S. cerevisiae strains were obtained from the culture collection of microbiol-

ogy laboratory in the Division of Post-Harvest Management, CISH (Central
Institute for Subtropical Horticulture). The yeast culture was maintained
on yeast potato dextrose agar (YPDA). Three yeast strains, viz., bakers’
yeast, industrial yeast, and S. cerevisiae were tested for anti-pathogenicity
90 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

for Colletotrichum gloeosporioides according to the standard protocol by

dual culture technique, and growth inhibition was measured.
Yeast strains were multiplied on YPDA medium, and the plates were
incubated at 30 ± 2°C for 3 days. The cells were collected using a cell
scrapper, and the final cell number was maintained @108 cells/mL in dis-
tilled water.
Green hard fruits of mangoes cv. Dashehari were harvested with stalks
of 8–10 mm; they were washed with water and dip treated for 10 minutes
with three yeast strains of S. cerevisiae, viz., T1, T2, T3, and T4 (control).
The fruits were surface dried, packed, and stored under ambient condi-
tions (34 ± 2°C and 85% to 90% RH). The observations of physiochemi-
cal parameters were recorded at an interval of 0, 4, 6, 8, and 10 days of
storage. The fruit weight was recorded at the time of packaging and subse-
quently at each withdrawal. The difference in weight was expressed as per-
cent weight loss. After withdrawal of fruits from each treatments, cell cfu/
mL microbial load were counted by serial dilution method with observed
effect on spoilage fruit. Firmness of the fruit was measured using a pene-
trometer (8 mm probe, USA) and expressed as kg/cm2. Total soluble solids
(TSS) was measured using a digital refractometer, model PAL-1 (Atago,
Tokyo, Japan). Titratable acidity (TA) was estimated by the methodology
propounded by Rangana (2000). Five grams of sample was diluted with 50
mL of distilled water and titrated with 0.1 mol/L NaOH solution, and the
results were expressed as percent citric acid.
Fruit pulp was macerated for the estimation of total carotenoids accord-
ing to the methods of Rangana (2000). Samples of 2 g each in triplicates
were extracted in 15 mL acetone thrice and filtered through cotton wool
in a conical flask. The extraction was done till colorless. Petroleum ether
(15 mL) was added to the extract and diluted with 2% (15 mL) sodium
chloride solution. All the extracts were transferred in a separating funnel
and washed with 10 mL of 2% sodium chloride. The nonaqueous layer
was extracted and collected in a 50-mL volumetric flask, and the volume
was made up with 3% acetone in petroleum ether; the observations were
recorded at 452 nm and expressed as mg/100 g.
The FRAP assay was done according to the methodology of Benzie
and Strain (1996). The reduction of a ferric–tripyridyltriazine complex to
its ferrous, colored form in the presence of antioxidants is the principle
Saccharomyces cerevisiae Postharvest DIP Treatment 91

of the assay. The FRAP agent contained 2.5 mL of a 10 mmol/L TPTZ

(2,4,6-tripyridy-s-triazine, Sigma) solution in 40 mmol/L HCL plus 2.5
mL of 20 mmol/L FeCl3 and 25 mL of 0.3 mol/L acetate buffer, pH 3.6 and
was prepared freshly and warmed at 37°C. Aliquots of 40 μL sample super-
natant were mixed with 0.2 mL distilled water and 1.8 mL FRAP reagent,
and the reaction mixture was incubated at 37°C for 10 min. Absorbance
measured spectrophotometrically at 593 nm. The standard solution used
was 1 mmol/L Trolox, and the final result was expressed as the concentra-
tion of antioxidants µmol trolox equivalent/g. If the FRAP value measured
was beyond the linear range of standard curve, then adequate dilutions
were made.
The 2,2-diphenyl-1-picrylhydrazyl (DPPH) estimation was done
according to the method of (Brand-Williams et al., 1995). DPPH was
weighed (24 mg) and dissolved in 100 mL methanol; this served as stock
solution and was stored at −20°C until needed. The working solution was
obtained by mixing 10 mL of stock solution with 45 mL methanol to get
an absorbance of 1.1 ± 0.02 units at 515 nm using the spectrophotometer.
Fruit extracts of 150 μL were allowed to react with 2850 mL of DPPH
solution for 24 h in the dark. Then, the absorbance was read at 515 nm.
The standard was a linear curve between 25 and 800 µM Trolox. Addi-
tional dilutions were made if the DPPH value measured was over the lin-
ear range of the standard curve.
Fruits were evaluated for spoilage according to McDonald et al. (1998)
by measuring the rotten area in relation to the total surface area and
expressed as percentage.
Surface microbial counts were monitored at 2 days intervals according
to the method of Collins and Lyne (1984).
All the analysis was carried out in triplicates, and the data recorded
during the course of investigation were subjected to statistical analysis by
SAS 9.3 and CD at the significance level of 0.05.

8.3 RESULTS AND DISCUSSION

Results from the dual culture assay showed that all antagonistic microor-
ganisms inhibited the mycelial growth of C. gloeosporioides with vary-
92 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

ing efficiencies. Yeast antagonists presented a moderate inhibition against

the majority of Colletotrichum isolates. Treatments of yeast significantly
exhibited strong antagonism against the isolates of C. gloeosporioides and
were found maximum growth inhibition by effect of baker yeast (12 mm),
industrial yeast (7 mm), S. cerevisiae (9 mm) compared with T4 as control
(18 mm), respectively.
The cumulative physiological loss in weight percent (CPLW %) dif-
fered significantly (p ≤ 0.5) among the treatments and the storage period
(Figure 8.1).
The CPLW percent increased with an increase in storage period. The
CPLW was maximum (13.08%) in control followed by T1 (12.87%) and
T2 (12.44%) and minimum in T3 (11.27%) on the 10th day of storage. The
increase in CPLW may be due to evapo-transpiration from the surface of
fruits. Fruit transpiration vary significantly according to climactic condi-
tions and characteristic of fruit that affects water balance; these findings
are in concomitance with the findings of Leonadi et al. (1999) in tomato.
There was a significant difference (p ≤ 0.5) in the firmness of fruits
among the treatments and the storage period (Figure 8.2).

FIGURE 8.1 (See color insert.) Effect of yeasts on the CPLW percent of mango cv.
Dashehari during storage. T1 (baker’s yeast), T2 (industrial yeast), T3 (Saccharomyces
cerevisiae), and T4 (control).
Saccharomyces cerevisiae Postharvest DIP Treatment 93

FIGURE 8.2 (See color insert.) Effect of yeasts on the firmness (kg/cm2) of mango
cv. Dashehari during storage. T1 (baker’s yeast), T2 (industrial yeast), T3 (Saccharomyces
cerevisiae), and T4 (control).

With the increase in storage period, the firmness of the fruits decreased.
The firmness was maximum in T3 (0.45 kg/cm2), followed by T1 and T2,
while minimum firmness was noted in T4 (0.32 kg/cm2) on the 10th day of
storage. The decrease in firmness may be due to rapid loss of cell structure
because of evapo-transpiration and water loss, causing ripening of fruits. As
the ripening stages advanced, fruit softening increased as reported by Raz-
zaq et al. (2013) in “Samar Bahisht Chaunsa” mangoes. Breakdown of cell
wall polymers occurs during fruit ripening, resulting in softening of mango
fruits (Zaharah and Singh, 2011). Firmness is affected by cell wall modifica-
tion, and polygalacturonase and pectin methyl esterase degradation activity
increased during ripening (Gonzalez-Aguilar et al., 2008). Similar reports
on firmness of “Ataulfo” mango fruits (Robies-Sanchez et al., 2009b).
The growth and development of microbes increased with an increase in
storage period (Table 8.1).
The spoilage of fruits increased with an increase in storage period and
varied significantly (p ≤ 0.5) among the treatments (Figure 8.3).
Maximum spoilage (15.33%) was observed in T4, followed by T2
(12.03%), T1 (11.33%), and minimum in T3 (7.99%) on the 10th day of stor-
94 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

TABLE 8.1 Effects of Yeasts on Microbial Growth during Storage and Ripening of
Fruits
Treatments Bacterial, cfu/gm Yeast, cfu/gm Fungus
0th day
Baker Yeast 45 8.1 × 106 0
Industrial Yeast 33 8.23 × 106 0
S. cerevisiae 20 9.16 × 10 6
0
Control 1.10 × 102 0 0
4th day
Baker Yeast 2.85 × 102 5.31 × 104 0
Industrial Yeast 36 9.52 × 10 4
0
S. cerevisiae 1.21 × 102 3.10 × 105 0
Control 8.51 × 102
12 0
6th day
Baker Yeast 5.62 × 105 6.25 × 103 0
Industrial Yeast 6.23 × 102
5.51 × 10 4
0
S. cerevisiae 2.13 × 104 7.36 × 103 0
Control 7.71 × 107
26 1
8th day
Baker Yeast 3.46 × 106 5.37 × 102 0
Industrial Yeast 2.41 × 103 4.35 × 103 0
S. cerevisiae 6.35 × 105
2.25 × 10 2
0
Control 8.31 × 108
65 4
10th day
Baker Yeast 9.74 × 105 8.35 × 102 5 – Ripening
Industrial Yeast 4.93 × 105 5.68 × 102 0 – Ripening
S. cerevisiae 2.57 × 106
3.63 × 10 2
1 – Ripening
Control 7.43 × 109 0 10 & spoilt

age. In control fruits, spoilage was noticed from the 6th day of storage, while
in T1 and T2, the spoilage of fruits was noticed on the 8th day of storage. The
spoilage was mainly due to anthracnose and stem end rot. S. cerevisiae has
been recognized as GRAS (generally recognized as safe) in Bio-safety level
1 in Europe (Murphy and Kavaragh, 1999). The main mode of action of
this yeast is competition for space and nutrients. Competition between yeast
and fungi was previously reported for grapes (Mc Laughlin et al., 1992),
Saccharomyces cerevisiae Postharvest DIP Treatment 95

FIGURE 8.3 (See color insert.) Effect of yeasts on the spoilage (%) of mango cv.
Dashehari during storage. T1 (baker’s yeast), T2 (industrial yeast), T3 (Saccharomyces
cerevisiae), and T4 (control).

apple (Filonow et al., 1996; Ippolito et al., 2000) and tomato (Kalogiannis et
al., 2006). The mechanism of action S. cerevisiae is by production of more
pseudohyphae and invasive growth than the food and industrial strains (de
Lianos et al., 2006). Mode of action of antagonistic yeasts are utilized for the
management of postharvest fungal diseases of fruits as reviewed by Liu et al.
(2013). Siderophore production was the antifungal pattern observed for the
consistent control of gray and sour rot with Saccharomyces as reported by
Nally et al. (2015). The production of volatile organic compounds (VOCs)
with in vitro and in vivo inhibitory effect on pathogen growth was observed
for S. cerevisiae as studied by Parafita et al. (2015).
The TSS and TA of the fruits significantly varied (p ≤ 0.5) among the
treatments (Figures 8.4 and 8.5).
With the increase in storage period, the TSS increased, while the TA
decreased among all the treatments. The TA was highest in T3 (0.10%),
while it was at par in T1, T2, and T4. TSS was highest in T1 (24.07 °B),
while it was at par in T2 and T4 at the end of the storage. TSS was lowest
in T3 (21.58 °B) on the 10th day of storage. Increase in sugar content and
decrease in TA were observed during ripening of mango cv. “Cogshall”
96 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 8.4 (See color insert.) Effect of yeasts on the TSS (°Brix) of mango cv.
Dashehari during storage. T1 (baker’s yeast), T2 (industrial yeast), T3 (Saccharomyces
cerevisiae), and T4 (control).

FIGURE 8.5 (See color insert.) Effect of yeasts on the titratable acidity (%) of mango
cv. Dashehari during storage. T1 (baker’s yeast), T2 (industrial yeast), T3 (Saccharomyces
cerevisiae), and T4 (control).
Saccharomyces cerevisiae Postharvest DIP Treatment 97

(Jacques et al., 2009). During ripening, the increase in TSS is attributed to

free sugar accumulation from the hydrolysis of starch (White, 2002).
There was a significant difference (p ≤ 0.5) in the total carotenoid con-
tent among the treatments and the storage period (Figure 8.6).
The total carotenoid content was maximum in T2 (7.22 mg/100 g), fol-
lowed by T3 (6.88 mg/100 g) and T1 (5.43 mg/100 g) and minimum in T4
(4.88 mg/100 g) on the 10th day of storage. Increase in carotenoid biosyn-
thesis in mango varieties is associated with the increase in respiration,
which is ethylene dependent (Saltveit, 1999).
Antioxidants varied significantly (p ≤ 0.5) among the treatments and
the period of storage. The antioxidants estimated by the FRAP method
(Figure 8.7) did not follow a particular trend as indicated by the carot-
enoid content of the fruits. It was maximum in T2 (7197.46 µmolar tro-
lox equivalent/g), followed by T3 (5011.75 µmolar trolox equivalent/g) on
the 8th day of storage; thereafter, it decreased. Antioxidants estimated by
DPPH (Figure 8.8) and expressed as percent inhibition was highest in T2
(77.44%), followed by T1 (72.62%) on the 8th day of storage; thereafter, it
decreased. Percent inhibition was highest in T3 (76.48 %) on the 10th day

FIGURE 8.6 (See color insert.) Effect of yeasts on the total carotenoids (mg/100
g) of mango cv Dashehari during storage. T1 (baker’s yeast), T2 (industrial yeast), T3
(Saccharomyces cerevisiae), and T4 (control).
98 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 8.7 (See color insert.) Effect of yeasts on the antioxidant FRAP (µmolar
TE/g) of mango cv. Dashehari during storage. T1 (baker’s yeast), T2 (industrial yeast), T3
(Saccharomyces cerevisiae), and T4 (control).

FIGURE 8.8 (See color insert.) Effect of yeasts on the antioxidant DPPH (% inhibition)
of mango cv. Dashehari during storage. T1 (baker’s yeast), T2 (industrial yeast), T3
(Saccharomyces cerevisiae), and T4 (control).
Saccharomyces cerevisiae Postharvest DIP Treatment 99

of storage. The antioxidant potential of mango varieties could be due to

complicated bioactive compounds and its synergistic actions contributing
to the antioxidant activity in mango (Liu et al., 2013). Total antioxidant
scavenging activities linearly increased up to 7 days and then decreased in
Chaunsa mango (Razzaq et al., 2013).

8.4 CONCLUSION

The CPLW was highest in untreated control and least in S. cerevisiae-

treated fruits on the 10th day of storage. The TSS increased and TA and
firmness of the fruits decreased during the course of fruit storage. The total
carotenoid contents were highest in yeast-treated fruits and minimum in
control fruits on the 10th day of storage. The antioxidant content estimated
was maximum in yeast-treated fruits on the 10th day of storage. The spoil-
age of the fruit was minimum in S. cerevisiae-treated fruits compared to
control fruits on the 10th day of storage. In vitro studies on the efficacy of
S. cerevisiae for controlling C. gloeosporioides (pathogen for anthracnose)
confirmed the efficacy of the bio-agent. Thus, S. cerevisiae may be used
as a potential bio-agent for improving the postharvest quality of mango.

KEYWORDS

•• antioxidants
•• carotenoids
•• post-harvest
•• quality
•• Saccharomyces cerevisiae
•• storability
100 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

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Gonzalez-Aguilar, G. A., Celis, J., Stotelo-Mundo, R. R., De La Rosa, Rodrigo-Garcia, J.,
& Alvarez-Parrilla, E., (2008). Physiological and biochemical changes of different
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Guo, J., Fang, W., Lu, H., Zhu, R., Lu, L., Zheng, X., & Yu, T., (2014). Inhibition of
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Ippolito, A. E. L., Ghaouth, A., Wilson, C. L., & Wisniewski, M., (2000). Control of post-
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Janisiewicz, W. J., & Korsten, L., (2002). Biological control of post harvest diseases of
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Kalogiannis, S., Tjamos, S. E., Stergiou, A., Antoniou, P. P., Ziogas, B. N., & Tjamos,
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CHAPTER 9

POSTHARVEST QUALITY
EVALUATION OF WINTER ANNUALS
SELLAM PERINBAN,1 BABITA SINGH,1 JAYOTI MAJUMDER,2 and
PUJA RAI1
1
Directorate of Floricultural Research, Indian Agricultural Research
Institute, Pusa Campus, New Delhi–110012, India,
E-mail: [email protected]
Bidhan Chandra Krishi Viswavidyalaya (BCKV), Kalyani,
2

West Bengal, India

CONTENTS

Abstract.................................................................................................. 103
9.1 Introduction................................................................................... 104
9.2 Materials and Methods.................................................................. 105
9.3 Results and Discussion................................................................. 107
9.4 Conclusion.....................................................................................113
Keywords................................................................................................114
References...............................................................................................114

ABSTRACT

Most of the winter annual flowers have very attractive flowers, and
these winter annuals are generally grown as land cover or border plants
in gardens. Due to their unique flower qualities, striking colors, and
seasonality, many of these flowers are sold in the retail markets as cut
104 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

flowers. Hence, this work was carried out at the Directorate of Flori-
cultural Research, IARI, New Delhi, during 2013–2014 to evaluate the
postharvest keeping quality of winter annuals. Five winter annual flow-
ers, namely antirrhinum, dimorphotheca, lupin, larkspur, and Sweet
William, were evaluated for vase life under five different preserva-
tives like sucrose, 8-hydroxyquinoline citrate (8-HQC), aminooxy ace-
tic acid (AOA), benzyl adenine (BA), and aluminum sulfate (Al2SO4)
in different combinations. From the results, it was observed that in
all flowers, the treatments with plant bioregulators (AOA & BA) and
aluminum sulfate significantly improved the vase life of flowers than
other treatments. Vase life of flowers treated with BA was maximum in
antirrhinum (9.3 days), dimorphotheca (8 days), and larkspur (8 days);
lupin treatment with 8-HQC improved the vase life up to 6 days, and
in Sweet William, the vase life of 9.67 days was observed after treat-
ment with AOA. Maximum stem elongation was observed in lupin (19
cm) after treatment with Al2SO4, whereas maximum flower diameter
of dimorphotheca was recorded in treatment with AOA. Other quality
parameters such as flower weight, percentage flower opening, and vase
solution uptake rate (VSUR) were significantly improved in treatments
with AOA, BA, and Al2SO4. Based on these results, it was evident that
among the annual flowers evaluated, antirrhinum and Sweet William
were found to be most suitable for commercial use as cut flower on the
basis of their vase life.

9.1 INTRODUCTION

Annual flowers in general are a group of herbaceous plants that grow

from seeds and complete their life cycle within 1 year or one season. In
particular, winter annuals provide a beautiful display of colors in the gar-
den during winter season in India. They enhance the decorative value of
a garden within a short span of time. But most of the annual flowers are
generally grown only for garden display purpose in various ways. These
flowers can be used as specialty cut flowers for their unique flowers and
distinct and attractive colors. Further, adding new specialty cut flowers
will improve the floriculture trade.
Postharvest Quality Evaluation of Winter Annuals 105

Vase life is an important criterion that determines the suitability of the

flower as a specialty cut flower. Earlier studies show that adding chemicals
to the vase solution will increase the vase life of cut flowers. Any vase
solution should contain an energy source and an antimicrobial compo-
nent. Sugar is the most commonly used energy source in vase solutions.
After harvest, the availability of sugar is limited in flowers. Adding sugars
mostly in the form of sucrose will delay the senescence and improve the
vase life of the flowers. But adding sucrose in the vase solution favors
the growth of microorganisms, which block the xylem vessels and reduce
the water uptake, thereby causing stem bending. Hence, biocides like
8-hydroxyquinoline citrate (8-HQC) (Ali and Hassan, 2014) and alumi-
num sulfate (Al2SO4) (Jowkar et al., 2012) are very much important to
reduce the microbial growth in the vase solution. Apart from these two
basic elements, ethylene produced during senescence of flowers acceler-
ates the senescence process that results in petal wilting, permeability of
petal cells, and degradation of membrane lipids. The senescence effects
can be reduced by inhibitors of ethylene biosynthesis like aminooxy acetic
acid (AOA) (Chaturaphat et al., 2003; Zuliana et al., 2008) and benzyl
adenine (BA) (Singh et al., 2008; Danaee et al., 2011).
This work was carried out with an objective to evaluate the postharvest
vase life of winter annual flowers to find their suitability as cut flowers.
For this study, five winter annuals, namely antirrhinum, dimorphotheca,
larkspur, lupin, and Sweet William, were selected as these flowers have
sturdy stem of more than 30 cm in length.

9.2 MATERIALS AND METHODS

The selected winter annuals were cultivated in the research field of the
Directorate of Floricultural Research, IARI, New Delhi, during 2013–
2014. The details of harvest stage for each flower are given in Table 9.1.
After harvest, the flower stems were trimmed to 40 cm and dipped in water
till they were placed inside the respective treatments.
Flower stems were held in centrifuge tubes filled with 50 mL dis-
tilled water (control) or other treatment vase solutions. All the chemi-
cals, viz., sucrose, 8-HQC, BA, and AOA in distilled water were used
106 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

TABLE 9.1 Details of Harvest Stage for Each Flower

Winter annual Stage of harvest Defoliation
Antirrhinum 3–4 flower open in the spike Defoliated
Dimorphotheca 9–10 mm Defoliated
Larkspur 3–4 flower open in the spike Not defoliated
Lupin 7–10 flowers open in a spike Defoliated
Sweet william 7–10 flowers open in a spike Non defoliated in the top near the
flower head.

as vase solution in different combinations [T0: control (distilled water),

T1: sucrose 2%, T2: sucrose (2%) + 8-HQC (200 ppm), T3: sucrose 2%
+ AOA (0.5 mM), T4: sucrose 2% + BA (0.22 mM), T5: sucrose 2% +
Al2SO4 (200 ppm)]. The tubes were covered with a cotton plug and
an aluminum foil to avoid evaporative losses and kept in a room with
natural light (12 h with 1600 lux light intensity). The average room tem-
perature and humidity during the study were recorded as 25 ± 2°C and
80% ± 5%, respectively. Each experiment was replicated three times
with three spikes per replication.
Weight of each spike was measured at 2 days interval. The relative
fresh weight of spikes was calculated as:
weight of flower spike at day t
Relative fresh weight (RFW) (%) = ×100
initial flower spike weight
where t = 0, 2, 4, 6, 8, 10 (He et al., 2006).
Difference in the stem/spike height was recorded at 2 days interval in
antirrhinum, larkspur, and lupin. Change in the volume of solution and
weight of tubes without spikes was recorded at 2 days interval. The fol-
lowing formulae were used to find the water relations and percent flower
opening:
Vase solution uptake rate (VSUR) (mL g−1 initial fresh weight (IFW)
day−1) is given by,

( St − 2 ) − ( St )
VSUR =
( IFW × 2 )

where t = 0, 2, 4, 6, 8, 10 (He et al., 2006).

Postharvest Quality Evaluation of Winter Annuals 107

Percentage ( No. of flowers at day t − Initial number of flowers )

       =
flower opening Initial number of buds

Percentage flower opening was recorded in antirrhinum, larkspur,

lupin, and Sweet William flowers. The flower diameter of dimorphotheca
was recorded at 2 days interval using a Vernier caliper.
Vase life of the flowers was characterized based on petal wilting,
flower drop, bud drop, stem bending and stem rotting. The average vase
life of flowers was assessed as terminated when 50% of the flowers were
senesced /dropped. The experiment was carried out in completely random-
ized block design (CRBD) with five treatments and three replications.

9.3 RESULTS AND DISCUSSION

9.3.1 CHANGE IN RELATIVE FRESH WEIGHT OF FLOWERS

From the results, it is evident that maximum increase in fresh weight was
observed in lupin (150.84%), and with respect to the treatments, maxi-
mum increase in weight during vase life was observed in treatment with
BA (T4) (Table 9.2).
In treatment with sucrose 2% (T2), there was an increase in RFW of
all flowers during initial 2 days of study. However, after that, the rate of
increase in fresh weight was reduced in all flowers, which could be due to
the microbial growth in the vase solution that might have caused physi-
cal plugging in the stems and blockage of xylem vessels (Danaee et al.,
2011). In lupin, the maximum RFW of 208.2% was recorded in treatment
with 8-HQC (T2) after 6 days of vase life. In treatments with HQC (T2) and
Al2SO4 (T5), both the agents acted as bactericides that reduced the growth
of stem plugging microorganisms and increased the water uptake for long
time than in T2 (Dole et al., 2009; Jowkar et al., 2012). In antirrhinum,
maximum RFW (160.6%) was recorded on day 6 in treatment with BA
(T4), whereas in Sweet William, it was recorded as 146.1% after 8 days
for the same treatment. Sakine et al. (2011) reported about delay in flower
opening of roses due to treatment with BA as cytokinins reported for its
negative effect on flower senescence.
 108

TABLE 9.2 Changes in Relative Fresh Weight and Vase Solution Uptake Rate of Winter Annuals during Vase Life
Treatments Relative Fresh Weight (%) Mean
Antirrhinum Dimorphotheca Larkspur Lupin Sweet willam
T0 (Control) 79.30 95.51 89.41 96.47 99.58 92.05
T1 (Sucrose 2%) 118.78 94.45 141.59 118.65 111.77 117.05
T2 (Sucrose 2%+ 8-HQC 200 ppm) 78.78 150.58 141.92 208.22 125.00 140.90
T3 (Sucrose 2%+ AOA 0.5mM) 123.46 160.68 123.12 133.09 133.41 134.75
T4 (Sucrose 2%+ BA 0.22 mM) 157.05 158.95 141.44 168.82 142.82 153.81
T5 (Sucrose 2%+ Al2SO4 200 ppm) 106.83 129.56 135.47 179.76 132.34 136.79
Mean 110.70 131.62 128.82 150.84 124.15
Vase solution Uptake rate (ml/g/flower)
T0 (Control) 0.10 0.28 0.82 0.85 0.41 0.49
T1 (Sucrose 2%) 0.25 0.23 1.06 1.09 0.36 0.59
T2 (Sucrose 2%+ 8-HQC 200 ppm) 0.21 0.33 1.04 1.12 0.32 0.60
T3 (Sucrose 2%+ AOA 0.5mM) 0.37 0.52 0.99 1.04 0.62 0.71
T4 (Sucrose 2%+ BA 0.22 mM) 0.59 0.39 1.01 1.06 0.81 0.77
T5 (Sucrose 2%+ Al2SO4 200 ppm) 0.31 0.34 1.08 1.09 0.57 0.68
Mean 0.30 0.35 1.00 1.04 0.51
Sustainable Horticulture Volume 2: Food, Health, and Nutrition
Postharvest Quality Evaluation of Winter Annuals 109

9.3.2 CHANGES IN VASE SOLUTION UPTAKE RATE (VSUR)

Maximum VSUR was observed in lupin (1.04 mL/g/day) and larkspur (1

mL/g/day), and the minimum uptake was observed in antirrhinum (0.30
mL/g/day) and dimorphotheca (0.35 mL/g/day). With respect to treatment,
maximum solution uptake (0.77 mL/g/day) was observed in treatment
with BA (T4).

9.3.3 CHANGES IN STEM ELONGATION, PERCENTAGE

FLOWER OPENING, AND FLOWER DIAMETER

Maximum stem elongation was recorded in lupin in treatment with

Al2SO4. Bending was not observed in lupin during vase life in all treat-
ments except control. In antirrhinum and larkspur, maximum stem elon-
gation was observed in treatment with BA (T4) (Figure 9.1), and the
finding is in accordance with Asil and Karimi (2010) who suggested
that BA at a lower concentration is required to delay flower senescence.
Maximum percentage flower opening in antirrhinum, larkspur and Sweet
William was observed in treatment with BA (T4), followed by treatment
with AOA (T3). This shows the ethylene sensitivity of flowers. The treat-
ments with AOA and BA increased the RFW of flower stems due to
delayed senescence, thus increasing the solution uptake and reduced tis-
sue transpiration and respiration (Keramat et al., 2012). In lupin, the
maximum percentage flower opening was observed in treatment with
HQC (T2) than in AOA and BA. This could be due the antimicrobial
activity of 8-HQC that prevented stem blocking and increased water
uptake, thereby reducing flower drop (Danaee et al., 2011). In dimor-
photheca, maximum flower diameter was observed in treatments with
AOA (T3) (90.17 mm) followed by BA treatment (T4) (87.73 mm) after
8 days of vase life. This clearly shows the ethylene sensitivity of the
flower and the ethylene inhibition properties of AOA and BA, which
improved the flower quality till 8 days in vase life when compared to the
other treatments (Figure 9.2 a,b).
110 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 9.1 (See color insert.) Effect of treatments on stem elongation and percent
flower opening of (a) antirrhinum, (b) dimorphotheca, (c) larkspur, (d) lupin, and (e) Sweet
William.

9.3.4 CHANGES IN VASE LIFE

In antirrhinum, the maximum vase life of 9.33 days was recorded in treat-
ment with BA (T4), followed by 8.33 days in AOA treatment (T3) (Figures
9.3 and 9.4).
In dimorphotheca, the maximum vase life of 8 days was recorded in
treatment with BA (T4), followed by AOA (T3). In larkspur, the maximum
vase life was observed in treatment with BA, whereas in lupin, it was
observed that the treatment with 8-HQC (T2) improved the vase life to 6
days as compared to other treatments. For Sweet William, the maximum
vase life was recorded in treatment with AOA (T3) (9.67 days), followed
Postharvest Quality Evaluation of Winter Annuals 111

FIGURE 9.2 (See color insert.) (a) Effect of treatments on flower diameter (mm) of
dimorphotheca and (b) effect of treatments on percent flower opening of Sweet William.
112 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 9.3 Effect of treatments on vase life of (a) antirrhinum, (b) dimorphotheca, (c)
larkspur, (d) lupin and (e) Sweet William.

by BA (9.33 days). From the results, it is evident that the plant bioregu-
lators AOA and BA significantly improved the vase life of antirrhinum,
dimorphotheca, Sweet William, and antirrhinum flowers. A significant
increase in vase life of flowers and the effect of AOA and BA on flower
Postharvest Quality Evaluation of Winter Annuals 113

FIGURE 9.4 (See color insert.) Effect of treatments on vase life of winter annuals.

senescence were reported by Zuliana et al. (2008) and Sakine et al. (2011).
Further, Sweet William and antirrhinum can be used as cut flower as they
have maximum cumulative vase life of 8.67 days and 7.72 days, respec-
tively, with respect to all treatments.

9.4 CONCLUSION

Winter annual flowers are not yet explored as cut flowers. The unique
flower features and colors make these flowers very attractive. Among
the five winter annual flowers evaluated in this study, antirrhinum and
Sweet William had minimum vase life of ~6 days and ~8 days in control
treatment itself. Adding sucrose to the vase solution improved the vase
life of larkspur from 4.33 days to 7.67 days; dimorphotheca vase life also
improved to 8 days by adding BA + sucrose in the vase solution. Based
on the vase life of flowers, antirrhinum and Sweet William can be used as
commercial cut flowers.
114 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

KEYWORDS

•• antirrhinum
•• dimorphotheca
•• larkspur
•• lupin
•• plant bioregulators
•• Sweet William
•• vase life
•• winter annuals

REFERENCES

Ali, E., & Hassan, F., (2014). Postharvest quality of Strelitzia reginae cut flowers in rela-
tion to 8-hydroxyquinoline sulphate and gibberellic acid treatments. Sci. Agri., 5(3),
97–102.
Asil, M. H., & Mahnaz, K., (2010). Efficiency of Benzyl adenine reduced ethylene pro-
duction and extended vase life of cut Eustoma flowers. Plant Omics Journal, 3(6),
199–203.
Chaturaphat, R., Saichol, K., Wouter, G., & Van Doorn., (2003). Effect of aminooxyacetic
acid and sugars on the vase life of Dendrobium flowers, Postharvest Biology and
Technology, 29, 93–100.
Danaee, E., Ruhangiz, N., Sepideh, K., & Ali, R. L. M., (2013). Evaluation the effect sali-
cylic acid and benzyl adenine on enzymatic activities and longevity of gerbera cut
flowers, Intl. Res. J., Appl. Basic. Sci., 7(5), 304–308.
Fard, E. S., Khodayar, H., & Ahmad, K., (2010). Improving the keeping quality and vase
life of cut alstroemeria flowers by pre and post-harvest salicylic acid treatments. Not.
Sci. Biol., 5(3), 364–370.
He, S., Joyce, D. C., Irving, D. E., & Faragher, J. D., (2006). Stem end blockage in cut
Grevillea ‘Crimson Yul-lo’ inflorescences. Postharvest Biol. Technol., 41, 78–84.
Jowkar, M. M., Mohsen, K. A. K., & Nader, H., (2012). Evaluation of aluminum sulfate
as vase solution biocide on postharvest microbial and physiological properties of
‘Cherry Brandy’ rose, Annals of Biological Research, 3(2), 1132–1144.
Kazemi, M., Hadavi, E., & Hekmati, J., (2012). Effect of salicylic acid, malic acid, citric
acid and sucrose on antioxidant activity, membrane stability and ACC-Oxidase activ-
ity in relation to vase life of carnation cut flowers. Journal of Agricultural Technol-
ogy, 8(6), 2053–2063.
Postharvest Quality Evaluation of Winter Annuals 115

Sakine, F., Roohangiz, N., & Oru, D. V. I., (2011). Effects of post harvesting on biochemi-
cal changes in gladiolus cut flowers cultivars (white prosperity). Middle East Journal
of Scientific Research, 9(5), 572–577.
Singh, A., Kumar, J., & Kumar, P., (2008). Effect of plant growth regulators and sucrose on
post harvest physiology, membrane stability and vase life of cut spikes of Gladiolus.
J., Plant Growth Regulators, 55, 221–229.
Sodi, A. M., & Antonio, F., (2005). Physiological changes during postharvest life of cut
sunflowers. Proc. VIIIth IS Postharvest Phys. Ornamentals, Acta. Hort., 669, 219–
224.
Zuliana, R., Boyce, A., Nair, H., & Chandran, S., (2008). Effects of aminooxyacetic acid
and sugar on the longevity of pollinated Dendrobium Pompadour. Asian Journal of
Plant Sciences, 7(7), 654–659.
CHAPTER 10

OPTIMIZING THE SHELF LIFE

OF WHOLE AND FRESH CUT
BREADFRUIT IN MAURITIUS
ROOP SOODHA MUNBODH
Food and Agricultural Research and Extension Institute (FAREI),
Reduit, Mauritius, Tel.: 59799325,
E-mail: [email protected]

CONTENTS

Abstract...................................................................................................117
10.1 Introduction..................................................................................119
10.2 Activity 1: Characterization and Evaluation of Quality
Attributes of Fresh Breadfruits at Mature and
Immature Stages.......................................................................... 120
10.3 Activity 2: Shelf Life Evaluation of Cling Filmed and
Non-cling Filmed Whole Breadfruit Stored at 13°C................... 125
10.4 Storage of Minimally Processed Cold Stored Breadfruit Slices.... 130
10.5 Conclusions................................................................................. 135
10.6 Future Work................................................................................ 136
Acknowledgments.................................................................................. 136
Keywords............................................................................................... 136
References.............................................................................................. 137

ABSTRACT

Breadfruit has a high potential for export as a fresh fruit, but the extreme
perishability of this fruit hampers its export trade. High postharvest losses
118 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

(25–30%) are incurred during the export transit, whereby the fruits ripen
on reaching export destination. Trials were carried out for the proper iden-
tification of optimum maturity stage for harvest of breadfruit and optimum
storage conditions to extend shelf-life for export. Fruits were harvested
from various sites, and fruit and peel physical characteristics were deter-
mined. For optimum shelf-life, the fruit should be pale green in color and
firm in texture with latex stains on uniform protruding polygonal segments
on the peel. The flesh of mature unripe breadfruit was observed to be ivory
white in color and had a starchy taste when boiled. Mature ripe breadfruit
had a yellowish green peel color, a cream flesh color with a soft texture,
and was sweet when boiled. Handling practices were of utmost impor-
tance to minimize mechanical damage as bruised fruits will soften rapidly,
causing reduction in shelf-life; thus, fruits were harvested early morning
to avoid heat build-up by using either a fruit picker with mesh bag or a har-
vesting pole with a cutting scythe and a collecting bag to avoid fruit drop.
The stems of the cut fruits of 3-4 cm length were pointed downward to
allow latex flow away from fruit surface and thus prevent brown staining
of the peel. Harvested fruits were placed in clean stackable plastic crates
and transported in covered vehicles that provided adequate ventilation and
protection (using a cover) to the harvested fruits. Storage trial showed
that when whole fruit was washed in sterilized water (4 mL of javel/L of
water), air-dried, cling filmed, and stored at 13ºC with a relative humid-
ity of 90–95%, breadfruit kept for 2 weeks, while noncling filmed fruits
remained marketable for only 7 days. Cumulative % weight loss was only
2.23 % +/– S.E 1.33 and 0.93% +/– S.E 0.24 for Pamplemousses experi-
mental station and Reduit crop research station, respectively, over 15 days
of storage. At 13ºC, cling filmed fruits retained their flavor and taste at 14
days of storage, while control fruits remained tasty for only 5–7 days. The
color of fruit peel was maintained as bright pale green when cling filmed
for 2 weeks as compared to noncling filmed fruits; the fruits assumed a
dull green brownish tinge. Cling filmed fruits maintained their firmness at
8–9 kg/m2. Postharvest disease incidence was observed after 2 weeks of
storage and was caused by pathogens Colletotrichum and Cladosporium
for cling filmed fruits at 5ºC. Under ambient conditions, noncling filmed
fruits ripened after 2 days, while cling filmed ones ripened after 4 days.
Storage trial of minimally processed breadfruit showed that fruit cut into
Optimizing the Shelf Life of Whole and Fresh Cut Breadfruit 119

longitudinal slices of 1.5 cm thickness, soaked in sodium metabisulfite

solution at 0.5 g/L for 5 minutes to prevent browning, air dried, and vac-
uum packed at 85% had a shelf-life of 12 days at 4 oC, wherein color, taste,
firmness, and flavor were maintained. Under ambient conditions, the fresh
cuts were observed to keep for only 2 days.

10.1 INTRODUCTION

Breadfruit is a climacteric fruit mostly grown in the tropics. The fruit is a

starchy staple, and mature, unripe fruit is cooked and eaten in much the
same way as tubers and root crops. Breadfruit has a high potential for export
as a fresh fruit. During the last 5 years, export of fresh breadfruits from
Mauritius to Europe has increased from 40 to196 tons, but the extreme per-
ishability of this fruit hampers its export trade. High postharvest losses are
incurred during the export transit. A large amount (25–30%) of exported
fruits lose their firmness and become ripe and soft before reaching their
retail outlet; as a result, there is an economic loss to the breadfruit export-
ers of Mauritius. Actually, in Mauritius, fresh breadfruit is currently being
exported at an attractive remunerable price. The main goal is therefore to
increase the amount of quality breadfruit for local and export markets and
thus to provide growers and exporters with improved postharvest manage-
ment practices to enable them to obtain quality breadfruits with longer mar-
ketable periods. There is therefore a need to devise strategies to minimize
postharvest losses of breadfruit along the supply chain and to improve the
actual packaging and storage conditions from the grower to the consumer
for the local and export markets. Reduced postharvest losses along the
export will definitely benefit the breadfruit growers and exporters eco-
nomically. Improved innovative packaging and storage conditions that will
lead to an increase in number of quality breadfruits from harvest to export
for local and export markets and lengthening of shelf-life of the minimally
processed breadfruit slices under different packing and storage conditions
will also encourage supermarkets to display fresh-cut breadfruit chunks on
the shelf, leading to local consumer acceptance of cut breadfruits. This will
help them for the easy preparation of breadfruit curry and other dishes.
Growers and exporters will also be able to tap new promising markets for
120 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

export and local markets. There will be a larger area of land devoted to
breadfruit plantation to satisfy the local demand, hence contributing to food
security. A rise in number of breadfruit growers and exporters will also
create more employment for the people in the business, including women
entrepreneurs, and also provide income generation. It is expected to receive
infrastructural government support as the breadfruit business expands.
Extended areas of breadfruit plantation will also contribute to a greener
Mauritius. Preliminary promising results will encourage the government to
devote funds (food security fund of the Ministry of Agro Industry and Food
Security) to develop the sector further for the benefit of vulnerable groups
and for food security. Breadfruit therefore has an important role to play in
food security, sustainable agriculture, and income generation and its profile
need to be raised at the national level.
Hence, trials were carried out to (a) characterize of local accessions
of breadfruit and to determine the optimum fruit maturity for storage and
export of breadfruit from local germplasm in two agro-climatic zones of the
island, (b) to evaluate the shelf-life of cling filmed and noncling filmed fresh
breadfruit stored at 13ºC, and (c) to evaluate the shelf-life of minimally pro-
cessed cold stored breadfruits under different packaging conditions.

10.2 ACTIVITY 1: CHARACTERIZATION AND EVALUATION

OF QUALITY ATTRIBUTES OF FRESH BREADFRUITS AT MATURE
AND IMMATURE STAGES

Breadfruits were harvested early in the morning from a sub-humid agro-

climatic zone at (a) Pamplemousses experimental station (ES) and from
a humid agro-climatic site; (b) Reduit crop research station (CRS) at (i)
mature green stage (young), characterized by light green skin and absence
of external latex with the fruit outer segments closely packed and having a
rough texture; and (ii) “fit” stage characterized by darker green skin with
some browning, external dried latex, and fully flattened outer segments
(Table 10.1). Fruits were harvested early morning to avoid heat build-up
by using either a fruit picker with mesh bag or a harvesting pole with a
cutting scythe and a collecting bag. Precautions were taken to avoid drop-
ping harvested fruits to the ground as bruised fruits will soften rapidly,
causing reduction in shelf-life. The stems of the cut fruits of 3–4 cm length
TABLE 10.1 Characterization of Breadfruits According to Site and Fruit Maturity
Site: Pamplemousses Immature Mature green Site Immature Mature green
Experiment Reduit
Station Crop research
Sub-humid Station.
Agro climatic zone Humid
Agro climatic zone
Fruit weight (g) 838 1287 Average Fruit size(g) 710 1360
Fruit color Light olive green 144C Pale green with Fruit color Olive green Pale green with
brown streaks 144B brown streaks
152D 144C
Type of fruit segment Close knit segments, Smooth surface Type of fruit segment Close knit seg- Smooth surface
rough surface with segments ments, rough with segments
flattened out surface flattened out
Fruit diameter (cm) 11.5 12.8 Fruit diameter 13.1 14.8
Fruit circumference (cm) 39.5 41 Fruit circumference 36.8 43.1
(cm)
Optimizing the Shelf Life of Whole and Fresh Cut Breadfruit

Fruit length (cm) 18.8 19.7 Fruit length 15 19.9

Fruit shape round oblong Fruit shape spherical oblong
Firmness (kg/m²) 7.5 7 Firmness 9.0 8.0
121
TABLE 10.1 (Continued) 122
Pulp color Ivory white 151D Cream, 155B Pulp color Ivory white Cream,155D
With slight browning No browning on 152D With No browning on
on cutting cutting slight browning cutting
on cutting
Core length 8.7 cm 8.66 cm Core length 7.15 cm 5.24
Flesh thickness (cm) 4.7 8.8 Flesh thickness (cm) 4.5 8.2
Sensory quality Watery bland Full starchy taste Sensory quality Watery bland starchy taste
Taste, no aroma. Taste, no aroma.
Sustainable Horticulture Volume 2: Food, Health, and Nutrition
Optimizing the Shelf Life of Whole and Fresh Cut Breadfruit 123

were pointed downward to allow latex flow away from fruit surface and
thus prevent brown staining of skin. Harvested fruits were placed in clean
stackable plastic crates to minimize handling damage. Fruits were loaded
in a transport vehicle that provided adequate ventilation and protection
(by using a cover) to the harvested fruits. Special care was taken that the
fruits (free from pest and disease attack) were not bruised and subjected to
mechanical injury by gently placing them in cushioned plastic crates and
brought to the postharvest laboratory within 1 hour. The fruits were stored
under ambient conditions in the postharvest laboratory and characterized
for the physical and sensory quality attributes at both the immature and
mature stages. A total of 25 fruits were characterized from each site repre-
senting the sub-humid and humid agro-climatic zones of the island.
Regarding harvest methods, Andrews and Mason (1992) reported that
the “pick and catch” method is preferred in the eastern Caribbean islands
from trees as much as 20 m tall.
The characterization process was carried out with the following instru-
ments:
(a) Fruit weights were recorded using a “Sartorius” electronic balance.
(b) Fruit firmness was measured using an “Atago” penetrometer and
the unit was kg/m².
(c) Firmness was measured as penetration force to depress 5 mm of
fruit flesh.
(d) Fruit pulp and peel color were recorded according to the color chart
of the Royal Horticultural Society.
(e) Flesh thickness and fruit diameter were recorded with a “caliper.”
(f) Fruit circumference was measured using a standard tailor measur-
ing tape.
(g) Sensory evaluation of boiled peeled chunks of mature and imma-
ture fruits was carried out using 10 untrained panelists on the sta-
tion, and they were given random boiled samples and were asked
to verbally describe the taste, texture, and overall appreciation of
the sample. Ten tasting sessions were informally carried out for
each maturity type. The descriptive terms used were:
(1) watery – starchy
(2) bland taste – strong breadfruit taste.
124 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

(3) No odor – strong aroma

(4) dislike – like
From the above results, it was observed that mature fruits are spheri-
cal to oblong shaped and have a higher mean weight, length, diameter,
and circumference values than immature fruits. However, immature fruits
are more firm and have higher mean flesh firmness values than mature
fruits. The outer peel color of immature fruits is characterized by light
green skin and absence of external latex with the fruit outer segments
closely packed and having a rough texture, while for mature fruits, the
“fit” stage was characterized by darker green skin with some browning,
external dried latex, and fully flattened outer segments. As reported by
Ragone (1977), fruits are globose to oblong, with yellowish green to yel-
low skin when mature with creamy white or pale yellow flesh.
Flesh color of immature fruits has been observed to be ivory white with
a strong tendency to brown compared to mature fruits where the flesh color
is light cream with little or no browning. On boiling the mature fruits, the
taste was good with a starchy, creamy texture. The boiling time was 25–30
minutes. For immature fruits, the boiled slice had a watery texture and the
taste was bland. Depending on the intended use of fruit as a vegetable for
curry, breadfruits should be harvested when the fruit is pale green in color
with latex stains on uniform protruding polygonal segments; free from
defects, sunscald, cracks and insect bites; and of uniform shape and firm in
texture. If used as desert and in cakes, breadfruits should be harvested when
fruit texture is softer and the polygonal segments are smooth with deeper
latex stains with a yellow green fruit color. The main internal indices of
breadfruit maturity are flesh color and starch content. The flesh of mature
unripe breadfruit is ivory white in color and starchy, whereas mature ripe
breadfruits have a cream color with a soft texture. The flesh of mature green
breadfruit when boiled for 20–25 minutes has a starchy taste, whereas the
taste of ripened fruits is sweeter, less starchy due to its conversion of starch
to sugar. As reported in the technical bulletin on postharvest care and mar-
ket preparation information on breadfruit from the New Guyana Marketing
Cooperation (May 2004), breadfruit should be harvested when green in
color and firm in texture if it is to be used as a starchy vegetable. The fruit
peel surface should have protruding segments on the surface that tends to
Optimizing the Shelf Life of Whole and Fresh Cut Breadfruit 125

be angular and ridged. If the fruit is to be used as dessert or for roasting or

baking, it needs to be harvested when the skin color turns yellow green in
color with brown latex stains and flattened expanded polygons of the fruit
peel. The principal internal indices are flesh color; the flesh of mature but
unripe fruit is white, starchy, and fibrous, while fully ripe fruit has a pale
yellow flesh color and is soft and fragrant.

10.3 ACTIVITY 2: SHELF LIFE EVALUATION OF

CLING FILMED AND NONCLING FILMED WHOLE
BREADFRUIT STORED AT 13°C

10.3.1 MATERIALS AND METHODS

Breadfruits (mature and mature green) were harvested early in the

morning from Pamplemousses ES and Reduit CRS at “fit” stage char-
acterized by darker green skin with some browning, external dried latex
and fully flattened outer segments as well as at mature green stage.
Special care was taken that the fruits (free from pest and disease attack)
were not bruised and subjected to mechanical injury. The fruits were
then brought to the postharvest laboratory. Fifteen fruits were used per
treatment. The fruits were washed under running tap water and then
soaked in sterilized javel water (5 mL/L water) for 10 minutes and then
drained; the dried fruits were then cling filmed and noncling filmed and
stored under ambient and at 13°C. Three batches of trials were carried
out for each site. The main objectives of this trial were to assess the
shelf-life of whole fresh breadfruits from local germplasm from Reduit
and Pamplemousses CRSs when cling filmed and non-cling filmed and
stored at 13°C. The parameters assessed and procedures carried out are
as mentioned in the following subsections.

10.3.1.1 Percentage Weight Loss

Percentage weight loss was calculated by subtracting the actual weight

from the initial weight of fruits per pack and dividing it by the initial
126 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

weight and multiplying by 100 using a “Sartorius” electronic balance to

two decimal places.

10.3.1.2 Disease Incidence

Disease incidence was observed on fruits showing any fungal or bacterial

growth. Identification of the disease and pathogen was carried out by the
Plant Pathology Division of FAREI (Food and Agricultural Research and
Extension Institute, Mauritius).

10.3.1.3 Color

Fruit pulp and peel color were recorded according to the color chart of the
Royal Horticultural Society.

10.3.1.4 Firmness

Fruit firmness was measured using an “Atago” penetrometer, and the

unit was kg/m². Firmness was measured as penetration force required to
depress 2.5 mm into the fruit using a penetrometer.

10.3.1.5 Sensory Evaluation

An informal sensory appraisal of the stored fruits by 10 untrained panelists

at the Wooton CRS was also carried out from randomly boiled samples.

10.3.3 RESULTS AND DISCUSSION

10.3.3.1 Shelf Life

Under ambient storage, breadfruits stored without any treatment were

observed to have ripened within 2 days. Such fruits remained marketable
for only 2–3 days, depending on the maturity stage of fruit. Those fruits
kept under cling filmed remained marketable for 3–4 days
Optimizing the Shelf Life of Whole and Fresh Cut Breadfruit 127

At 13°C, fruits packed under cling film remained marketable for 14–15
days, while those untreated or soaked in water remained marketable for
only 7–10 days for both sites, Reduit CRS and Pamplemousses ES Table
10.2 summarizes the results obtained from the trials.
At 26–28°C, under ambient conditions, the storage life was only
2–3 days (Thompson et al., 1974c; Maharaj and Sankat, 1990a) after
which skin browning is noted as well as fruit ripening and softening. As
reported in the Journal of National Tropical Botanical Garden, Bread-
fruit Institute (2012), packaging the fruit in sealed polyethylene bags
or plastic wrap can improve shelf life and fruit quality for 5–7 days.
The optimum storage temperature is 12–16°C for unpacked fruits, and
the shelf-life ranges from 7–10 days. If the fruit is packaged, it can be
stored for up to 2 weeks. Packaging influenced positively the shelf-life
of stored breadfruits. Storage life in sealed 150-gage polyethylene bags
was significantly greater than for unwrapped fruits at both high and low
temperatures (Thompson et al., 1974).

10.3.3.2 Weight Loss

Cling filmed fruits lost weight more rapidly under ambient storage than at
13°C for both sites as shown in the table above. Cling filmed fruits from
Reduit CRS under ambient storage had an average cumulative weight loss
% of 1.7% at 25°C at 3 days storage, whereas those fruits from Pample-
mousses ES had a value of 3.1%; control noncling filmed fruits stored
under ambient conditions had a cumulative % weight loss of 6–6.1% for

TABLE 10.2 The Results Obtained from the Trials

25ºC 13ºC 25ºC 13ºC
cling film cling film no cling film no cling film
Shelf life 3 14 2 7-10
Flesh color Light cream Ivory white Light cream cream
Taste starchy starchy starchy starchy
Days to 3-4 14-15 2 7
ripening
128 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

both sites. Sankat and Maharaj (1993) noted that fruits stored under ambi-
ent conditions demonstrated rapid ripening through weight (2.64%) and
volume losses (3.46%) (Figure 10.1). As reported by Worrell and Sean
Carrington (1997), the use of polyethylene wraps and bags in conjunction
with low temperature has proved to be beneficial in maintaining quality and
shelf-life for at least 2 weeks, particularly with LDPE (low-density poly-
ethylene) films and HDPE (high-density polyethylene) 40- and 60-micron
films. Packaging and refrigeration therefore successfully slow the rate of
water loss from the fruits by reducing the water holding capacity of the sur-
rounding air, slowing rates of diffusion, and providing a physical barrier to
air currents. Huang and Scott (1985) and Wong et al. (1991) reported that by
packing fruit in plastic containers and overwrapping with a semi-permeable
membrane, fruit desiccation was reduced with minimum condensation.
Under 13°C storage, the cling filmed fruits had a very low cumulative
% weight loss of 2.23% ± SE 1.9 and 0.93% ± SE 1.6 for Pamplemousses
ES and Reduit CRS, respectively, over 15 days of storage. Refrigerated
storage extends the postharvest life of most produce (Kader, 1992), and

FIGURE 10.1 (See color insert.) Chart showing the % cumulative weight loss of cling
filmed and noncling filmed breadfruits from Pamplemousses ES stored at 13°C.
Optimizing the Shelf Life of Whole and Fresh Cut Breadfruit 129

breadfruit being no exception, 12–13°C was found to be optimal (Thomp-

son et al., 1974; Maharaj and Sankat, 1990).

10.3.3.3 Sensory Appraisal

At 13°C, the fruits under cling film retained their flavor and taste at 14 days
of storage, while control fruits remained tasty for only 5–7 days. Under
ambient conditions, taste of boiled fruits (cling filmed and control) were
maintained for only 2–3 days after which they had a slight fermented taste.

10.3.3.4 Color

Color of fruit peel was maintained as bright pale green when cling filmed
for a longer period (2 weeks at 13°C) as compared to non-cling filmed
fruits, when the fruits assumed a dull green color with brownish tinge after
7 days of storage. Wills et al. (1981) stated that the green color of the fruit
is due to the presence of chlorophyll, a magnesium organic compound
that degrades in storage principally by pH changes, oxidative systems,
and chlorophyllases; along with chlorophyll disappearance, the synthesis
of carotenoids is responsible for yellow pigments in fruits. Flesh color of
mature green cling filmed fruits were maintained as an ivory cream over
the storage time at 13°C. As the fruits ripened, the flesh color changed
from ivory to cream.

10.3.3.5 Fruit Firmness

The cling filmed fruits maintained their firmness and ripened gradually
over a longer storage period compared to noncling filmed fruits at 13°C.
Fully ripened fruits had a lower firmness value (1.5 kg/m2) compared to
nonripened mature fruits at 8–9 kg/m2. Loss in fruit firmness is caused by
the enzyme polygalacturonase (a cell wall-degrading enzyme) that cata-
lyzes upon ripening the hydrolysis of glucosidic linkages of pectic sub-
stances in the cell wall and results in weakened texture of fruits (Wills et
al., 1981).
130 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

10.3.3.6 Postharvest Disease Incidence

The pathogens Colletotrichum and Cladosporium were identified by the

Plant Pathology Division on diseased, cold-stored cling filmed fruits after
2 weeks of storage. As reported by Worell and Carrington (1997), all
films delayed both the onset of softening and skin browning by at least a
week compared to unwrapped controls. At 13°C, controls began to soften
within a week, while wrapped fruit remained firm even after 21 days of
storage. One drawback of film wrapping was microbial growth observed
from the second week of ambient storage and internal flesh discoloration
at 13°C storage. As reported in the technical bulletin on postharvest care
and market preparation information on breadfruit from the New Guyana
Marketing Cooperation (May 2004), physical damage is the major cause
of postharvest decay of unripe fruits, which may be incurred by rough
handling, by improper packaging, or during transport. Wounds such as
punctures, cuts, abrasions, and cracks provide potential points of entry
for decay organisms. Postharvest decay can be adequately controlled by a
regular sanitation program involving application of preharvest fungicides
and careful harvesting and handling practices and storing at 12.5°C.
No chilling injury was detected in stored fruits at 13°C, but noncling
filmed fruits had the fruit peel turning a dull brown color after 7 days
of storage. Worell and Carrington (1997) suggested that breadfruit skin
browning may be a water loss problem, because water loss from epidermal
cells causes cell damage which in turn brings soluble phenolics into con-
tact with polyphenol oxidase, causing browning and ultimately cell death.
Recommendations were made to carefully select the fruits at harvest and
to minimize damage during handling.

10.4 STORAGE OF MINIMALLY PROCESSED COLD STORED

BREAD FRUIT SLICES

10.4.1 INTRODUCTION

Minimal processing involves fruits and vegetables that have been

trimmed, peeled, washed, cut, washed, dried, and packed. The purpose
Optimizing the Shelf Life of Whole and Fresh Cut Breadfruit 131

of minimal processing is to provide the consumer with a fresh product

with an extended shelf life and also ensure food safety and maintain
nutritional and sensory quality. In order to produce minimal processing
products that are safe, high quality with a long shelf life, the processor
has to apply Good Manufacturing Practice in his processing plant. The
product flow should be in one direction from the receiving area to the
finished packed product to avoid cross-contamination from raw vegeta-
bles and fruits to ready-to-eat minimally processed produce. Cleanliness
and sanitation are two important factors in quality control and safety
for minimally processed products. The main requirements for minimal
processing of fruits and vegetables are (a) good quality raw material, (b)
efficient and fast operations of cutting, slicing, and trimming, (c) ade-
quate disinfection of the working area and equipment, (d) use of sharp
disinfected equipment, (e) provision of clean potable water for washing
of produce, (f) maximum draining of produce before packing, (g) proper
labeling of packed produce, and (h) provision of optimum temperature
(4–5°C) to maintain quality and shelf-life. Special treatments in minimal
processing include the use of antibrowning agents such as citric acid
and sodium metabisulfite. The quality of minimally processed products
should be visually acceptable, have an appealing and fresh appearance,
be of consistent quality throughout the package, and be free of defects.
Improved shelf-life of fresh-cut breadfruit will encourage local work-
ing consumers to adopt breadfruit more often in their routine cooking
recipes. The hotel industry will also use this product for exotic tropical
cuisine dishes.

10.4.2 MATERIALS AND METHODS

The breadfruits of sound health, free of any disease and pest attack were
harvested early in the morning at Reduit CRS and Pamplemousses ES.
The fruits were placed in plastic crates and immediately transported to
the postharvest laboratory at Wooton CRS. The fruits were washed under
running tap water and then peeled to remove the outer skin layer and then
placed on a sterilized chopping board and cut longitudinally into regular
slices of 2 cm thickness. The cut peeled portions were then washed in
132 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

sanitized chlorine water (2.0 mL/L of water) to reduce microbial load

and rinsed again in tap water (washing after peeling and cutting removes
microbes and tissue fluid, thus reducing microbial growth and enzymatic
oxidation during storage). The microbiological quality of the washing
water must be good, and its temperature should be low. After slicing,
soaked in Sodium metabisulfite solution(0.2 g/L of water) for 5–7 min-
utes and then drained and dried under a fan. The dried segments were
then packed in clip-on punnets (100 g per punnet) and also packed under
vacuum (85%) in vacuum bags and stored at 5°C. The main objectives
were therefore to assess the (a) shelf-life of packed minimally processed
treated breadfruit under two packaging at 5°C and (b) to study the physi-
ological disorders and pathological diseases observed in the minimally
processed breadfruit during storage trial.

10.4.3 RESULTS AND DISCUSSION

10.4.3.1 Shelf Life

Under clip-on packaging and storage at 5°C, the cut slices packed in
clip-on and dipped in sodium metabisulfite retained their color and
freshness for 5–7 days irrespective of the site (Reduit CRS and Pample-
mousses ES) after which they turned rancid and fermented with the
presence of leachate. When vacuum packed at 85% vacuum, the cut
slices remained marketable for 18 days irrespective of the site. Vacuum
packaging was more advantageous for fresh-cut breadfruits than clip-on
barquettes.

10.4.3.1.1 Color

The cut slices maintained ivory white color before turning to a dull yel-
low color after 5 days storage at 5°C for packed samples in clip-on and
sodium metabisulfite. Ivory white color of the cut slices was maintained
for 18 days under vacuum packing. Browning of fresh cut was prevented
using the anti-browning dip sodium metabisulfite. As reported by Whita-
ker and Lee (1995), enzymatic browning requires four different compo-
Optimizing the Shelf Life of Whole and Fresh Cut Breadfruit 133

nents: oxygen, an enzyme, copper, and a substrate. In order to prevent

browning, at least one component must be removed from the system.
Polyphenol oxidase (PPO)-catalyzed browning of fruits and vegetables
can be prevented by factors such as (a) heat or reaction inactivation of
the enzyme, (b) exclusion or removal of one or both of the substrates
(oxygen and phenols), (c) lowering the pH to two or more units below the
optimum, and (d) adding compounds that inhibit PPO or prevent melanin
formation.

10.4.3.1.2 Weight loss

Cumulative weight loss % was observed to be minimal for cut slices stored
under cold storage at 4–5°C and treated with food-grade sodium metabi-
sulfite, irrespective of the site. Breadfruit slices when vacuum packed at
85% and stored at 5°C were observed to have a lower cumulative % weight
loss over storage time compared to those stored in clip-on barquettes.
Type of packaging influenced the rate of weight loss of fruits over stor-
age time due to their permeability properties. Sealed packaging type as
vacuum-packed products allows for a lower volume of air space compared
to clip-on packaging, and hence for a decreased rate of respiration, flow of
gases, and water vapor from the vacuum packed produce. For the clip-on
barquettes, more freedom of gas exchange and water vapor was allowed
through the thin air space between the closed sides of the pack compared
to the vacuum-sealed plastic bags. This explains the significantly higher
% cumulative weight loss of fresh-cut breadfruit slices (Tables 10.3 and
10.4) when stored in clip-on barquettes over storage time compared to
those that were vacuum packed (Figure 10.2).

10.4.3.2 Sensory Appraisal

Boiled samples of stored cut slices were randomly presented at weekly

intervals to panelists who affirmed that starchiness, breadfruit taste, tex-
ture, and aroma were maintained throughout the storage period until the
end of shelf-life.
134 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

TABLE 10.3 The Cumulative % Weight Loss of Breadfruit Slices Over 7 Days of
Storage in Clip-on Packs at 4–5°C
Site/storage days 3d 7d S.D S.E
Reduit Crop 0.455 1.332 0.620 +/– 0.219
Research
Station
Pamplemousses 0.433 1.161 0.515 +/– 0.143
Experiment
Station

TABLE 10.4 The Cumulative % Weight Loss of Breadfruit Slices Over 18 Days Storage
from Pamplemousses ES when Vacuum Packed and Stored at 4–5°C
Storage 7d 9d 11d 14d 18d S.D S.E
period (days)
% cumulative 0.151 0.332 0.599 0.791 0.875 0.305 +/– 0.088
weight loss

FIGURE 10.2 Chart showing the % cumulative weight loss of fresh-cut vacuum-packed
breadfruit slices stored at 5°C.
Optimizing the Shelf Life of Whole and Fresh Cut Breadfruit 135

10.4.3.3 Postharvest Disease Incidence

Postharvest fungi Rhizopus was detected on cut slices stored in clip-on

barquettes after 7 days of storage. According to Willcox et al. (1994), high
humidity and the large number of cut surfaces can provide ideal conditions
for the growth of microorganisms. No disease incidence was detected in
vacuum-packed samples. Presence of leachate in stored samples at the
end of storage life was due to physiological deterioration of cellular tis-
sues. Because minimally processed fresh fruits and vegetables are not heat
treated, regardless of additives and packaging, they must be handled and
stored at refrigerated temperatures at 5°C or under in order to achieve suf-
ficient shelf-life and microbial safety.

10.5 CONCLUSIONS

The maturity stage at which breadfruits are harvested is of crucial importance

for the fruit storability and marketability, and it will depend on immediate
end use of the fruit (a) whether for storage for export purposes, (b) for imme-
diate sale in the local markets for curries, or (c) for use for baking or deserts.
For export purposes, it is advised to harvest “three quarter” mature green
fruits with pale green color with white to brown latex stains but with polygo-
nal segments still tightly bound together. The flesh of mature unripe bread-
fruit was observed to be ivory white in color and had a starchy taste when
boiled. Mature ripe breadfruits had a yellowish green peel color, a cream
flesh color with a soft texture, and was sweet to taste when boiled. Handling
practices were of utmost importance to minimize mechanical damage as
bruised fruits will soften rapidly causing reduction in shelf life for export;
thus, fruits should be harvested early morning to avoid heat build-up by
using either a fruit picker with mesh bag or a harvesting pole with a cutting
scythe and a collecting bag to avoid fruit drop. The stems of the cut fruits
of 3–4 cm length should be pointed downward to allow latex flow away
from fruit surface and hence prevent brown staining of the peel. Harvested
fruits should be placed in clean stackable plastic crates and transported in
covered vehicles that provide adequate ventilation and protection (using a
cover) to the harvested fruits. Use of a controlled temperature vehicle will
be advantageous as it will pre-cool the fruit by removing field heat from the
136 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

harvested fruit prior to export. It is advised to wash the fruits in sanitized

water to remove dust particles and to thoroughly dry them before they are
cling filmed prior to storage at 13°C as the fruits retain their peel and flesh
color with minimum weight loss as well as maintain firmness taste and fla-
vor over a storage time of 14 days. For increasing marketing opportunities
of fresh breadfruit and providing easy access to the working consumer, it
is recommended to be vacuum packed at 85% cut slices of breadfruit. That
should be dipped in sodium metabisulfite solution (0.2 g/L of water) before
being stored at 5°C, resulting shelf life of the cut slices of 18 days during
which the color, freshness, flavor, and taste is maintained.

10.6 FUTURE WORK

Future research could involve (a) evaluating the shelf-life of commercial

hydro-cooled cling filmed breadfruits in Mauritius, (b) shelf life evalua-
tion of breadfruits using biodegradable type packaging to minimize envi-
ronmental pollution, and (c) evaluating the use of surface coating based
on polysaccharide and sucrose ester-based coatings on cold-stored bread-
fruits to minimize fruit desiccation.

ACKNOWLEDGMENTS

Grateful to the CEO of FAREI, Mr. Rajkumar, for giving permission to

present this paper; Principal Research Scientist, Fruit Division, Mrs Ram-
burn and staff for their help and cooperation in editing this paper.

KEYWORDS

•• breadfruit
•• fresh cut
•• sensory quality
•• shelf life
•• storage
Optimizing the Shelf Life of Whole and Fresh Cut Breadfruit 137

REFERENCES

Adel, A. K., (2012). Breadfruit: Recommendations for Maintaining Postharvest Quality,

Department of Plant Sciences, University of California, Davis CA 95616.
Barbados, Grenada, St Lucia, Dominica, & St Vincent, (2003). Breadfruit-Postharvest
Guidelines.
Breadfruit Sector Consortium supported by PAEPARD project, Mauritius 2012.
Breadfruit, (2004), Postharvest Care and Market Preparation, Ministry of Fisheries, Crops
and Livestock, New Guyana Marketing Corporation, National Agricultural Research
Institute, with assistance of United States Agency for International Development,
Technical Bulletin, No. 24.
Laurila, E., & Ahvenainen, R., (2004). Minimal processing in practice Fresh fruits and
vegetables, Minimal Processing Technologies in the Food Industry.
Maharaj, R., & Sankat, C. K., (1990). The shelf life of breadfruit stored under ambient and
refrigerated conditions. Acta Horticulturae, 269:411-424.
Maharaj, R., & Sankat, C. K., (2007). A review of postharvest storage technology of bread-
fruit. Acta Horticulturae, 757:183-192.
Marita Cantwell, (2004). Postharvest Handling Systems: Minimally Processed Fruits and
Vegetables. University of California Cooperative Extension, Vegetable Research and
Information Center. http://vric.ucdavis.edu/selectnewtopic.minproc.htm. April.
National Tropical Botanical garden-Breadfruit Institute, (2012). Tropical Plant Research,
Education and Conservation- Breadfruit Institute. http://ntbg/breadfruit/uses/table1.
php.
Ragone, D., (1997). Breadfruit: Artocarus Atilis (Parkinson) Fosberg. International Plant
Genetic Resources Institute, Rome.
Ragone, D., (2011). Farm and forestry production and marketing profile for breadfruit,
Speciality Rops for Pacific Island Agroforestry (http://agroforestry.net/scps).
Thompson, A. K. B. O., & Perkins, C., (1974b). Storage of fresh breadfruit. Tropical Agri.
(Trinidad), 51, 407–415.
Wills, R. B. H., Lee, T. H., Graham, D., McGlasson, W. B., & Hall, E. G., (1981). Posthar-
vest: an Introduction to the Physiology and Handling of fruits and Vegetables, New
South Wales University Press, Sydney.
Worell, D. B., & Sean, C. C. M., (1997). Breadfruit, In: Mitra, S. K., (ed.), Post Harvest
Physiology and Storage of Tropical and Sub Tropical Fruits, edn., Cab International.,
Oxford, UK, pp. 347–363.
CHAPTER 11

STANDARDIZATION OF
DEHYDRATION TECHNIQUES OF
SOME ORNAMENTAL FOLIAGES
MOUMITA MALAKAR, SUKANTA BISWAS,
and PINAKI ACHARYYA
Department of Horticulture, University of Calcutta,
51/2, Ballygunge Circular Road, Kolkata–700019, India,
E-mail: [email protected]

CONTENTS

Abstract.................................................................................................. 139
11.1 Introduction................................................................................. 140
11.2 Materials and Methods................................................................ 141
11.3 Results and Discussions.............................................................. 142
11.4 Conclusion.................................................................................. 146
Keywords............................................................................................... 149
References.............................................................................................. 149

ABSTRACT

The eco-friendly dehydrated foliages and plant parts secured much pop-
ularity among users and have become key components in floriculture
industry. Foliages with highly variable keeping quality are used as filler
element in flower vase. Dehydration of foliages has not been studied at
large. This investigation was carried out with ornamental foliages of three
140 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

species, viz., Araucaria cunninghamii, Thuja orientalis, and Juniperus

chinensis. White sand, silica gel, and boric acid were used as embedding
materials, and two drying conditions of microwave oven and room dry-
ing were adopted for three durational treatments, viz., 10, 20 and 30 s
and 4, 8, and 16 days, respectively. In both Araucaria and T. orientalis,
silica gel + microwave oven combination for 30 and 20 s, respectively,
exhibited best results in respect of moisture loss (49.23% and 58.33%)
and quality. White sand + room condition also caused 61.41% moisture
loss in T. orientalis when treated for 16 days. In J. chinensis, white sand
+ microwave oven and silica gel + room condition for 20 s and 16 days
showed moisture loss of 44.26% and 50.16%, respectively. Boric acid
as embedding materials was also found to be effective in dehydration of
these species.
All the three species were treated with glycerin:water of 1:1 and 1:3
(vol/vol) for 24, 48, and 96 h, followed by drying in hot air oven at 70–80°C
for 5 h and open air of room condition for 24 h. A significant moisture loss
of 60.56% to 62.56% was recorded in T. orientalis when dehydrated in hot
air oven for 96 h.

11.1 INTRODUCTION

Drying and preserving flowers, foliages, and plant materials are a form
of artistic expression that was very popular during the Victorian age and
has once again gained much popularity in the modern age. The decorative
value of dehydrated plant parts led to the use of vase decoration, bouquets,
and arrangements of gifts as well as ceremonial decoration of both home
and working places. Dried plant materials with decorative value have now
been accepted globally as natural, eco-friendly, long lasting, and inexpen-
sive. In the context of international trade of floriculture of India, dried
flowers and plant parts are the key segment and constitute 70% of total
share of floriculture products exported from this country (Sheela, 2008).
The reasons for the development of dry flower industry in India is possi-
bly due to easy availability of wide range of materials throughout the year
and manpower for such labor-intensive craft. Zizzo and Fascella (1997)
opined that the dried materials can be enjoyed whole year by arranging
Standardization of Dehydration Techniques 141

them in vases, creating arts like candle holders, and in other home deco-
rations. Possibly, the most common use of dried materials is in a wreath
or floral arrangement and also in ornament gift packages, masks, hats,
and lamp shades. Dried materials often embellish stationery or are used to
create unique pictures. Dried plant materials are extra special as they pos-
sess the characteristics of novelty, eco-friendly, esthetically near to fresh
flowers, flexibility, and year-round availability (Joyce, 1998). In the situ-
ation of climatic abnormalities in different parts of the world, which are
not congenial for keeping fresh-cut flowers in vases, dried flowers have
established tremendous potentiality, which is very much observed during
the last decades (Dhatt et al., 2007).
Moisture holding of dried flowers and foliages influence their quality
and longevity (Singh et al., 2003). Gill et al. (2002) studied the efficacy
of various methods of drying, viz., microwave oven, embedded in a desic-
cant, solar drying, press drying, and preservation in glycerin for drying
fern asparagus (Asparagus sp.) and silver oak (Grevillea robusta) leaves
for commercial and domestic purpose, and observed that embedding in
silica gel for 30 h was the best method for drying of ferns for commercial
purposes, whereas drying by embedding in a silica gel for 36 h and press
drying for 60 h was best for domestic purpose. In the light of the above,
the present investigation was undertaken with three perennial species to
standardize suitable method for developing dry leaves of these species.
Cut leaves of these species are widely used for vase decoration and other
decorations. Different methods, viz., embedding in different desiccants
and drying either in microwave oven or in room condition was attempted
along with preservation in glycerin.

11.2 MATERIALS AND METHODS

Fresh leaves of three different ornamental species of conifers, viz., Arau-

caria cunninghamii, Sweet (Hoop pine), Juniperus chinensis, Linn. (Chi-
nese juniper) belonging to the family Aracariaceae, and Thuja orientalis,
Linn., (Chinese thuja) of the family Cupressaceae were used for this study.
Mature fresh young green foliages with more or less uniform in size and
shape were collected in the morning hours followed by bagging in poly-
142 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

ethylene bag to avoid further desiccation. Fresh weight and size of foliages
were recorded. The experiment was conducted in the laboratory of the
Department of Horticulture, Institute of Agricultural Science, University
of Calcutta.
Foliages were embedded in pure drying media, viz., silica gel, boric
acid, and white sand followed by drying in microwave oven (600 wt) for
10, 20, and 30 s and open air in room condition for 4, 8, and 16 days. Leaf
samples were placed horizontally on desiccants at the bottom of micro-
wave-resistant glass containers followed by covering of about a depth of
2 inches with the same media to avoid further moisture absorption from
air, and the lid was tightened. Post drying characteristics were recorded
2 h after taking out from the microwave oven condition and keeping in
ambient condition. For glycerin preservation, foliages were submerged in
glycerin:water solution of 1:1 and 1:3 for 24, 48, and 96 h using boiling
water in which glycerin was added slowly, gently, and then dried in hot air
oven at 70–80°C for 5 h and open air of room condition for 24 h. To dis-
card excess glycerin:water solution, the materials after taking out of from
the glycerin were kept in hanging position.
In both the experiments, each treatment was replicated thrice consider-
ing one specimen as replication and the average data of each parameter
are presented. The data were analyzed using completely randomized block
design with factorial concept (Panse and Sukhatme, 1985).

11.3 RESULTS AND DISCUSSIONS

Results revealed (Table 11.1) that accelerated moisture loss is proportional

with the increase of duration in all three desiccants and species.
Highest moisture loss was recorded in silica gel. T. orientalis and A.
cunninghamii foliage materials recorded moisture loss of 58.33% in 20
s and 49.23% in 30 s under microwave oven condition. Tandon (1982),
Bhutani (1990), and Datta (1999) established silica gel as the best embed-
ding material followed by borax and sand. J. chinensis, on the other hand,
recorded 44.2% moisture loss in white sand + microwave oven condition
for 20 s treatment (Table 11.1). Electronically produced microwaves liber-
ate moisture from organic substances by agitating the water molecules;
TABLE 11.1 Effect of Different Embedding Media in Drying of Three Different Foliages under Different Drying Conditions
Embed- Dura- Average Aver- Chlo- Average Aver- Chlo- Average Aver- Chlo-
ding Mate- tion of moisture age size rophyll moisture age size rophyll moisture age size rophyll
rials treat- loss (%) reduc- content loss (%) reduction content loss (%) reduction content
ments tion (mg/gm.) (%) (mg/gm.) (%) (mg/gm.)
(sec.) (%)
Araucaria cunninghamii Thuja orientalis Juniperus Chinensis
Microwave oven condition
Silica gel 10 39.48c 2.64c 0.26a 56.66a 1.10b 0.35a 35.40c 1.20b 0.44a
20 46.37b 3.32b 0.20b 58.33a 1.30a 0.30a 40.28b 1.50b 0.37b
30 49.23a 3.88a 0.16c 57.07a 1.40a 0.23b 41.45a 2.10a 0.24c
White sand 10 23.47c 3.82b 0.13a 29.02c 1.20b 0.35a 40.44b 3.58b 0.43a
Standardization of Dehydration Techniques

20 32.30b 4.01b 0.07b 45.63b 2.19a 0.29b 44.26a 4.51a 0.35b

30 39.53a 5.92a 0.05b 49.91a 2.23a 0.16c 43.83a 4.64a 0.22c
Boric acid 10 29.19c 2.67c 0.14a 35.04c 1.07b 0.38a 33.60b 0.12b 0.38a
20 34.80b 4.02b 0.09b 53.92a 3.75a 0.28b 35.75b 0.13b 0.29b
30 40.88a 4.89a 0.06b 50.86b 4.27a 0.17c 40.40a 3.33a 0.21c
Room condition
Silica gel 4 49.72a 5.90a 0.40a 44.07b 3.27a 0.25a 31.48b 2.23c 0.36a
8 53.40a 6.03a 0.25b 57.86a 3.85a 0.22a 33.09b 3.53b 0.31a
16 56.16a 6.19a 0.21b 60.36a 3.86a 0.21a 50.16a 4.42a 0.28b
143
TABLE 11.1 (Continued) 144
Embed- Dura- Average Aver- Chlo- Average Aver- Chlo- Average Aver- Chlo-
ding Mate- tion of moisture age size rophyll moisture age size rophyll moisture age size rophyll
rials treat- loss (%) reduc- content loss (%) reduction content loss (%) reduction content
ments tion (mg/gm.) (%) (mg/gm.) (%) (mg/gm.)
(sec.) (%)
Araucaria cunninghamii Thuja orientalis Juniperus Chinensis
White sand 4 27.64b 4.50a 0.37a 22.58c 3.14b 0.23a 20.39c 1.71c 0.24a
8 50.09a 5.50a 0.28b 48.73b 3.26b 0.20a 26.16b 2.27b 0.15b
16 50.21a 5.53a 0.22b 61.41a 4.22a 0.19b 45.90a 3.11a 0.11b
Boric acid 4 12.38c 2.06a 0.30a 19.75b 1.04b 0.21a 16.47c 2.00c 0.29a
8 31.59b 2.15a 0.24b 56.20a 3.04a 0.18b 36.72b 2.93b 0.23b
16 37.04a 2.62a 0.18c 60.68a 3.90a 0.16b 49.07a 4.90a 0.23b
Sustainable Horticulture Volume 2: Food, Health, and Nutrition
Standardization of Dehydration Techniques 145

this is the principle underlying the quickest microwave oven drying (Bhu-
tani, 1990a). In all the three species, the effect of white sand and boric acid
as desiccants was almost at par. Size reduction of leaves was increased
with the increase in treatment duration. Bhalla et al. (2006) observed that
average foliage size reduction in chrysanthemum was more with white
sand and boric acid than silica gel, but maximum size reduction of flowers
was noted while embedded in silica gel and dried in microwave condi-
tion. In Araucaria cunninghamii and Juniperus chinensis size reduction
of 5.92% and 4.64%, respectively, was recorded while drying under white
sand + microwave for 30 s. In T. orientalis, a reduction of size by 4.27%
was recorded in boric acid +microwave for 30 s.
Dried leaves with better chlorophyll content could be obtained with
minimum duration of treatment, as it is the main pigment content of foliages
and play a vital role for the determination of dried plant part’s attractive-
ness. In regard to chlorophyll restorability, silica gel-embedded materials
showed maximum chlorophyll recovery with 0.26 and 0.44 mg/g in 10 s
treatment in A. cunninghamii and J. chinensis. Boric acid represented as
best desiccant for T. orientalis with the maximum chlorophyll recovery of
0.38 mg/g in 10 s treatment. Meman and Barad (2009) observed higher
pigment reduction at higher temperature, which was consistent with our
result. No marked variation in the shape and texture of post-drying foli-
ages was observed.
In room drying, increased duration caused increased moisture reduc-
tion. The activity of all three desiccants was more or less the same under
room drying, but among them, silica gel showed good result by reduc-
ing 56.16% of moisture in 16 days treatment, followed by white sand in
A. cunninghamii. Moisture reduction of T. orientalis leaves was similar
under all embedding materials with an extent of losing 60.82% moisture.
Silica gel and boric acid caused 50.16% and 49.07% moisture loss from J.
chinensis leaves, respectively, but white sand failed to show any effective
result. Misra (2002) established the fact that dried flowers and leaves with
a specific moisture level can be stored for a very long period without los-
ing their appearance and decorative value.
With regard to size reduction of leaves through room drying, the
reduction varied with the desiccants and species. Size reduction was little
with boric acid in A. cunninghamii and T. orientalis (2.06% and 1.04%,
146 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

respectively). Color retention was much better in room drying than in

microwave oven + sand media. It is noteworthy to mention that the main
principle of drying is to diminish moisture content to a point at which
the biochemical changes are minimized but maintaining cell structures,
pigment level, and actual shape of flowers or foliages (Dana and Larner,
2001).
In another part of experiment, foliages of the three species were
allowed to absorb in two concentrations of glycerin:water solution, viz.,
1:1 and 1:3 for 24, 48, and 96 h. Lee et al. (2003) exclaimed that dye with
glycerin gave the flowers and foliages a more vibrant color in addition to
its soft and pliable feel (Table 11.2).
It is evident from Table 11.2 that moisture loss from materials
increased with the imbibition time of glycerin:water, irrespective of dry-
ing techniques. Highest moisture loss was recorded with leaves of all three
species treated for 96 h in both combination of glycerin used. Insignificant
variation of moisture loss from leaves due to the use of different concen-
trations of glycerin were also observed. Among three species, T. orientalis
leaves showed a moisture loss of 62.5% under hot air oven drying after 96
h absorption in 1:1 glycerin:water solution, while the same trend was also
noted under room drying. Minimum size reduction of leaf was recorded
with 1:1 glycerin:water solution (24 h) in T. orientalis and J. chinensis
leaves both under hot air oven and room drying. In A. cunninghamii foli-
ages, the 1:3 glycerin:water solution showed a reduction of 1.20–2.04%
in different drying conditions. Chlorophyll deterioration trend was promi-
nent with the increase in treatment duration of all the species. Campbell et
al. (2001) and Sohn et al. (2003) experimented and proved glycerol as an
appropriate preservative for Eucalyptus cinerea and Magnolia grandiflora
for retaining natural appearance as it replenishes the natural moisture of
the leaf with a substance that maintains leaf fall, texture, color, and sugar
level.

11.4 CONCLUSION

Dehydration is an important post-harvest technology for enhanc-

ing ornamental keeping quality of flowers and foliages as it quickly
TABLE 11.2 Effect of Glycerin Preservation of Three Different Foliages under Different Drying Conditions
Treatments Duration of Average Foli- Chlo- Average Foliage Chlo- Average Foliage Chlo-
treatments moisture age size rophyll moisture size re- rophyll mois- size re- rophyll
(hrs.) loss (%) reduction content loss (%) duction content ture duction content
(%) (mg/gm.) (%) (mg/gm.) loss (%) (%) (mg/gm.)
Araucaria cunninghamii Thuja orientalis Juniperus Chinensis
Hot-air-oven condition
Glycerin- 24 13.81c 2.01b 0.24a 24.48c 3.44c 0.25a 26.79c 2.05c 0.36a
water (1:1) 48 24.62b 2.90a 0.17b 42.23b 4.02b 0.21a 32.12b 2.38b 0.25b
96 33.33a 3.06a 0.12b 62.50a 4.47a 0.15b 33.95a 4.23a 0.20c
Glycerin- 24 22.98b 1.20c 0.25a 24.40c 3.49a 0.22a 24.60c 3.19c 0.31a
water (1:3) 48 23.44b 2.62b 0.16b 36.81b 3.51a 0.19b 30.50a 4.20b 0.22b
Standardization of Dehydration Techniques

96 30.08a 2.79a 0.11b 60.56a 3.59a 0.13b 31.50a 5.30a 0.18c

Room condition
Glycerin- 24 15.10c 2.56c 0.22a 27.20b 0.10b 0.23a 13.49c 1.33a 0.30a
water (1:1) 48 25.50b 2.86b 0.16b 28.30b 0.11ab 0.16b 20.50b 1.48a 0.22b
96 34.40a 3.38a 0.14b 29.60a 0.12a 0.13b 22.80a 1.56a 0.15c
Glycerin- 24 23.50c 2.04b 0.20a 26.50b 0.14b 0.21a 10.00c 2.36b 0.28a
water (1:3) 48 25.50b 3.08a 0.14b 27.60ab 0.20a 0.14b 19.07b 2.58ab 0.21b
96 31.50a 3.18a 0.09c 28.50a 0.23a 0.12b 21.65a 2.92a 0.13c
147
148 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

reduced the moisture content to a point at which there is little biochemi-

cal change, while keeping cell structure and shape unaffected. Dry foli-
ages can be stored for unlimited period if they are well secured from
the damage of atmospheric high humidity. Since the last three decades,
scientists are putting their efforts to standardize the dehydration tech-
niques for flowers and foliages that require special care to protect their
shape, sizes, and color, while other plant parts like branches, cones,
and barks can be dried with little care. With this aim, this investigation
succeeded to point out the efficacy of various desiccants and drying
conditions for dehydrating the selected foliages. Drying in microwave
oven showed quicker result as compared to room drying, as increased
rate of moisture loss due to more conduction and convection of heat to
foliage tissue and water evaporation from the surface might have caused
rapid drying in microwave oven (Singh and Dhaduk, 2005). Irrespec-
tive of media, moisture reduction was accelerated with the increased
duration in all species of plants. Silica gel caused maximum mois-
ture loss by attracting moisture following the phenomenon known as
physical absorption and capillary condensation irrespective of drying
condition, followed by boric acid and white sand. In microwave oven
drying, Araucaria sp. foliages reduced maximum in size, while silica
gel reduced maximum size in room condition. Sand media in micro-
wave drying proved best for J. chinensis. Foliage color in all species
were bleached to some extent, while the fading effect of boric acid has
also been reported by Pamela (1992) and subsequently confirmed by
Singh and Dhaduk (2005). Embedding methods were found to be appro-
priate for maintaining shape of foliages as it recommended for dry-
ing to get three dimensional views (Singh and Dhaduk, 2005). It was
observed through our experiment that glycerin uptake was increased
with the increase in absorption duration. Highest moisture loss percent-
age was obtained in T. orientalis leaves in hot air oven drying after 96
h absorption in 1:1 glycerin:water solution. Similar findings were also
reported by Joyce and Dubious (1992). Thus, owing to great availability
of variety of plants, flowers, and other artistic raw materials, India has
got enormous scope to propagate this field, as dry flower market is still
small in comparison to that of the fresh flower market, which might be
due to its more recent introduction in the floricultural trade.
Standardization of Dehydration Techniques 149

KEYWORDS

•• dehydrated
•• eco-friendly
•• embedding
•• foliage
•• moisture

REFERENCES

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techniques of Chrysanthemum (Dendranthemum grandiflorum Tzvelev). Journal of
Orna- mental Horticulture, 9(3), 159–163.
Bhutani, J. C., & Tandon, R. K., (1982). Sukhe phoolon se sajawat kijiye. Phal-Phool.,
5(3), 3–5.
Bhutani, J. C., (1990). Dry rose craft. Rose News, 9(11), 8–9.
Buzarbarua, A., (2000). A Textbook of Practical Plant Chemistry. S., Chand and Company,
New Delhi, pp. 83–87.
Campbell, S. J., Ogle, H. J., & Joyce, D. C., (2001). Glycerol Uptake Preserves Cut Juve-
nile Foliage of Eucalyptus Cinera-School of Land and Food, 3rd edition., Australia,
vol. 15, pp. 492.
Dana, M. N., & Larner, B. R., (2002). Preserving Plant Materials- University Cooperative
Extension Service. 2nd edition., West Lafayette. Department of Horticultural Flowers,
vol. 19, pp.102.
Datta, S. K., (1999). Dehydrated flowers, foliage and floral craft. In: Floriculture and
Landscap-ing. Naya Prakash, Kolkata, pp. 696–703.
Dhatt, K. K., Singh, K., & Kumar, R., (2007). Studies on methods of dehydration of rose
buds. Journal of Ornamental Horticulture, 10(4), 264–267.
Gill, S., Bakshi, S., & Arora, S., (2002). Standardization of drying methods for certain cut
flowers. Journal of Ornamental Horticulture, 9(3), 159–163.
Joyce, D. C., & Dubious, P., (1992). Preservation of fresh cut ornamental plant material
with glycerol. Postharvest Biology and Technology, 2(2), 145–153.
Joyce, D. C., (1998). Dried and preserved ornamental plant materials- not new but often
over-looked and underrated. Acta. Horticulture, 45(4), 133–145.
Lee, W. Y., Mijeong, Y., Chunho, P., & Beyounghwa, K., (2003). Effects of various drying
methods for wild flowers. Korean Journal of Horticulture Science and Technology,
21(1), 50–56.
Meman, A., & Barad, V., (2009). Study on dry leaf production of asparagus. Journal Hor-
ticulture and Forestry, 11(3), 43–47.
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Pamela, W., (1992). The Complete Flower Arranger. Annes Publishing Limited, London,
pp. 255.
Panse, V. G., & Sukhatme, P. V., (1985). Statistical Methods for Agricultural Workers.
ICAR, New Delhi, pp. 55–70.
Ranjan, J. K., & Mishra, S., (2002). Dried flowers: A way to enjoy their beauty for a long
period. Indian Horticulture, 10, 32–33.
Sharma, G. K., Semwal, A. D., & Arya, S. S., (2000). Effect of processing treatments on
carotenoids composition of dehydrated carrots. Journal of Food Science Technology,
37(2), 196–200.
Sheela, V. L., (2008). Dry flowers: a profitable floriculture industry. In: Flower for Trades.
3rd edition., New India Publishing Agency, New Delhi., vol. 10, pp. 65–67.
Singh, A., Dhaduk, B. K., & Shah, R. R., (2003). Effect of dehydration on post harvest life
and quality of zinnia flowers. Journal of Ornamental Horticulture, 6(2), 141–142.
Singh, A., Dhaduk, B. K., & Shah, R. R., (2005). Effect of dehydration techniques of some
selected flowers. Journal of Ornamental Horticulture, 6(2), 155–156.
Sohn, K., Kwon, H., & Kim, E., (2003). A optimum drying temperature to maintain sea
sand dry color of roses. Korean Journal of Horticulture Science and Technology,
21(2), 141–145.
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culture, 10(2), 45–49.
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51–60.
CHAPTER 12

DEPENDENCE ON NON-TIMBER
FOREST PRODUCTS FROM A
COMMUNITY FOREST AS A SAFETY
NET FOR LIVELIHOOD SECURITY
AMONG THE VILLAGERS OF MAMIT
DISTRICT, MIZORAM
K. LALHMINGSANGI and U. K. SAHOO
Department of Forestry, School of Earth Sciences and Natural
Resource Management, Mizoram University, Aizawl–796004, India,
E-mail: [email protected]

CONTENTS

Abstract.................................................................................................. 151
12.1 Introduction................................................................................. 152
12.2 Materials and Methods................................................................ 154
12.3 Results and Discussion .............................................................. 156
12.4 Conclusions................................................................................. 170
Keywords............................................................................................... 172
References.............................................................................................. 172

ABSTRACT

Non-timber forest products (NTFPs) provide an important source of liveli-

hood security for the inhabitants living adjacent to forest areas. A survey
152 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

was conducted in five villages under Mamit district of Mizoram, and it was
found that NTFPs act as safety nets by collecting available NTFPs from
the community forest (Village Forest Development Committee (VFDC)
plantation areas). The main aim of the study is to focus on the dependence
of NTFPs from the community forest of the surveyed villagers. For this,
participatory rural appraisal (PRA) was done along with questionnaires, per-
sonal interviews, and group discussion with the villagers. The weaker sec-
tion of the society gets benefited by providing food security in times of
unavailability of agricultural cash crops, which was one of the basic needs
for their livelihood. Besides the home consumption of NTFPs, they were
sold to meet the cash requirements especially by the widow and landless
farmers in all the surveyed villages. Such is the case; it gave them a kind
of natural insurance and security for future needs. Bamboo pole, broom
stick, fruits, wild foods, and fuelwood are the main NTFPs that they have
collected from the community forest area. Among the various NTFPs,
fuelwood (56%) has the highest percentage of household involved in har-
vesting alone among the studied villages, followed by wild food (46%),
broomstick (35%), bamboo pole (31%), fruits (8%), and medicinal plants
(5%). The potential of NTFPs in community forest are well benefited by
fulfilling their daily food requirements, building materials, meeting cash,
and saving money by exploiting NTFPs and reduce the yearly expenditure.

12.1 INTRODUCTION

Non-timber forest products (NTFPs) are considered to be important for

sustaining rural livelihood, reducing rural poverty, biodiversity conserva-
tion, and facilitating rural economic growth (Global NTFP partnership,
2005). Several opportunities for improved rural development are linked
to NTFPs (Adepoju et al., 2007) as the villagers are traditionally depen-
dent on forest resources for their food, shelter, and income through col-
lection and marketing of NTFPs (Sahoo et al., 2010). If the villagers
were not extracting wild food plants from the forest, their needs of cash
income would double to maintain their daily meal. Household may trade
in NTFP’s in times of hardship, with this activity forming an important
safety net (Takasaki et al., 2004).
Dependence on Non-Timber Forest Products 153

In recent years, with the advent of globalization, there are indications

of a rapid increase in the extraction mostly from natural populations. In
India alone, it is estimated that over 50 million people are dependent on
NTFPs for their subsistence and cash income (National Centre for Human
Settlements and Environment, 1987; Hegde et al., 1996). Recent studies
estimated that 275 million poor rural people in India equal to 27% of the
total population depend on NTFPs for at least part of their subsistence
and cash livelihoods (Bhattacharya and Hayat, 2009; Malhotra and Bhat-
tacharya, 2010). The women of mountainous regions have maximum work
inputs for agricultural activities and domestic responsibilities, particularly
in rural areas (Salam et al., 2005).
Forest resources are also important to battle the global warming and
climate change problems such as carbon sink (IIPCC, 2001). It has also
been realized that this will not be possible without active co-operation of
the inhabitants living adjacent to forest areas. This situation demands a
more coherent system for sustainable development of forest resources in
the Joint Forest Management program, which should be based on regu-
lating management for capacity building of community. Introduction of
forestry practices for maximum production of fuel wood, fodder, fruits,
timber, fiber, etc., must be encouraged (Rawat and Sharma, 2010). How-
ever, when there is no market for the NTFPs, the methods of evaluation
are not as straight forward. While environmental economists and ecologi-
cal economists have done extensive research on hypothetical markets, the
techniques they have developed are not always suitable to estimate the
value of nonmarketed NTFPs. In some communities, NTFPs might be
exchanged for marketed products, rather than sold directly. NTFPs that
are consumed rather than sold in the market can be considered income in
kind rather than in cash. Thus, ignoring the role of NTFPs consumption
in the livelihoods of rural populations gives a very distorted view of the
importance of NTFPs and of their economic values (Chopra, 1993).
The Joint Forest Management program is a co-management regime
for protection, regeneration, and development of degraded forests where
the role of NGOs’ has been very useful in bridging the gap between state
and the people dependent on forests. Under this program, the forest area
under consideration is protected and managed jointly by the local com-
munity and forest department, while the ownership of the forest lies with
154 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

the government. The people get in turn the forest products free of cost
from the community-managed forests. This paper analyzes the extent of
dependence on NTFPs by the people from the jointly managed forest area.

12.2 MATERIALS AND METHODS

12.2.1 STUDY SITE

The study was carried out in five villages of Mamit district, viz., Dapch-
huah, Chhipui, Chungtlang, Tuahzawl, and Lengte Village Forest Devel-
opment Committee (VFDC) plantation areas during the year 2013–2015
(Figure 12.1). Even though the occupation of the people from these five
villages is mainly jhum cultivation, they are exploiting NTFPs from
VFDC plantation areas to supplement the agricultural crops and meet
their daily needs. These VFDC plantation areas are mainly situated 2–5
km away from the village in the community land areas and are monitored
by the villagers along with the Forest Department, which provide a sense
of ownership to the villagers.

12.2.2 METHODOLOGY

Participatory rural appraisal (PRA) was adopted for the field study. Primary
and secondary data were collected using questionnaire, personal interview,
and interaction with the villagers. Semi-structured questionnaire was given
to approximately 10% of the household from each village to provide infor-
mation on to what extent they were involved in the exploitation of NTFPs
as well as to know the different benefits they got through the community
forest, i.e., VFDC plantation areas. A small group interaction was done along
with the village leaders including females to avoid gender division and to
get a real picture discussing the positive and negative aspect in harvesting,
processing, and marketing of NTFPs from the plantation sites. Socioeco-
nomic survey, land use pattern, value addition of NTFP, and dependency
on forest as well as marketing strategy were also taken into consideration.
Major emphasis was given on the collection of data by direct interaction
with those individuals who were actually engaged in utilization of NTFPs.
Dependence on Non-Timber Forest Products 155

FIGURE 12.1 (See color insert.) Map of Mamit district showing the study sites.
156 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

12.3 RESULTS AND DISCUSSION

12.3.1 SOCIOECONOMIC PROFILE OF THE STUDIED

VILLAGE

The occupation of most of the villagers is agriculture in which most of

them practice shifting cultivation, and approximately 9–11% of them
have horticulture farm, 1–2% hold government jobs, and 1–2% are
involved in trading. NTFPs are exploited from the community forest of
the VFDC plantation area and from other community forests nearby their
villages. Most of them are part-time NTFP exploiter since these NTFP
are seasonal; besides marketing, they harvest and exploit those NTFPs
as a substitute for their cash crop. As shown in the table, among all the
studied villages, the population was highest in Dapchhuah village with
1130 total population and 235 households, followed by Chhippui village
with 950 total population and 186 households, Lengte village with 530
population and 135 households, Tuahzawl village with 520 population
and 100 households, and the least was in Chungtlang village with 476
total population and 100 households. Literacy rate was high in all the
studied villages and varied between 97% and 99%. Tuahzawl village has
the highest literacy rate of 99%, followed by Chhippui, Chungtlang, and
Lengte village constituting 99% literacy rate and the least was in Dapch-
huah village at 97%.

12.3.2 INFRASTRUCTURE

As shown in Table 12.1, Anganwadi centers and primary and middle

schools were located in all the studied villages, whereas high school
was present only in Dapchhuah and Chhippui villages; these schools
are mainly run by the state government. Health subcenters and bank
facilities were absent in all the five villages, and the villagers, in case of
health emergency, have to go to their nearby town. One community hall
was present in Dapchhuah, Tuahzawland, and Lengte village each. Four
to five churches as well as spring water sources were present in each
village.
Dependence on Non-Timber Forest Products 157

TABLE 12.1 Socioeconomic Profile of the Five Surveyed Villages under Mamit district
in Mizoram
Attributes Surveyed VFDC Villages
Dapchhuah Chhippui Tuahzawl Chungtlang Lengte
No. of house- 235 186 100 100 135
hold
Male 510 550 288 271 250
Female 620 400 232 205 280
Total popula- 1130 950 520 476 530
tion
Matriculation 15 36 41 20 39
Graduate 2 20 20 7 15
Literacy rate % 97 98 99 98 98
Average land 1 1 1 1 1
holding (ha)
Horticulture 30 15 12 10 27
garden (HH)
Jhum cycle 7 6 8 6 7
(year)
Anganwadi 3 4 3 2 3
Centre
Primary school 1 1 1 1 1
Middle school 1 1 1 1 1
High school 1 1 - - -
Health Sub - - - - -
centre
Bank - - - - -
Community 1 - 1 - 1
hall
Church 4 5 5 4 5
Spring water 4 4 5 4 4

12.3.3 USE OF VARIOUS NTFPS

From the surveyed villages, 31% of household are involved in harvest-

ing and processing of bamboo pole, whereas 35.4% household in broom
grass, 6% in fruits, 45.8% in wild foods, 56.4% in fuel wood and 4.6%
in medicinal plants. Fuel wood has the highest percentage of household
158 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

involvement among all the NTFPs mainly because it is in high demand by

most of the households all throughout the year. Least involvement of the
villagers in the selected NTFPs was found in medicinal plants due to less
abundance in all the areas, and youths are more interested in other medi-
cines and therefore are lesser drawn to it (Table 12.2).

12.3.3.1 Bamboo Pole

The most common bamboo species used for bamboo pole in the studied
villages are Bambusa tulda (Rawthing), Melocanna baccifera (Mau-
tak), Dendrocalamus longispathus (Rawnal), Schizostachylum dullooa
(Rawthla), Bambusa vulgaris (Vai rua), Dendrocalamus giganteus (Raw-
pui), Schizostachyum fuchsiamum (Raw te), Dendrocalamus hookeri
(Rawlak), Schzostachyum mannii (Raw te), and Dendrocalamus hamil-
tonii (Phul rua), which meet the requirement of bamboo poles; however,
people do also harvest bamboo poles from the adjoining forest areas. People
have a strong believe that for all species of bamboo, harvesting mature culms
and in the right season sustains their productivity. The number of household
using bamboo pole is high in most of the villages, with the highest in
Chhippui at 60%, followed by Chungtlang at 30%, Dapchhuah at 30%,
Lengte at 25%, and the least in Tuahzawl at 10%.
Most of them harvest for their own consumption, except in Chungtlang
and Chhippui villages. Dendrocalamus longispathus is mainly harvested
for weaving local carrier (em), and for this one, mature bamboo of 15 ft is
sufficient to complete the carrier (Figure 12.2 and 12.3). Bambusa tulda is
preferred for weaving local winnowing fan, and for this, 1 m bamboo with
four nodes is usually harvested. M. baccifera is used for making different
instrument handles. The local carrier made from D. longispathus is in high
demand all throughout the year, especially in towns and villages. Weaving
is done according to the demands and order.

12.3.3.2 Broom Grass (Thysanolaena maxima)

The inflorescence of broom grass is harvested on maturity when the pan-

icles become tough and their color change to light green or red during
TABLE 12.2 Use of Different NTFPs in the Five Studied Villages
NTFPs Surveyed villages
Dapchhuah Chhippui Tuahzawl Chungtlang Lengte
Bamboo Poles % of HHs involved/year 30 60 10 30 25
Quantity harvested (kg/hh/yr) 100 200 20 50 30
Home consumption (kg/hh/year) 45 70 20 50 30
Quantity sold (Kg/hh/year) 55 120 - - -
Income generated (Rs/hh/year) 300 1400 - - -
Broom Grass % of HHs involved/year 7 80 50 20 20
Quantity harvested (kg/hh/yr) 28 20 10 10 5
Home consumption (kg/hh/year) 10 20 10 10 5
Quantity sold (Kg/hh/year) 18 - - - -
Dependence on Non-Timber Forest Products

Income generated (Rs/hh/year) 280 - - - -

Fruits % of household involved/year 15 3 9 - 3
Quantity harvested (kg/hh/yr) 12 5 5 - 5
Home consumption (kg/hh/year) 12 5 5 - 5
Quantity sold (Kg/hh/year) - - - - -
Income generated (Rs/hh/year) - - - - -
159
TABLE 12.2 (continued) 160

NTFPs Surveyed villages

Dapchhuah Chhippui Tuahzawl Chungtlang Lengte
Wild food % of household involved/year 60 70 20 50 29
Quantity harvested (kg/hh/yr) 104 149 30 35 50
Home consumption (kg/hh/year) 20 120 10 30 10
Quantity sold (Kg/hh/year) 84 29 20 5 40
Income generated (Rs/hh/year) 1050 240 200 50 650
Fuel Wood % of household involved/year 57 60 50 60 55
Quantity harvested (kg/hh/yr) 66 50 35 45 30
Home consumption (kg/hh/year) 16 50 35 45 30
Quantity sold (Kg/hh/year) 50 - - - -
Income generated (Rs/hh/year) 150 - - - -
Medicinal plants % of household involved/year 6 9 4 2 2
Quantity harvested (kg/hh/yr) 5 4 4 3 3
Home consumption (kg/hh/year) 5 4 4 3 3
Quantity sold (Kg/hh/year) - - - - -
Income generated (Rs/hh/year)
Sustainable Horticulture Volume 2: Food, Health, and Nutrition
Dependence on Non-Timber Forest Products 161

FIGURE 12.2 Weaving of local carrier (Pai em) from Dendrocalamus longispathus.

FIGURE 12.3 Different weaving products of Dendrocalamus longispathus.

winter season from January to March. It is harvested by hand pulling of

the culms. It is further processed by exposure to sunlight for few days and
then cleaning off the seeds. It is then wrapped and cut in a proper man-
ner to add the value. Thatch grass is harvested in the month of November
to April when the grass reach its highest length and maturity and further
processed by exposure to direct sunlight for about a month. Even though
broom grass is used by all the households, harvesting is done mainly
for their own consumption and not for market purpose. Further, 80% of
households are involved in harvesting of broom grass in Chhippui, fol-
162 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

lowed by 50% in Tuahzawl, 20% in Chungtlang and Lengte, and the least
7% in Dapchhuah. Even though Dapchhuah has the least percentage of
household involvement in harvesting, they are the only one who market
broom grass to local markets.

12.3.3.3 Wild Fruits

Garcinia lanceifolia Roxb, Artocarpus heterophyllus, and Emblica offici-

nalis fruits are harvested by the villagers. They harvest only for their own
consumption. Children below the age of 15 years are mostly involved in
harvesting of these fruits. Some adult exploiters when they cross by the
plant also harvested it. Highest consumption of wild fruits was seen in
Dapchhuah village constituting 15% of household; this is mainly because
the community forest is easily accessible to them, and they need to cross
that area frequently. This was followed by Tuahzawl at 9%, Chhippui and
Lengte at 3%, and Chungtlang village at 0%. There are no market benefits
from the fruits harvested from the studied villages (Figure 12.4).

FIGURE 12.4 Garcinia lanceifolia fruit harvest in Tuahzawl village.

Dependence on Non-Timber Forest Products 163

12.3.3.4 Wild Food

Amorphophallus paeonifolius, Homalomena aromatica, mushroom,

Solanum torvum Sw, bamboo shoots (M. baccifera and D. longispathus),
Eurya acuminatea, and Amomum dealbatum Roxb. are the main foods
harvested from the community forest of the studied villages (Figure
12.5). In the case of elephant foot yam (A. paeonifolius), only the male
tuber weighing about 4–5 kg is harvested by digging the ground with
spade or any other equipment without harming the tuber. Value addition
is done by slicing the tuber and then boiling it with water for 1 h; subse-
quently, the water is drained, the peel is taken off, and the tuber is boiled
again with local-made sodium carbonate. It is then smashed and wrapped
in a banana leaves. Solanum torvum young fruits are also harvested and
cooked along with other vegetables and serve as important vegetables. A
common medicinal plant Homalomena aromatic leaves is harvested and

FIGURE 12.5 (1) Amorphophallus paeonifolius before processing. (2) Amomum

dealbatum in local market. (3) Bamboo shoots (Dendrocalamus longispathus) in local
market. (4) Eurya acuminatea in its habitat.
164 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

consumed by the villagers of Chhippui villages as a vegetable. Trem-

ellomycetes fuciformis (pa sawntlung), Schizzophylum commune (pasi),
and other varieties of mushrooms are available for consumption during
the month of July to October and are harvested by direct hand pluck-
ing, which provide good income. However, mushroom once available in
plenty are very scanty now due to the death of bamboo (owing to bamboo
flowering) a few years back and because of insufficient decomposition
of bamboo base that produce mushroom (Figure 12.6). Lesser people are
involved in harvesting of mushroom because mushrooms need a good
timing to harvest them and also because it is less abundant in the planta-
tion area. The marketing rate of mushroom is more or less high in all the
villages. Local market price of mushroom was 100–150 Rs/kg. Bamboo
shoots are one of the most important vegetables and have a high market
demand as well. The shoots of M. baccifera and D. longispathus are
mainly consumed by the villagers. They are consumed in fresh form,
and the cooked shoots are even sun dried and preserved for off season.
Bamboo shoots are one the most important substituents of agricultural
cash crops and constitute the highest percentage of household involved
among all the foods collected from the community forest. Eurya acumi-
natea leaves are also harvested and used as vegetables from the surveyed
villages. Amomum dealbatum young shoots and buds are cooked and
used as vegetables. Dapchhuah at 60% and Chhippui at 70% have the
highest percentage of household involved in harvesting different avail-
able wild foods from the community forest areas and also the highest
income as compared to the other villages (Chungtlang 50% and Lengte
29% and Tuahzawl 20%).

FIGURE 12.6 Processing of bamboo shoot at a roadside market.

Dependence on Non-Timber Forest Products 165

12.3.3.5 Fuel Wood

In all the studied villages, all the households are using fuelwood in addi-
tion to LPG. Consumption of fuelwood is high in most of the villages;
the more interior the place, the higher is consumption of fuelwood. Even
though most of them have an LPG connection, they cannot fully rely on
that due to scarcity of gas and the high cost. They are more comfortable
using fuelwood; therefore, all the household somehow harvest fuelwood
from the nearby forest as well as from the VFDC plantation area. Com-
mon fuelwood species harvested by the villagers are Quercus pachyphylla,
Anoglissus acumulata, Mesua ferrea, Schima wallichi, Bischofia javanica,
and Callicarpa arborez. M. baccifera is also harvested as a substitute to
these species in times of scarcity of fuelwood. A total of 56.4% of house-
hold are involved in harvesting fuelwood from the community forest, and
the remaining households collect fuelwood from their own land. Among
all the NTFPs, household participation in the collection of fuelwood from
the community forest is highest in fuel wood as compared to other NTFPs.
Chungtlang and Chhippui villages have the highest density of fuelwood
species (Figure 12.7), covering 50% of the plantation area. Most of them
are naturally grown. The highest consumption of fuelwood from the area
is also from these two villages, with 60% of the households involved in
the harvesting of these fuelwood species from the plantation area. This
was followed by Dapchhuah at 57%, Lengte at 55% and Tuahzawl at 55%.

FIGURE 12.7 Fuelwood in Dapchhuah village.

166 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Besides the density of fuelwood species, the distance of community forest

from the village also matters; the nearer the community forest, the more is
the collection seen in most of the villages. Dapchhuah is the only village
that sells the fuelwood and gets income from it. This is mainly because
Dapchhuah is located on national highway, which gives a good chance of
marketing the fuelwood.
In ancient Mizo culture, every family is expected to have a fuelwood
stock. This is mainly because when some people pass away within the vil-
lages, the Young Mizo Association (YMA) (which was one of the biggest
nongovernmental organization (NGO) in Mizoram) used to collect 1–2
fuelwood for that family as a helping hand. This is still practiced in all the
studied villages, which make them to increase the rate of harvesting from
the plantation areas as well as from the nearby forest besides their own
consumption.
Mature Gmelina arborea is also used for making the wheels of local-
made vehicle (Figure 12.8), which is one of the most important means of
transportation, especially for carrying goods from the field to home, for
carrying water, etc., within the village.

12.3.3.6 Medicinal Plants

There is no specific season for harvesting of medicinal plants. The villag-

ers harvest different medicinal plants and consume them in fresh form only
when needed. Leaves are mostly harvested. Litsea monopetala (Roxb.)
(Figure 12.9) Pers leaves are harvested by the villagers of Chhippui for

FIGURE 12.8 Local-made vehicle with wheels made of mature Gmelina arborea.
Dependence on Non-Timber Forest Products 167

FIGURE 12.9 Litsea monopetala, an important medicinal plant in the area.

the treatment of cattle sores. They add the leaves along with the cattle
food so as to heal the sores. Leaves of Mikania micrantha Kunth, a quick
growing climber, is commonly used as an antiseptic and applied to fresh
cuts. Clerodendrum colebrookianum Walp. leaves are also harvested by
the villagers for reducing high blood pressure, in addition to being used as
vegetables. Plant medicines remain one of the most affordable and easily
accessible sources of treatment in primary healthcare in the studied vil-
lages. The beneficiaries at Chhippui village (9% household) are making
the best use of the medicinal plants from the plantation area, followed by
Dapchhuah village (6% house hold) while least utilization on medicinal
plants was found on Chungtlang and Lengte village (2% household each).

12.3.4 MARKET FLOW OF NTFPS

In the present study, from the five villages, the localities did all the harvest-
ing, processing and even marketing of NTFPs. Marketing of selected NTFPs
are mainly done in their own village, neighboring villages and towns, road-
side, junction selling points, and markets in the nearest urban centers and cit-
ies. Most of the NTFP exploiters are part-time exploiter as they are seasonal
and occasional. However, approximately 3% household among the NTFP
producers are full time with varying products and locations. Medicinal plants
168 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

are not sold by the villagers; they use it only for their own consumption.
Few years back, Homalomena aromatica rhizomes were harvested in a large
amount and were sold in national market through a middleman from Chhip-
pui village; the rhizomes are now preserved, and collection of such plants in
a large amount was prohibited by the local leaders (Figure 12.10–2.15).

FIGURE 12.10 (See color insert.) Percentage of household involved in harvesting of

NTFPs.

FIGURE 12.11 (See color insert.) Quantity of NTFPs harvested in the studied villages
(kg/hh/year).
Dependence on Non-Timber Forest Products 169

FIGURE 12.12 (See color insert.) Amount of NTFPs used for home consumption in
different villages (kg/hh/year).

For a period of time, prohibition of harvesting of bamboo shoots by

the Forest Department created a problem in marketing of bamboo shoots.
Broom grass is the only NTFP which have its marketing channel at
national level. A number of stakeholders are involved in the marketing of
broom grass as it is a lengthy channel. Though, local trade has an advan-
tage which allow participation by the poor.

12.3.5 SOCIAL BENEFITS FROM ENTRY POINT ACTIVITIES

(EPA)

Various incentive works of community development was done with

maximum benefits to the village people. This led to draw the attention
and attitude of village inhabitants toward the importance of conservation
and propagation of the forest. Minor infrastructural benefits like con-
struction of community information center, community hall, public uri-
nal, bazaar set, vegetable warehouse, public water tank, approach road,
step, and funds for playground were also obtained. Distribution of torch
light (for aged persons), pressure cooker, and LPG gas connection was
also done to reduce the dependence of the villagers on fuelwood alone,
which is a means of conservation of forest resources. They even got an
170 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 12.13 (See color insert.) Amount of NTFPs sold in different villages.

FIGURE 12.14 (See color insert.) Estimated income generated from NTFPs in different
villages.

opportunity to work in the community forest and received payment from

the Forest Department (Figure 12.16).

12.4 CONCLUSIONS

It is important to explore potential of different NTFPs, which can bring

in more economic benefits to local communities if their potential is har-
Dependence on Non-Timber Forest Products 171

FIGURE 12.15 (See color insert.) Mean of all the NTFPs uses in the five surveyed
villages.

nessed properly. The NTFPs provides a security net by providing the

basic requirements for the villagers in one way or the other. Hence, it
is very important to protect them and harvest in a sustainable way, and
more emphasis has been placed on this. While unsustainable and sustain-
able practices are seen in harvesting forest fruits, it is important to have
awareness among the villagers who are always in touch with the forest
resources. Villagers have to put extra efforts to bridge a wide gap between
the large demand and less supply of NTFP resources. Efforts should be
made to introduce the quick profitable crops (cash crops, agricultural,
horticultural, herbal, etc.) and multipurpose tree species for fulfilling the
basic requirements of the villagers, which will reduce the pressure on
natural resources.

FIGURE 12.16 Entry point activities (EPA) in Lengte and Tuahzawl village.
172 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

KEYWORDS

•• bamboo
•• broom grass
•• forest product
•• fuel wood
•• livelihood
•• wild fruits

REFERENCES

Anderson, A., (1992). ‘Land use strategies for successful extractive economies in Amazo-
nia.’ Advances in Economic Botany, 9, 67–77.
Chaluvaraju, Singh, D. B., Rao, M. N., Ravikanth, G., Ganeshaiah, K. N., & Uma S. R.,
(2001). Conservation of bamboo genetic resources in Western Ghats: status, threats
and strategies. In: Forest Genetic Resources: Status, Threats and Conservation Strat-
egies, New Delhi, India: Oxford-IBH Publications, pp. 97–113.
Chopra, K., (1993). The value of non-timber forest products: an estimation for tropical
deciduous forests in India. Economic Botany, 47(3), 251–257.
Hegde, R., Suryaprakash, S., Achoth, L., & Bawa, K. S., (1996). Extraction of NTFPs
in the forests of BR Hills 1. Contribution to rural income. Economic Botany, 50,
243–250.
Homma, A., (1992). ‘The dynamics of extraction in Amazonia: A historical Perspective.’
Advances in Economic Botany, 9, 23–31.
Maikhuri, R. K., & Gangwar, A. K., (1991). Fuel wood use by different tribal and nontribal
communities in North East India. Natural Resources Forum, 15, 162–165.
National Centre for Human Settlements and Environment, (1987). Documentation of For-
est and Rights. vol. 1. New Delhi, India, National Centre for Human Settlements and
Environment.
Prasad, S., Chellam, R., & Krishnaswamy, J., (2001). Fruit removal patterns and dispersal
of Emblica officinalis (Euphorbiaceae) at Rajaji National Park, India. In: Tropical
Ecosystems: Structure, Diversity and Human Welfare, New Delhi, India. Oxford-
IBH Publications, 513–516.
Salam, M. A., Noguchi, T., & Koike, M., (2005). Factors influencing the sustained par-
ticipation of farmers in participatory forestry: a case study in central Sal forests in
Bangladesh. J., of Environment and Management, 74, 43–51.
Takasaki, Bahram, B. L., & Coomes, O. T., (2004). Risk coping strategies in tropical for-
ests: floods, illness and resource extraction. Environment and Development Econom-
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Uma Shaanker, R., Ganeshaiah, K. N., & Rao, M. N., (2001). Genetic diversity of medici-
nal plant species in deciduous forest of South India: impact of harvesting and other
anthropogenic pressures. Journal of Plant Biology, 28, 91–97.
Yashwant, S., Rawat, Chandra, M.,& Sharma. (2010). Sustainable development and man-
agement of forest resources: a case study of site specific microplan preparation and
joint forest management (JFM) Implementation in district rudraprayag, Central
Indian Himalaya. Eviron. We. Int. J., Sci. Tech., 5, 1–12.
CHAPTER 13

EFFECTS OF BLACK ROT ON THE

ANTIOXIDANT PROPERTIES OF
MORRIS AND SARAWAK PINEAPPLE
(ANANAS COMOSUS)
HASVINDER KAUR BALDEV SINGH, VICKNESHA
SANTHIRASEGARAM, ZULIANA RAZALI,
and CHANDRAN SOMASUNDRAM
Institute of Biological Sciences, Faculty of Science and the Centre
for Research in Biotechnology for Agriculture (CEBAR),
University of Malaya, 50603, Kuala Lumpur, Malaysia,
E-mail: [email protected]

CONTENTS

13.1 Introduction................................................................................. 176

13.2 Materials and Method................................................................. 177
13.3 Results and Discussion............................................................... 179
13.4 Conclusion.................................................................................. 183
Acknowledgment................................................................................... 183
Keywords............................................................................................... 184
References.............................................................................................. 184

Black rot is a postharvest disease that is currently affecting the pineapple

industry of Malaysia. The pineapple cultivars that are susceptible to this
postharvest disease include Morris and Sarawak. The objective of this
study was to determine the antioxidant properties of healthy and infected
176 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Morris and Sarawak pineapples. The methanolic extract of the pineapples

was assessed for total polyphenol content, total flavonoid content, total
antioxidant capacity, and DPPH radical scavenging activity. The total
polyphenol content was assayed using the Folin-Ciocalteu method. The
total flavonoid content was detected using the aluminum chloride colori-
metric method. Antioxidant activities were determined using the total anti-
oxidant capacity and DPPH radical scavenging assay. Both the infected
pineapple cultivars were found to contain a higher content of polyphenols
and flavonoids. Similarly, both the antioxidant assays conducted showed
a higher level of antioxidant activity in infected pineapples. The increased
level of antioxidant properties in pineapples infected by black rot is related
to the secondary response of the plant defense mechanism.

13.1 INTRODUCTION

Agriculture plays an important role in the local and export industries of

Malaysia. Pineapple is one of the industry that contributes to the country’s
economic status. Based on statistics, Malaysia’s production of pineapples
was estimated to be around 309,331 metric tons, 314,405 metric tons, and
315,977 metric tons in the year 2011, 2012, and 2013 respectively (FAO,
2016). Pineapple is a short, herbaceous perennial plant that is grown on
peat and mineral soils. Ranking as the third tropical fruit in the world,
pineapple is the only edible member of the Bromeliaceae family and can
be consumed fresh, cooked, or preserved (Bartholomew et al., 2003; Chan,
2010; Kudom and Kwapong, 2010).
Pineapple has an extraordinary flavor, aroma, and juiciness. This queen
of fruits also contains a rich amount of nutrients such as vitamin C, cal-
cium, potassium, magnesium, phosphorus and crude fiber. Some minerals
of pineapple such as manganese is essential for the activation of certain
enzymes in the human body as well as for the formation of bones. Other
beneficial trace elements like copper helps to promote iron absorption and
aids in regulating heart rate and blood pressure. Besides these benefits,
pineapple is used as a nutritional supplemented fruit for health purpose as
it contains a high amount of ascorbic acid, sugar, and moisture content.
The high level of vitamin C in pineapple is helpful in preventing the risk
Effects of Black Rot on the Antioxidant Properties 177

of gastrointestinal cancer. Pineapple also contains bromelain, which is a

proteolytic enzyme that acts as an anti-inflammatory and digestive agent.
Pineapple fruit also has a rich amount of antioxidants such as flavonoids,
ascorbic acid, and other types of phenolic compounds that are associated
with antioxidant activities and are significant indicators of fruit standards
in the global market (Hemalatha and Anbuselvi, 2003; Brat et al., 2004;
Mhatre et al., 2009; Sabahelkhier et al. 2010; Debnath et al., 2012).
In Malaysia, the most prominent pineapple cultivars are Morris, Sar-
awak, Gandul, Maspine, and Yankee. Josapine and N36 pineapple hybrids
are also produced in Malaysia. However, as reported by the Malaysian
Agricultural Research and Development Institute (MARDI), the pineapple
industry of Malaysia is facing difficulties with the Morris and Sarawak
cultivars as they are susceptible to a destructive pineapple disease called
black rot.
The pineapple black rot disease is caused by a facultative parasitic
fungus called Thielaviopsis paradoxa (De Seyen.). This postharvest dis-
ease is a common fresh fruit problem and usually occurs when the fruit
is wounded during harvest. This fruit disease is characterized by a water-
soaked lesion and browning of the infected tissue. During an early infec-
tion, the tissue becomes soft and watery. When the fruit further decays,
the tissue will be enclosed with a black coating composed of macrospores
of the fungus near to the core of the fruit (Bratley & Mason, 1939; Paull
& Reyes, 1996; Mardi, 2015). Thus far, there are no studies done to eval-
uate the changes in antioxidant activities in pineapples infected by the
black rot disease. Hence, the objective of this study was to determine
the antioxidant properties of healthy and infected Morris and Sarawak
pineapples.

13.2 MATERIALS AND METHOD

13.2.1 PLANT MATERIAL

Morris and Sarawak pineapples were used as samples for the experiments.
Fresh and infected fruit were obtained from the local market located 3.7
km from the Postharvest Physiology and Biotechnology Laboratory, Uni-
versity of Malaya. Both the Morris and Sarawak pineapple were selected
178 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

visually by picking those that have the same size and harvest time. The
fresh fruits were ensured not to have any defects as indicated by the firm-
ness and physical appearance; the infected fruits were picked based on
external defects.

13.2.2 SAMPLE EXTRACTION

The pineapple peels were removed, and the flesh was cut into small chunks.
The small chunks were then homogenized using liquid nitrogen with a
mortar and pestle set. The extraction method was performed according
to Xu et al. (2008) with slight modifications. Equal parts of the homog-
enized sample (10 g) was mixed with 10 mL of 80% methanol to purify the
sample (ratio 1:1). The mixture was placed in a shaking incubator (Shellab
Orbital Shaking Incubator S14, OR, USA) at 250 rpm for 30 min at room
temperature and was then centrifuged. The supernatant was used for the
analysis of antioxidant activity.

13.2.3 DETERMINATION OF ANTIOXIDANT PROPERTIES

13.2.3.1 Total Polyphenol Content

Total polyphenol content of sample extracts was determined using the

Folin-Ciocalteu assay (Singleton et al., 1965) modified to a microscale
(Bae et al., 2007). A standard curve of gallic acid (y = 0.00566x, r2 =
0.9955) was prepared, and the results were reported as milligrams of gal-
lic acid equivalent (GAE) per 100 mL sample extract.

13.2.3.2 Total Flavonoid Content

Flavonoid content of the samples was determined using a colorimet-

ric method described by Sakanaka et al. (2005). A standard curve of
(+)-catechin (y = 0.0135x, r2 = 0.9943) was prepared, and the results
were reported as milligrams of catechin equivalent (CE) per 100 mL
sample extract.
Effects of Black Rot on the Antioxidant Properties 179

13.2.3.3 Total Antioxidant Capacity

Total antioxidant capacity of the samples was determined using the phos-
phomolybdenum method described by Prieto et al. (1999). A standard
curve of ascorbic acid (y = 0.0018x, r2 = 0.9981) was prepared, and the
results were reported as micrograms of ascorbic acid equivalent (AAE)
per milliliter sample extract.

13.2.3.4 DPPH Radical Scavenging Activity

DPPH assay was carried out as described by Oyaizu (1986) and Bae and
Suh (2007). A standard curve of ascorbic acid (y = 10.145x, r2 = 0.9907)
was prepared, and the results were reported as micrograms of ascorbic
acid equivalent (AAE) per milliliter of the extract. The radical scavenging
activity was calculated accordingly:

% DPPH inhibition = (Acontrol – Asample/Acontrol) × 100

where Acontrol is absorbance reading of control and Asample is absorbance

reading of the sample.

13.2.4 STATISTICAL ANALYSIS

Statistical analysis was carried out using SPSS 19.0 software (SPSS Inc.,
IBM), and the data were represented by mean values ± standard deviation
(SD). The differences found between the mean values of the samples were
obtained through analysis of variance (one-way ANOVA) by referring to
Tukey’s honestly significant difference (HSD) test. This was done at p <
0.05 significance level.

13.3 RESULTS AND DISCUSSION

In this study, four antioxidant assays were performed on Morris and Sar-
awak pineapples. The antioxidant assays were total polyphenol content,
180 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

total flavonoid content, total antioxidant capacity, and DPPH radical scav-
enging activity (Figure 13.1).
As can be seen in the results above, Figure 13.1 showed that the infected
Morris and Sarawak pineapples possessed highest total polyphenol content of
64.46 ± 3.16 mg GAE/mL and 29.46 ± 5.89 mg GAE/mL, respectively. The
healthy Morris and Sarawak pineapples had much lower polyphenol content
of 44.11 ± 1.64 mg GAE/mL and 24.82 ± 9.41 mg GAE/mL than the infected
pineapples (Figure 13.2). Higher values for the total polyphenol content were
observed in both healthy and infected Morris pineapples than those observed
in Sarawak pineapples. Nevertheless, a previous study reported three-fold
higher increase in polyphenol content in the infected tissue of plum leaves. It
is reported mentioned that the polyphenols can act as defense-related metab-
olites in the infected plants and that antioxidative substances are also related
to the stress factors of the plants (Favali and Pressacco, 2000).
In plants, flavonoids act as a physiological regulator and chemical
messenger other than having a role in the plant flower coloration. Fla-
vonoids also defend plants against the attack of herbivores and patho-
gens. Figure 13.2 above shows the total flavonoid content estimated
using a catechin equivalent. The infected Morris pineapple showed the
highest total flavonoid content with 20.00 ± 0.92 mg CE/100 mL, while

Total polyphenol content (mg GAE/ml)

80
a
70
60
50
a
40 b
30 b
20
10
0
Healthy Infected

Morris Sarawak

FIGURE 13.1 Total polyphenol content of Morris and Sarawak pineapples. 1Values
followed by different letters are significantly different (p < 0.05) (n = 9).
Effects of Black Rot on the Antioxidant Properties 181

Total flavanoid content (mg CE/100ml)

25
a
20

15

10 a
b
5 b

0
Healthy Infected

Morris Sarawak

FIGURE 13.2 Total flavonoid content of Morris and Sarawak pineapples. 1Values
followed by different letters are significantly different (p < 0.05) (n = 9).

the infected Sarawak pineapple showed total flavonoid content of 5.63

± 1.08 mg CE/100mL. Both the healthy pineapples of Morris and Sar-
awak were found to have a lower amount of total flavonoid content than
the infected pineapples. Healthy Morris pineapple had 7.80 ± 1.28 mg
CE/100 mL flavonoid content, while the healthy Sarawak pineapple had
a lower flavonoid content of 3.22 ± 0.49 mg CE/100 mL. The increase
in flavonoids is associated with its function as a secondary antioxidant
defense system when a plant is exposed to the conditions of abiotic and
biotic stresses. The results were supported by several studies showing the
protective effects of flavonoids against pathogenic infections (Rice-Evans
et al., 1995; Cook and Samman, 1996; Kumar et al., 2003; Pandey, 2007;
Agati et al., 2012).
Total antioxidant capacity was determined according to Prieto et al.
(1999). In Figure 13.3, the antioxidant capacity was found to be highest
in the infected Morris and Sarawak pineapples. However, the total anti-
oxidant capacity was higher in the infected Sarawak pineapple (1132.2
± 50.22 µg AAE/mL) than in the infected Morris pineapple (1099.40 ±
20.55 µg AAE/mL). Both the healthy Morris and Sarawak pineapples
182 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Total anoxidant capacity (µg AAE/ml)

1400
b a b
1200
1000
a

800
600
400
200
0
Healthy Infected

Morris Sarawak

FIGURE 13.3 Total antioxidant capacity of healthy and infected fruits of both Morris
and Sarawak pineapples. 1Values followed by different letters are significantly different
(p < 0.05) (n = 9).

had a lower total antioxidant capacity of 928.90 ± 22.50 µg AAE/mL and

1072.20 ± 37.88 µg AAE/mL, respectively.
DPPH is a stable free radical that is commonly used to determine the
radical scavenging activity of natural compounds. With the reduction by
an antioxidant, the absorption of DPPH reduces because of the forma-
tion of DPPH-H, which is a nonradical form. Figure 13.4 shows that the
infected pineapples of Morris and Sarawak had significantly higher radi-
cal scavenging activity than the healthy pineapples. In comparison, the
infected Morris pineapple had higher amount of radical scavenging activ-
ity of 6.01 ± 0.09 µg AAE/mL than the infected Sarawak pineapple, which
had only 3.03 ± 0.22 µg AAE/mL. Similarly, the healthy Sarawak pine-
apple showed a higher radical scavenging activity of 4.2 ± 0.08 µg AAE/
mL than healthy Morris pineapple, which had only 2.42 ± 0.39 µg AAE/
mL. In a study done by Nahiyan and Matsubara (2012), the infected plant
showed an increase in DPPH radical scavenging activity, polyphenols, and
ascorbic acid content. Thus, the disease tolerance is linked to the antioxi-
dative substance of a plant.
Effects of Black Rot on the Antioxidant Properties 183

DPPH radical scavenging acvity (µg AAE/ml)

7
a
6
5 a
4
b
3 b
2
1
0
Healthy Infected

Morris Sarawak

FIGURE 13.4 DPPH radical scavenging activity of healthy and infected fruits of both
Morris and Sarawak pineapples. 1Values followed by different letters are significantly
different (p < 0.05) (n = 9).

13.4 CONCLUSION

Results obtained revealed a significant increase in antioxidant proper-

ties of the Morris and Sarawak cultivars during the infection of black
rot. Overall, Morris pineapple showed a higher total polyphenol content,
total flavonoid content, and radical scavenging activity than the Sarawak
pineapple. The Sarawak pineapple only showed higher amount in total
antioxidant activity than Morris pineapple. All the four assays showed
increase in antioxidant properties during the infection. More studies are
needed to investigate in detail the biochemistry and physiology of these
actions.

ACKNOWLEDGMENT

The authors would like to thank the University of Malaya and MOSTI
E-Science grant 02-01-03-SF1019 for supporting this research.
184 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

KEYWORDS

•• antioxidants
•• black rot
•• flavonoid
•• morris
•• pineapple
•• polyphenol
•• Sarawak

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 PART II

PROTECTION OF
HORTICULTURE CROPS
CHAPTER 14

ESSENTIAL OILS AS GREEN

PESTICIDES FOR PLANT PROTECTION
IN HORTICULTURE
MURRAY B. ISMAN and RITA SEFFRIN
Faculty of Land and Food Systems, University of British Columbia,
Vancouver V6T1Z4, Canada

CONTENTS

Abstract.................................................................................................. 190
14.1 Introduction ................................................................................ 190
14.2 Biological Activity of Essential Oils:
A Historical Perspective.............................................................. 192
14.3 Medicinal Plants Used to Produce Essential Oil as
Pesticides and Their Constituents............................................... 194
14.4 Essential Oils Synergy Versus Isolated Constituents and
Their Effects on Agricultural Pests............................................. 203
14.5 Essential Oil-Based Pesticides: From the Data to the Crop
Field After 40 Years of Research................................................ 205
14.6 Advantages and Disadvantages of Essential Oil-Based
Pesticides for Pest Management in Horticulture......................... 207
14.7 Conclusion.................................................................................. 210
Keywords................................................................................................211
References...............................................................................................211
190 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

ABSTRACT

Interest in, and research on, bioactivities of plant essential oils to insects
has exploded in the past 15 years, according to a recent bibliometric analy-
sis. However, commercial exploitation of this knowledge is being realized
much more slowly although essential oil-based pesticides have begun to
establish a market presence at least in the USA. The volatility of essential
oils makes them especially suitable as fumigants in protected environments
and for protection of stored products, but they also have demonstrated util-
ity for protection of horticultural crops. Many essential oils and their major
constituents, monoterpenes and sesquiterpenes, have contact toxicity to
insects and mites, but their utility is broadened owing to their sublethal
behavioral effects as deterrents and repellents. These bioactivities result
from neurotoxicity of the terpenes, with at least two distinct mechanisms
of action identified thus far. One intriguing aspect of the toxicity of some
essential oils in insects is synergy among particular terpenes within an oil,
thus enhancing bioactivity. Although many essential oils display bioactiv-
ity against insects when tested in the laboratory, only a few commodity
oils – those used extensively in the flavor and fragrance industries – have
been developed for use as pesticides. These include certain oils from the
families Lamiaceae, Lauraceae, Myrtaceae, and Poaceae. Their potential
as protectants for horticultural crops is discussed.

14.1 INTRODUCTION

Our global population has doubled in the last 45 years. If the present
growth rate of 1.3% per year persists, the population will double again
within a mere 50 years according to World Watch Magazine (2004).
There are huge pressures to provide food at low cost. Synthetic pes-
ticides have been an important component of industrialized agricul-
ture throughout the world since the 1950s. The “second generation”
pesticides were very effective at killing pests and thus boosting crop
yields, and being relatively inexpensive, their use quickly spread over
the globe. Over time, we discovered that many of these chemicals were
extremely pervasive in our environment as a result of their widespread
Essential Oils as Green Pesticides for Plant Protection 191

and repeated use and, in some cases, their environmental persistence.

Some chemicals take an extremely long time to degrade, such that even
those banned decades ago, including dichlorodiphenyltrichloroethane
(DDT) and its secondary products, can still be found in the environ-
ment today. Organochlorine, organophosphate, carbamate, and pyre-
throid pesticides were introduced between the late 1940s and the mid of
1970s, and they helped to usher in industrial agriculture or the “Green
Revolution.” More recently, other types of pesticides (e.g., neonicoti-
noids) have been introduced into the world market and industrial agri-
culture has come to rely more and more on the use of synthetic chemical
pesticides to protect crops from pests.
In recent decades, there has been a steady increase in the amount
of pesticides used in agriculture. In the European Union alone, more
than 200,000 tons of pesticides (active ingredients) are used annually
(Eurostat Statistical, 2007). In developing countries, the effects of acute
poisoning due to exposure to dangerous pesticides in food are far more
severe than in industrialized countries. For example, an estimated 70,000
tons of active ingredient of pesticides per year are imported annually
in Central America, and the main groups of insecticide include organo-
phosphates, carbamates and pyrethroids (Castillo et al., 2010). Pesticides
can be hazardous to humans and their residues can accumulate in food
chains, damaging birds, fish, and other forms of wildlife. In many cases,
these side effects are not immediately apparent, but may show up later,
for example, in the abnormal eggs of birds that have eaten pesticide-
laded insects.
The recent increase in organic farming practices in Europe dem-
onstrates that farming without synthetic pesticides is feasible, scalable,
economically viable, and environmentally safe. Land under organic culti-
vation increased from 5.7 million hectares in 2002 to 9.6 million hectares
in 2011, and includes horticultural and orchard crops as well as animal
sectors (European Commission, 2013). Investigators have long searched
for new, highly selective, and biodegradable pesticides to solve the prob-
lem of long-term toxicity to mammals and, environmental persistence of
pesticides and develop techniques that can be used to reduce overall pesti-
cide use while maintaining crop yields. Plant essential oils have been used
since antiquity for many purposes, including pest control in agriculture and
192 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

against nuisance pests such as mosquitoes, flies, and ticks. In recent years,
consumers have increasingly expressed interest in purchasing organically
grown foods as well as using natural and naturally derived materials to
eradicate pests in their lawns, gardens, and homes. Pesticides based on
plant essential oils or their constituents have demonstrated efficacy against
many agricultural pests. In fact, pesticides derived from plant essential
oils can have several important benefits. Because of their volatile nature,
there is a much lower level of risk to the environment than with current
synthetic pesticides. Predator, parasitoid, and pollinator insect populations
will be less impacted because of the minimal residual activity, making
essential oil-based pesticides compatible with integrated pest management
programs. Additionally, resistance to essential oil-based pesticides may
develop more slowly if at all, owing to the complex mixtures of constitu-
ents that characterize many of these oils.

14.2 BIOLOGICAL ACTIVITY OF ESSENTIAL OILS:

A HISTORICAL PERSPECTIVE

Interest has increased in the use of oils to replace conventional chemical

pesticides in pest control. Many plant essential oils show a broad spectrum
of activity against pest insects owing to their repellent, fumigant, insecti-
cidal, attractant, antifeedant, oviposition deterrent, and growth regulatory
activities. These oils also have a long tradition of use in the protection of
stored products. Citronella oil, discovered in 1901, was the most widely
used personal repellent before the 1940s, and is still used today in many
formulations (Brown and Hebert, 1997). The fumigant toxicity of essential
oils owing to their high volatility may have first attracted the attention of
researchers. The fumigant toxicity of essential oils and their main constitu-
ents, the volatile monoterpenes, was first reported in the 1960s (Smely-
anets and Kuznetsov, 1968).
Many studies of the insecticidal activity of essential oils were pub-
lished in the 1970s, especially against stored product pests. Patchouli,
Pogostenmon heyneanus (Solanaceae) and sweet basil, Ocimum basilicum
(Lamiaceae), essential oils showed insecticidal activity against Sitophilus
oryzae (Coleoptera: Curculionidae), Stegobium paniceum (Coleoptera:
Anobiidae), Tribolium castaneum (Coleoptera: Tenebrionidae) and Bru-
Essential Oils as Green Pesticides for Plant Protection 193

chus chinensis (Coleoptera: Bruchidae) (Deshpande et al., 1974; Desh-

pande and Tipnis, 1977). Pulegone, linalool, and limonene were effective
fumigants against rice weevil, Sitophilus oryzae, while Mentha citrata oil
containing linalool and linalyl acetate also exhibited significant fumigant
toxicity to these weevils (Singh et al., 1989). Oviposition inhibition and
ovicidal activities have been reported for carvacrol, carveol, geraniol,
linalool, menthol, terpineol, thymol, verbenol, carvones, fenchone, men-
thone, pulegone, thujone, verbenone, cinnamaldehyde, citral, citronellal,
and cinnamic acid against house fly, M. domestica eggs (Rice and Coats,
1994). Cinnamyl alcohol, 4-methoxy-cinnamaldehyde, cinnamaldehyde,
geranylacetone, and α-terpineol are attractive to adult corn rootworm bee-
tles, Diabrotica sp. (Hammack, 1996; Petroski and Hammack, 1998) and
have been used in traps for these pests.
According to Regnault-Roger (1997), the examination of patents
involving essential oils showed that a majority of the inventions focused
on household uses, and several formulations were proposed to control
mosquitoes and flies; some of them in combination with pyrethroids
(Liang, 1988; Kono et al., 1993). Isman (2000) mentioned several investi-
gations confirming that certain plant essential oils not only repelled insects
but had contact and fumigant insecticidal actions against specific pests.
As part of an effort aimed at the development of reduced-risk pesticides
based on plant essential oils, toxic and sublethal effects of some essen-
tial oil terpenes and phenols were investigated using the tobacco cutworm
(Spodoptera litura) and the green peach aphid (Myzus persicae) as model
pest species.
Survey of the scientific literature on the biopesticidal potential of essen-
tial oils from the year 2000 onwards indicates that plants of the families
Myrtaceae, Lamiaceae, Asteraceae, Apiaceae, and Rutaceae are important
sources for natural pesticides effective against pests in the insect orders
Lepidoptera, Coleoptera, Diptera, Isoptera, and Hemiptera.
The mode of action of essential oils also received attention from the
research community. Enan (2001, 2005) has provided evidence that many
essential oil constituents poison insects by blocking octopamine receptors.
Octopamine, synthesized from tyramine, is a neurotransmitter and neuro-
modulator in arthropods and may have neurohormonal influences as well.
The rapid action against some pests is indicative of a neurotoxic mode-of-
194 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

action and there is some evidence for interference with gamma-aminobu-

tyric acid (GABA)-gated chloride channels as well (Priestley et al., 2003).
According to Isman and Machial (2006), the majority of studies on
essential oil bioactivity to insects between 2005 and 2008 were aimed at
stored products pests with 78 essential oils tested on Sitophilus oryzae and
22 on Acanthoscelides obtectus with fumigant toxicity evaluated. They
also noted that since 2005, essential oils from 30 plant families were tested
against coleopterans pests of stored grain. Essential oils from 22 species
belonging to the family Lamiaceae, 17 species of Asteraceae, and 10 spe-
cies of Myrtaceae were shown to provide control of coleopteran pests of
the families Bostrichidae, Bruchidae, Chrysomelidae, Cucujidae, Curcu-
lionidae, Dermestidae, and Tenebrionidae (Pérez et al., 2010).
Much research of this type has focused on greenhouse experiments
(Regnault-Roger, 2012). The repellence of essential oils from fruit skins
of laranja pera (Citrus sinensis Osbeck var. pera) and laranja lima (Citrus
aurantium L.) to the two-spotted spider mite, Tetranychus urticae Koch,
on string was evaluated in greenhouse experiments. Although both oils,
rich in d-limonene, showed similar repellent effects in laboratory bioas-
says, lima oil prevented the movement of mites between plants across oil-
treated strings for 1 week in the greenhouse (Camara et al., 2015). In 2011,
around 60,500 pounds of d-limonene (the dominant monoterpene from
Citrus peels) were applied as a pesticide in California, although 71% of
that total was for non-agricultural uses, primarily for structural pest control
(Isman, 2014).

14.3 MEDICINAL PLANTS USED TO PRODUCE ESSENTIAL OIL

AS PESTICIDES AND THEIR CONSTITUENTS

Not all terrestrial plants produce essential oils, but many aromatic plants,
including some used as culinary herbs and spices, have been used since
antiquity as folk medicine and as preservatives in foods (Christaki et al.,
2012). Certain of these source plants have been traditionally used for the
protection of stored commodities, especially in the Mediterranean region
and in Southern Asia, but interest in the oils for pest control was renewed
with demonstrations of their fumigant and contact insecticidal activities to
a wide range of pests in the 1990s (Isman, 2000).
Essential Oils as Green Pesticides for Plant Protection 195

Many herbs and spices, including rosemary, oregano, sage, thyme,

peppermint, and garlic, can be found worldwide, with many originating
from the Mediterranean area (Bampidis et al., 2005; Kadri et al., 2011).
They contain a wide range of chemical substances such as polyphenols,
alkaloids, polypeptides, or their oxygen-substituted derivatives (Cowan,
1999; Perumalla and Hettiarachchy, 2011). Most spices belong to the fam-
ilies Lamiaceae, Lauraceae, Myrtaceae, and Poaceae. Common members
from which essential oils are obtained are described in more detail in the
following sections.

14.3.1 LAMIACEAE FAMILY

Rosmarinus officinalis, commonly known as rosemary, is a garden plant

grown around the world. Aerial parts of rosemary are used as flavoring
agent in foods, beverages, and cosmetic preparations and have various
traditional uses in ethnomedicine including analgesic, anti-inflammatory,
anti-rheumatic, and spasmolytic applications (Minaiyan et al., 2011).
Native to the Mediterranean, rosemary grows freely in large areas of
southern Europe and is cultivated worldwide. Leading regions of rose-
mary production are the Mediterranean countries, Northern Africa, Eng-
land, Mexico, and the USA. For commercial purposes, there are three main
types based on their predominant chemical constituent and geographical
origin: camphor-borneol (Spain), 1,8-cineole (Tunisia), and verbenone
(France). To obtain essential oil of the highest quality, plants should be
in bloom and only the flowering tops should be harvested for distillation.
With mechanical harvesting, it is better to cut frequently because yields are
higher from rapid regrowth. Isman et al. (2008) explored the relationship
between chemical composition and insecticidal activity of 10 commercial
samples of rosemary oil, based on laboratory bioassays with two agricul-
tural pests, the armyworm Pseudaletia unipuncta Haworth (Noctuidae)
and the cabbage looper Trichoplusia ni. Hübner (Noctuidae). Nine major
terpenoid constituents of rosemary oil were quantified in the samples
by gas chromatography–mass spectrometry (GC-MS). The major con-
stituents were 1,8-cineole, α-pinene, β-pinene, and camphor; on average
1,8-cineole made up 52% of the oil by weight. Rosemary oil repels female
Thrips tabaci (Koschier and Sedy, 2003) and is toxic to the two-spotted
196 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

spider mite Tetranychus urticae (Miresmailli et al., 2006). In a fumigant

toxicity study on stored grain pests, both rosemary oil and linalool proved
to be highly effective in controlling the lesser grain borer, Rhyzopertha
dominica (Shaaya et al., 1991).

14.3.1.1 Mentha × Piperita

The three main types of mint are peppermint, spearmint, and cornmint. While
peppermint and spearmint are used as flavorings in their own right, corn-
mint is primarily used as a source of menthol. The plant is aromatic, stimu-
lant, and used for relieving chest and nasal congestion. Its oil is one of the
most widely used essential oils in food products, cosmetics, pharmaceuticals,
dental preparations, mouthwashes, soaps, and alcoholic liquors (Sujana et
al., 2013). Over the past 20 years, India has come to dominate the global
production of mint oils. Peppermint and spearmint command higher prices
than cornmint. Major constituents of mint oil are menthol and menthone and
vary according to species. Koschier and Sedy (2003) tested repellence and
oviposition effects of mint oil, along with several other essential oils, against
onion thrips (Thrips tabaci). Mint oil concentrations of 0.1% and 1% signifi-
cantly reduced egg laying by females on leaves (0.8 eggs per leaf disc treated
with 1% mint oil compared to 7.2 eggs per leaf disc on untreated controls).
The volatile components of peppermint oil are primarily menthol (29–48%),
menthone (20–31%), menthyl acetate (31%), menthofuran (1–7%), and lim-
onene (Khan and Abourashed, 2010). Peppermint was included in a study
by Choi et al. (2003) that tested the efficacy of 53 essential oils on 3 life
stages of the greenhouse whitefly Trialeurodes vaporariorum. Peppermint
oil was ranked as one of the eight most toxic oils on all three life stages. In
a separate study by Choi et al. (2004), peppermint oil was considered highly
toxic in a diffusion bioassay to two-spotted spider mites (Tetranychus urti-
cae). Harwood et al. (1990) reported that peppermint monoterpenes resulted
in reduced growth and molting abnormalities in cutworms. Peppermint oil
was also highly effective on the red flour beetle (Tribolium castaneum) as
a fumigant in a study evaluating toxicity of essential oils and constituents
against four stored-grain pests. Of the constituents tested, 1, 8-cineole, which
is commonly found in peppermint oil and is also known as eucalyptol, was
one of the most toxic components (Shaaya et al., 1991).
Essential Oils as Green Pesticides for Plant Protection 197

14.3.1.2 Thymus vulgaris

Common thyme, is a shrubby, woody plant native throughout the Medi-

terranean region (Spain, France, and Italy) (Stahl-Biskup and Sáez, 2002).
It is also cultivated in some European and New World countries such as
Brazil. It is a very popular aromatic herb used as a condiment in many
dishes (Jakiemiu et al., 2010). Oil of thyme is the important commercial
product obtained by distillation of the fresh leaves and flowering tops of T.
vulgaris. Thyme oil consists mainly of the phenols carvacrol and thymol
(20–80%), with thymol typically being the dominant compound present.
In some varieties, up to 51% monoterpene hydrocarbon content has been
reported, being made up largely of p-cymene and γ-terpinene. Alcohols
such as linalool, α-terpineol and thujan-4-ol are also present (Khan and
Abourashed, 2010). Choi et al. (2003) tested efficacy against greenhouse
whitefly adults, nymphs, and eggs. At the highest rate of 9.3 × 10−3, thyme
oil produced 100% mortality to adults and 88% mortality to eggs. In a
study by Shaaya et al. (1991), thyme oil had high fumigant toxicity against
the stored-grain pest Oryzaephilus surinamensis. Among constituents
tested in the same study, carvacrol, linalool, and α-terpineol were also
highly toxic. Machial et al. (2010) tested 17 essential oils for toxicity to
two lepidopteran species, the oblique-banded leafroller (Choristoneura
rosaceana), and the cabbage looper (Trichoplusia ni), in which thyme oil
was the second most toxic to 1st instar C. rosaceana larvae.

14.3.2 LAURACEAE FAMILY

14.3.2.1 Cinnamomum Species

Cinnamomum is a large genus, many species of which yield a volatile

oil on distillation. The most important Cinnamomum oils in world trade
are those from C. verum (cinnamon bark and leaf oils), C. cassia and
C. camphora. Cinnamon is one of the most important spices used daily
by people all over the world. Cinnamon oil primarily contains cinnam-
aldehyde, cinnamic acid, and cinnamic alcohol. In addition to being an
antioxidant, anti-inflammatory, antidiabetic, antimicrobial, and lipid-
198 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

lowering compound, cinnamon has also been reported to have activi-

ties against neurological disorders, such as Parkinson’s and Alzheimer’s
diseases (Visweswara Rao and Hua Gan, 2014). Given the large num-
ber of Cinnamomum species that exist, their widespread distribution in
Asia, and the number still not characterized in terms of essential oil con-
tent and composition, the genus has much potential for providing new
tree crops in developing countries. Cinnamomum camphora is native
to China south of the Yangtze River, Taiwan, southern Japan, Korea,
and Vietnam, and has been introduced to many other countries. The top
four cinnamon-producing countries are Indonesia, China, Sri Lanka,
and Madagascar. In a study conducted by Eun-Jeong et al. (2008), cin-
namon oil was found to have an LD50 value of 0.016 mg/cm2 on rice
weevil (Sitophilus oryzae), a common destructive pest of stored grains.
This study also tested fumigant activity of (E)-cinnamaldehyde and 41
structurally related compounds, of which allyl cinnamate was the most
toxic, with 83% mortality using the closed container fumigant method
at a 0.013 mg/cm3 dosage.

14.3.3 POACEAE FAMILY

Cymbopogon species are commonly known as lemongrass. Some spe-

cies are commonly cultivated as culinary and medicinal herbs by people
in many countries because of their scent, resembling that of lemons. In
Brazil, for example, the tea, infusion, and extracts of C. citratus, which
are prepared with fresh or dry leaves, are often used in popular medi-
cine as a restorative, digestive, effective drug against colds, and as an
analgesic, antihermetic, anticardiopatic, antithermic, and anti-inflamma-
tory (Negrelle and Gomes, 2007). Citronella oil is one of the essential
oils obtained from the leaves and stems of different species of Cymbo-
pogon. The oil is used extensively as a source of perfumery chemicals
such as citronellal, citronellol, and geraniol. Supply is dominated by India
and Guatemala. There are two main types of citronella oil, referred to as
Ceylon and Java. Both types contain citronellal, citronellol, and geraniol
as the major components, but the proportions of these vary greatly depend-
ing on source and type, with Java having a higher percentage made up
Essential Oils as Green Pesticides for Plant Protection 199

of these 3 components. The Ceylon type contains a higher percentage of

monoterpenes than the Java type, and West Indian lemongrass oil contains
65–85% citral, whereas Cameroonian C. citratus contains geranial as its
major component, comprising 33% of the oil. Other compounds that may
be present in lemongrass oil include myrcene (12–20%), dipentene, meth-
ylheptenone, β-dihydropseudoionone, neral, β-pinene, linalool, methyl-
heptenol, α-terpineol, geraniol, nerol, farnesol, citronellol, and volatile
acids such as isovaleric, geranic, caprylic, citronellic and others (Khan
and Abourashed, 2010).
Lemongrass oil is also known for its insecticidal properties and was
included, among 52 other essential oils, in a test against greenhouse white-
fly by Choi et al. (2003). The oil was the most effective on the egg stage
(98% mortality). A fumigant toxicity study of essential oils and constit-
uents on four stored-product beetle species showed that lemongrass oil
had little to no toxic effect. However, linalool and terpinen-4-ol, found in
lemongrass oil as minor constituents, were found to be highly effective
(Shaaya et al., 1991). In a study by Machial et al. (2010) on the toxicity of
essential oils on two lepidopteran species, lemongrass oil was the second
most toxic to 1st instar cabbage looper larvae, with 53% mortality at a
concentration of 5 μL/mL, an LC50 of 7.2 μL/mL, and an LD50 of 60.5 μL/
mL. A GC-MS analysis found the major constituents of this lemongrass
oil sample to be citral (47.1%), trans-verbenol (32.1%), and camphene
(10.7%).

14.3.4 MYRTACEAE FAMILY

Clove (Syzygium aromaticum) is one of the most valuable spices that has
been used for centuries as a food preservative and for many medicinal
purposes. Clove is native to Indonesia but is now cultivated in several
parts of the world including Brazil. This plant represents one of the richest
source of phenolic compounds and has great potential for pharmaceuti-
cal, cosmetic, food, and agricultural applications (Cortés-Rojas, 2014). A
major component of clove taste is imparted by the chemical eugenol. It
is widely used in agricultural applications to protect foods from micro-
organisms during storage, which might have an effect on human health,
200 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

and as a pesticide and fumigant (Kamatou et al., 2012). The production of

all types of clove oil – leaf, stem, and bud – is dominated by Indonesia.
Madagascar, the largest exporter of cloves, also exports some oil, typically
clove leaf oil. Clove bud oil typically contains 60–90% eugenol, 2–27%
eugenol acetate, and 5–12% β-caryophyllene. Constituents found in clove
leaf and clove stem oils are very similar, but may differ in ratio. Naphtha-
lene may be present in leaf and stem oils, but does not occur in the bud oil
(Khan and Abourashed, 2010). In Choi et al. (2003), clove oil provided
98% mortality to greenhouse whitefly nymphs at 9.3 × 10−3 concentration,
and clove (bud) oil produced 94% mortality of eggs at the same concentra-
tion. Among the 53 essential oils included in this study, only 8 (including
clove), were considered highly effective against all three stages (adult,
nymph, and egg) of greenhouse whitefly. Eugenol is a phenylpropanoid
found in many essential oils, but in highest concentrations in clove and
cinnamon oils. Eugenol is known for its herbicidal, insecticidal, and anti-
fungal activity. Enan (2005) determined eugenol LD50 values of 1.9 µg/
insect for fruit fly (Drosophila melanogaster). Isman (2000) compared
the toxicity of eugenol with α-terpineol and terpinen-4-ol. Eugenol was
7–9 times more toxic than the two terpenes in the western corn rootworm
beetle (Diabrotica virgifera virgifera).

14.3.4.1 Eucalyptus Species

Some Eucalyptus species have attracted attention from horticulturists,

global development researchers, and environmentalists because of desir-
able traits such as fast-growing sources of wood, and production of oil that
can be used for cleaning and as a natural insecticide.
Eucalyptus oil is the generic name for distilled oil from the leaf of
Eucalyptus, a genus of the plant native to Australia and cultivated world-
wide. Eucalyptus oil has a history of wide application. The oil is antiseptic
and is used in infections of the upper respiratory tract and certain skin
diseases (Kumar et al., 2013). Production of eucalyptus oil is dominated
by China. A range of secondary sources include Brazil, India, Australia,
and South Africa. There are a range of eucalyptus oil types – medicinal
(1,8-cineole-rich); perfumery (citronellal-type); industrial (piperitone-
type) – and care is therefore needed in interpreting prices and volumes.
Essential Oils as Green Pesticides for Plant Protection 201

The insecticidal effects of Eucalyptus dundasii Maiden essential oil was

studied on the adults of the lesser grain borer, Rhyzopertha dominica (F.),
and the saw-toothed grain beetle, Oryzaephilus surinamensis (L.). Chemi-
cal analysis indicated that 1,8-cineole (54.15%), p-cymene (12.41%),
α-thujene (11.37%), and β-caryophyllene (6.7%) were the major constitu-
ents. E. dundasii essence was repellent for both insects (Aref et al., 2005).
Jemâa et al. (2012) investigated seasonal variation in chemical composi-
tion of essential oils isolated from leaves of Eucalyptus camaldulensis, E.
astringens, E. leucoxylon, E. lehmannii, and E. rudis and assessed their
fumigant activity against three stored-date moth pests: Ephestia kuehni-
ella, Ephestia cautella, and Ectomyelois ceratoniae. The five essential oils
contained 1,8-cineole, α-pinene, and α-terpineol as the major constituents.
Of the other major constituents, β-pinene and p-cymene were only pres-
ent in E. rudis essential oil. In addition, o-cymene was specific only to E.
camaldulensis and E. rudis essential oils. Results demonstrated that fumi-
gant toxicity varied with season, insect species, essential oil concentration
and exposure time. E. camaldulensis essential oil was more toxic against
E. cautella and E. kuehniella. LC50 values were, respectively, 11.07 and
26.73 mL/L air. However, for E. ceratoniae, E. rudis essential oil was
more effective, with an LC50 of 31.4 mL/L air.

14.3.5 GERANIACEAE FAMILY

The essential oil of geranium is extracted through steam distillation of

stems and leaves of the geranium plant Pelargonium spp. Essential oils
derived from these aromatic plants have demonstrated biological prop-
erties and can be used to prevent and treat human systemic diseases,
including infectious diseases (Carmen and Hancu, 2014). Production of
geranium oil is dominated by Egypt and China. Very small quantities
of “rose” geranium oil (bourbon oil) are produced in a number of Afri-
can countries – South Africa, Madagascar, Rwanda – and this oil com-
mands a significant price premium. Geranium oil consists of 60–70%
alcohols, primarily citronellol and geraniol, with linalool and phenethyl
alcohol also present. The esters geranyl tiglate, geranyl acetate, citro-
nellyl formate and citronellyl acetate comprise 20–30% of the oil (Khan
202 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

and Abourashed, 2010). Geranium oil was tested for toxicity against
three life stages of greenhouse whitefly, among 53 other essential
oils by Choi et al. (2003). All three stages were effectively controlled
(>90% mortality) by geranium oil, but at the highest dose only. Joen et
al. (2009) found that the acaricidal effects of geraniol on storage food
mites (Tyrophagus putrescentiae) were more effective than the industry
standard benzyl benzoate, with LD50 values of 1.27 µg/cm3 and 1.95 µg/
cm3, respectively.

14.3.6 AMARYLLIDACEAE FAMILY

With a history of human use of over 7,000 years, Allium sativum,

commonly known as garlic, is native to central Asia. Garlic has been
cultivated for around 4,000 years and has not only been used in food
preparation but also as a medicine and crop protection product. Garlic
is known for its positive effects on health, particularly the prevention of
cardiovascular diseases and certain digestive cancers (Lalla et al., 2013).
This long history gave garlic a head start in recent efforts to gain EU reg-
ulatory approval for its use in agriculture. A vast body of literature was
available to drawn upon when compiling the required data, for example,
data on toxicity, residues, ecotoxicity and fate and behavior in the envi-
ronment. Most published literature cites that diallyl disulfide is the main
compound in garlic oil, at 60%, with diallyl thiosulfinate, allylpropyl
disulfide, diallyl disulfide, and diallyl trisulfide also being major com-
ponents. Non-sulfur compounds that may be found in garlic oil include
citral, geraniol, linalool, and β-phellandrene (Khan and Abourashed,
2010). Machial et al. (2010) reported garlic oil to be the most effective
of 17 tested essential oils on 1st instar cabbage looper larvae (Tricho-
plusia ni), with LC50 = 3.3 μL/mL and LD50 = 22.7 µg/insect. It ranked
5th out of 17 in terms of its toxic effect to the oblique-banded leaf roller
(Choristoneura rosaceae), with 22% mortality of 1st instar larvae at 5
μL/mL. The major constituents of the garlic oil sample used in that study
were 35.2% diallyl disulfide, 26.2% di-2-propenyl trisulfide, 20.7% 3,3
thiobis-1-propene, 6.6% methyl 2-propenyl trisulfide, and 4.5% methyl
1-propenyl disulfide.
Essential Oils as Green Pesticides for Plant Protection 203

14.4 ESSENTIAL OILS SYNERGY VERSUS ISOLATED

CONSTITUENTS AND THEIR EFFECTS ON AGRICULTURAL PESTS

Essential oils have received attention in recent years, in part owing to

concerns about synthetic pesticides and for their potential to reduce the
development of resistance to pesticides. They possess various biological
properties. The wide range of biological activities showed by an essen-
tial oil can be related to its qualitative and quantitative composition. They
can consist of terpenoids (monoterpenes, sesquiterpenes, and diterpenes
in the form of hydrocarbons, alcohols, aldehydes, ketones, ethers, esters,
peroxides, and phenols), aromatic compounds (C6-C3 and C6-C1 com-
pounds; less frequent but characteristic of certain essential oils), and low-
molecular-weight aliphatic compounds (hydrocarbons, alcohols, acids,
aldehydes, esters, and lactones) with different physical, chemical, and
pharmacological properties, responsible for the activity of whole essential
oil (Blázquez, 2014). Many formulations include a blend of essential oils
to create a product with a broad spectrum of action and multiple modes
of action. In fragrances and flavors, mixing essential oils or blending is
considered part art and part science. When a formulation is prepared, it is
necessary to consider: (1) the chemistry of the oil to determine its volatil-
ity, viscosity, and other physical properties and (2) the desired action as
blending correctly allows for a synergistic effect within the blend. This
means the action of the oil is increased by mixing several oils together; in
some cases, the sequence in which the oil is blended can also be a factor.
Changing the sequence can change the properties.
Isolated compounds from essential oils can also be effective as pesti-
cides, and in some cases, higher toxicity is achieved when the compound
is removed from the context of the parent oil. Certain essential oils, com-
monly used in fragrances and as flavors, are exempt from pesticide reg-
istration in the USA. Specifically, those essential oils (and constituents)
on FIFRA (Federal Insecticide, Fungicide, and Rodenticide Act) List
25B (“Exempt Active Ingredients”) have been used to create insecticides,
fungicides, and herbicides since 1998. The following essential oils and
compounds are exempt from pesticide registration in the USA: cinnamon,
citronella, clove, garlic, geranium, lemongrass, mint, peppermint, rose-
204 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

mary, thyme, eugenol, and geraniol. Australia has a similar list of exempt
products that includes many essential oils.
Considerable research has been conducted in Canada to evaluate
synergy and compare toxicity of individual compounds to their parent
essential oils. Contact and fumigant toxicities of thymol, citronellal,
eugenol, and rosemary oil were tested on the wireworm Agriotes obscu-
rus. Thymol was the best contact toxin (LD50 = 196.0 µg/Larva), whereas
citronellal and eugenol were less toxic (LD50 = 404.9 and 516.5 µg /
larva, respectively). Rosemary oil did not show any significant contact
toxicity, even at 1,600 µg /larva. In terms of fumigant toxicity, citronel-
lal was the most toxic to wireworm larvae (LC50 = 6.3 µg /cm3) followed
by rosemary oil (LC50 = 15.9 µg /cm3), thymol (LC50 = 7.1 µg /cm3),
and eugenol (LC50 = 20.9µg /cm3) (Waliwitiya et al., 2005). Monoter-
penoids (terpenes and biogenically related phenols) commonly found in
plant essential oils were tested for acute toxicity via topical application
to tobacco cutworms (Spodoptera litura Fab.), the most toxic among 10
compounds being thymol (LD50 = 25.4 µg /larva) from garden thyme,
Thymus vulgaris. The compounds were then tested for sublethal effects,
specifically inhibition of larval growth after topical application of low
doses. Because minor constituents in complex essential oils have been
suggested to act as synergists, binary mixtures of the compounds were
tested for synergy vis à vis acute toxicity and feeding deterrence. Trans-
anethole synergized with thymol, citronellal, and α-terpineol in terms of
both acute toxicity and feeding deterrence. On the basis of these find-
ings, several complex mixtures were developed and tested as leads for
effective control agents. Candidate mixtures demonstrated good syner-
gistic effects (Hummelbrunner and Isman, 2001). Bioassays of rosemary
(Rosmarinus officinalis L.) essential oil and blends of its major constitu-
ents were conducted using host-specific strains of the two-spotted spider
mite, Tetranychus urticae Koch, on bean and tomato plants. Two constit-
uents tested individually against a bean host strain and five constituents
tested individually against a tomato host strain accounted for most of
the toxicity of the natural oil. Toxicity of blends of selected constituents
indicated a synergistic effect among the active and inactive constituents,
with the presence of all constituents necessary to equal the toxicity of the
natural oil (Miresmailli et al., 2006). Tak et al. (2015) reported a strong
Essential Oils as Green Pesticides for Plant Protection 205

synergistic interaction between 1,8-cineole and camphor, the major con-

stituents of rosemary oil, against the cabbage looper Trichoplusia ni and
the mechanism of synergy is through enhanced penetration of the cuticle
by camphor when admixed with 1,8-cineole (Tak and Isman, 2015).

14.5 ESSENTIAL OIL-BASED PESTICIDES-FROM THE DATA TO

THE CROP FIELD AFTER 40 YEARS OF RESEARCH

A literature search encompassing the past 40 years with the keywords

“essential oil” and “insects” yielded no less than 2,000 scientific papers.
Most papers document the immediate effects as acute toxicity or repel-
lence on arthropods (Regnault-Roger et al., 2012). In spite of widespread
public concern for long-term health and environmental effects of synthetic
pesticides, especially in Europe and North America, natural pesticides of
plant origin have not had much impact in the marketplace thus far. Many
conventional insecticides upon which growers had depended for decades
(e.g., organophosphates and carbamates) in the USA. were dramatically
restricted in use by the Food Quality Protection Act of 1996. In turn, this
act created a market opportunity for alternative products, in particular
“reduced-risk” pesticides that are favored by the Environmental Protec-
tion Agency. Some American companies took advantage of the regulatory
exemption and have been able to bring essential oil-based pesticides to
market in a far shorter time period than would normally be required for a
conventional pesticide.
The most attractive aspect of using essential oils and/or their constitu-
ents as crop protectants (and in other contexts for pest management) is their
favorable mammalian toxicity (Isman, 2000). Since 2000, EcoSMART
Technologies Inc. has been a world leader in essential oil-based pesti-
cides. An insecticide/miticide containing rosemary and peppermint oils
as active ingredients was introduced for use on horticultural crops under
the name EcoTrol EC. These and other EcoSMART pesticides have also
been approved for use in organic food production. They are currently
sold for home and garden use under the EcoSMART brand name (a divi-
sion of Kittrich Corporation, Pomona, CA, USA), and for crop protection
as Ecotec EC (Brandt Consolidated, Springfield, IL, USA) and Ecotrol
206 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

EC (KeyPlex, Winter Park, FL, USA). Several smaller companies in the

USA and the UK developed garlic-oil based pest control products and in
the USA. There are also consumer insecticides for home and garden use
that contain mint oil as the active ingredient. Menthol was approved for
use in North America for the control of tracheal mites in beehives, and a
product produced in Italy (Apilife VARTM) containing thymol and lesser
amounts of 1,8-cineole, menthol and camphor has been used to control
Varroa mites in honeybee hives since 1996. CinnamiteTM insecticide/miti-
cide was developed for greenhouse use by Mycotech Corporation in 1999.
Based on cinnamon oil, this pesticide was labeled for use against mites
and aphids, but is no longer produced.
Although several plant essential oils are exempt from registration in
the USA, many more oils are not, and few other countries currently pro-
vide for such exemptions. Accordingly, regulatory approval continues to
be a barrier to commercialization and will likely continue to be a barrier
until regulatory systems are adjusted to better accommodate these prod-
ucts (Isman and Machial, 2006).
Requiem®, an essential oil-based pesticide, was registered in USA in
2008. It was the first botanical registered in the USA since 1990 (Chias-
son et al., 2004) and is based on terpene constituents of Chenopodium
ambrosioides. It is currently marketed by Bayer Cropscience, both in the
USA and EU. Plants of the genus Chenopodium, notably the species C.
quinoa, C. album, and C. ambrosioides, have traditionally been used in
agriculture as food staples or as natural insecticides (Quarles, 1992). Eco-
oil® is an Australian botanical miticide/insecticide designed to control a
wide range of insects such as scale, mites, aphids, whitefly, and leafmin-
ers. It is a blend of essential oils and contains 2% tea tree oil (Melaleuca
alternifolia), eucalyptus oil, and canola oil.
TopiaTM insecticide was registered in 2009. It is the first completely
organic aerosol product from FMC, utilizing geraniol as the active ingre-
dient to eliminate a wide variety of household pests including ants, bed
bugs, cockroaches, silverfish, and stink bugs. Even more recently (2014),
the EPA approved Captiva®, an insecticide based on Capsicum oleoresin
and garlic oil, produced by Ecoflora Agro in Colombia, and Prev-Am®
(orange peel oil) was approved in the European Union. Other formula-
tions based on garlic oil (Biorepel®) or mixed with clove oil (Pest Out®;
Essential Oils as Green Pesticides for Plant Protection 207

SaferGro) are miticide/insecticides that provide control of mites, thrips,

and aphids.
Some recently introduced botanical pesticides based on essential oils
were registered in Mexico (Isman, 2014), including Akabrown based on
cinnamon, oregano, mint, and clove oils and Ebioluzion with 5% terpe-
noids. EcoVia™ EC (Rockwell Labs Ltd), based on thyme oil and rose-
mary oil, was developed for the control of mosquitoes, ticks, sucking
insects as well as numerous other flying and crawling insect pests on flow-
ering ornamental plants, trees and turf.

14.6 ADVANTAGES AND DISADVANTAGES OF ESSENTIAL OIL-

BASED PESTICIDES FOR PEST MANAGEMENT IN HORTICULTURE

The environmental problems caused by overuse of synthetic pesticides have

been the matter of concern for both scientists and the general public in recent
decades. Their high toxicity and residues in soil, water resources, and crops
that affect public health led researchers to search for new highly selective
and biodegradable pesticides. Natural products are one viable alternative to
synthetic pesticides as a means to reduce negative impacts to human health
and the environment. The move toward green chemistry and the continu-
ing need for developing new crop protection tools with novel modes of
action makes discovery and commercialization of natural products as green
pesticides an attractive and profitable pursuit that is commanding attention
(Koul et al., 2008). The concept of “Green Pesticides” refers to all types of
nature-oriented and beneficial pest control materials that can contribute to
pest population suppression and increased food production. They are more
compatible with the environmental than many synthetic pesticides (Isman
and Machial, 2006). Use of essential oils or their components are consistent
with this natural concept owing to their volatility and limited persistence
under field conditions, and several are exempt from registration in the USA.
In general, essential oil-based pesticides are considered safe, and they offer
some advantages when compared with synthetic pesticides:
1. Essential oils have been widely used for medicinal and clinical
proposes and in addition, they show low toxicity to other verte-
208 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

brates including fish and birds because they do not persist in soil
and water (Isman, 2000).
2. Essential oils can be applied as tank mixtures with synthetic insec-
ticides that can lessen the quantities of synthetic pesticides used.
Further, essential oils can be applied in rotation with conventional
products to mitigate the development of insecticide resistance
in pest population or for early season application in conjunction
with augmentative biological control when pest pressures are
low. Nattudurai et al. (2013) tested two synthetic volatile com-
pounds (benzaldehyde and propionic acid) and two volatile oils
(camphor and eucalyptus). They were screened individually and
in combinations against different life stages of Tribolium castaneum.
The individual treatments of camphor and eucalyptus oils were less
effective, but combinations of benzaldehyde–camphor oil were
found to be effective. Benzaldehyde–propionic acid combination
recorded 99.3% adult mortality inside a 1 m3 wooden cage after 15
days, and this mixture can be used as a fumigant in store houses.
3. They can be compatible with biocontrol because of the lack of foliar
residues. The essential oils from leaves of Schinus molle var. areira,
Aloysia citriodora, Origanum vulgare, and Thymus vulgaris have
shown potential as insecticides against the green stink bug Nezara
viridula. Their toxicological and behavioral effects on the parasitoid
Trissolcus basalis, a biological control agent of this pest insect, were
also evaluated. The essential oils from O. vulgare and T. vulgaris
proved to be highly selective when used as fumigants and did not
change parasitoid behavior. After 1 week, the residues of these oils
were harmless and did not show sublethal effects against T. basalis.
Based on these results, essential oils have potential applications for
the integrated management of N. viridula (Werdin González, 2013).
4. Owing to their volatility, the oils and their constituents are envi-
ronmentally non-persistent, with outdoor half-lives of <24 h on
surfaces, in soil and in water. There has not been any report of bio-
magnification of essential oils through the food chain (Regnault-
Roger et al., 2012).
5. Limited toxicity to pollinators. Although essential oils have fewer
nontarget effects on natural enemies, direct contact on beneficial
Essential Oils as Green Pesticides for Plant Protection 209

insects such as pollinating bees can cause mortality. White et al.

(2009) showed that wintergreen oil applied as a fumigant on crop
pollinators, such as Osmia cornifrons (Radoszkowski) (Hymenop-
tera: Megachilidae) to control parasitic mites, required >2,473.5
ppm to cause bee mortality. However, when wintergreen oil was
topically applied to bees, 353.4 ppm of wintergreen oil caused bee
mortality within 10 min. On the other hand, Ebert et al. (2007)
focused on Apis mellifera adult toxicity when testing 10 prod-
ucts: 1,8-cineole, clove oil, formic acid, marjoram oil, menthol,
oregano oil, oxalic acid, sage oil, thymol, and wintergreen. Each
product was tested at several concentrations in a sugar syrup fed
to bees over several days, and dead bees were counted daily. Men-
thol and 1,8-cineole had mortality levels no different from controls
fed plain syrup after 8 days of treatment. At 14 days of treatment,
wintergreen oil was the least toxic. These results indicate that the
tested products could all be used safely for treating bees orally if
dose is carefully managed in the hive.
6. In terms of green pesticide technology, using oil-in-water micro-
emulsions as a nano-pesticide delivery system to replace tradi-
tional emulsifiable concentrates (oil), in order to reduce the use of
organic solvents and increase dispersion, wettability and penetra-
tion of the droplets is being developed. The advantages of using
pesticide oil-in-water microemulsions for improving the biological
efficacy and reducing the dosage of pesticides would be a useful
strategy in green pesticide technology (Koul et al., 2008).
7. No toxic residues remain on harvested produce. Many oils are
obtained from medicinal herbs and have been used as food preser-
vation and flavor for centuries. They are also safe for applicators
and field workers.
8. They can be cost competitive and some are approved for organic
agriculture in the USA and EU.
Despite many advantages, the market for essential oil-based insecti-
cides has a number of major challenges, and is very small at present. How-
ever, at least based on data from California, it is growing at a much faster
rate than that for synthetic insecticides.
210 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

1. Since essential oils tend to evaporate quickly from surfaces, the

spice-based pesticides need to be applied to crops more frequently
than conventional pesticides. Some last only a few hours, com-
pared to days or even months for conventional pesticides. Because
these natural pesticides are much less toxic than conventional pes-
ticides, they will likely be applied in higher concentrations. There-
fore, coverage is important, and reapplications may be necessary to
achieve acceptable control.
2. Some essential oils can be phytotoxic if very high rates are applied
although phytotoxicity depends on plant species’ susceptibility. No
phytotoxic symptoms were observed on grape leaves treated with cit-
rus essential oils, and low phytotoxicity was caused by the essential
oils of lavender, thyme-leaved savory, and mint, whereas the highest
phytotoxicity was observed when basil oil was used (Karamaouna et
al., 2013). Vapors of lavender oil, lemon balm, oregano, and thyme
caused desiccation of cayenne plants at 2 μL/L, and the same concen-
tration of oregano killed broad bean plants (Digilioa et al., 2008).
3. Many essential oils have best efficacy against small, soft bodied
pests (mites, thrips, aphids, mealybugs) and as fumigants for cole­
opteran stored product pests, but less efficacy against more robust
foliar-feeding lepidopterans and coleopterans.

14.7 CONCLUSION

In recent years, consumers have increasingly expressed interest in pur-

chasing organically grown foods, as well as using natural and naturally
derived materials to eradicate pests in their lawn, garden and homes. Also,
there has been a steady increase in the amount of pesticides marketed for
organic food production, demonstrating that farming without synthetic
pesticides is entirely feasible, scalable, economically profitable, and envi-
ronmentally safe. Pesticides based on plant essential oils or their constitu-
ents have demonstrated efficacy against many agricultural pests. Due to
their volatile nature, there is a much lower level of risk to the environ-
ment than with current synthetic pesticides, and they can be compatible
with integrated pest management programs. The volatility of essential oils
makes them especially suitable as fumigants in protected environments
Essential Oils as Green Pesticides for Plant Protection 211

and for protection of stored products, but they also have demonstrated
utility for protection of horticultural crops.

KEYWORDS

•• essential oil
•• fumigant
•• green pesticide
•• medicinal plants
•• plant protection
•• toxicity

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214 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

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Sustainable Horticulture Volume 2: Food, Health, and Nutrition A

FIGURE 6.1 Changes in anthocyanin content during storage of blended jamun-aonla

ready-to-serve beverages.

FIGURE 6.2 Changes in phenolics in blended jamun–aonla ready to-serve beverages

during storage.
B Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 6.3 Sensory evaluation of blended jamun–aonla ready-to-serve beverages

during storage.

160
Ascorbic acid (mg/100ml)

120

80 0 Month
3 Month
6 Month
40

0
Aonla- Aonla- Aonla- Aonla-
Litchi I Litchi II Grape I Grape II

FIGURE 7.1 Changes in ascorbic acid content of blended aonla squash during storage.
Sustainable Horticulture Volume 2: Food, Health, and Nutrition C

1000

800
Total phenolics (mg/100ml)

600 Aonla-Litchi I
Aonla-Litchi II
Aonla-Grape I
400 Aonla-Grape II

200

0
0 Month 3 Month 6 Month
Months of storage

FIGURE 7.2 Changes in total phenolic content of blended aonla squash during storage.

9
Anthocyanins (mg/100ml)

6
Aonla-Grape I
Aonla-Grape II

0
0 Month 3 Month 6 Month
Months of storage

FIGURE 7.3 Changes in anthocyanin content of aonla–grape squash during storage.

9
Sensory score (out of 9)

6
0 Month
3 Month
6 Month
3

0
Aonla-Litchi I Aonla-Litchi II Aonla-Grape I Aonla-Grape II

FIGURE 7.4 Changes in sensory scores of blended aonla squash during storage.
D Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 8.1 Effect of yeasts on the CPLW percent of mango cv. Dashehari during
storage. T1 (baker’s yeast), T2 (industrial yeast), T3 (Saccharomyces cerevisiae), and T4
(control).

FIGURE 8.2 Effect of yeasts on the firmness (kg/cm2) of mango cv. Dashehari during
storage. T1 (baker’s yeast), T2 (industrial yeast), T3 (Saccharomyces cerevisiae), and T4
(control).
Sustainable Horticulture Volume 2: Food, Health, and Nutrition E

FIGURE 8.3 Effect of yeasts on the spoilage (%) of mango cv. Dashehari during storage.
T1 (baker’s yeast), T2 (industrial yeast), T3 (Saccharomyces cerevisiae), and T4 (control).

FIGURE 8.4 Effect of yeasts on the TSS (°Brix) of mango cv. Dashehari during storage.
T1 (baker’s yeast), T2 (industrial yeast), T3 (Saccharomyces cerevisiae), and T4 (control).
F Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 8.5 Effect of yeasts on the titratable acidity (%) of mango cv. Dashehari during
storage. T1 (baker’s yeast), T2 (industrial yeast), T3 (Saccharomyces cerevisiae), and T4
(control).

FIGURE 8.6 Effect of yeasts on the total carotenoids (mg/100 g) of mango cv Dashehari
during storage. T1 (baker’s yeast), T2 (industrial yeast), T3 (Saccharomyces cerevisiae), and
T4 (control).
Sustainable Horticulture Volume 2: Food, Health, and Nutrition G

FIGURE 8.7 Effect of yeasts on the antioxidant FRAP (µmolar TE/g) of mango cv.
Dashehari during storage. T1 (baker’s yeast), T2 (industrial yeast), T3 (Saccharomyces
cerevisiae), and T4 (control).

FIGURE 8.8 Effect of yeasts on the antioxidant DPPH (% inhibition) of mango cv.
Dashehari during storage. T1 (baker’s yeast), T2 (industrial yeast), T3 (Saccharomyces
cerevisiae), and T4 (control).
H Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 9.1 Effect of treatments on stem elongation and percent flower opening of (a)
antirrhinum, (b) dimorphotheca, (c) larkspur, (d) lupin, and (e) Sweet William.

FIGURE 9.2 (a) Effect of treatments on flower diameter (mm) of dimorphotheca and (b)
effect of treatments on percent flower opening of Sweet William.
Sustainable Horticulture Volume 2: Food, Health, and Nutrition I

FIGURE 9.4 Effect of treatments on vase life of winter annuals.

Chart showing the % cumulative weight loss of cling filmed and non cling filmed breadfrutis
from Pamplemousses E.S. stored at 13 oC.

18.000

16.000

14.000

12.000
% weight loss

10.000
cling film
no cling film
8.000

6.000

4.000

2.000

0.000
2d 3d 6d 7d 9d 13d 15d
Days of storage

FIGURE 10.1 Chart showing the % cumulative weight loss of cling filmed and noncling
filmed breadfruits from Pamplemousses ES stored at 13°C.
J Sustainable Horticulture Volume 2: Food, Health, and Nutrition

MAMIT
DISTRICT

FIGURE 12.1 Map of Mamit district showing the study sites.

Sustainable Horticulture Volume 2: Food, Health, and Nutrition K

% of hh involved in harvesng of NTFPs 90

80
70
60
50 Dapchhuah
40 Chhippui
30 Tuahzawl
20 Chungtlang
10
Lengte
0
Bamboo Broom grass Fruits Wild foods Fuelwood Medicinal
pole plants

NTFPs

FIGURE 12.10 Percentage of household involved in harvesting of NTFPs.

250
Quanty of NTFPs harvest (kg/hh/year)

200

150
Dapchhuah
Chhippui
100
Tuahzawl

50 Chungtlang
Lengte
0
Bamboo Broom Fruits Wild foods Fuelwood Medicinal
pole grass plants
NTFPs

FIGURE 12.11 Quantity of NTFPs harvested in the studied villages (kg/hh/year).

140
Home consumpon (kg//hh/year)

120

100

80 Dapchhuah

60 Chhippui
Tuahzawl
40
Chungtlang
20 Lengte

0
Bamboo Broom grass Fruits Wild foods Fuelwood Medicinal
pole plants
NTFPS

FIGURE 12.12 Amount of NTFPs used for home consumption in different villages (kg/
hh/year).
L Sustainable Horticulture Volume 2: Food, Health, and Nutrition

140

120
NTFPs sold (Kg/hh/year)

100

80 Dapchhuah
60 Chhippui

40 Tuahzawl
Chungtlang
20
Lengte
0
Bamboo Broom grass Fruits Wild foods Fuelwood Medicinal
pole plants
NTFPs

FIGURE 12.13 Amount of NTFPs sold in different villages.

1600
1400
Income generate d from

1200
NTFPs(Rs/hh/year)

1000
Dapchhuah
800
Chhippui
600
Tuahzawl
400
Chungtlang
200
Lengte
0
Bamboo Broom Fruits Wild foods Fuelwood Medicinal
pole grass plants
NTFPs

FIGURE 12.14 Estimated income generated from NTFPs in different villages.

900
800
700
600
household

500
400 % of household involved/year
300 Quanty harvested(kg/hh/year)
200
Home consumpon(kg/hh/year)
100
0 Quanty sold(kg/hh/year)
Income(Rs/hh/year)

NTFPs

FIGURE 12.15 Mean of all the NTFPs uses in the five surveyed villages.
Sustainable Horticulture Volume 2: Food, Health, and Nutrition M

FIGURE 18.1 In vitro effect of plant extracts on the mycelial growth of Alternaria solani.

FIGURE 20.1 Antifungal efficacy (%) of the test samples against C. capsici – A graphical
representation.
N Sustainable Horticulture Volume 2: Food, Health, and Nutrition

100 50 25 Standard
40
35
Zone of Inhibition (mm±S.E.)

30
25
20
15
10
5
0
-5 E.coli P. vulgaris S. typhi K. pneumoniae P. aeruginosa
Concentration (ug/ml)

FIGURE 22.1 Effect of essential oil showing zone of inhibition against bacterial
pathogens.

120
Average % igrowth nhibition

100

80

60

40

20

0
2.5 1.25 0.625 0.3125 0.156

Concentration (mg/ml)

E.coli P. vulgaris S. typhi K. pneumoniae P. aeruginosa

FIGURE 22.2 Average percent growth inhibition of peppermint oil against bacterial
pathogens.
Sustainable Horticulture Volume 2: Food, Health, and Nutrition O

FIGURE 24.1 Map of Morocco and the study area.

FIGURE 26.2 EDXRF spectra of major elements in Solanum nigrum linn and Spilanthes
acmella.
P Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 26.3 ED-XRF spectra of minor elements in Solanum nigrum linn and Spilanthes
acmella.

FIGURE 26.4 ED-XRF spectra of earth elements in Solanum nigrum linn and Spilanthes
acmella.
CHAPTER 15

INTEGRATED HEALTH MANAGEMENT

IN HORTICULTURAL CROPS
C. CHATTOPADHYAY1 and AJANTA BIRAH2
1
Uttar Banga Krishi Viswavidyalaya, Pundibari,
Coochbehar–736165, West Bengal, India,
E-mail: [email protected]
ICAR-National Research Centre for Integrated Pest Management,
2

New Delhi–110012, India

CONTENTS
Abstract ................................................................................................. 217
15.1 Introduction ................................................................................ 218
15.2 Integrated Health Management .................................................. 219
15.3 Conclusion ................................................................................. 227
Keywords............................................................................................... 227
References.............................................................................................. 228

ABSTRACT

Plant protection is one of the key issues in the overall gamut of Indian
agriculture. In view of indiscriminate use of chemical pesticides, environ-
mental safety vis-á-vis sustaining crop yields, threats to farm bio-secu-
rity, and crop health in the era of globalization, the situation has become
rather challenging. Integrated pest management (IPM) follows the prin-
ciples of understanding of the crop, pest, and the environment and their
interrelationships to enable advanced planning with emphasis on routine
monitoring of crop and pest conditions and balancing of cost/benefits of
218 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

all management practices. Agriculture and allied activities are the main
source of livelihood for the people of northeastern region of our coun-
try, and any attempt to reduce poverty as well as to place the region in
developmental paradigm should have a system-based eco-regional plan-
ning for agricultural development. IPM is a complex process, and farmers
lack understanding of the biological processes of pests, their predators,
and methods of application of new components. There are a number of
IPM practices that work best when applied by the entire community and
in a synchronized mode. Thus, an integrated decision support system for
crop protection services may be required to be devised centrally to moni-
tor the pest dynamics through e-pest surveillance, analysis of pest risks,
and provision of pest forecasts along with mobile-based dissemination
of advisories keeping in view prevailing weather and changes in climate.
This would necessitate networking of all stakeholders so that they could
contribute effectively in a cohesive manner.

15.1 INTRODUCTION

Crop yield losses in India due to pests (includes all biotic stresses, viz.,
weeds, insect pests, diseases, nematodes, rodents, etc.) range 15–25%
(varied estimates by different sources). Losses due to damages caused by
pests in terms of quantity are almost to an extent such that if that loss could
be saved, India could meet its needs of 2020 domestically even with the
present levels of crop productivity as well as keeping in view the stagna-
tion in yields for certain crops vis-á-vis impacts of climate change. Lack
of knowledge/awareness about eco-friendly methods of pest management
apart from unavailability of inputs required for the same makes the job fur-
ther difficult. However, efforts of researchers in crop protection have been
able to make sizable dent on reducing losses due to biotic stresses with
the use and up-scaling of eco-friendly technological packages, thereby
improving crop productivity and livelihood of farming community. There
has been quite some time since we faced any acute epidemic or epizootic,
and relevant technologies have also played their roles in minimizing such
incidences. There has been considerable head way in the area of technol-
ogy development and its implementation at the field level – for instance,
the National Food Security Mission has a rich component of plant protec-
tion in overall implementation of the scheme at the national level. There-
Integrated Health Management in Horticultural Crops 219

fore, plant protection currently holds the key to redeem much of the losses
due to pests through convergence of stakeholders.

15.2 INTEGRATED HEALTH MANAGEMENT

IPM technology envisages to integrate different management techniques

to keep crop plant stress due to different deleterious organisms below eco-
nomic injury/threshold level and thereby targets to maintain good crop
health. The components of host resistance (manipulation of crop to with-
stand or tolerate pests) include use of clean certified seed/indexed plant-
ing material of the variety recommended for the targeted region, cultural
management (to optimize growing conditions for the crop or anything that
increases a crop’s competitive edge to result in increased tolerance to pests
often resulting in reduced pesticide use or create unfavorable conditions
for the pest), sanitary management that avoids introducing pest to crop
field (clean field equipment, removal of the affected plant/its part), and
natural and biological management that enhances beneficial organisms
or releases predators, parasites, etc. Economic injury level (EIL) is the
cost of control that equals value of damage caused by the pest, which is
determined through extensive research and is the information necessary
to develop an economic threshold (ETL), which is used by crop advisors.
The northeast (NE) region comprising eight states, viz., Assam,
Arunachal Pradesh, Meghalaya, Manipur, Mizoram, Nagaland, Tripura,
and Sikkim, has a total geographical area of 262180 km2, which is nearly
8% of the total area of the country with more than 39 million people. The
total area under horticultural crops is around 822.5 thousand hectares,
which is around 3.14% of the total geographical area of the region, which
provides a production of 6818.4 thousand tons. Out of the total area under
different fruit crops in the NE region, ~60.6 thousand hectares is only under
banana cultivation. Among the biotic stresses, various fungal, bacterial, and
viral diseases starting from nursery to postharvest stage cause consider-
able loss. As the diseases are either planting material-borne or soil-born,
they are very difficult to manage. Only early diagnosis and IPM practices
can be effective in managing these problems. Food grain production and
requirement scenario indicates a deficit existing in all the NE states, vary-
ing from 10% for Tripura/Sikkim to 69% for Mizoram, except Nagaland
that has some surplus food grains. While planning this, the strength of the
220 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

farming system approach to judicious utilization and conservation of natu-

ral resources of the region with concurrent policy and research support to
increase production, add value to the produce, and their disposal/sale man-
agement shall be of paramount importance (Vision 2030, ICAR NEH).
Green revolution, one of the greatest success stories of India, was due
to intensive agriculture (IA). However, factors involved in IA like genetic
uniformity of crops, dense plant population, mono-cropping, higher fer­
tilization and irrigation, inappropriate cropping systems, practices (chem­
icalization) with immediate profit motives favoured insect pests and diseases
incidence. IPM and the use of biotic agents to minimize the unsystematic
and indiscreet use of chemical pesticides will be the fundamental theory
of plant protection. Biopesticides represent only 2.89% (as on 2005) of the
overall pesticide market, with an expected annual growth rate of 2.3% (as
in 2006). Neem-based pesticides, Bt, NPV (Ha/Sl), and Trichoderma, are
the major biopesticides produced and used in India. Although only 0.6 kg/
ha chemical pesticide is used in India compared to 14 kg/ha in China and
12 kg/ha in Japan, the use is highly injudicious in some pockets that have
led to several issues of pest resurgence, development of resistance in pests
to chemical pesticides apart from human health problems [Chemical pesti-
cides were responsible for 49% lower sperm count in men eating raw fruits
(Sheiner et al., 2003); they are among the factors responsible for neurologi-
cal problems (Ascherio et al., 2006; Baldi et al., 2010), neurodevelopmen-
tal disorders (Beseler et al., 2008; Jurewicz and Hanke 2008), birth defects
(Winchester et al., 2009). Organochlorine pesticides were linked to preterm
deaths, reduced baby weight, and ovarian cancer in north India (Mustafa
et al., 2015; Sharma et al., 2015; Tyagi et al., 2015), and around 2.2 mil-
lion people die annually of cancer related to chemical pesticide poisoning
(McCauley et al, 2006; Gilden et al., 2010). Large use of chemical pesticides
leads to several diseases (April 7, 2015, Times of India; WHO, CSAUAT).
Chemical pesticides are linked to increased risk of diabetes, and exposure
to them significantly increases the risk of type 2 diabetes by nearly 60%
(Fotini Kawoura, September 25, 2015, Medscape Medical News, UK).] and
disturbance to environment and biodiversity. This also brought significant
shift in the insect population dynamics and change in the status of several
insect pests. The cost of plant protection on various crops ranged 7–40% of
the total crop production cost. Though IPM has been advocated for the past
two decades, the number of farmers who adopted IPM practices in various
Integrated Health Management in Horticultural Crops 221

crops in India is debatable. IPM research in the past decade brought out
changes in the farmers’ attitude in pest management, which resulted reduc-
tion in pesticide use in different crops. To be more effective, readdressing the
policies for encouraging eco-friendly options and strengthening extension,
involving farmers should be considered as high priority.
In most cases, IPM consists of scouting with timely application of a
combination of strategies and tactics. These may include site selection and
preparation; utilizing resistant cultivars; altering planting practices; modi-
fying the environment by drainage, irrigation, pruning, thinning, shading,
etc.; and applying pesticides, if necessary. But in addition to these tradi-
tional measures, monitoring environmental factors (temperature, moisture,
soil pH, nutrients, etc.), disease forecasting, and establishing economic
thresholds are important to the management scheme. These measures
should be applied in a coordinated and harmonized manner to maximize
the benefits of each component. For example, balancing fertilizer applica-
tions with irrigation practices helps promote crop health.
India has successfully reduced pesticide consumption without
adversely affecting the agricultural productivity. This was facilitated by
appropriate policies that discouraged pesticide use and favored IPM appli-
cation. Despite such efforts, adoption of IPM is low owing to a number of
socioeconomic and other constraints. Though many technology programs
are based on community approach, they do not have any proper policy to
sustain the group approach. The IPM policy should also provide incentives
to farmers to adopt IPM as a cardinal principle of plant protection.
The IPM is knowledge intensive on crop, pest(s), environment, and
their inter-relationships; holistic in approach; and requires expert advice,
timely decision-making, and immediate actions for solutions to pests.
Basic principles of IPM involve advanced planning, balancing of cost and
benefit from any management interventions, and routine monitoring of
crop and pest conditions. Farmers needs involve pest (including patho-
gen, weed, etc.) identification (diagnostics) during crop surveillance, pest
surveillance, monitoring preferably accomplished by e-pest surveillance
at the national/global level, pest forecasting and dissemination of expert
information on pest management for quick action to solution. Availabil-
ity of appropriate inputs of IPM also impedes following principles of the
same. Thus, an important need of the hour is to improve levels of aware-
ness on IPM among field functionaries and farmers.
222 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Since 1985, the Government of India (GoI) has enabled farmers to adopt
IPM practices to bring down losses due to pests and also provide ways to
reduce use of chemical pesticides. In view of the same, the National Research
Centre for Integrated Pest Management (NCIPM) was set up in VII Plan
(1988) under the Indian Council of Agricultural Research. In the National
Agricultural Policy announced by the GoI in 2001, para 24 emphasizes IPM
and use of biological control agents to minimize indiscriminate and injudi-
cious use of chemical pesticides as a cardinal principle for crop protection.
During >25 years of existence, NCIPM has achieved successes in validat-
ing and harmonizing IPM technologies in different crops, viz., Astha vil-
lage (Maharashtra) for cotton, Bambawad (UP) in basmati rice, etc. In 1992,
Central Integrated Pest Management Centres (CIPMCs) were established
by the GoI by merging all Central Plant Protection Stations (CPPS), Cen-
tral Surveillance Stations (CSS), and Central Biological Control Stations
(CBCS). Presently, there are 31 CIPMCs in 28 states and 1 UT, who are
engaged in pest monitoring and field release of biological control agents,
conduct farmers’ field schools (FFSs), and train extension officers and mas-
ter trainers. CIPMCs are in turn linked with 98 state biocontrol laboratories.
To facilitate popularizing the IPM approach among farming community
under the Central Sector Scheme “Promotion of Integrated Pest Manage-
ment” of the Department of Agriculture and Cooperation, GoI, an informa-
tion system for IPM has been created, which helps in efficient reporting and
dissemination of information on pest surveillance, rearing of host culture,
production and release of biological control agents in the field, conserva-
tion of naturally occurring biological control agents for the control of crop
pests, and transfer of innovative IPM skills/methods/techniques to exten-
sion workers and farmers through conduct of training and FFSs in all states
by CIPMCs of the Directorate of Plant Quarantine and Storage, Faridabad.
Biological control is also a very effective component of crop protection.
Due to public awareness about the hazards related to the use of chemical pes-
ticides, there has been a lot of interest generated for the use of eco-friendly
strategies targeted at the management of crop pests. For this purpose, biopes-
ticides could be a cost-effective, eco-friendly, and sustainable option, when
proven source of host resistance/tolerance against several pests is not avail-
able. However, the quality, quantity, application method, and timeliness play
a significant role in determining the level of success of biological control.
There are several success stories of biological control doing a commend-
Integrated Health Management in Horticultural Crops 223

able job in the field of crop protection. Successful biological management of

papaya mealy bug and sugarcane wooly aphid alone have saved >Rupees 2.5
thousand crore (>4100 m US$) in 2 years for the nation. The dreaded weed
Mikania micrantha is being successfully managed in southwest and NE
India by Puccinia spegazzinii (rust fungus). Credibility of the bioformula-
tions Kalisena [technology developed by Indian Agricultural Research Insti-
tute (IARI), New Delhi and Indian Council of Agricultural Research (ICAR),
and transferred for marketing in Asia, Africa, North and South America to
M/s Cadila Pharmaceuticals Ltd.] has been established with the farmers and
their advisors, which is an excellent example of translational research. Garlic
bulb aqueous extract (2% w/v) has also been adopted by farmers and the
Government of Rajasthan in managing pests of Indian mustard. Use of qual-
ity strains of Trichoderma, Pseudomonas fluorescens, etc. in recommended
quantity even as seed treatment has been found to be very successful in man-
aging dreaded diseases of different field and horticultural crops, which could
safeguard from high yield losses. When they are combined with soil applica-
tion and/or foliar spray, they result in even better impact not only in reducing
pests and increasing yields and economic benefits but also in safeguarding
the environment from dangerous chemical pesticide load.
During the last 28 years, NCIPM has successfully launched IPM strate-
gies for different crop sectors across the country; has been successful in
promoting the IPM concept and in developing state-of-the-art technologies
and strategies concepts like those in Astha village (Maharashtra), eight dis-
tricts of Punjab and Jind (Haryana) models for cotton cultivation, IPM in
rice (Bambawad, UP; Chhajpur, Haryana; Hooghly, WB), pulses (Gulbarga
and Bidar, Karnataka; Jabalpur, MP; Mirzapur, UP), groundnut (Udaipur,
Rajasthan; Kadiri, Andhra Pradesh), mustard (Alwar, Rajasthan; Mohin-
dergarh, Haryana), vegetable (Anantpura, Rajasthan; onion at Singohi-Sin-
goha-Rambha and Bitter Gourd at Padhana, Haryana), and fruit (mango at
Navsari, Gujarat; citrus at Abohar, Punjab) crops. Other ICAR institutions
have shown success with grapes at Nashik, Maharashtra, and pomegranate
at Solapur, Maharashtra. NCIPM has also played a vital role in strengthen-
ing the capacity and capability in IPM in the country by organizing 55 pro-
grams to train 1582 participants, which include 476 KVK (Krishi Vigyan
Kendra) personnel among existing 643 KVKs from all eight zones of India.
Recently, NCIPM in active collaboration with ICAR Complex for the
North Eastern Hill (NEH) region and State Department of Agriculture,
224 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Government of Tripura, launched a project to uplift the socioeconomic sta-

tus of tribal farmers. The critical IPM input kits consisting Trichoderma,
Trichocards, yellow sticky, and pheromone traps were distributed among
the farmers. Demonstrations on the use of the inputs were also provided.
Mode of operation and establishment of infrastructure for the launch of
e-pest surveillance in Tripura has been planned. Potato grown in Tripura
offer a great potential for organic cultivation as the application of chemi-
cal pesticides is minimal. The productivity of potato is low due to various
biotic stresses of late blight, bacterial blight, common scab, and viral dis-
eases. Under these circumstances, validation of IPM modules developed
by NCIPM was undertaken at Barkhatal, Hazamara block, West Tripura.
Similarly, tomato is also one of the important vegetable crops cultivated in
Tripura as a postmonsoon crop. The crop is affected by several diseases like
bacterial wilt, tomato leaf curl, etc. A field trial was conducted to evaluate
the eco-friendly management practices, viz., staking, spray of neem oil,
application of bleaching powder and combination thereof; it was observed
that the combination provided significant reduction in incidence of diseases
in tomato. Accordingly, these management strategies were discussed with
the farmers, and efforts were undertaken to implement the same.
A few states have been more progressive in encouraging biological con-
trol of crop pests, viz., Gujarat, Tamil Nadu, West Bengal, etc. Safeguarding
intellectual property on strains of bioagents is an important issue in the pres-
ent era. Accordingly, there is a need to have DNA barcode data of all such
strains in order to sustain IPR. There is a need to undertake a specific policy
to encourage biopesticides, streamlining their label claim issues, simplifi-
cation of process of registration for biopesticides with strict and adequate
quality check from government departments (CIPMCs, SAUs (State Agri-
cultural Universities), etc.), increased support to biopesticide industry for
scaling up of production as a matter of government policy (viz., subsidies
to biopesticides, higher taxes on chemical pesticide industries, etc.), which
shall also enable generation of employment for small/micro-industries at
the village level in line with concepts of model bio-village.
Precision pest management to reduce indiscriminate use of chemi-
cal pesticides could plan use of state-of-the-art technology through inno-
vative and strategic research to enable devise integrated decision support
system (IDSS) for crop protection services that suggests operational focus,
research priorities, and evolution in a phased manner. Surveillance is the
Integrated Health Management in Horticultural Crops 225

foundation of plant protection for early alert. But it is missing in most of

the developing countries. In the recent past, the information communication
technology (ICT)-based system of real time pest surveillance has played an
important role in our country in collection and transfer of data from remote
villages to main station through the Internet. The information is compiled
and displayed on the website in tabulated and graphical form and that can
be directly accessed by SAUs for issue of advisory through the State Agri-
culture Department by SMS to farmers and extension workers for imple-
mentation in farmers’ fields. There is dramatic reduction in outbreak of any
major pest on selected crops since the inception of ICT activity in different
states. As the farmers are getting regular SMSs for IPM interventions, there
is much awareness about IPM. Holistic planning provides farmers with the
management tools they need to manage biological complex farming systems
in a profitable manner. A successful IPM program requires time, money,
patience, short- and long-term planning, flexibility, and commitment.
One of the most common approaches that have been adopted by many
countries is pre-sowing treatment of seed. Seed treatment is defined as
chemical or biological substances applied to seed or vegetatively propa-
gated material to manage diseases organisms, insect pests, etc. Seed treat-
ment pesticides include bactericides, fungicides, and insecticides. Most
seed treatments are applied to true seeds such as corn, wheat, or soybean,
which have a seed coat surrounding the embryo.
An IPM plan should identify important pests, determine pest manage-
ment options, and blend them together to achieve the goals listed above. To
use seed treatments effectively, it is important to understand the purposes
of seed treatment, alternatives, or supplements to seed treatments, and the
various advantages and disadvantages of seed treatments. Natural enemy
cum beneficial fauna population such as coccinellids, spiders and Chrysop-
erla, pollinators, and honey bee remain unharmed due to seed treatment.
Bridging the gap between policy and practice is a herculean task in
plant protection in the wake of extensive diversity (of crops, regions, pest
problems, and practices) variegated resources (knowledge, experts, qual-
ity inputs, machinery, molecules, and financial support), require highly
motivated and focused yet effective efforts (toward research, education,
extension) to tackle the issues in plant protection. However, through well-
coordinated efforts involving multistakeholders – policy makers, admin-
226 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

istrators, researchers, educationists, extension machinery, industry, media,

etc. visible contributions can be made in this important area.
Although GoI has put IPM as part of its National Agricultural Policy,
there is possibly a need to undertake a specific policy to push IPM by pro-
viding credits for greener pesticide molecules, streamlining label claim
issues, simplification of process of registration for biopesticides with strict
and adequate quality check from government departments (CIPMCs, SAUs,
etc.), increased support to biopesticide industry for scaling up of production
as a matter of government policy (viz., subsidies to biopesticides, higher
taxes on chemical pesticide industries, etc.), which shall also enable gen-
eration of employment for small/micro-industries at the village level in line
with concepts of model bio-village. This shall bring a shift in the chemical
pesticide industries and transform them toward producing biopesticides.
KVKs and NGOs need to play a vital role in improving awareness
levels of field functionaries and farmers in IPM, apart from fast-tracking
of crop protection advisories. IPM happens to be a knowledge-intensive
holistic approach wherein advanced planning, good agricultural prac-
tices (GAP) toward environment-safety linked to maximum residue
limit (MRLs) of chemical pesticides and cost-benefit management, crop
monitoring coupled with accurate diagnosis of the problem (pest), expert
advice, timely decision-making, and quick action make the real difference
to tackle unforeseen pest outbreaks.
Mankind has always been eager to know the unknown of the future
to enable plan suitably for the same. With the generation of knowledge,
the disease triangle and tetrahedron was considered involving the interac-
tion among host and pathogen in a given environment over time. Plant
disease forecasting is a management system used to predict the occur-
rence or change in the severity of plant diseases. At the field scale, these
systems are used by growers to make economic decisions about disease
management. The environment is usually the factor that controls whether
disease develops or not, the vulnerability of the host, and the presence of
the pathogen in a particular season through their effects on processes such
as overseasoning or ability of the pathogen to cause disease. In these cases,
a disease forecasting system attempts to define when the environment will
be conducive to disease development.
Integrated Health Management in Horticultural Crops 227

The NEH region also has tremendous potential for IPM. Unfortunately,
not a single IPM module has been developed for any crop for any part
of the NEH region so far. As IPM is location-specific, the IPM modules
developed for one region may not work in another. The NEH region is
unique and different from the rest of the country in many ways. This is also
true in the wider context of agriculture and more specifically, pests and
natural enemies abundant therein. For example, the common blister beetle
is a minor pest in the rest of the country, but in the NEH, it is a major pest.
Certain pests are found here, which may not be found in the mainland. This
is true for natural enemies too. The natural enemy diversity and abundance
is more in the NEH region; therefore, natural control is very high here.

15.3 CONCLUSION

Today’s challenge is to “produce more from less.” The big questions are,
do we have enough information on the fitness-cost of different pests of
major crops, their injury profiles vis-á-vis attainable yields, and are we
prepared enough to face the impacts of climate change in general and spe-
cifically on the pest scenario evolving as a result of the same? Now, the
challenge is to bring continuous improvement in productivity, profitabil-
ity, stability, and sustainability of major farming systems, wherein scien-
tific management of plant pests holds a pivotal role. Crop loss models,
representing a dynamic interaction between pests and host, are essential
for forecasting losses thereof.

KEYWORDS

•• biopesticide
•• horticulture
•• integrated
•• intensive agriculture
•• IPM
•• management
228 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

REFERENCES

Ascherio, A., Chen, H., Weisskopf, M. G. O.,’Reilly, E., McCullough, M, L., Calle, E. E.,
Schwarzschild M. A., & Thun, M. J., (2006). Pesticide exposure and risk for Parkin-
son’s disease. Ann Neurol., 60, 197–203.
Baldi, I., Lebailly, P., Mohammed-Brahim, B., Letenneur, L., Dartigues, J. F., & Brochard,
P., (2010). Neurodegenerative diseases and exposure to pesticides in the elderly. Am.
J., Epidemiol. 157, 409–414.
Beseler, C. L., Stallones, L., Hoppin, J. A., Alavanja, M. C., Blair, A., & Keefe, T., (2008).
Depression and pesticide exposures among private pesticide applicators enrolled in
the Agricultural Health Study. Environ. Health Perspect, 116, 1713–1719.
Gilden, R. C., Huffling, K., & Sattler, B., (2010). Pesticides and health risks. Journal of
Obstetric, Gynecologic, and Neonatal Nursing. 39(1), 103–110.
Jurewicz, J., & Hanke, W., (2008). Prenatal and childhood exposure to pesticides and neu-
robehavioral development: review of epidemiological studies. International Journal
of Occupational Medicine and Environmental Health, 21, 121–32.
McCauley, A., Linda Anger, K. W., Keifer, M., Langley, R., & Robson, G. M., (2006).
Studying health outcomes in farm worker populations exposed to pesticide. Environ
Health Perspec, 114, 6–8.
Mustafa, M., D, Garg Neha, Banerjee, B. D., Sharma, T., Tyagi, V., Ahmad, D. S., Guleria,
K., Rafat, S., Ahmad, Vaid Neelam, & Tripathi, A. K., (2015). Inflammatory-medi-
ated pathway in association with organochlorine pesticides levels in the etiology of
idiopathic preterm birth, Reproductive Toxicology, 15, 345–49.
Sharma, T., Banerjee, B. D., Mazumdar, D., Tyagi, V., Thakur, G., Guleria, K., Rafat, S.,
Ahmed, & Tripathi, A. K., (2015). Association of organochlorine pesticides and risk
of epithelial ovarian cancer: A case control study, Journal of Reproductive Health
and Medicine, 65, 1–7.
Sheiner, E. K., Sheiner, E., Hammel, R., D, Potashnik, G., &Carel, R., (2003). Effect of
occupational exposures on male fertility: literature review. Ind Health, 41(2), 55–62.
Tyagi, V., Garg, N., Mustafa, M. D., & Banerjee, B. D., Guleria, K., (2015). Organochlo-
rine pesticide levels in maternal blood and placental tissue with reference to preterm
birth: a recent trend in North Indian population, Environ Monit Assess, 187, 471–479.
Winchester, P. D., Huskins, J., & Ying, J., (2009). Agrichemicals in surface water and birth
defects in the United States. Acta Paediatr, 98, 664–669.
CHAPTER 16

TECHNOLOGY FOR DIAGNOSIS OF

PLANT VIRUSES IN HORTICULTURAL
CROPS IN INDIA
BIKASH MANDAL
Advanced Centre of Plant Virology, Division of Plant Pathology,
Indian Agricultural Research Institute, New Delhi–110012, India,
E-mail: [email protected]

CONTENTS

Abstract.................................................................................................. 229
16.1 Introduction................................................................................. 230
16.2 Immunodiagnosis of Plant Virus................................................. 230
16.3 Nucleodiagnosis of Plant Viruses .............................................. 232
16.4 Concluding Remarks................................................................... 233
Keywords............................................................................................... 233
References.............................................................................................. 233

ABSTRACT

Plant viruses are significant constraints in horticultural crops in India.

Vegetables, fruits, and ornamental crops are known to be affected by sev-
eral viruses in India. Identification of viruses based on visual observation
of symptoms alone is not reliable. Molecular diagnostic kits based on the
immunological and nucleic acids properties of viral protein and genome
are commercially available for the detection of plant viruses in Europe and
230 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

the USA. In India, the plant virus diagnostic kits are generally imported,
and they are highly expensive. Of late, studies conducted in many Plant
Virology laboratories in India on the viral genomics have facilitated devel-
oping molecular assays for several plant viruses occurring in horticultural
crops in India. Engineered antigens and antibodies and the key reagents for
immunodiagnosis of viruses have prepared, and diagnostic kit prototypes
have been developed for several viruses infecting horticultural crops. For
rapid and on-site detection, lateral flow assay (LFA) was developed for the
selected viruses. Single, duplex, and multiplex polymerase chain reaction
(PCR) methods have been developed for viruses affecting many important
vegetables. The diagnostic reagents and procedures developed are suitable
for up-scaling and commercial applications.

16.1 INTRODUCTION

Diagnosis of plant virus is a prerequisite for developing strategies for their

management, ensuring biosecurity from the introduction of exotic viruses
in a country, and providing credible phytosanitary certificate. Further, virus
diagnostic methods are necessary for the identification and classification
of viruses, generating virus-free planting materials, breeding for selective
virus resistance, monitoring spread of viruses, and studying infection pro-
cess. The diagnostic methods are broadly based on the properties of viral
protein and nucleic acids, which are the essential constituents of viruses.
There are several techniques available for the detection of plant viruses,
among which enzyme-linked immunosorbent assay (ELISA)- and poly-
merase chain reaction (PCR)-based diagnoses are commonly utilized.

16.2 IMMUNODIAGNOSIS OF PLANT VIRUS

Immunological assay is the widely used method for plant virus diagnosis,
which requires virus-specific antibody, the key reagent of diagnosis. Of
the several immunoassays are available, ELISA is the most popularly used
method for the detection of plant viruses. The immunoassays or nucleic
acid-based assays are laboratory-based techniques that require technical
manpower and expensive reagents and equipments. Currently lateral flow
Technology for Diagnosis of Plant Viruses 231

assay (LFA) or dip-stick assay has gained popularity in plant virus detec-
tion. Anyone without technical expertise can detect a specific virus within
10–15 min by using LFA.

16.2.1 RECOMBINANT ANTIGEN

To prepare antibody, traditionally plant virus is purified from the plant

tissues and used as an antigen to immunize animals. Purification of ade-
quate quantity of virus from plant tissues is tedious, and often the purified
preparation contains plant proteins and other impurities, which influence
the quality of antibody. Further, many plant viruses multiply in low rate,
and some are difficult to propagate in suitable hosts. Therefore, adequate
and renewable supply of high quality plant virus antigen is the major
limitation in generating antibody to the specific virus. Both polyclonal
and monoclonal antibodies are widely used for immunodiagnosis of plant
viruses, and these are prepared traditionally through cumbersome meth-
odology. In the alternative approach, instead of culturing viruses on plant
and purifying from the plant tissues, the full-length or partial coat protein
(CP) gene of virus is cloned and overexpressed as a recombinant protein
in bacteria (Escherichia coli). The expressed viral protein is purified from
bacterial cells and used as a recombinant antigen for raising antibody. The
procedure of generation of recombinant antigen is suitable for up-scaling,
and any quantity of antibody can be raised as and when is required.
The bacterial expressed recombinant antigen has been used for raising
antibody against several individual plant viruses at the Advanced Center of
Plant Virology (ACPV), Indian Agricultural Research Institute (IARI), New
Delhi (Jain et al., 2005; Rani et al., 2010; ; Vijayanandraj et al., 2013; Pha-
neendra et al., 2014; Soumya et al., 2014 ). We have further demonstrated
that the portion of CP of plant viruses from diverse genera can be combined
into a fusion construct, and cocktail antibody can be generated, which can be
used to detect mixed infections in plants (Mandal et al., 2011; Kapoor et al.,
2013a,b). The dual (cucumber mosaic virus [CMV] (genus Cucumovirus) +
papaya ringspot virus [PRSV] (genus Potyvirus) and Potato virus X + potato
virus Y) and triple (CMV + PRSV + groundnut bud necrosis virus [GBNV]
(genus Tospovirus) fusion proteins were successfully overexpressed in E.
232 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

coli. The cocktail antibodies raised from these fusion proteins were shown
to detect the target viruses (Kapoor et al., 2013a,b).

16.2.2 RECOMBINANT ANTIBODY

Polyclonal antibody (PAb) and monoclonal antibody (MAb) are the key
reagents for immunodiagnosis. Traditionally, both types of diagnostic anti-
bodies are produced in an animal system, which requires elaborate arrange-
ment and precaution of raring experimental animals that often invites ethical
issues. Traditionally, PAb is produced by immunizing live animals with the
viral antigen, whereas MAb is produced in an animal tissue culture. Immu-
noglobulin G (IgG) is a major antibody in the serum, which is composed
of heavy chain (H) and light chain (L) peptides. The N termini of both the
peptides are highly variable (V). The terminal regions contain paratopes that
recognize the epitopes of an antigen. The VH and VL peptides can be directly
overexpressed as recombinant antibody fragments (Fab, Fv, and scFv) in
different hosts like bacteria, insect, yeast, plant, and mammalian cells. At the
Advanced Center of Virology, IARI, New Delhi, we have generated recom-
binant antibodies to GBNV and PRSV (Yogita et al., 2015a,b). The VH and
VL of 351 and 360 nucleotides, respectively, of PRSV were expressed indi-
vidually as ~14 kDa proteins in E. coli. Both the antibody fragments indi-
vidually or together detected PRSV efficiently in the crude sap (Maheshwari
et al., 2015a). In another study, we combined 372 nucleotides of VH and 363
nucleotides of VL of GBNV into a single chain variable fragment (scFv).
The E. coli cell-expressed recombinant ScFV antibody to GBNV detected
the virus in the field samples of cowpea, groundnut, mungbean and tomato
and differentiated watermelon bud necrosis virus, a close relative of GBNV
in the serogroup-IV tospovirus (Maheshwari et al., 2015b).

16.3 NUCLEODIAGNOSIS OF PLANT VIRUSES

The partial or complete genome sequence information of alexivirus, bad-

navirus, begomovirus, carlavirus, cucumovirus, closterovirus, ilarvirus,
macluravirus, mandrivirus, potexvirus, potyvirus, tobamovirus, and tospo-
virus has been generated at ACPV, IARI, New Delhi. The sequence infor-
Technology for Diagnosis of Plant Viruses 233

mation has been utilized for designing degenerate and specific primers for
the detection of these viruses infecting several horticultural crops. Duplex
and multiplex PCR methods were developed for important viruses affect-
ing cucurbits and potato. The multiplex reverse transcription-polymerase
chain reaction (RT-PCR) could simultaneously detect six RNA viruses
infecting potato in India.

16.4 CONCLUDING REMARKS

The approaches to prepare the antigen and antibody have evolved with
the application of molecular techniques, and recombinant antigen and
antibodies are increasingly used in immunodiagnosis of plant viruses. At
ACPV, IARI, New Delhi, the antigen and antibody engineering approach
has been utilized for generating the key reagents for immunodiagnosis
of plant viruses (Mandal and Jain, 2010, 2012). The technology for virus
diagnostic kits based on ELISA and PCR have been developed and vali-
dated, which can be exploited for commercial applications.

KEYWORDS

•• diagnosis
•• horticulture
•• immunological
•• plant virus
•• recombinant

REFERENCES

Jain, R. K., Pande, A., & Mandal, B., (2005). Immonodiagnosis of groundnut and water-
melon bud necrosis virus using polyclonal antiserum to recombinant nucleocapsid
protein of Groundnut bud necrosis virus. J., Virol. Methods, 130, 162–164.
Kapoor, R., Mandal, B., Paul, B. K., & Jain, R. K., (2013b). Simultaneous detection of
potato viruses Y and X by DAC-ELISA using polyclonal antibodies raised against
234 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

fused coat proteins expressed in Escherichia coli. Journal of Plant Biochemistry and
Biotechnology, 10.1007/s13562-013-0251-5.
Kapoor, R., Mandal, B., Paul, P. K., Phaneendra, C., & Jain, R. K., (2013a). Production of
cocktail of polyclonal antibodies using bacterial expressed recombinant protein for
multiple virus detection. Journal of Virological Methods, 196, 7–14.
Maheshwari, Y., Verma, H. N., Jain, R. K., & Mandal, B., (2015a). Engineered antibody
fragments for immunodiagnosis of papaya ringspot virus. Molecular Biotechnology,
57, 644–652. doi: 10.1007/s12033-015-9854-5.
Maheshwari, Y., Vijayanandraj, S., Jain, R. K., & Mandal, B., (2015b). Engineered single-
chain variable fragment antibody for immunodiagnosis of groundnut bud necrosis
virus infection. Arch Virol. D., OI 10.1007/s00705-015-2345-y.
Mandal, B., & Jain, R. K., (2010). ELISA kit for tospoviruses detection. ICAR News, 16(3),
13.
Mandal, B., & Jain, R. K., (2011). Molecular Diagnosis of Plant Viruses: production of
cocktail antibody to CMV and PRS. VNAIP Annual Report, pp.74–75.
Mandal, B., & Jain, R. K., (2012). Diagnostic kits for of some chronic and emerging plant
viruses in India. Virus Research News, 1(1), 2–3.
Mandal, B., Kumar, A., Rani, P., & Jain, R. K., (2012). Complete genome sequence, phy-
logenetic relationships and molecular diagnosis of an Indian isolate of Potato virus
X. J., Phytopathol, 160, 1–5.
Phaneendra, C., Sambasiva Rao, K. R. S., Kapoor, R., Jain, R. K., & Mandal, B., (2014).
Fusion coat protein of Pumpkin yellow vein mosaic virus with maltose binding
protein: applications in immunodiagnosis of begomoviruses. Virus Disease, 25(3),
390–393, D., OI 10.1007/s13337-013-0189-1.
Rani, P., Pant, R. P., & Jain, R. K., (2010). Serological detection of Cymbidium mosaic and
Odontoglossum ring spot viruses on orchids with polyclonal antibodies produced
against their recombinant coat proteins. J., Phytopath., 158, 542–545.
Soumya, K., Yogita, M., Prasanthi, Y., Anitha, K., Kavi-Kishor, P. B., Jain, R. K., & Man-
dal, B., (2014). Molecular characterization of Indian isolate of Peanut mottle virus
and immunodiagnosis using bacterial expressed core capsid protein. Virus Disease,
25(3), 331–337.
Vijayanandraj, S., Yogita, M., Das, A., Ghosh, A., & Mandal, B., (2013). Highly efficient
immunodiagnosis of large cardamom chirke virus using the polyclonal antiserum
against Escherichia coli. D., OI 10.1007/s13337-013-0159-7.
CHAPTER 17

ACARICIDAL ACTIVITY OF
PETROLEUM ETHER EXTRACT FROM
SEED OF CUSTARD APPLE, ANNONA
SQUAMOSA L. (ANNONACEAE)
AGAINST RED SPIDER MITE,
OLIGONYCHUS COFFEAE (NIETNER)
INFESTING TEA
BIPLAB TUDU and SIBDAS BASKEY
Regional Research Station (Hill Zone), North Bengal Agriculture
University, Kalimpong, Darjeeling, West Bengal – 734301, India,
Tel. 03552255606, E-mail: [email protected]

CONTENTS

17.1 Introduction................................................................................. 236

17.2 Objectives................................................................................... 237
17.3 Materials and Methods................................................................ 237
17.4 Results and Discussion............................................................... 242
17.5 Summary and Conclusion........................................................... 246
Acknowledgment................................................................................... 248
Keywords............................................................................................... 248
References.............................................................................................. 249
236 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

17.1 INTRODUCTION

Tea, Camellia sinensis (L) O. Kuntze, is one of the most popular bev-
erages in the world. The crop suffers from the attack of a number of
pests and pathogens, causing significant yield losses. The red spider
mite (RSM) Oligonychus coffeae (Nietner) is one of the major pests of
tea plantation, causing 5–15% crop loss in India. This mite is character-
ized by a high reproductive capacity, which leads to high population
levels in a short period of time, thereby causing important economic
damage (Das, 1959, 1960). Nymphs and adults of O. coffeae (Nietner)
normally infest the upper surface of mature tea leaves and lacerate cells,
producing minute characteristic reddish brown marks, and when sever-
ity of infestation increases, they move even to the lower surface of older
leaves and tender tea shoots. Severe infestation ultimately leads to defo-
liation (Selvasundaram and Muraleedharan, 2003). The feeding activity
of this mite on the leaves of tea induce drastic reduction in the level of
the major photosynthetic pigment chlorophyll. Chlorophyll is an essen-
tial element of photosynthesis, and its content in plant leaves indicates
their photosynthetic capacity (Jayakrishnan and Ramani, 2015). The
modern usages of various synthetic chemicals for the control of pests
have led to various environmental concerns. Botanical insecticides have
long been touted as an attractive alternative strategy for pest manage-
ment, because botanicals reputedly pose little threat to the environment
or to human health. Annonaceae is the largest plant family in the order
Magnoliales (Westra and Maas, 2012) and comprises around 2,500 spe-
cies and 130 genera (Pirie et al., 2005). Except for two related North
American genera (Asimia and Deeringothamnus), the family is entirely
tropical (Thomas and Doyle, 1996). Among terrestrial plant families,
Annonaceae has drawn considerable attention since the 1980s, owing
to the presence of acetonins, a class of natural products with broad-
spectrum insecticidal bioactivities. Crude extract from the plant spe-
cies of Annonaceae have been extensively studied in recent years for
bioactivity to insect pests and related arthropods worldwide. Asimina
triloba, Annona muricate, and Annona squamosa are the species that
have been most frequently tested for their insecticidal activities (Isman
and Seffrin, 2014; Ocampo and Ocampo, 2006). The acetogenins are
Acaricidal Activity of Petroleum Ether Extract from Seed 237

found in the leaves and branches and, predominantly, in the seeds of

annonaceous plants. In this context, the present study was undertaken at
Bidhan Chandra Krishi Viswavidyalaya (State Agriculture University),
Mohanpur, Nadia, West Bengal, India, during 2010–2011 to evaluate
acaricidal properties of petroleum ether extracts from the custard apple
seed (CASE) Annona squamosa L. (Annonaceae) on red spider mite
infesting tea plantations.

17.2 OBJECTIVES

1. To study the acaricidal activity of petroleum ether extract from cus-

tard apple seed on tea red spider mites.
2. To study the persistent toxicity of petroleum ether extract from cus-
tard apple seed on tea red spider mites.
3. To study the phytotoxicity on tea bushes, if any.

17.3 MATERIALS AND METHODS

17.3.1 COLLECTION AND EXTRACTION OF PLANT

MATERIAL

The mature fruits of custard apple were collected from Bankura district
under Red and Lateritic Zone of West Bengal. The seed materials were
separated from fruits, dried in shade, and made into powder by using an
electric grinder or blender. The seed extracts were prepared by the Soxhlet
extraction method using petroleum ether as the solvent at its boiling range
of 60 ˚C to 80 ˚C temperature. The powdered seed material and solvent
were taken for the extraction, keeping the ratio of 1:5 (v/v). After 8 hours
of extraction to obtain the desirable alkaloids present, if any in the seed
material. The extract was filtered using a Whatman No. 1 filter paper.
Later, they were evaporated to obtain concentrated slurry and kept in the
refrigerator as stock solution. Further dilution was done with the distilled
water to get the desired doses/concentrations during the spraying (Dwivedi
and Venugopalan, 2001).
238 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

17.3.2 TEST SPECIES USED

The tea red spider mite O. coffeae Nietner (Acari: Tetrancychidae) was
obtained from the nucleus culture maintained in the Acarology Laboratory,
Department of Agricultural Entomology, Bidhan Chandra Krishi Viswavidy-
alaya (State Agriculture University), Kalyani, Nadia, West Bengal, India.

17.3.2.1 Maintenance of Mite Population

Standard modified leaf disc technique of mass rearing of mite was fol-
lowed. With the help of binocular, the last instar female mite and one
adult male were picked up with the help of camel brush and were trans-
ferred to a freshly washed tea leaf. The same step was repeated for a
number of sets. The leaves were placed with the bottom side up on the
wet cotton sponge in which the petiole was also cushioned. One of the
healthy sets was selected and was successfully proliferated for conduct-
ing experiments.

17.3.2.2 Stage of the Specimens Used

Egg, one-day old larva, and adult stages of tea red spider mites were used
for the experiment.

17.3.3 TEST CONDITION

The test was conducted in laboratory at 25–28°C temperature and 55–80%

relative humidity (RH).

17.3.4 TREATMENT DOSAGE FOR BIOASSAY

A series of concentrations ranging from 0.1% to 0.5% of botanical extract

with three replications were evaluated separately against egg, larval, and
adult stages of the tea red spider mite and an untreated control (water
spray) as a check to obtain a concentration Probit mortality curve. All
Acaricidal Activity of Petroleum Ether Extract from Seed 239

purpose suspension agent (APSA) at the rate of 0.33 mL/L of water was
added during treatment.

17.3.4.1 Bioassay Method

Fresh tea leaves of 2 cm × 2 cm area were cut into pieces. Three replica-
tions were taken for each treatment, and every replication was composed
of five leaf discs. For different experiments, in disc, 20 numbers of adult,
nymph, and egg stages were treated separately. For eggs, firstly adults
were released and given 24 hours to lay sufficient eggs. On the next day,
the adults were removed, and spraying was done on eggs. In all the repli-
cations, Petri dishes were turn upside down, where absorbent cotton pieces
were placed and soaked in water. The leaf discs were placed facing the bot-
tom of the disc on the surface of water soaked cotton with dorsal surface
of the leaf disc facing upward and sprayed with the desired concentration
of the botanical solution by using a glass atomizer at a distance of 1.5 ft.

17.3.4.2 Post Treatment Observation

Observation with respect to adult and larval stages was recorded at the 3rd,
5th, and 7th days after treatment (DAT) and in case of egg stages at the 8th
DAT. Moribund mites (adult and larvae) and eggs turned black, collapsed,
or shriveled were considered as dead.

17.3.5 TREATMENT DOSE FOR THE STUDY OF PERSISTENT

TOXICITY

For the study of persistent toxicity, tea bushes were sprayed with 0.58%
concentration of botanical extract with a high volume sprayer @ 400 l
spray fluids/ha.

17.3.5.1 Site of Experiment

The field experiments were conducted at the eco-friendly tea garden,

Gayeshpur, Nadia, West Bengal, and the laboratory experiments were car-
240 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

ried out at the Acarology Laboratory, Bidhan Chandra Krishi Viswavidya-

laya (State Agriculture University), Kalyani, Nadia, West Bengal.

17.3.5.2 Methods of Persistent Toxicity Study

Tea bushes were sprayed separately using a high volume Knapsack sprayer,
taking care to avoid drip. Only one spraying with the high volume sprayer
@ 400 L/ha was done as high volume spraying. The leaves were collected
from the treated plants at the 0, 1st, 3rd, 5th, 7th, and 10th DAT starting from
2 hours from treatment and brought into the laboratory.
Three replications were taken for each treatment, and every replication
comprised five leaf discs. For different treatments, in each dish, 20 num-
bers of adult and larval stages of mites were treated separately.

17.3.5.3 Post Treatment Observation

The mortality count was taken after the 7th DAT for adult and larval stage
of mites. In case of egg stage, if the egg was laid, the reading was taken
on the 8th DAT.

17.3.6 EXPERIMENT ON PHYTOTOXICITY

17.3.6.1 Site of Experiment

The phytotoxicity study was conducted at the eco-friendly tea garden,

Gayeshpur, Nadia, West Bengal.

17.3.6.2 Treatment Dosages for Study of Phytotoxicity

The phytotoxicity study of the botanicals was carried out at the recom-
mended dose and two and three times higher than the recommended
dose of botanical extract, i.e., 0.58%, 1.16%, and 1.74% with three rep-
lications.
Acaricidal Activity of Petroleum Ether Extract from Seed 241

17.3.6.3 Methods of Phytotoxicity Study

The total quantity of each botanical extract for particular treatment was
measured for spraying, and plot size was then marked. The calculated
amount of botanical extract for each replicated plot was diluted with water,
and they were sprayed separately with the help of the high volume Knap-
sack sprayer, taking care to avoid drip. Only one spraying with the Knap-
sack sprayer @ 400 L/ha was done as high volume spraying.

17.3.6.4 Posttreatment Observation

Phytotoxicity symptoms were recorded continuously for the first 15 days

by the observing plots in each treated plot for any adverse effect on plants,
like leaf chlorosis, leaf tip burning, leaf necrosis, leaf epinasty, leaf hypo-
nasty, vein clearing, wilting and resetting, etc., according to Central Insec-
ticides Board, Government of India guidelines. Phytotoxicity Scale: (No
phytotoxicity = 0 scale; 1% to 10% phytotoxicity = 1 scale, 11 to 20%
phytotoxicity = 2 scale ……….91 to 100% phytotoxicity = 10 scale).

17.3.7 Statistical Methods Utilized

The data obtained in percentage were first subjected to angular transforma-

tion (Fisher and Yate, 1938). The percent mortality observed in different
treatments was corrected using modified Abbott’s formula (Abbott, 1925).

P (%) =
( P / −C ) ×100
(100 − C )
where P = percentage corrected mortality, Pl = percentage observed mortality;
C = percentage control mortality.
To calculate the LC50 values, data obtained on mortality were subjected
to Probit analysis (Finney, 1971). Relative toxicity of the botanicals was cal-
culated on the basis of LC50 value of one of the higher LC50 values as unity.
For persistent toxicity, the rate of mortality (percent) of the red spi-
der mites to different doses at various time intervals after treatment was
242 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

recorded. The average residual toxicity, index of persistent toxicity, and

the order of relative efficacy were calculated by the criterion elaborated by
Pradhan and Venkataraman (1962). To calculate the PT50 values, the data
were subjected to Probit analysis as in case of LC50 values.

17.4 RESULTS AND DISCUSSION

17.4.1 ACARICIDAL ACTIVITY

It is evident from studies that the LC50 value of CASE was the lowest, i.e.,
0.10% on one-day old larval stage and highest against the egg stage with
LC50 value of 0.29% (e.g., see Table 17.1). This indicates that CASE was
most toxic to the one-day old larval stage followed by adult and egg stage
of RSM. The LC50 values for crude extracts from the leaves of A. senega-
lensis and A. squamosa were 0.67 and 0.64 µg/mL, respectively, for Culex
quinquefaciatus (Magadula et al., 2009). Begum et al. (2010) investigated
the toxic effects of ethanol extracts of seeds of A. squamosa and Calot-
ropis procera (Asclepiadaceae) against different developmental stages of
the housefly Musca domestica L. (Diptera: Muscidae). LC50 values for the
extracts of C. procera and A. squamosa seeds were 870 and 345 mg/L,
respectively. An ethanolic extract of A. squamosa leaves showed potent
activity against the rice weevil S. oryzae. The extract produced significant
knockdown (KDT50) at 1% (23.1 min) and 5% w/v (11.4 min). Complete
mortality was achieved at 39.6 ± 1.4 and 14.5 ± 1.1 min for 1% and 5%
w/v, respectively (Kumar et al., 2010). Kamaraj et al. (2011) assessed the
larvicidal activities of hexane, chloroform, ethyl acetate, acetone, and
methanol dried leaf and bark extracts of A. squamosa, Chrysanthemum
indicum, and Tridax procumbens against 4th instar larvae of the malaria

TABLE 17.1 Acaricidal Activity of CASE on Tea Red Spider Mites after 72 h of
Treatment
Stage of mites d.f. χ2 Regression equation Fiducial LCSO(%)
limit
Egg 7 21.78 Y=55+1.02X –0.54 ± 0.04 0.29
Larva 7 1.03 Y=6.40+1.38X –1.02 ± 0.04 0.10
Adult 7 2.83 Y=5.59+0.64X –0.92 ± 0.04 0.12
Acaricidal Activity of Petroleum Ether Extract from Seed 243

vector, Anopheles subpictus Grassi, and the Japanese encephalitis vec-

tor Culex tritaeniorhynchus Giles (Diptera: Culicidae). All plant extracts
showed moderate effects after 24 h of exposure; however, the most toxic
were the methanolic bark extract of A. squamosa, leaf ethyl acetate extract
of C. indicum, and leaf acetone extract of T. procumbens against the lar-
vae of A. subpictus (LC 50 = 93.80, 39.98, and 51.57 mg/L, respectively)
and methanolic bark extract of A. squamosa, leaf methanol extract of C.
indicum, and leaf ethyl acetate extract of T. procumbens against the larvae
of Cx. tritaeniorhynchus (LC50 = 104.94, 42.29, and 69.16 mg/L, respec-
tively). Attia et al. (2011) reported that garlic juice at a concentration of
7.49% showed LD50 value against Tetranychus urticae Koch. Grzybowski
et al. (2013) tested crude ethanolic extracts of A. muricata L. seeds and
Piper nigrum L. fruits against A. aegypti larvae. The LC50 value for A.
muricata was 93.48 μg/mL and for P. nigrum 1.84 μg/mL. The index of
persistent toxicity (PT) of CASE when applied @ 0.58% on the egg stage
was 230.19, followed by one-day old larva and adult at 99.34 and 90.27,
respectively. Further, the average residual toxicity (T), i.e., 39.04, was
highest in the egg stage than in other stages and persisted for period (P) of
5 days (e.g., see Table 17.2). PT50 values of this botanical extract on egg,
one-day old larva, and adults of tea red spider mite were 1.01, 0.27, and
0.22 day, respectively (e.g., see Table 17.3).

17.4.2 BIOEFFICACY OF CASE ON THREE DEVELOPMENTAL

STAGES OF RSM

The performance of petroleum ether fractions of custard apple seed @

0.10%, 0.15%, and 0.20% caused 80.60%, 89.14%, and 97.09% larval
mortality at the 7th DAT, respectively, and all the higher dosages, i.e.,
0.25%, 0.30%, 0.35%, 0.40%, 0.45%, and 0.50% at the 7th DAT showed
100% larval mortality. Lowest dose, i.e., 0.10% at the 3rd DAT caused
54.30% larval mortality. Eggs of Oligonychus coffeae were successfully
killed up to 77.67% at concentration of 0.50% of CASE. Results also
revealed that petroleum ether extract of custard apple seed at concentra-
tion of 0.25% at the 7th DAT recorded 84.65% mortality and at concentra-
tion of 0.10%, 0.15%, and 0.20% caused mortality of 77.37%, 79.13%,
and 80.52%, respectively, of adult stage of O. coffeae. At the highest dose,
244 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

TABLE 17.2 Persistent Toxicity of CASE at a Concentration of 0.58 % on Tea Red

Spider Mites
Stage of % mortality at various days interval (days) P T PT
mites 0.083 1 3 5 7 10
Egg 91.64 83.64 5.64 3.23 N/A N/A 5 46.04 230.19
Larva 84.61 11.47 3.26 N/A N/A N/A 3 33.11 99.34
Adult 78.86 9.14 2.26 N/A N/A N/A 3 30.09 90.27
P = Period in days; T = Average residual toxicity; PT = Index of persistent toxicity.

TABLE 17.3 PT50 Values of CASE on Tea Red Spider Mites

Stage of mites d.f. χ2 Regression equa- Fiducial limit PT50
tion (days)
Egg 4 128.63 Y=5.01–2.23X 0.01±0.03 1.01
Larva 4 0.99 Y=3.88–1.97X –0.57±0.03 0.27
Adult 4 0.37 Y=3.72–1.92X –0.67±0.03 0.22

i.e., 0.5%, 93.47% adult mortality was observed at the 7th DAT (e.g., see
Table 17.4). Similarly, Pavela (2009) reported 100% mortality of Tetrany-
chus urticae Koch by spraying pongam oil at 1% and 3% concentration.
Lin et al. (2009) tested the cold pressed oil from the seeds of A. squamosa.
The oil was effective in controlling the Kanzawa spider mite Tetraanychus
kanzawai Kishida (Acari: Tetranychidae) on soybean leaves. The high
level of mortality of head louse, Pediculus humanus capitis, with petro-
leum ether extract from Indian neem, A. indica, and a seed of A. squamosa
at concentration of 0.1%, 1%, and 10% have been reported by Kosalge
and Fursule (2009). The acetone, chloroform, hexane, petroleum ether,
and ethanol extracts of A. squamosa foliage were studied against the early
4th instar larvae of A. aegypti, Anopheles stephensi, and Culex quinque-
fasciatus. Larval mortality was observed after 24 h exposure. All extracts
showed moderate larvicidal effects; however, the greatest larval mortality
was obtained with a petroleum ether extract (Kumar et al., 2011). Gonza-
lez Esquinca et al. (2012) used water and ethanolic extracts to determine
the activity of stem and leaf extracts of A. muricata L., A. diversifolia
Saff., and A. lutescens Saff. against larvae of Anastrepha ludens (Mexican
fruit fly). Extracts of the three Annona species showed time-dependent
larvicidal activity against A. ludens, with variable mortality rates at 72 h
Acaricidal Activity of Petroleum Ether Extract from Seed 245

TABLE 17.4 Effect of CASE on Tea Red Spider Mites (Mean of Two Applications and
Three Replications)
Sl. Concen- % mortality of mites at various days after treatment
No. tration Larval Adult Egg
3rd 5th 7th 3rd 5th 7th 8th
DAT DAT DAT DAT DAT DAT DAT
1. 0.1% 54.30 56.84 80.60 51.06 56.60 77.37 38.28
(47.47) (48.93) (63.87) (45.61) (48.79) (61.59) (38.22)
2. 0.15% 58.60 58.91 89.14 53.21 61.49 79.13 39.46
(49.95) (50.13) (70.76) (46.84) (51.64) (62.82) (38.91)
3. 0.20% 64.80 69.53 97.09 54.97 69.19 80.52 42.11
(53.61) (56.50) (80.25) (47.85) (56.28) (63.81) (40.46)
4. 0.25% 71.16 75.87 100.00 55.13 69.95 84.65 45.07
(57.52) (60.58) (90.00) (47.94) (56.76) (66.94) (42.17)
5. 0.30% 74.47 79.21 100.00 56.57 73.58 86.21 45.77
(59.65) (62.88) (90.00) (48.77) (59.07) (68.20) (42.57)
6. 0.35% 79.66 81.65 100.00 58.01 75.86 86.53 47.12
(63.20) (64.63) (90.00) (49.61) (60.57) (68.47) (43.35)
7. 0.40% 81.20 82.71 100.00 64.02 80.21 88.34 47.44
(64.31) (65.43) (90.00) (53.15) (63.58) (70.04) (43.53)
8. 0.45% 83.36 84.20 100.00 65.75 82.78 90.96 58.10
(65.93) (6.58) (90.00) (54.18) (65.49) (72.51) (49.66)
9. 0.50% 84.13 86.00 100.00 69.74 84.56 93.47 77.67
(66.52) (68.02) (90.00) (56.63) (66.86) (75.20) (61.80)
10. Untreated 8.29 11.97 19.31 6.56 8.33 11.91 10.07
(16.73) (20.24) (26.07) (14.83) (16.77) (20.19) (18.50)
C. D. at 5% 0.87 0.55 0.83 0.62 0.47 0.52 0.46
level
N.B.: Figure in the parentheses is angular transformed values.

of exposure as follows: A. lutescens 87–94%, A. diversifolia 70–90%, and

A. muricata 63–74%. Radhakrishnan and Prabhakaran (2014) reported
that aqueous extracts of Allamanda catharitica and Conyza bonariensis
showed 100% and 80% adult mortality, respectively, at 5% concentration
after 96 h of observation under laboratory condition. Mech et al. (2015)
found that methanolic extract from Parthenium hysterophorus leads to
highest mortality against the adults of O. coffeae followed by petroleum
ether and chloroform extracts. The LC50 value of methanol extract against
the adult mite was 0.12% (48 h).
246 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

17.4.3 PHYTOTOXIC EFFECT OF BOTANICAL EXTRACT

None of the treated concentrations, i.e., 0.58%, 1.16%, and 1.74% of

CASE showed phytotoxicity on tea leaves (e.g., see Table 17.5). Selva-
sundaram (2004) reported that Exodus, a new herbal product derived from
Sophora flavescens, was tested against the red spider mite Oligonychus
coffeae under laboratory and field conditions. Field studies conducted
from January to May 2004 in Anamallais, Tamil Nadu, India, indicated
that spraying the herbal product at 250 and 500 mL/ha reduced the number
of red spider mites. Application of this product did not result in phytotox-
icity to tea at the tested concentrations. Acaricidal and ovicidal activities
of Clerodendrum viscosum Ventenat (Verbenaceae), a common weed of
India, were investigated on tea red spider mite, Oligonychus coffeae Niet-
ner (Acarina: Tetranychidae). Different solvent extracts (water, methanol,
acetone, and petroleum ether) of C. viscosum at different concentrations
(1, 2, 4, and 8%) were used. No phytotoxic effect (score, 0–5% and grade
1) was observed in the field from tea bushes sprayed with different doses
of extracts of C. viscosum (Roy et al., 2011). Aqueous seed extract of
Melia azedarach (L.) was evaluated against the tea red spider mite O. cof-
feae (Nietner), in relation to mortality of adult mites, viability of eggs and
subsequent adult emergence and oviposition deterrence in the laboratory,
and the extract underwent field evaluation in terms of percent reduction
of the mite population. No phytotoxic effect (score 0–5% and grade 1)
was observed in the field when tea bushes were sprayed with different
doses of aqueous seed extract of M. azedarach (Roy and Mukhopadhyay,
2012).

17.5 SUMMARY AND CONCLUSION

Finally, it may be concluded that the LC50 value of CASE was the low-
est (0.10%) on one-day old larval stage and highest against the egg stage
(0.29%). This indicates that CASE was most toxic to the one-day old lar-
val stage, followed by adult and egg stage of RSM. The index of persistent
toxicity (PT) of CASE when applied @ 0.58% on egg stage was 230.19,
followed by one-day old larva and adult at 99.34 and 90.27, respectively.
TABLE 17.5 Evaluation of Different Treatment Schedules of CASE for Phytotoxicity on Tea Bushes during March–April 2010 at
Gayeshpur Tea Garden, West Bengal (Based on One Application and Ten Replications)
Sl. Treat- Dose Visual rating (phytotoxicity) in 0 – 10 scale of grading
No. ment (%) 0 1 2 3 4 5 6 7 8 9 10
0.0% 1– 11– 21– 31– 41– 51– 61– 71– 81– 91–
10% 20% 30% 40% 50% 60% 70% 80% 90% 100%
1. CASE 0.58 0 0 0 0 0 0 0 0 0 0 0
2. CASE 1.16 0 0 0 0 0 0 0 0 0 0 0
3. CASE 1.74 0 0 0 0 0 0 0 0 0 0 0
4. Untreated N/A 0 0 0 0 0 0 0 0 0 0 0
Acaricidal Activity of Petroleum Ether Extract from Seed

N.B.: Observations were based on 1, 3, 7 and 15 days after spraying on necrosis, epinasty, hyponasty, leaf tip injury, leaf surface injury, wilting, vein clearing
etc.
247
248 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Further, the average residual toxicity (T), i.e., 39.04, was highest in the
egg stage than in other stages and persisted for period (P) of 5 days. PT50
values of this botanical extract on egg, one-day old larva and adults of tea
red spider mite were 1.01, 0.27, and 0.22 day, respectively. The present
investigation revealed that petroleum ether extract from the custard apple
seed Annona squamosa L. (Annonaceae) has good acaricidal activity. The
results of this study indicate the plant-based compounds may be an effec-
tive alternative to conventional acaricides for the management of RSM.
Plant allelochemicals may be quite useful in increasing the efficacy of
biological control agents because plant produces a large variety of com-
pounds that increase their resistance to insect attack (Senthil Nathan et
al., 2005). Hence, by using extracts of custard apple seed in their field,
tea planters may reduce the incidence of RSM in tea plantations and may
find scope in integrated pest management system of Oligonychus coffeae
(Nietner). However, further investigation is necessary for optimization of
the bioactive compound.

ACKNOWLEDGMENT

We are in great pleasure to express our deepest sense of gratitude to late

Dr. A. K. Somchoudhury, Professor and former Dean, Post Graduate Stud-
ies, Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, West Bengal, for
his learned and affectionate guidance, helpful criticism, and encourage-
ment during the entire course of investigation.

KEYWORDS

•• acaricidal
•• bioassay
•• custard apple
•• red spider mite
•• tea
•• toxicity
Acaricidal Activity of Petroleum Ether Extract from Seed 249

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Das, G. M., (1960). Occurrence of red spider Oligonychus coffeae (Nietner) on tea in North
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Dwivedi, S. C., & Venugopalan, S., (2001). Evaluation of leaf extracts for their ovicidal
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Breceda, S. F., & Gerardo, P. M., (2012). In vitro larvicidal evaluation of Annona
muricata L. A., diversifolia Saff. and A., lutescens Saff. extracts against Anastrepha
ludens larvae (Diptera, Tephritidae). Interciencia, 37, pp. 264–289.
Grzybowski, A., Tiboni, M., Silva, M. A., Chitolina, R. F., Passos, M., & Fontana, J. D.,
(2013). Synergistic larvicidal effect and morphological alterations induced by etha-
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Isman, M. B., & Seffrin, R., (2014). Natural insecticides from Annonaceae: a unique
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Jayakrishnan, T. V., & Ramani, N., (2015). Reduction of major photosynthetc pigments
induced by oligonychus coffeae (Nietner) (Acari: Tetranychidae) infesting camellia
sinensis (L) Kuntze, O. International Journal of Recent Scientific Research., 6(5),
pp. 3947–3950.
Kamaraj, C., Bagavan, A., Elango, G., Zahir, A. A., Rajakumar, G., Marimuthu, S., San-
thoshkumar, T., & Rahuman, A. A., (2011). Larvicidal activity of medicinal plant
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CHAPTER 18

EVALUATION OF SOME PLANT

EXTRACTS AGAINST EARLY BLIGHT
OF TOMATO UNDER NET HOUSE
CONDITION
MANISHA DUBEY,1 T. S. THIND,1 S. K. JINDAL,2 and R. K. DUBEY3
1
Department of Plant Pathology, 2Department of Vegetable
Science and 3Department of Floriculture and Landscape,
Punjab Agricultural University, Ludhiana–141004, India,
E-mail: [email protected]

CONTENTS

18.1 Materials and Methods................................................................ 253

18.2 Results and Discussion............................................................... 256
18.3 Effect of Plant Extracts on Early Blight in a Net House............. 256
Acknowledgment................................................................................... 258
Keywords............................................................................................... 258
References.............................................................................................. 258

The antifungal effect of extracts of three plants, viz., Allium sativum, Euca-
lyptus chamadulonsis, and Azadirachta indica against Alternaria solani,
the cause of early blight of tomato, was evaluated under in vitro and field
conditions. Effect of plant extracts and the chemical fungicide Indofil
M-45 (as standard check) at various concentrations on mycelial growth of
A. solani was determined by the poisoned food method. The leaf extract
of E. chamadulonsis and A. indica at 20% concentration caused highest
reduction of mycelia growth of A. solani (73.3% and 63.2%, respectively),
252 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

while A. sativum at 20% concentration caused the lowest inhibition of

fungal growth. In net house experiments, the highest reduction of disease
severity was achieved by the fungicide (Indofil M-45) at 79.3%, followed
by extracts of E. chamadulonsis and A. sativum at 20% concentration.
Fungicide and plant extracts increased the fruit yield by 80.0%, 71.4%,
66.7%, and 60.0% compared to untreated control. All treatments of plant
extracts and fungicide (Indofil M-45) significantly reduced the early blight
disease as well as increased the yield of tomato under field condition.
Tomato (Solanum lycopersicon L.) is one of the most important “pro-
tective food” because of its special nutritive value and its wide production.
Among the fungal diseases affecting tomato crop, early blight caused by
Alternaria solani (Ell. & Martin) causes considerable yield losses. Most
tomato cultivars are susceptible to this disease and depending upon age
of the plant, season, and environmental factors at the time of infection,
and yield losses range between 50% and 86% (Mathur and Shekhawat,
1986). The fungus Alternaria can cause the disease on all parts of the plant
(leaf blight, stem lesions, and fruit lesion at the pedicel end) that results in
severe damage during all stages of plant development (Abada et al., 2008).
Control of early blight disease has been accomplished primarily by the
application of chemical fungicides (Jones et al., 1991). Several effective
fungicides have been recommended for use against A. solani, but these are
not considered to be long-term solutions due to concerns of expense, expo-
sure risks, fungicide residues and other health and environmental hazards.
In an attempt to ameliorate this condition, some alternative methods of
control have been adopted. Natural products isolated from various plants
appear to carry minimal environmental impact and danger to consumers in
contrast to synthetic pesticides (Varma and Dubey, 1999).
The use of plant extracts has been shown to be ecofriendly and effec-
tive against many plant pathogens (Bowers and Lock, 2004; Saadabi,
2006). A number of plant species have been reported to possess natural
substances that are effective against several plant pathogenic fungi (Gous-
sous et al., 2010). Sallam (2011) evaluated the effect of different plant
extracts on mycelial growth of A. solani and found that leaf extracts of
some plants, i.e., Tamarix aphylla and Salsola bayosma, totally inhibited
the growth of the pathogen. Obagwu (1997)9 reported that garlic extracts
significantly reduced the early blight disease of tomato. The present study
Evaluation of Some Plant Extracts Against Early Blight 253

was conducted to determine the efficacy of leaf extracts of E. chamadulon-

sis and A. indica and bulb extract of Allium sativum for the control of early
blight of tomato under net house and field conditions. The treatments were
compared with the commonly used fungicide Indofil M-45.

18.1 MATERIALS AND METHODS

The causal fungus was isolated from naturally infected tomato leaves and
fruits showing blight symptoms. Pathogenicity tests of Alternaria solani
isolates were carried on tomato plants grown under net house of the Veg-
etable Department, PAU, Ludhiana. The inoculum was prepared by scrap-
ing A. solani colony surface in sterilized water. The resulting conidial
and mycelia fragment suspension was adjusted to 5 × 106 cfu/mL using a
hemocytometer and used for inoculation of 15 tomato plants in pots in net
house. After inoculation, the net house was covered with a polythene sheet
for 48 h to maintain high humidity conditions. After 48 h, the polythene
sheet was removed. The pots were arranged in a completely randomized
design under net house conditions. Disease severity was recorded 2 weeks
after inoculation. Ten plants were randomly selected and scored individu-
ally using 0–5 rating scale based on leaf area, stem and fruit covered by
blight symptoms following the rating scale described by Pandey et al.
(2003).
Percentage disease index (PDI) was calculated as follows:

Sum of all rating

PDI = ×100
Total no.of observation × Maximum ratingg growth

The mean value of the PDI from 10 individual plants was calculated
for each of the observations at 90 and 120 days after transplanting and
averaged.

18.1.1 PREPARATION OF EXTRACTS

Leaves of two plants namely Eucalyptus chamadulonsis and Azadirachta

indica and bulbs of Allium sativum were collected from the field of Punjab
254 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Agricultural University, Ludhiana, Punjab and used for preparing extracts.

Plant leaf extract was prepared according to Sallam (2011). Ten grams of
fresh leaf material of each plant species was collected, washed with water,
and crushed in a grinder by adding sterile distilled water at the rate of 10
mL/gm of plant tissue, and the homogenates were centrifuged at 10,000
× g for 15 min and the supernatant solutions were collected. The plant
extract was diluted further to obtain 10% and 20% concentration (v/v).
These fractions were sterilized using 0.22-µm Millipore filters and used
for assay of antimicrobial activity as described below.

18.1.2 IN VITRO SCREENING OF PLANT EXTRACTS

The filtrate of each plant extract was mixed with autoclaved potato dex-
trose agar (PDA) medium @ 10% and 20% concentration. The plant
extract supplemented medium was poured in sterilized Petri plates and
allowed to solidify. These Petri plates were inoculated at the center with a
5-mm agar disc obtained from the 7-days-old fungal culture of Alternaria
solani. In the control, a Petri plates containing PDA medium with the req-
uisite amount of sterilized water instead of a plant extract was inoculated
with the test pathogen. Indofil M-45 (mancozeb), the standard fungicide
recommended for control of early blight of tomato, @ 0.2% was used as
a positive control for comparison. The inoculated Petri plates were then
incubated at 25+2°C for 7 days. The diameter of the fungal colony was
measured using a meter rule along two diagonal lines drawn on the reverse
side of each petri plate 24 h after inoculation. Each treatment was repli-
cated three times with three plates per replication. Percentage inhibition of
mycelial growth was calculated, using the formula:
dc − dt
% growth inhibition = ×100 × 100
dc
where dc= average diameter of fungal colony in control plates; dt= aver-
age diameter of fungal colony in treated plates.
Three plates per each treatment were used as replications. The diameter
of the fungal colony was measured using a meter rule along two diagonal
lines drawn on the reverse side of each Petri plate 7 days after inoculation.
Each treatment was replicated three times with three plates per replication.
Evaluation of Some Plant Extracts Against Early Blight 255

18.1.3 EFFICACY OF THE TESTED PLANT EXTRACTS

AGAINST A. SOLANI IN A NET HOUSE

Plant extract treatments at 20% concentrations were applied as foliar appli-

cation on 7-weeks-old tomato plants and every 15 days up to 60 days. The
fungicide Indofil M-45 (mancozeb, 75%) @ 0.2% was applied as the stan-
dard check. Tomato plants were inoculated with 20 mL of A. solani sus-
pension containing 5 × 106 cfu/mL. The first spray was carried out as soon
as the first symptom of early blight was seen in the net house. Two weeks
after inoculation, disease severity was recorded. Ten plants were randomly
selected and scored individually using the 0–5 rating scale (Table 18.1,
Figure 18.1) based on leaf area, stem, and fruit covered by blight systems
following the rating scale described by Pandey et al. (2003).
Observations on disease severity were recorded 2 weeks after the last
application. Percent disease severity (PDI) was calculated by the standard
formula as follows

Sum of all rating

PDI = ×100
Total no.of observation × Maximum ratingg growth

Statistical analysis of the data was done using completely randomized

block design (CRD).

TABLE 18.1 In Vitro Effect of Plant Extracts on the Mycelial Growth of Alternaria solani
Treatments Concentration (%) Percent of mycelial
growth reduction (%)
Eucalyptus chamadulonsis 10 59.13
20 73.30
Allium sativum 10 53.30
20 63.00
Azadirachta indica 10 56.10
20 63.20
Indofil M-45 (mancozeb 75%) 0.2 80.00
Control (water) - 0.00
CD - 7.20
256 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 18.1 (See color insert.) In vitro effect of plant extracts on the mycelial growth
of Alternaria solani.

18.2 RESULTS AND DISCUSSION

18.2.1 EFFECT OF PLANT EXTRACTS ON MYCELIAL

GROWTH OF A. SOLANI

Three plant species and the fungicide Indofil M-45 were evaluated for the
antifungal activity against A. solani. Leaf extracts of Eucalyptus chamad-
ulonsis and Azadirachta indica and bulb extract of Allium sativum at 10%
and 20% concentration were effective in inhibiting the radial growth of A.
solani as compared to untreated control. The leaf extract of E. chamadu-
lonsis and A. indica at 20% concentration caused highest reduction of
mycelial growth of A. solani (73.3% and 63.2%, respectively). Bulb
extract of Allium sativum at 20% concentration caused the lowest inhibi-
tion of mycelial growth of the pathogen. Overall, Indofil M-45 at 2.0 g/L
caused the highest reduction of the pathogen by 80.0% (Table 18.2).

18.3 EFFECT OF PLANT EXTRACTS ON EARLY BLIGHT IN A NET

HOUSE

The protective action of the plant extracts relative to the recommended

fungicide Indofil M-45 against early blight in tomato plant that evaluated
Evaluation of Some Plant Extracts Against Early Blight 257

TABLE 18.2 Effect of Plant Extracts on Early Blight of Tomato in a Net House
Treatments Phytotoxicity Disease sever- Disease re- Fruit yield
ity (%) duction (%) (% increase)
Eucalyptus NO 19.50 68.30 71.40
chamadulonsis
(20%)
Allium sativum NO 22.50 63.40 66.70
(20%)
Azadirachta indica NO 24.90 59.40 60.00
(20%)
Indofil M-45 NO 12.70 79.30 80.00
(0.2%)
Control (water) - 61.40 0.00 -
CD - - 5.20
NO = Not observed.

under net house conditions is shown in Table 18.2. The results showed that
Indofil M-45 was the most effective treatment against A. solani followed
by Eucalyptus chamadulonsis, Allium sativum and A. indica, respectively,
at 20% concentration. There was no phytotoxicity of the tested plant
extracts observed on tomato.
The plant extracts of E. chamadulonsis, A. sativum, and A. indica
caused significant reduction in the linear growth of A. solani. This reduc-
tion was gradually increased by increasing concentration of extracts in the
growth medium. Indofil M-45 was more effective than the plant extracts.
Similar effects of various other plant products effective against Alternaria
spp have been reported by several workers (Dushyant and Bohra, 1997;
Bowers and Lock, 2004; Latha et al., 2009). The bulb extracts of A. sati-
vum, leaf extract of Aegle marmelos, and flower extract of Catharanthus
roseus inhibited the spore germination and mycelia growth of A. solani
(Vijayan, 1989). Garlic and neem products have also shown some antimi-
crobial properties and have been used in the control of fungal pathogens
(Stoll, 1998). Methanolic extracts of peppermint (15%) and eucalyptus
(15%) were reported as the best in preventing the spore germination of
Alternaria sesame (Zaker, 2013).
258 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

ACKNOWLEDGMENT

The authors are thankful to the Head, Department of Vegetable Science,

for providing necessary facilities to conduct the experiment, and Depart-
ment of Science and Technology New Delhi, for sponsoring the study
under the WOS B scheme.

KEYWORDS

•• Alternaria solani
•• early blight
•• fungicide
•• plant extracts
•• tomato

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Dushyent, G., & Bohra, A., (1997). Effect of extracts of some halophytes on the growth of
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Jones, J. B., Jones, J. P., Stall, R. E., & Zitter, J. A., (1991). Infectious diseases : Diseases
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Latha, P., Anand, T., Raghupati, N., Prakasam, V., & Samiyappan, R., (2009). Antimicro-
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Evaluation of Some Plant Extracts Against Early Blight 259

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extracts. Plant Pathology Journal, 10(4), 187–191.
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CHAPTER 19

BIO-EFFICACY OF ECO-FRIENDLY
PESTICIDES ON THE MANAGEMENT
OF SPODOPTERA LITURA FAB. ON
CABBAGE
R. MANDI and A. PRAMANIK
Department of Agricultural Entomology, Bidhan Chandra Krishi
Viswavidyalaya, Mohanpur, Nadia, 741252, West Bengal, India,
E-mail: [email protected]

CONTENTS

Abstract.................................................................................................. 261
19.1 Introduction................................................................................. 262
19.2 Materials and Methods................................................................ 263
19.3 Results and Discussion............................................................... 264
Keywords............................................................................................... 276
References.............................................................................................. 276

ABSTRACT

The bio-efficacy of eco-friendly pesticides (botanicals: Azadirachtin,

10000 ppm, neem seed kernel extract (NSKE), Mittimax; microbials:
Bacillus thuringiensis 5% WP, Beauveria bassiana, Spinosad 45% SC;
and IGR (insect growth regulator): diflubenzuron + deltamethrin 22% SC)
was evaluated for use in managing cabbage head borer (Spodoptera litura
262 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Fab.) population. An experiment was conducted at the Central Research

Farm, Gayeshpur, Bidhan Chandra Krishi Viswavidyalaya, Nadia, West
Bengal, during the rabi season of 2007–2010. The experiment was carried
out in randomized block design (RBD) with three replications. Cabbage
(cv. Green express) seedlings were transplanted in the plot of 4 m × 3 m
area with 50 cm × 50 cm spacing. The counting of S. litura population
was made one day before as pretreatment and 1, 3, 7, and 10 days after
treatment. All the insecticides were superior in controlling the Spodoptera
population in comparison to untreated control. Among the different treat-
ments, diflubenzuron + deltamethrin 22% SC recorded the highest 75.81%
population reduction and proved to be the most effective treatment fol-
lowed by B. bassiana @ 0.5% and NSKE, with population reduction of
60.72% and 60.35%, respectively. The percent increase of yield over con-
trol ranged from 11.41% to 50.13% in pesticide treatments with the high-
est yield in Spinosad 45% SC as against 32.79 t/ha in untreated control.
The research indicated that all the eco-friendly pesticides were effective
against cabbage head borer.

19.1 INTRODUCTION

India is mostly an agro-based country, and agriculture is a major compo-

nent of the Indian economy; more than 75% of Indian people have their
livelihood as agriculture and agriculture-oriented works (Thenmozhi and
Thilagavathi, 2014). Among the cruciferous vegetables, Cabbage (Brassica
oleracea L. var. capitata) is the most popular and grown throughout India.
It is used as salad, boiled, and dehydrated vegetable as well as in cooked
curries and pickles. The main edible part of cabbage is head/card, i.e., leaf
is a good source of protein 1.6%; vitamins A, B1, B2, and C; sulfur; amino
acids; minerals (calcium, iron, magnesium, phosphorus, and potassium);
low amount of calories 2.4%; fat 0.2%; carbohydrate 4.8%; and substan-
tial amount of b-carotene (Hanif et al., 2006). Insect pests, diseases, and
weeds are the major constraints limiting agricultural productivity growth.
It is estimated that herbivorous insects eat about 26% of the potential food
production (Sing and Sharma, 2004). The cabbage borer Spodoptera litura
Fabricius (Noctuidae: Lepidoptera) is an economically important, regular,
multivoltine, gregarious, polyphagous pest that seriously harms cabbage,
Bio-Efficacy of Eco-Friendly Pesticides on the Management 263

cauliflower, other vegetables, soybeans, cotton, and cash crops. It is widely

distributed throughout tropical and temperate Asia, Australia, and Pacific
islands (Kranthi et al., 2002). Indiscriminate use of chemical pesticides to
manage this pest has resulted in resistance to chemical pesticides, resur-
gence, harmful effects on nontarget organisms, environmental pollution,
and human health hazards (Vastrad et al., 2003, 2004; Haseeb et al., 2004).
With the increasing concern on the health hazards, environmental pollu-
tion, and pest resistance due to over use of synthetic pesticides, the usage
of eco-friendly pesticides (botanicals, microbials, IGRs) in the pest con-
trol is gaining importance, which can minimize the use of these synthetic
chemicals. Botanical pesticides (Nathan and Kalaivani, 2005), microbial
pesticides (Asi et al., 2013), and IGRs (Deb Prasad et al., 2012) are highly
effective, safe, and ecologically acceptable. In the present investigation, the
bio-efficacy of eco-friendly pesticides was evaluated against an economi-
cally important agricultural devastating pest S. litura.

19.2 MATERIALS AND METHODS

The field experiment was conducted with cabbage var. “Green express” in
the experimental field of Central Research Farm, Bidhan Chandra Krishi
Viswavidyalaya, Gayeshpur, Nadia, West Bengal, during the rabi season
of 2007–2008, 2008–2009, and 2009–2010. The experiment was carried
out in randomized block design (RBD) with three replications and nine
treatments including untreated control. The seedlings were transplanted in
the plot of 4 m × 3 m area with 50 cm × 50 cm spacing during the last week
of December for the 3 seasons.
The pesticides evaluated were one bacterial pesticide T1= Bacillus
thuringiensis - 5% WP @ 0.2% a.i., one fungal pesticide T2= Beauvaria
bassiana – 2 × 109 spore/g @ 0.5% a.i. and T3= B. bassiana – 2 × 107
spore/g @ 1.0% a.i., two neem-based pesticides T4= azadirachtin 10,000
ppm @ 0.002% a.i. and T5= neem seed kernel extract (NSKE) @ 5% a.i.,
one IGR T6= diflubenzuron + deltamethrin 22% SC @ 0.022% a.i.; one
soil actinomycetes T7 = spinosad 45% SC @ 0.01% a.i., one organic pesti-
cide T8 = mittimax @ 0.15% a.i., and T9 = untreated control.
When the cabbage head borer population was evenly distributed, the test
pesticides were applied as foliar spray by the back pack hydraulic sprayer
264 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

(Aspee, Mumbai) with a hollow cone nozzle, with the spray fluid of 500–
600 L/ha depending on the stage of the crop growth twice at 15 days interval
except in the untreated control plots. The counting on the larval population
of S. litura was made one day before as pretreatment and 1, 3, 7, and 10
days after treatment on 10 randomly selected plants in each plot. The percent
efficacy was calculated on the basis of mortality of larvae at the above inter-
vals after treatments. Reduction percent of different pests was calculated by
applying a correction factor given by Henderson and Tilton (1995).

Percent reduction over control = 100 [1–{(Ta × Cb)/(Ta × Ca)}]

where, Ta = number of insects after treatment; Tb = number of insects

before treatment.; Ca = number of insects in untreated plot after treatment;
Cb = number of insects in untreated plot before treatment. The cabbage
card harvested from each plot was recorded and calculated as tons/ha.
The critical difference (CD) at 0.05% level of significance were worked
out from the data of percent reduction population of replication before
treatment and various days interval after treatment of two consecutive
sprays per year. The data analyzed in RBD were subjected to Duncan’s
multiple range test (DMRT) at 5% level after making necessary transfor-
mation wherever needed.

19.3 RESULTS AND DISCUSSION

Field experiment of eco-friendly pesticides on S. litura Fab. and its effect

on yield parameters was observed during three consecutive years, 2008,
2009, and 2010, and are described as follows:

19.3.1 EFFECT OF ECO-FRIENDLY PESTICIDES ON LARVAL

POPULATION OF CABBAGE HEAD BORER DURING 2008–2010

19.3.1.1 Effect of Eco-Friendly Pesticides on S. litura Infesting

Cabbage during 2008

The results achieved on the bio-efficacy of different botanical, microbial, and

chemicals against S. litura (Fab.) on cabbage during 2008 are presented in
Bio-Efficacy of Eco-Friendly Pesticides on the Management 265

Table 19.1. On the first day of first and second spraying, it was found that
the highest (67.67% and 70.50%, respectively) reduction in larval population
was recorded in NSKE @ 5.0%. The highest population reduction on the
third day after first and second spraying (91.67% and 90.50%, respectively)
was obtained in diflubenzuron + deltamethrin 22% SC @ 0.022%. After the
seventh day of first spraying, the maximum population reduction (85.17%)
was recorded in Mittimax @ 0.15%, which was significantly at par with
82.33% in diflubenzuron + deltamethrin − 22% SC @ 0.022%. On the sev-
enth day after the second treatment, maximum (85.17%) population reduc-
tion was recorded in diflubenzuron + deltamethrin 22% SC @ 0.022% and
was significantly at par with that of (85.00%) B. bassiana @ 1.0%. On the
10th day after first and second spraying, maximum larval mortality of 90.00%
and 88.67%, respectively, was recorded in B. bassiana @ 1.0%. Considering
the overall population reduction during the year 2008, the highest population
reduction of 77.46% was recorded in diflubenzuron + deltamethrin 22% SC
@ 0.022% and the lowest population reduction of 35.60% was obtained in
Mittimax @ 0.15%. Considering the overall mean population reduction dur-
ing the year 2008 the highest population reduction of 77.46% was recorded in
diflubenzuron + deltamethrin 22% SC @ 0.022% and the lowest population
reduction of 35.60% was obtained in Mittimax @ 0.15%.

19.3.1.2 Effect of Eco-Friendly Pesticides on S. litura Infesting

Cabbage during 2009

The results of 2009 are presented in Table 19.2. A more or less similar
trend was also observed during rabi season of 2009; from the overall mean
population reduction after 1st to 10th day of two spraying, highest larval
population reduction (76.63%) was recorded in diflubenzuron + deltame-
thrin 22% SC @ 0.022% and lowest population reduction (35.25%) was
obtained in Mittimax @ 0.15%.

19.3.1.3 Effect of Eco-Friendly Pesticides on S. litura Infesting

Cabbage during 2010

Filed experiment of eco-friendly pesticides on the cabbage head borer in

cabbage during 2010 is depicted in Table 19.3. On the first and third day
 266

TABLE 19.1 Effect of Ecofriendly Pesticides on Spodoptera litura (Fab.) Infesting Cabbage During 2008
Treat- Dosage Pre treat- Mean percent efficacy (% Reduction Over Control) at different days after spraying Overall
ments ment larval After 1st Spray After 2nd Spray mean of
population/ st post spray
plant 1 day 3rd day 7th day 10th day 1st day 3rd day 7th day 10th day count
T1 0.2% 4.55 27.67 54.00 65.67 82.67 35.00 49.17 66.50 81.33 57.75
bc c c b cd c c b
(31.99) (47.58) (54.45) (65.81) (36.55) (44.81) (54.95) (64.78)
T2 0.5% 4.50 30.00 56.00 63.33 84.00 42.50 61.83 74.00 80.00 61.46
bc c cd b b b b b
(33.40) (48.73) (53.08) (66.82) (40.98) (52.17) (59.75) (63.80)
T3 1.0% 4.61 17.33 40.67 71.67 90.00 20.33 39.50 85.00 88.67 56.65
e e b a e d a a
(24.97) (39.91) (58.17) (72.06) (27.09) (39.22) (67.63) (70.81)
T4 0.002% 5.78 33.67 51.00 61.00 66.33 38.67 50.83 59.33 60.33 52.65
b d d d bc c d d
(35.77) (45.86 (51.66) (54.85) (38.74) (45.77) (50.67) (51.26)
T5 5.0% 5.44 67.67 79.00 63.00 43.00 70.50 60.00 38.00 45.50 58.33
(55.74)a (63.08)b (52.83)cd (41.26)e (57.42)a (51.06)b (38.34)e (42.71)e
T6 0.022% 5.17 65.00 91.67 82.33 74.00 67.00 90.50 85.17 64.00 77.46
(54.03)a (73.80)a (65.55)a (59.70)c (55.26)a (72.55)a (67.76)a (53.44)c
Sustainable Horticulture Volume 2: Food, Health, and Nutrition
TABLE 19.1 (continued)

Treat- Dosage Pre treat- Mean percent efficacy (% Reduction Over Control) at different days after spraying Overall
ments ment larval After 1st Spray After 2nd Spray mean of
population/ st rd th th
post spray
plant 1 day 3 day 7 day 10 day 1st day 3rd day 7th day 10th day count
T7 0.01% 4.05 19.33 31.00 43.17 64.00 19.00 40.33 60.00 63.67 42.56
(26.43)de (34.14)g (41.36)e (53.43)d (26.19)e (39.72)d (51.06)d (53.24)c
T8 0.15% 5.67 23.67 37.33 85.17 24.67 33.33 39.00 19.67 22.00 35.60
(29.44)cd (37.95)f (67.76)a (30.09)f (35.57)d (38.94)d (26.66)f (28.30)f
T9 0.00 7.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
f h f g f e g
(4.05) (4.05) (4.05) (4.05) (4.05) (4.05) (4.05) (4.05)g
SEm± 1.32 0.48 0.76 0.74 0.81 0.94 0.96 0.53
CD at 3.94 1.43 2.26 2.23 2.43 2.83 2.87 1.58
0.05%
* Figures in parentheses are angular transformed values.
* Means followed by common letter are not significantly different by DMRT (p = 0.05).
Bio-Efficacy of Eco-Friendly Pesticides on the Management
267
 268

TABLE 19.2 Effect of Eco-Friendly Pesticides on Spodoptera litura (Fab.) Infesting Cabbage During 2009
Treat- Dosage Pre treat- Mean percent efficacy (% Reduction Over Control) at different days after spraying Overall
ments ment After 1st Spray After 2nd Spray mean of
larval post spray
popula- 1st day 3rd day 7th day 10th day 1st day 3rd day 7th day 10th day count
tion/plant
T1 0.2% 5.55 42.67 57.17 69.83 82.83 34.83 55.17 67.00 79.00 61.06
c c bc b d cd c b
(41.07) (49.41) (57.01) (65.93) (36.47) (48.26) (55.24) (63.08)
T2 0.5% 4.78 46.33 57.00 71.33 84.00 37.17 59.50 66.00 77.50 62.35
c c b b c bc c b
(43.18) (49.32) (57.95) (66.86) (37.86) (50.79) (54.63) (62.04)
T3 1.0% 4.56 25.00 43.67 83.33 91.33 19.50 45.00 72.00 82.00 57.73
(30.31)f (41.65)d (66.33)a (73.48)a (26.56)g (42.42)de (58.37)b (65.30)a
T4 0.002% 6.67 36.33 52.67 63.83 69.83 37.83 51.83 57.00 66.17 54.44
(37.36)d (46.81)c (53.34)cd (57.05)c (38.25)c (46.34)cd (49.31)e (54.75)cd
T5 5.0% 5.61 73.50 82.00 50.00 45.67 69.67 68.50 61.33 44.17 61.85
(59.37)a (65.28)b (45.29)e (42.80)d (56.90)a (56.54)b (51.85)d (41.94)e
T6 0.022% 4.28 66.00 90.67 82.00 70.17 67.17 88.00 80.67 68.33 76.63
(54.64)b (73.02)a (65.46)a (57.21)c (55.35)b (70.25)a (64.30)a (56.07)c
T7 0.01% 6.39 23.17 37.83 59.00 67.67 22.67 32.67 42.67 64.00 43.71
f e d c f f f d
(29.05) (38.24) (50.49) (55.66) (28.76) (35.15) (41.07) (53.43)
Sustainable Horticulture Volume 2: Food, Health, and Nutrition
TABLE 19.2 (continued)

Treat- Dosage Pre treat- Mean percent efficacy (% Reduction Over Control) at different days after spraying Overall
ments ment st nd mean of
After 1 Spray After 2 Spray
larval post spray
popula- 1st day 3rd day 7th day 10th day 1st day 3rd day 7th day 10th day count
tion/plant
T8 0.15% 5.33 29.67 40.18 18.33 24.67 28.00 38.33 81.00 21.83 35.25
(33.27)e (39.63)de (25.64)f (30.02)e (32.27)e (38.54)ef (64.55)a (28.19)f
T9 00 6.39 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
(4.05)g (4.05)f (4.05)g (4.05)f (4.05)h (4.05)g (4.05)g (4.05)g
SEm± 0.94 0.96 1.44 1.06 0.36 2.19 0.43 0.55
CD at 2.82 2.89 4.31 3.18 1.09 6.55 1.27 1.66
0.05%
* Figures in parentheses are angular transformed values.
* Means followed by common letter are not significantly different by DMRT (p = 0.05).
Bio-Efficacy of Eco-Friendly Pesticides on the Management
269
 270

TABLE 19.3 Effect of Eco-Friendly Pesticides on Spodoptera Litura (Fab.) Infesting Cabbage During 2010
Treat- Dosage Pre treat- Mean percent efficacy(% Reduction Over Control) at different days after spraying Overall
ments ment st nd mean of
After 1 Spray After 2 Spray
larval st rd th th
post spray
popula- 1 day 3 day 7 day 10 day 1st day 3rd day 7th day 10th day count
tion/plant
T1 0.2% 3.67 29.00 38.33 62.75 83.08 36.50 54.50 66.67 83.17 56.75
(32.86)d (38.54)d (52.69)c (66.12)c (37.45)d (47.87)c (55.05)c (66.18)b
T2 0.5% 4.39 32.67 42.92 61.58 85.33 40.83 52.50 72.17 78.83 58.35
c c cd b c c b c
(35.16) (41.21) (51.99) (67.89) (39.99) (46.72) (58.49) (62.96)
T3 1.0% 4.78 16.17 18.25 67.25 90.48 19.00 41.83 84.50 90.50 53.50
f f b a e e a a
(24.03) (25.66) (55.41) (72.54) (26.19) (40.59) (67.22) (72.55)
T4 0.002% 4.45 33.67 36.67 52.25 69.28 36.33 48.50 63.67 65.83 50.78
c d e d d d c d
(35.77) (37.56) (46.58) (56.66) (37.36) (44.43) (53.27) (54.54)
T5 5.0% 4.17 69.17 73.00 59.48 42.33 72.50 81.17 44.33 45.00 60.87
a a d f a b d e
(56.59) (59.03) (50.76) (40.88) (58.71) (64.65) (42.03) (42.42)
T6 0.022% 4.44 62.33 67.92 79.38 69.42 67.83 89.50 84.50 65.83 73.34
b b a d b a a d
(52.44) (55.81) (63.36) (56.74) (55.76) (71.57) (67.22) (54.53)
Sustainable Horticulture Volume 2: Food, Health, and Nutrition
TABLE 19.3 (continued)

Treat- Dosage Pre treat- Mean percent efficacy(% Reduction Over Control) at different days after spraying Overall
ments ment After 1st Spray After 2nd Spray mean of
larval post spray
popula- 1st day 3rd day 7th day 10th day 1st day 3rd day 7th day 10th day count
tion/plant
T7 0.01% 3.00 18.17 20.50 37.62 63.65 20.83 32.83 40.50 64.17 37.28
f f f e e f d d
(25.59) (27.27) (38.12) (53.22) (27.50) (35.27) (39.82) (53.53)
T8 0.15% 3.39 21.17 33.58 79.48 21.00 33.83 80.50 85.83 22.00 47.17
e e a g d b a f
(27.74) (35.72) (63.43) (27.56) (35.87) (64.16) (68.31) (28.29)
T9 0.00 2.67 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
(4.05)g (4.05)g (4.05)g (4.05)h (4.05)f (4.05)g (4.05)e (4.05)g
SEm± 0.69 0.55 0.52 0.57 0.72 0.57 0.79 0.45
CD at 2.06 1.65 1.57 1.70 2.17 1.72 2.27 1.35
0.05%
* Figures in parentheses are angular transformed values.
* Means followed by common letter are not significantly different by DMRT (p = 0.05).
Bio-Efficacy of Eco-Friendly Pesticides on the Management
271
272 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

after first spraying, the highest population reduction (69.17% and 73.00%,
respectively) was obtained in NSKE @ 5.0%, and on the third day after
second spraying, the highest (89.50%) population reduction was recorded
in diflubenzuron + deltamethrin- 22% SC @ 0.022%.
Considering the overall population reduction after 1st to 10th day of
two spraying in 2010, the highest population reduction of 73.34% was
recorded in diflubenzuron + deltamethrin 22%SC @ 0.022% and the low-
est population reduction (37.28%) was recorded in spinosad 45%SC @
0.01%.

19.3.1.4 Comparative Efficacy of Eco-Friendly Pesticides on

S. litura Infesting Cabbage (2008, 2009, and 2010)

The pooled data (Table 19.4) indicated that all the insecticidal treatments
were significantly superior over control in reducing the larval population
of S. litura at 1, 3, 7, and 10 days after pesticide application. The treatment
with diflubenzuron + deltamethrin 22% SC was found significantly supe-
rior as compared to other pesticides. The highest population reduction of
83.42 and 81.24% in first spraying and 89.33 and 83.44% in second spray-
ing was recorded at 3 and 7 days after application, respectively.
The results of the present studies are in conformity with the earlier
findings of Hussain et al. (2002), Zaz (1989), and Venkadasubramanian
and David (1999) who also observed that the microbial pesticide B.
thuringiensis at 500 mL/ha and 1000 mL/ha was effective against S. litura
on cabbage. B. thuringiensis at 500 mL/ha and 1000 mL/ha was superior
to untreated control in causing larval decline of S. litura with a mean per-
cent reduction of 28.46% to 70.40% after spraying (Hussain et al., 2002).
According to Zaz (1989), B. thuringiensis caused 5.0–72.5% larval mor-
tality in S. litura. Venkadasubramanian and David (1999) reported that
B.t. products and other botanicals recorded 20.37–65.55% mortality of S.
litura on cabbage crop. The efficacy of spinosad (Tracer 45 SC) and B.
bassiana against S. litura was studied in Bapatla, Andhra Pradesh, India,
during the rabi of 2002–2003. Spinosad and B. bassiana gave 61.8%, and
62.3% larval mortality, respectively (Srinivas and Nagalingam, 2005), and
it supports the present investigation.
TABLE 19.4 Comparative Efficacy of Eco-Friendly Pesticides on Spodoptera Infesting Cabbage (2008, 2009 and 2010)
Treat- Dosage Pre treat- Mean percent efficacy (% Reduction Over Control) at different days after spraying Overall
ments ment larvae st nd mean of
After 1 Spray After 2 Spray
population/ post spray
plant 1st day 3rd day 7th day 10th day 1st day 3rd day 7th day 10th day count
T1 0.2% 4.59 33.11 49.83 66.08 82.86 35.44 52.94 66.72 81.17 58.52
(35.31)d (45.18)d (54.72)c (65.95)b (36.82)e (46.98)d (55.08)d (64.68)b
T2 0.5% 4.55 36.33 51.97 65.42 84.44 40.17 57.94 70.72 78.78 60.72
(37.25)c (46.42)c (54.34)c (67.19)b (39.61)c (49.89)c (57.63)c (62.93)c
T3 1.0% 4.65 19.50 34.19 74.08 90.61 19.61 42.11 80.50 87.06 55.96
(26.44)f (35.74)g (59.97)b (72.69)a (26.61)g (40.74)e (64.41)b (69.55)a
T4 0.002% 5.63 34.56 46.78 59.03 68.48 37.61 50.39 60.00 64.11 52.62
(36.30)cd (43.41)e (50.53)e (56.19)d (38.12)d (45.51)d (51.08)b (53.52)e
T5 5.0% 5.07 70.11 78.00 57.49 43.67 70.89 69.89 47.89 44.89 60.35
a b e b a b g b
(57.23) (62.46) (49.63) (41.65) (57.67) (57.42) (44.07) (42.35)
T6 0.022% 4.63 64.44 83.42 81.24 71.19 67.33 89.33 83.44 66.06 75.81
(53.71)b (67.54)a (64.79)a (57.88)c (55.45)b (71.46)a (66.43)a (54.68)d
T7 0.01% 4.48 20.22 29.78 46.59 65.11 20.83 35.28 47.72 63.94 41.18
(27.03)f (33.21)h (43.32)f (54.10)e (27.48)g (36.71)f (43.98)g (53.40)e
Bio-Efficacy of Eco-Friendly Pesticides on the Management

T8 0.15% 4.80 24.83 37.03 60.99 23.44 31.72 52.61 62.17 21.94 39.34
e f d g f d e g
(30.15) (37.77) (52.28) (29.22) (34.57) (47.21) (53.17) (28.26)
T9 00 5.35 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
(4.05)g (4.05)i (4.05)g (4.05)h (4.05)hj (4.05)g (4.05)h (4.05)h
SEm± 0.59 0.40 0.57 0.47 0.38 0.82 0.44 0.30
CD at 1.67 1.15 1.62 1.34 1.09 2.32 1.24 0.84
0.05%
* Figures in parentheses are angular transformed values.
273

* Means followed by common letter are not significantly different by DMRT (p = 0.05).
274 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

19.3.2 CABBAGE FRESH YIELD AND PERCENT INCREASE

IN YIELD OVER CONTROL DURING 2008, 2009, AND 2010

The results of different treatments were reflected in the yield of cabbage.

Treatment-wise average saleable yield (tons/ha) and percent yield increase
over control in 2008, 2009, and 2010 and pooled mean value are presented
in Table 19.5.
All the treatments provided higher yields than that of control. In 2008,
the maximum yield of cabbage (48.03 tons/ha) was found in diflubenzuron
+ deltamethrin 22% SC followed by (47.78 tons/ha) in Spinosad 45% SC.
A similar trend was observed in the next 2 years, 2009 and 2010. In 2009,
the highest cabbage yield (49.05 tons/ha) was obtained in diflubenzuron+
deltamethrin 22% SC which was at par with that of (47.60 tons/ha) in
Spinosad 45% SC. In 2010, the maximum cabbage yield (50.61 tons/ha)
was obtained in diflubenzuron + deltamethrin 22% SC, which was sta-
tistically at par with that of (50.18 tons/ha) Spinosad 45% SC. From the
3-year pooled mean value, the highest cabbage yield (49.23 tons/ha) was
obtained in diflubenzuron+ deltamethrin 22% SC followed by (48.52 tons/
ha) Spinosad 45% SC and the lowest yield (32.79 tons/ha) was recorded
in untreated control.
The treatment diflubenzuron + deltamethrin 22% SC exhibited
39.92% increase in the yield of cabbage over control followed by
39.18% in Spinosad 45% SC. A similar trend was observed in the 2nd
and 3rd year (2009 and 2010). In 2009, the highest increase in cabbage
yield of 55.43% was noticed in diflubenzuron + deltamethrin 22% SC,
which was statistically at par with that of 50.85% yield increase in Spi-
nosad 45% SC. In 2010, diflubenzuron+ deltamethrin 22% SC showed
55.77% increase in yield of cabbage over control followed by 54.43%
in Spinosad 45% SC. From the pooled mean value, highest percent
increase in yield (50.13%) was obtained in diflubenzuron + deltame-
thrin 22% SC followed by 47.96% in Spinosad 45% SC and 34.06% in
NSKE. Higher yield (42.4 tons/ha) of cabbage from B.t. (Halt)-treated
plot was earlier reported by Ghosh et al. (2001), which is in conformity
with the present studies.
TABLE 19.5 Cabbage Fresh Yield and % Increase in Yield Over Control During 2008, 2009 and 2010 and Pooled
Treat- Dosage Cabbage yield (tons/ha)
ments 2008 2009 2010 Pooled
Fresh % increase Fresh Yield % increase Fresh Yield % increase Fresh % increase
Yield over control over control over control Yield over control
T1 0.2% (41.93)bc 22.14 (42.05)b 33.25 (43.80)bc 34.81 (42.59)c 29.89
a a a a
T2 0.5% (47.78) 39.18 (47.60) 50.85 (50.18) 54.43 (48.52) 47.96
T3 1.0% (40.38c)d 17.63 (40.16)c 27.25 (42.84)cd 31.83 (41.13)d 25.41
d c cd d
T4 0.002% (39.26) 14.35 (39.45) 25.01 (42.22) 29.94 (40.31) 22.92
T5 5.0% (43.10)b 25.54 (43.17)b 36.79 (45.63)b 40.42 (43.96)b 34.06
a a a a
T6 0.022% (48.03) 39.92 (49.05) 55.43 (50.61) 55.77 (49.23) 50.13
T7 0.01% (35.97e)f 4.79 (35.30)d 11.85 (38.34)e 17.99 (36.54)f 11.41
de c d e
T8 0.15% (37.96) 10.57 (38.47) 21.90 (40.71) 25.28 (39.04) 19.06
f e f g
Bio-Efficacy of Eco-Friendly Pesticides on the Management

T9 00 (34.33) (31.56) (32.49) (32.79)

SEm± 0.81 0.63 0.74 0.24
CD at 2.43 1.88 2.22 1.20
0.05%
* Figures in parentheses are angular transformed values.
* Means followed by common letter are not significantly different by DMRT (p = 0.05).
275
276 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

KEYWORDS

•• cabbage head borer

•• eco-friendly pesticides
•• Spodoptera litura

REFERENCES

Asi, M. R., Bashir, M. H., Afzal, M., Zia, K., & Akram, M., (2013). Potential of entomo-
pathogenic fungi for biocontrol of Spodoptera litura fabricius (Lepidoptera: Noctui-
dae). J. Anim. Plant Sci., 23(3), 913–918.
Ghosh, S. K., Choudhuri, N., Ghosh, J., Chattarjee, H., & Senapati, S. K., (2001). Field
evaluation of pesticides against the pest complex of cabbage under terai region of
West Bengal. Pestology, 25(2), 95–97.
Hanif, R., Iqbal, Z., Iqbal, M., Hanif, S., & Rasheed, M., (2006). Use of vegetables as
nutritional food: role in human health. Journal of Agricultural and Biological Sci-
ence, 1, 18–22.
Haseeb, M., Liu, T. X., & Jones, W. A., (2004). Effects of selected insecticides on Cotesia
plutellae (Hymenoptera: Braconidae), an endolarval parasitoid of Plutella xylostella
(Lepidoptera:Plutellidae). Biocontrol, 49, 33–46.
Henderson, L. F., & Tilton, E. W., (1995). Tests with acaricides against brown wheat mite.
Journal of Economic Entomology, 48(2), 152–154.
Hussain, M. A., Pachori, R., & Choudhary, B. S., (2002). Population dynamics of Spodop-
tera litura Fab. on cabbage in Jabalpur, Madhya Pradesh. JNKVV-Research Journal,
36(1/2), 106–107.
Kranthi, K. R., Jadhav, D. R., Kranthi, S., Wanjari, R. R., Ali, R. R., & Russell, D. A.,
(2002) Insecticide resistance in five major insect pests of cotton in India. Crop Pro-
tection, 21, 449–460.
Nathan, S., & Kalaivani, K., (2005). Efficacy of nucleopolyhedrovirus (NPV) and azadi-
rachtin on Spodoptera litura Fabricius (Lepidoptera :Noctuidae). Biological Control,
34, 93–98.
Ray, D. P., Dutta, D., Srivastava, S., Kumar, B., & Saha, S., (2012). Insect growth regula-
tory activity of Thevetia nerifolia Juss. against Spodoptera litura (Fab.). J. Appl. Bot.
Food Qual., 85, 212–215.
Singh, A., & Sharma, O. P., (2004). Integrated pest management for sustainable agricul-
ture. Pratap, S., Birthal, O. P., Sharma, (Eds) proceedings 11 In: Integrated Pest Man-
agement in Indian Agriculture. NCAP and NCIPM. New Delhi, India, pp. 11–24.
Srinivas, M. S., & Nagalingam, B., (2005). Influence of certain microbial agents on
Spodoptera litura (Hubner) and coccinellids in groundnut. Journal of Entomological
Research, 29(1), 31–34.
Bio-Efficacy of Eco-Friendly Pesticides on the Management 277

Thenmozhi, S., & Thilagavathi, P., (2014). Impact of agriculture on Indian economy.
IRJARD, 3(1), 96–105.
Vastrad, A. S., Lingappa, S., & Basavana Goud, K., (2004). Ovicides for managing resis-
tant populations of diamondback moth, Plutella xylostella, L. Resistant Pest Man-
agement Newsletter, 14, 16–17.
Vastrad, A. S., Lingappa, S., & Basavanagoud, K., (2003). Management of insecticide
resistant populations of diamondback moth, Plutella xylostella (L.) (Yponomeutidae:
Lepidoptera). Pest Management in Horticultural Ecosystem, 9(1), 33–40.
Venkadasubramanian, V., & David, P. M. M., (1999). Insecticidal toxicity of commercial
Bacillus thuringiensis (Berliner) products in combination with botanicals to Spodop-
tera litura (Fabricius) and Heliciverpa armigera (Hubner). Journal of Biological
Control, 13, 85–92.
Zaz, G. M., (1989). Relative effectiveness of Bacillus cereus Frankland and frankland,
Bacillus thuringiensis Benliner and endosulfan against Spodoptera litura (Fab.) on
cauliflower. Indian Journal of Plant Protection, 18(1), 85–88.
CHAPTER 20

FOLIOSE LICHEN SPECIES:

A POTENTIAL SOURCE FOR
BIO-CONTROL AGENT AGAINST
COLLETOTRICHUM CAPSICI
M. CHINLAMPIANGA,1 A. C. SHUKLA,1 A. R. LOGESH,1
and D. K. UPRETI2
Department of Horticulture, Aromatic and Medicinal Plants,
1

Mizoram University, Aizwal – 796004, India

2
Lichenology Laboratory, Plant Biodiversity, Systematics and
Herbarium Division, CSIR-National Botanical Research Institute,
Lucknow – 226001, Uttar Pradesh, India

CONTENTS

Abstract.................................................................................................. 279
20.1 Introduction................................................................................. 280
20.2 Materials and Methods................................................................ 282
20.3 Results and Discussion............................................................... 285
20.4 Conclusion.................................................................................. 287
Acknowledgments.................................................................................. 287
Keywords............................................................................................... 288
References.............................................................................................. 288

ABSTRACT

Colletotrichum capsici Butler & Bisby is an important phytopathogenic

fungi causing many diseases (anthracnose or die back in chili, capsicum,
280 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

betel vine, turmeric, ashwagandha, black pepper, cucurbits, etc.) on vari-

ous horticultural crops; it not only reduces the quality and yield of the
infected crops but also enhances the cost of production. The objective
of the present study was to determine the antifungal efficacy of foliose
macrolichen species, viz., Parmotrema reticulatum and Everniastrum
cirrhatum against C. capsici, so that an alternative to the synthetic anti-
fungals can be explored. Keeping these views in mind, an in vitro anti-
fungal study of the acetone extracts of P. reticulatum and E. cirrhatum
(at 5% concentration) was investigated against C. capsici. The observa-
tions recorded that efficacy of both the lichens at low doses were not
much effective; however, as the doses increased, efficacy also increased.
Further, it was recorded that the acetone extracts of P. reticulatum and
E. cirrhatum (5% at 50 mL/L concentration) showed cidal efficacy and
having similar efficacy to that of the synthetic antifungal “Mancozeb.”
This was determined by using the modified spore germination inhibition
technique (MSGIT).
Findings of the present investigation show that after detailed in vitro
investigations, the active constituents of lichens can be used as a potential
substitute of synthetic fungicides.

20.1 INTRODUCTION

Phytopathogens, particularly, fungi are responsible for poor establishment

and stand loss in a variety of commercial horticultural crops. Colletotri-
chum capsici diseases account for marked reduction in productivity and
economic return in various horticultural crops including chilies. Anthrac-
nose (both pre- and postharvest) is one of the most important diseases in
chili. It results in yield loss (up to 50%) and deterioration of fruit qual-
ity. The disease is known to be caused by various species of Colletotri-
chum such as C. capsici, C. acutatum, C. gloeosporiodes, C. coccodes,
and C. dematium. Among these, C. capsici is the one of the important
phytopathogens, causing enormous losses in horticultural crops (Than et
al., 2008; Narasimhan et al., 2012; Susheela, 2012; Masoodi et al., 2013).
Although there are number of chemicals that are widely used for the
management of plant diseases, they are not beneficial in many respects
Foliose Lichen Species 281

such as high cost, breakdown of resistance, residual problem, and del-

eterious effect on non-target organisms including humans (Cowan, 1999;
Poorniammal and Sarathambal, 2009). Therefore, search for alternatives
to control plant diseases has been a challenging task for the plant scientist/
researchers, since the last few years.
Plants and their associated epiphytic plants, including lichens, serve
mankind as an important source of food, nutraceuticals, and medicine
from ancient time. They are also used for the management of diseases
caused by biotic agent/pathogens such as bacteria, fungi, mycoplasma,
actinomycetes, and nematodes, as they possess some defense mechanisms
against enemies, due to some unique substances synthesized through the
metabolic pathway.
Natural products such as plant-based formulations, solvent, and cow
urine extracts of plants, lichens and their metabolites, etc., have been
reported to possess potential antifungal effect against various types of
phytopathogenic fungi. Moreover, these agents are nontoxic and are easily
decomposed (Halama and Haluwin, 2004; Goel et al., 2011; Dileep et al.,
2013; Kambar et al., 2013; Rakesh et al., 2013; Vivek et al., 2013).
Lichens are symbiotic associations of a fungus (mycobiont) and a pho-
tosynthetic partner (photobiont) that can be either green algae or cyano-
bacteria (Ahmadjian, 1995). Lichens occurs in different growth forms, viz.,
crustose, foliose, fruticose, leprose, and squamulose and are well known to
grow on rocks and soil as well as epiphytes on the trees, leaves, and twigs.
Lichens synthesize a wide variety of secondary metabolites. Lichens are
well known for their prolific sources of biologically active natural prod-
ucts, as they produce a diverse range of secondary metabolites that are not
reported in any other plant groups. Slow growth and often-harsh living
make production of protective metabolites a necessity to lichens, and these
metabolites are believed to have antigrowth, anti-herbivore, and antimi-
crobial activity (Hale, 1983; Rankovic et al., 2008). The Eastern Himala-
yan region in northeast India and Western Ghats in south India represents
luxuriant and diverse growth of a number of lichen species. So far, only
a few lichen species from India have been screened for their antifungal
activities (Shahi et al., 2001, 2011; Tiwari et al., 2011). Owing to the rich
diversity of lichens in the country, there is a lot of scope for such studies.
Therefore, in the present study, an attempt has been made to evaluate the
282 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

antifungal efficacy of lichen Parmotrema reticulatum especially against

phytopathogenic fungi, which are responsible for the huge loss of agricul-
tural and horticultural yield throughout the world.

20.2 MATERIALS AND METHODS

20.2.1 COLLECTION AND IDENTIFICATION OF LICHENS

Lichens species were collected in September 2014 from two areas of the
Murlen National Park, Champhai district, Mizoram, India. P. reticulatum
(Taylor) Choisy (LWG Acc. No. 14-019172), corticolous habit, was col-
lected from the western part of the study area, while Everniastrum cirrha-
tum (Fr.) Hale ex Sipman (LWG Acc. No. 14-031427), corticolous habit,
was collected from the eastern part of the study area. The lichens were
identified on the basis of morphology, anatomy, and chemical tests. Color
tests were carried out on cortex and medulla of lichens by using 25%
potassium hydroxide (K), Steiner”s stable paraphenylene-diamine solu-
tion (P), and calcium hypochlorite solution (C). Secondary metabolites
were detected by thin layer chromatography (TLC) using solvent system A
(toluene 180 mL: 1-4 dioxane 60 mL: acetic acid 8 mL) following Walker
and James (1980). The lichen specimens were authenticated and depos-
ited at the Lichenology Laboratory, Plant Biodiversity, Systematics and
Herbarium Division, CSIR-National Botanical Research Institute (NBRI),
Lucknow, Uttar Pradesh.

20.2.2 EXTRACTION

The lichen samples/materials (whole thallus) used for in vitro antifungal

activity collected from the study area were sorted, cleaned/washed, and
then air dried at room temperature. Before extraction, the dried samples
were pulverized to powder form. Each powdered form of lichen sample
was stored in a sterile glass bottle in the refrigerator. A stock solution of
acetone extract was prepared by macerating 10 g of lichen material in 20
mL of acetone by using a pestle and mortal, followed by filtering through
a muslin cloth and a Millipore filter (pore size 0.22 mm). The solvent
Foliose Lichen Species 283

extraction was carried out at the specific boiling temperature at 56°C for
48 h for complete extraction of secondary compounds. The 10 g portion
of sieved powder was added to 100 mL of each solvent and left for 3
days at room temperature. The crude extract was prepared by decant-
ing, followed by filtration through a muslin cloth, and further filtered
with a Whatman No. 1 filter paper to obtain a clear filtrate. The filtrates
were sterilized by membrane filtration using 0.45-µm pore size filters.
The extracts were then evaporated to dryness under reduced pressure and
again dissolved in respective solvents to attain the required concentra-
tions of 5% for antifungal bioassay. The lichen extracts were kept at 4°C
till used.

20.2.3 ANTIFUNGAL ACTIVITY OF LICHEN EXTRACTS

The test fungus Colletotrichum capsici Butler & Bisby (MTCC 8473) was
procured from IMTECH, Chandigarh, India. The antifungal activity of
lichen extracts was determined using the modified spore germination inhi-
bition technique (MSGIT) of Shahi et al. (1997) with slight modification
of Shukla (2011). Potato dextrose broth was prepared and amended with
penicillin G (5 mg/L) and streptomycin sulfate (5 mg/L) in the medium
at 40°C in order to prevent bacterial growth, as suggested by Gupta and
Banerjee (1997). Culture discs containing spores (5 mm diameter) cut out
from the 7-day-old cultures grown in petri dishes were transferred asepti-
cally in flasks (100 mL) containing the broth and shaken thoroughly for
homogenous distribution of spores. The numbers of spores were counted
per microscopic field using “Modified Cytometer Technique” (MCT)
(Shahi et al., 1997). The diameter of microscopic field was measured by
micrometer, and the area and volume of the microscopic field was calcu-
lated using the following formula:

AMF = πr2

VMF = (AMF) h

where AMF = area of microscopic field; VMF = volume of microscopic

field; h = thickness of medium (in between slide and cover glass) 0.1 mm.
284 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

The number of spore (average count value of five microscopic fields)

was counted by eliminating the overlapped spores. The number of spores
in the volume of microscopic fields (NSV) was calculated using the for-
mula:

NSV = ANS/VMF

where ANS = average number of spores in the microscopic field.

The volume of liquid medium (VLM) per microscopic field was calcu-
lated by the formula:

VLM = 2rh

The total inoculums density (TID) was calculated in the initial volume
of medium by using the formula:

TID = (NSV/VLM) IVM

where IVM = initial volume of medium.

The effective concentration of acetone extract was determined by dis-
solving requisite quantity of extracts in acetone (2% of the required quan-
tity of the seed medium) and mixing it with the standardized inoculum
suspension in the culture tubes. In control, distilled water was used in
place of the lichen extracts. Besides, a comparison of the lichen extracts
with a synthetic fungicide “Mancozeb” (75%) was also made. Culture
tubes thus prepared were incubated at 27°C ± 1°C, and the observations
were recorded at the interval of 24 h up to 96 h by counting the number
of germinated spores. Percentage of zone of inhibition/spore germination
(SGI) by the extracts against test fungal culture was calculated using the
formula:

SGI (%) = (Gc − Gt)/Gc × 100

where Gc = number of germinated spore in control; Gt = number of ger-

minated spore in treatment.
The experiments were repeated twice and each of the tests was made
in triplicates.
Foliose Lichen Species 285

20.2.4 STATISTICAL ANALYSIS

The results recorded in Table 20.2 are the mean values.

20.3 RESULTS AND DISCUSSION

The details of color tests and TLC of P. reticulatum and E. cirrhatum are
recorded in Table 20.1. The antifungal efficacy of the 5% acetone extracts
of Parmotrema and Everniastrum lichen species against C. capsici are
recorded in Table 20.2 and Figure 20.1.
The observations show that an inhibition of >50% was recorded in both
the extracts at 5% concentration of 10 mL/L, and it was cidal at 50 mL/L,
respectively (Table 20.2 and Figure 20.1). Further, it was also observed

TABLE 20.1 Observations of Color Tests and TLC of Parmotrema and Everniastrum spp.
S. No. Lichen Colour test TLC
1 E. cirrhatum (Parmeli- Cortex K + yellow; medulla K+ Atranorin,
aceae) yellow turning red, C-, KC-, P+ salazinic and
orange. protoliches-
terinic acid
2 P. reticulatum (Parmeli- Medulla K+ yellow then red,C-, Salazinic acid
aceae) KC-, PD+ orange – red and consala-
zinic acids

TABLE 20.2 Antifungal Activity of Extract of P. reticulatum and E. cirrhatum against

C. Capsici and Their Comparison with Synthetics
5% Conc Control Spore germination inhibition (%) Synthetic Antifungal
(ml/L) P. reticulatum E. cirrhatum ‘Mancozeb’ (2.6 mg/L)
00.05 00 9.00 6.00 13.00
00.10 00 12.03 13.03 26.25
02.00 00 24.05 25.05 39.50
05.00 00 43.11 46.10 57.40
10.00 00 55.02 52.08 78.15
20.00 00 87.00 87.00 100.00c
50.00 00 100.00c 100.00c 100.00c
s
indicates static, indicates cidal in nature; Conc. indicates concentration.
c
286 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

FIGURE 20.1 (See color insert.) Antifungal efficacy (%) of the test samples against C.
capsici – A graphical representation.

that the activity of the two extracts and the synthetic fungicide “Manco-
zeb” (75%) exhibited more or less similar inhibitory efficacy against C.
capsici (Figure 20.1).
Further, diagrammatic representation of the findings, as recorded in
Table 20.2, is shown in Figure 20.1.
Literature revealed that the antifungal efficacy of 50% ethanolic extract
from some macrolichens, Parmelia tintorum, Ramalina sp., Teloschistes
flavicans, and Usnea undulate had been tested by Dikshit (1991) against
the pathogenic fungi Aspergillus flavus. Acetone extract of Stereocaulon
sp. was found to be more effective than Ramalina sp. against plant patho-
genic fungi, viz., Alternaria alternata, Aspergillus flavus, and Penicillium
italicum (Shukla et al., 2011). Three Parmotrema species, viz., P. tincto-
rum, P. grayanum, and P. praesorediosum were reported as bio-control
agents against C. capsici (Kekuda et al., 2014). Further, in another study,
Preeti et al. (2014) reported the activities of solvent extracts of Parmo-
trema reticulatum against some phytopathogenic fungi.
Foliose Lichen Species 287

Similarly, aqueous extract of Heterodermia leucomela was found to

exhibit significant inhibition of germination of spores of some phyto-
pathogenic fungi (Shahi et al., 2001). A lichen-forming fungus isolated
from Heterodermia sp. was found to exhibit strong inhibitory activity
against Pythium spp. (Hur et al., 2003). Similarly, acetone extracts of
Evernia prunastri and Hypogymnia physodes were found effective in
inhibiting Pythium ultimum, Ustilago maydis, and Phytophthora infes-
tans. Lichenic acids, viz., evernic acid, usnic acid (±), protolchesterinic
acid, and atranorin were reported to have strong inhibitory activity
against two phytopathogenic fungi (Kekuda et al., 2011, 2012). Metha-
nol extract of Parmelia sulcata, Flavoparmelia caperata, and Evernia
prunastri were shown to possess antifungal activity against a panel of
human and phytopathogenic fungi (Mitrovic et al., 2011). However, in
the present study, antifungal efficacy of acetone extracts of P. reticula-
tum and E. cirrhatum against C. capsici was recorded cidal in nature
(5% concentration at 50 mL/L), respectively (Table 20.2 and Figure
20.1).

20.4 CONCLUSION

The acetone extracts of P. reticulatum and E. cirrhatum showed fungicidal

efficacy against C. capsici (5% concentration at 50 mL/L). To the best
of our knowledge, this is the first report on inhibitory effect of Evernias-
trum species and Parmotrema species from Mizoram, India, against the
test fungi. Therefore, based on these findings as well as detailed in vivo
investigations, lichen-based formulations can be effective agents for the
management of the disease caused by the fungal pathogen C. capsici in
various commercial horticultural crops and can be an alternative source
for developing natural fungicides.

ACKNOWLEDGMENTS

The authors are thankful to the Director, CSIR-National Botanical

Research Institute, Lucknow, and the authorities of Mizoram University,
Aizawl, for the necessary facilities as well as to the Council of Scientific
288 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

and Industrial Research, New Delhi, for financial support [CSIR project
No. 38(1376)/ 13/ EMR-II; dated. 1.10.2013)].

KEYWORDS

•• anthracnose
•• antifungal
•• Colletotrichum capsici
•• everniastrum
•• modified spore germination inhibition technique
•• parmotrema

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Kambar, Y., Vivek, M. N., Manasa, M., Kekuda, P. T. R., & Nawaz, N. A. S., (2013).
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CHAPTER 21

FUNGAL DISEASES OF MEDICINAL

AND AROMATIC PLANTS AND THEIR
BIOLOGICAL MANAGEMENT
RAMESH S. YADAV,1 D. MANDAL,2 and AMRITESH C. SHUKLA2
1
Department of Plant Pathology, Sardar Vallabhbhai Patel University
of Agriculture & Technology, Meerut–250110, India
2
Department of Horticulture, Aromatic and Medicinal Plants,
Mizoram University, Aizawl–796004, India,
E-mail: [email protected]

CONTENTS

Abstract.................................................................................................. 292
21.1 Introduction................................................................................. 292
21.2 Biocontrol Products in Medicinal Plants Disease
Management................................................................................ 295
21.3 Microbial and Biocontrol Product Formulations ....................... 295
21.4 Shelf-Life of Formulation........................................................... 300
21.5 Mechanisms of Action................................................................ 300
21.6 Delivery of Biocontrol Products Under Field Conditions.......... 306
21.7 Conclusion.................................................................................. 308
Acknowledgment................................................................................... 308
Keywords............................................................................................... 308
References.............................................................................................. 309
292 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

ABSTRACT

Biological control of fungal diseases constitutes a very broad category of

control, consisting of a wide variety of different organisms, mechanisms,
interactions, and processes. It has enormous potential to supplement and
complement existing disease control strategies. However, biological con-
trol also has different properties, requirements, and constraints than pre-
vious conventional controls and needs to be properly implemented and
integrated with current production strategies. Biological control depends
on the effective functioning of the appropriate antagonist strains within
each particular plant-pathogen ecosystem. Identifying the appropriate
antagonist strains is generally the first step in this process. Understand-
ing how, where, when, and why the biocontrol works may also be crucial
to successful development of the biological control system. Because of
complexity of soil microbial communities and the role of the biocon-
trol organism within these communities, an ecological approach to the
development of the biological control system is recommended. Evaluat-
ing the ecological interactions of the biocontrol organism with the patho-
gen, host plant, surrounding microbial community, and the environment
will be useful in developing the best strategies for the implementation
and management of the biological control system. For maximum effec-
tiveness, biocontrol organisms that are locally adapted to the particular
environments and patho-systems where they are suited may need to be
developed.

21.1 INTRODUCTION

Over the centuries, the use of medicinal plants and their products have been
an important part of daily life despite the progress in modern medicine and
pharmaceutical research. Approximately 3000 plant species are reported
in India to have medicinal properties (Prakash, 1998). The Rigveda (3700
BC) mentions the use of medicinal plants. Our traditional system of medi-
cine, viz., Ayurveda, Yunani, Siddha, Homeopathy, etc., uses herbs or herb
products for the treatment of various diseases of human beings and ani-
mals. It is estimated that 80% of the world population depends directly
Fungal Diseases of Medicinal and Aromatic Plants 293

on plant-based medicine for their healthcare (WHO, 2003). During the

last 20 years, there has been a complete transformation of the medicinal
system in the world and especially in those countries of the West that were
totally dependent on modern medicine. Because of the realization of toxic-
ity associated with the use of antibiotics and synthetic drugs, the Western
society is obsessed with the idea that drugs from medicinal herbs are safer
than synthetics. The importance of plants as a major source of clinical
agents is much more in developing countries of Asia, Africa, and South
America, where traditional medicine is practiced widely. Considerable
emphasis has also been laid in the research programs of academic institu-
tions/scientific organizations and drug companies during the last 10 years
to trap the efficacy of the plant kingdom for treatment of such diseases
for which the modern medicine does not have any effective treatment,
including virus diseases like AIDS and herpes, cancer, arthritic disorder,
liver ailments, among others (Bhakuni and Jain, 1995). In India, medicinal
plants offer low cost and safe healthcare solution.
Medicinal plants also suffer from various kinds of diseases that are
similar to those affecting animals and man. Plant growth and yield depend
on protecting the plants from diseases. Anything that affects the health of
medicinal plants is likely to affect their growth and yield and also seriously
reduce their usefulness to mankind. Plant diseases are thus important to
man because they not only damage plants but also the plant products. Plant
diseases are caused by fungi, bacteria, viruses, phytoplasma, spiroplasma,
viroids, nematodes, algae, protozoa, and parasitic higher plants apart from
environmental and physiological factors.
Fungi are small, generally microscopic, eukaryotic, usually filamen-
tous, branched, spore-bearing organisms that lack chlorophyll. Fungi have
cell wall that contains chitin and glucans (but no cellulose) as the skeleton
components. These are embedded in a matrix of polysaccharides and gly-
coprotein. Fungal cells have distinct nucleus that contain genetic mate-
rial and is surrounded by a special envelope called nuclear membrane.
Fungi are unicellular or multicellular and can reproduce sexually or asexu-
ally. The unicellular forms of fungi are larger than bacteria. They obtain
nourishment from soil, water, animal, or plant hosts. Vast number of fungi
inhabit on the Earth in which more than 10,000 species of fungi can cause
disease in plants (Agrios, 2005).
294 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

There are several kinds of fungal diseases affecting the medicinal

plants, and each plant can be affected by a number of fungal plant patho-
gens. Disease conditions in the medicinal plants are recognized according
to the symptoms produced by the pathogens. The usual disease symptoms
are root rots, wilts, collar rot, stem rot, leaf spots, blights, canker, anthrac-
nose, rust, mildews, smuts, damping-off seedling, etc. Fungal diseases of
medicinal plants, thus, can be identified by the symptoms and presence of
the pathogens especially on the site of infection.
Although there are number of synthetic fungicides available in the
market for the control of fungal diseases of medicinal plants, concerns
about the health, safety, and environmental effects of chemicals in our
water, soil and food require that these inputs be minimized. In addition,
biological control may especially important for use in patho-systems in
which chemical controls are not economical or ineffective. Biological
control may also lessen other problems associated with certain chemical
controls, such as the development of pathogen resistance to chemicals,
reduction in beneficial organisms’ populations, and the creation of bio-
logical vacuums (Cook and Baker, 1983). Biological control generally
has more specific effects, with only the target pathogen organism(s) being
adversely affected, leaving other beneficial organisms and a diverse soil
microbial community intact to provide for healthier plants and roots.
Thus, biological control can be safer for humans, the plants, and the
environment. Biological control has the potential to be more stable and
longer lasting than some other controls and is compatible with the con-
cept and goals of integrated plant disease management and sustainable
agriculture.
The foundation for the development of contemporary biological con-
trol was laid by the two landmark books by Cook and Baker (1983) and
Baker and Cook (1974). In addition, there have been numerous books,
reviews, and research papers on more recent development in biological
control concepts, product formulations, and implementations (Cambell,
1994; Chet, 1994; Wilson and Wisniewski, 1994; Lumsden et al., 1995;
Mukherjee and Mukhopadhyay, 1995; Fravel and Larkin, 1996; Mukho-
padhyay, 1996, 2003; Fravel et al., 1997; Mishra et al., 1998, 2000; Butt et
al., 2001; Gnanamanickam, 2002; Singh et al., 2004; Khilari et al., 2008,
2009; Yadav et al., 2009, 2010). The objective of this chapters is to high-
Fungal Diseases of Medicinal and Aromatic Plants 295

light the current status of biological control work including product for-
mulations and delivery systems and to optimize biocontrol products with
organic substrates (farm yard manure (FYM), vermicompost, and press
mud) at the grower field for the control of fungal pathogens of medicinal
plants particularly in Indian context.

21.2 BIOCONTROL PRODUCTS IN MEDICINAL PLANTS

DISEASE MANAGEMENT

Most of the efforts on biocontrol of plant diseases with antagonistic fungi

and bacteria were diverted against pre- and postemergence seedling,
root rot and wilt diseases, and only a few reports deals with the foliar
pathogens. Major pathogens targeted are Rhizoctonia solani, Sclerortium
rolfsi, Fusarium (particularly F. oxysporum), Sclerotinia, Macrophomina,
Pythium, Phytophthora, etc. Almost all present generation fungicides are
static in their action. Therefore, they are unable to kill completely rest-
ing structures like sclerotia, chlamydospores, rhizomorph, etc. formed by
many soil-borne pathogens. Higher doses and more frequent application
of fungicides to soil disturb ecological balance. Therefore, biological con-
trol alone or in combination with cultural and other practices seems to be
the only practical answer for these types of pathogens. A critical review of
the literature reveals that soil-borne diseases such as pre- and post-emer-
gence damping off, root rots, wilts, collar rots, stem rots, etc., under field
condition were successfully suppressed by applying biocontrol products
(Table 21.1).

21.3 MICROBIAL AND BIOCONTROL PRODUCT

FORMULATIONS

A wide variety of microorganisms including fungi, bacteria, and actinomy-

cetes have been shown to have biocontrol activity against various fungal
pathogens or the disease they cause and have been studied as biocontrol
agents in several patho-systems. Because of high reproductive ability,
short generation time, targeted host specificity, and capability of surviving
in adverse conditions as either saprobes or resistant structures, scientist
296 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

TABLE 21.1 Microbial species with Demonstrated Success as Biocontrol Products

Plants Diseases Target Biocontrol product Mode of
pathogen(s) with (source) application
of products
Chlorophy- Anthrac- Colletotricum BioJect Spot-Less (Pse- Root dip treat-
tum borivil- nose sp. domonas aureofaciens) ment, Soil
ianum (Safed Trichodex (Trichoderma treatment
musli) harzianum)
Planttago Damping- Pythium ulti- Companion (Bacil- Seed treat-
ovata (Isab- off mum, Rhizoc- lus subtilis str. GB03) ment Soil
gol-Indian tonia solani Gliogard (Gliocladium treatment
Psyllium) virens GL-21)
Wilt Fusarium HiStick N/T (Bacillus Soil treatment
oxysporum subtilis str. MB1600)
Trichodex (Trichoderma
harzianum)
Powdery Erysiphae AQ10 Biofungicide Spray
mildew cichoracearum (Ampelomyces quisqua-
lis M-10)
Papaver Powdery Erysiphae AQ10 Biofungicide Spray
somniferum mildew polygoni (Ampelomyces quisqua-
(Opium lis M-10)
Poppy) Root rot Macrophomina Ecofit (Trichoderma Soil treatment
phaseolina, viride) Kodiak (Bacillus
Rhizoctonia subtilis str. GB03); Con-
bataticola, tans WG (Coniothyrium
Sclerotinia minitans)
sclerotiorum
Wilt Fusarium Fusaclean-Nonpatho- Soil treatment
oxysporum genic Fusarium oxys-
porum
Rauolfia Wilt F. oxysporum f Biotox C- Nonpathogen- Soil treatment
serpentine sp. rauvolfii ic Fusarium oxysporum
(Surpgandha) Root Shield (Tricho-
derma harzianum)
Cantharan- Foot rot Sclerotium Trichoderma 2000 Soil treatment
thus roseus rolfsii (Trichoderma harzia-
(Periwinkle) num)
Cassia Damping- Macrophomina Promote (Trichoderma Soil treatment
angustifolia off at phaseolina, harzianum and T. viride)
(Senna) seedling Rhizoctonia
stage bataticola
Fungal Diseases of Medicinal and Aromatic Plants 297

TABLE 21.1 (Continued)

Plants Diseases Target Biocontrol product Mode of
pathogen(s) with (source) application
of products
Atropa Root rot Pythium T-22 (Trichoderma Soil treatment
belladonna butleri harzianum)
(Belladona) Collar rot Sclerotium Trichoderma 2000 Soil treatment
rolfsii (Trichoderma harzia-
num)
Cinchona Damping Pythium Chaetomium globosum Soil treatment
ledgeriana off vexans
(Cinchona) Collar rot Sclerotium Trichoderma 2000 Soil treatment
rolfsii (Trichderma harzianum)
Coleus Leaf Rhizoctonia Conquer (Pseudomonas Spray
forskohlli blight solani fluorescens) Deny (Bur-
(Coleus- kholderia cepacia)
Pathar Chur)
Datura Root rot Sclerotium Trichoderma 2000 Soil treatment
stramonium rolfsii (Trichoderma harzia-
(Datura) num)
Digitalis Anthrac- Colletotrichum Trichoject (T. harzia- Soil treatment
purpurea nose fuscum num and T.viride )
(Digitalis)
Dioscoria Tuber rot Aspergillus HiStick N/T (Bacillus Tuber treat-
dettoidea niger subtilis str. MB1600) ment Soil
(Dioscoria) treatment
Panax Anthrac- Colletotrichum BioJect Spot-Less (P. Soil treatment
ginseng (Gin- nose dematium aureofacens)
seng)
Powdery Erysiphae AQ10 Biofungicide Spray
mildew panax (Ampelomyces quisqua-
lis M-10)
Damping- Pythium sp., Deny (Burkholderia Soil treatment
off Rhizoctonia cepacia)
solani
Gycyr- Root rot Rhizoctonia Gliogard (Gliocladium Rhizome
rhiza glabra bataticola virens GL-21) treatment
(Liquorice- Wilt Fusarium sp. Chaetomium globosum Soil treatment
Mulathi)
Withania Damping Pythium sp. Root Shield (T. harzia- Soil treatment
somnifera off num)
(Ashwa- Seedling Alternaria Kodiak (Bacillus subti- Soil treatment
gandha) rot alternata lis str. GB03)
298 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

TABLE 21.1 (Continued)

Plants Diseases Target Biocontrol product Mode of
pathogen(s) with (source) application
of products
Mentha spp. Stolon rot Macrophomina Ecofit (Trichoderma Stolen treat-
(Mint) phaseolina viride) ment Soil
treatment
Verticil- Verticillium Verticillium biguttatum -
lium wilt albo-atrum
Powdery Erysiphae AQ10 Biofungicide Spray
mildew cichocearum (Ampelomyces quisqua-
lis M-10)
Chrysanthe- Damping- Pythium sp. Polygandron (Pythium Soil treatment
mum ciner- off oligandrum)
ariifolium Verticil- Verticillium Talaromyces flavus -
(Pyrethrum) lium wilt albo-atrum
Curcuma Rhizome Pythium Intercept (Burkholderia Rhizome
longa (Tur- and root aphanider- cepacia) BINAB-T WP treatment Soil
meric) rot matum P. (Trichoderma. harzia- treatment
graminicolum num and T.polysporum)
Carica Stem or Pythium Deny (Burkholderia Seed treat-
papaya (Pa- foot rot aphaniderma- cepacia) BINAB-T WP ment Soil
paya) tum (Trichoderma. harzia- treatment
num and T. polysporum)
Musa sapien- Panama Fusarium Biotox C- Nonpatho- Sucker treat-
tum (Banana) wilt oxysporum f. genic (Fusarium oxys- ment Soil
sp. cubense porum) Root Shield (T. treatment
harzianum)
Lycopersicon Damping- Pythium sp. Trichopel (Tricho- Soil treatment
esculentum off derma. harzianum and Seed treat-
(Tomato) T. viride) ment
Fusarial Fusarium oxy- Ecofit (Trichoderma Soil treatment
wilt sporum f. sp. viride) HiStick N/T Seed treat-
lycopersici (Bacillus subtilis str. ment
MB1600

manipulated the ability of microorganisms for the control of diseases of

crop plants and developed several formulations.
Formulations are an important component in the development of an
effective biological control system (Lewis and Papavizas, 1991). Their
Fungal Diseases of Medicinal and Aromatic Plants 299

development and application can have a major impact on the success of

a biocontrol agent (Vidhyasekaran and Muthamilan, 1995). Formulations
must deliver the antagonist to the appropriate location in sufficient quan-
tities and in the proper form. The activity and growth of the biocontrol
agent depend greatly on the endogenous nutritional status of the formu-
lated propagule and the energy source provided in the formulation. An
effective formulation provides nutrients for germination and sporula-
tion of the biocontrol organism, withstands harsh environmental condi-
tions, interacts with other organisms to the advantage of the biocontrol
agent, can be integrated with cultural practices, and can be applied with
existing machinery or methods. The formulation must also be inexpen-
sive, easy to produce, and have an appropriate shelf-life (Lumsden et al.,
1995). Currently, there are over 40 different formulations of biocontrol
agents commercially available around the world for the control of vari-
ous plant diseases (Fravel et al., 1997). The composition of formulation
varies greatly with the intended use (Fravel et al., 1997). Fungi and acti-
nomycetes are currently on the market as pellets, wettable powders, gran-
ules, micro-granules, water dispersible granules, and dusts. Fungi are also
impregnated in sticks and produced on grain. Actinomycetes have been
specifically produced as powders for drenches or sprays or added through
irrigation systems. Bacteria have been formulated as aqueous suspensions
of fermenter biomass to spray, liquid suspension for drench or for drip
irrigation, dry powders and wettable powders. Bacterial and fungal prod-
ucts can also be applied to seed with a sticker and a peat and clay carrier
(Fravel et al., 1997).
In recent years, many small and medium entrepreneurs have entered
into commercial production of biocontrol agents, resulting in several prod-
ucts into Indian market, and to date, approximately 80 biocontrol prod-
ucts/biopesticides have been developed so far (Singh et al., 2004).
At present, the market share of biopesticides is around 2.5% of total
pesticide market compared to 1% in 2001 (Singh et al., 2004). This increase
is due to utilization of biopesticides mainly in medicinal plants and veg-
etable crops, sugarcane, cotton and paddy. The projected share of these
biopesticides by the end of 2015 is 15% of the total pesticide market. They
are now well accepted in “organic farming” and are major part of private
market mainly in Punjab, Haryana, western Uttar Pradesh, Uttarakhand,
300 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Madhya Pradesh, Maharashtra, and Tamil Nadu. The Indian biopesticide

industry is the umbrella of biopesticides products available in the country
today (Table 21.2). Many products are being exported from India now.
India has over 400 biopesticide production units with 8–10 major players
(Singh et al., 2004). Technologies for joint ventures in foreign countries
are also available. Commercialization of biopesticides is a multistep pro-
cess involving a wide range of activities (Singh et al., 2004).

21.4 SHELF-LIFE OF FORMULATION

Shelf-life of a biocontrol agent plays a crucial role in storing a formulation.

In general, the antagonist multiplied in an organic food base has greater
shelf life than the inert or inorganic food bases (Vidhyasekaran and Muth-
amilan, 1995; Fravel et al., 1997). Shelf-life of Trichoderma harzianum in
coffee husk was more than 18 months. Talc, peat, lignite, and kaolin-based
formulation of Trichoderma viride had a shelf-life of 4 months. Shelf-life
of this species in gypsum-based formulation was 4 months (Fravel et al.,
1997).

21.5 MECHANISMS OF ACTION

Biocontrol mechanism involving antagonistic microbial species/strains

operates by way of competition, antibiosis, parasitism, and induced resis-
tance.

21.5.1 COMPETITION

Competition has been defined as the active demand in excess of the imme-
diate supply of material on the part of two or more organisms (Clarke,
1965). The result is a restriction on population size or microbial activity
of one or more of the competitors (Paulitz and Baker, 1987). Competi-
tion between microorganisms generally refers to competition for nutrients
such as available carbon, nitrogen, iron, or trace elements or competition
for space such for colonization or infection sites on the root or seed sur-
Fungal Diseases of Medicinal and Aromatic Plants 301

TABLE 21.2 Microbial Agents Registered and Commercially Marketed as Biocontrol

Products
Biocontrol Microbial Developing agency
Products
Antagon- TV Trichoderma. viride Green Tech Agro Products, Coim-
batore
Biocon T. viride Tocklai Experimental Station, Tea
Research Association, Jorhat, Assam
Bioderma T. viride + T. harzianum Biotech International Limited, New
Delhi
Bioguard T. viride Krishi Rasayan Export Pvt. Ltd.,
Solan (H.P.)
Ecoderma T. viride + T. harzianum Margo Biocontrol Pvt. Ltd., Bangalore
Ecofit Trichoderma viride Hoechst and Schering AgroEvo Ltd.,
Mumbai
Funginil T. viride Crop Health Bioproduct Research
Centre, Ghaziabad
Kalisena SD
Kalisena SL Aspergillus niger AN-27 Cadilla Pharmaceuticals Ltd., Ahmed-
abad
Pant Biocontrol T. harzianum G.B. Pant University of Agriculture
Agent-1 Technology, Pantnagar
Sun-Derma T. viride Sun agro Chemicals, Chennai
Trichoguard T. viride Anu Biotech International Ltd.,
Faridabad
Tricho T. viride Excel Industries Ltd., Mumbai
Antifungus Trichoderma spp. Grondortsmettingen De Cuester,
Belgium
AQ10 Ampelomyces quisqualis Ecogen, Inc. Israel
isolate M-10
Aspire Candida oleophila I-182 Ecogen, Inc. Israel
Bas-derma T. viride Basarass Biocontrol Res. Lab., India
Biact/Paecil Paecilomyces lilacinus Technological Innovation Corporation
Pty. Ltd., Australia
BINAB T Trichoderma harzianum Bio-Innovation Eftr AB UK Bredhol-
(ATCC 20476) and Tricho- men, Sweden
derma polysporum (ATCC
20475)
Bioderma Trichoderma viride/ T. Biotech International Ltd., India
harzianum
302 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

TABLE 21.2 (Continued)

Biocontrol Microbial Developing agency
Products
Biofungus Trichoderma spp. Grondortsmettingen deCuester n. v.,
Belgium
Bio-Trek 22G Trichoderma harzianum Bio works, Inc. of Geneva, NY
Blue Circle Burkholderia cepacia ecoScience Corp., USA
Coniothyrin Coniotyrium minitans Russia
Contans Coniotyrium minitans Prophyta Biologischer Pflanzenschutz
GmbH’, Germany
BioJect Spot- Peudomonas aureofaciens Eco soil systems, USA
Less
Fusaclean Fusarium oxysporum Natural Plant Protection, France
(Nonpathogenic)
Kali sena Aspergillus niger Cadilla Phrma., India
Polyversum Pythium oligandrum Biopreparaty Ltd., Czech Republic
Polyversum/ Pythium oligandrum Biopreparaty Ltd., Czech Republic
Polygandron
Prestop, Prima- Gliocladium catenulatum Kemria Agro. Oy, Finland
stop
Root Pro. Trichoderma harzianum Bioworks Inc., USA
Root shield, Trichoderma harzianum Bioworks Inc., USA
Plant Shield, Rifai strain KRL-AG (T-22)
T-22 Planter
Box
Rootstop Phlebia gigantean Kemira Agro. Oy, Finland
Gliogard Gliocladium Virens strain Thermo Trilogy, USA
GL-21
Supresivit Trichoderma harzianum Borregaard and Reitzel, Czech Re-
public
Trichoderma Trichoderma sp. Myocontrol Ltd., Israel
2000
Trichodex, Trichoderma harzianum Makhteshim Chemical Works Ltd.,
Trichopel USA
Trichopel, Trichoderma harzianum and Agrimm Technologies Ltd., New
Trichoject, Trichoderma viride Zealand
Trichodowels,
Trichoseal
Tri-control Trichoderma spp. Jeypee Biotechs, India
Fungal Diseases of Medicinal and Aromatic Plants 303

TABLE 21.2 (Continued)

Biocontrol Microbial Developing agency
Products
Trieco Trichoderma viride Ecosense Labs Pvt. Ltd., Mumbai,
India
TY Trichoderma sp. Myocontrol, Israel
Vaminoc Mycorrhizal Fungi Biological Crop Protection Ltd., USA
Biofox C Fusarium oxysporum SIAPA, Italy
Companion Bacillus subtilis GB03 Growth Products, USA
Deny/Intercept Burkholderia cepacia Stine Microbial Products, Shwance,
KS
HitStick N/T Bacillus subtilis str. MicroBio Group, UK
MB1600
Kodiak Bacillus subtilis str. GB03 Gustaffson Inc., USA
Conquer Peudomonas fluorescens Mauri Foods, Australia
Koni Coniothyrium minitans BIOVED Ltd, Hungary
Victus Peudomonas fluorescens Sylvan Spawn Laboratory, USA

face. Nutrients from roots and seeds support microbial growth and other
activities in the spermosphere and rhizosphere (Paulitz, 1990). Nutrients
from roots and seeds are derived from several sources including exudates,
secretion lysates, and mucilages. These nutrients are chemically diverse
and include carbohydrates, amino acids, peptides organic acids, and other
plant metabolites (Curl and Truelove, 1986).
Competition can be an effective biocontrol mechanism when the
antagonist organism is present in sufficient quantities at the correct
time and location and can utilize limited nutrients or other resources
more efficiently than the pathogen. An example of this interaction was
observed in Fusarium wilt suppressive soils in France (Alabouvette et
al., 1993). High populations of saprophytic strains of Fusarium oxys-
porum effectively competed with the pathogenic strains of F. oxyspo-
rum for reduced carbon sources in these soils, resulting in an inhibition
of pathogen propagule germination, reduced saprophytic growth of the
pathogen, and low levels of disease. Competition for iron between bio-
control bacteria and plant pathogens has been well documented (Leong,
1986). Iron is required for growth by microbes, but is typically limited in
304 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

availability in soil. Strains of certain biocontrol bacteria, such as Pseu-

domonas fluorescens, produce siderophore, which have a high affinity
for soluble ferric iron (Fe+3). Siderophores bind iron, allowing these bio-
control bacteria to effectively compete with pathogens for iron. Produc-
tion of siderophores and the limited availability of iron in soil have been
associated with disease suppression by several bacterial antagonists
(Buysens et al., 1996).

21.5.2 ANTIBIOSIS

Antibiosis refers to the inhibition or destruction of the pathogen by a

metabolite product of the antagonist, such as the production of specific
toxin, antibiotics, or enzymes. This interaction can result in suppression
of activity of pathogen or destruction of pathogen propagules. To be effec-
tive, antibiotics must be produced in situ in sufficient quantities at the
precise time and place where they will interact with the pathogen. The pro-
duction of antibiotics by various strains of P. fluorescens, which include
phenazine compounds and 2,4-diacetylphloroglucinol has been shown to
be important in biocontrol through mutational analysis. Mutant strains of
P. fluorescens that do not produce these antibiotics have reduced efficacy
as biocontrol agents (Keel et al., 1992).
Antibiosis has also been shown to be important in biocontrol inter-
actions between fungal biocontrol agents and fungal plant pathogens.
Gliocladium virens, an important biocontrol fungus, has activity against
several soil-borne plant pathogens, including Pythium ultimum and Rhi-
zoctonia solani (Papavizas and Lewis, 1989). Different strains of G.
virens produce a variety of metabolites, including gliotoxin and glioviri-
din, which are toxic or inhibitory to several fungal pathogens (Wilhite et
al., 1994). In addition, gliotoxin has been detected in a number of soils
colonized by G. virens, and quantities of gliotoxin in these soils have
been correlated with disease suppression (Lumsden et al., 1992). Glio-
toxin is not found in any commercial formulations, but when the spores
of G. virens begin to grow in the soil, they produce the antibiotic. Further-
more, gliotoxin is sensitive to oxidation and probably poses no health risk
because of rapid degradation.
Fungal Diseases of Medicinal and Aromatic Plants 305

Trichoderma spp. secrete diverse secondary metabolites with anti-

biotic properties, including polyketides, terpenoids, polypeptides, and
metabolites derived from alpha-amino acids (Taylor, 1986). T. harzianum
produces trichorzianins, trichkindins, trichozins, and harzianic acids that
exhibit antifungal activity (Corley et al., 1994). Furthermore, a novel anti-
fungal protein from T. viride “tricholin” causes cessation of growth and
uptake of amino acids and is effective against Rhizoctonia solani (Lin et
al., 1994).

21.5.3 PARASITISM

Parasitism occurs when the antagonist feeds on or within the pathogen,

resulting in the direct destruction or lysis of propagules and structures.
When one fungus parasitizes another fungus, it is called mycoparasitism
(Lumsden, 1992). Trichoderma, Chaetomium, and Gliocladium spp. are
well-known parasites of a wide variety of pathogenic fungi. Biocontrol
fungi coil around and parasitize mycelia of the fungal pathogen. This
complex process involves the activities of cell wall hydrolytic enzymes
such as glucanases, chitinases, proteases, and lipase produced by biocon-
trol fungi (Schirmbock et al., 1994; Haran et al., 1995). Glucanse and
chitinase activities have been detected in sterile soil containing mycelium
of Sclerotium rolfsii, Rhizoctonia solani, and Pythium aphanidermatum
inoculated with T. harzianum (Elad et al., 1982). Cell wall-degrading
enzymes produced by T. harzianum and G. virens inhibit conidia germi-
nation and germ tube elongation of Botrytis cinerea in vitro (Lorito et al.,
1993).

21.5.4 INDUCED RESISTANCE

Induced resistance, when an antagonist induces defense responses within

the host plant; resulting in resistance to diseases through reducing, restrict-
ing, or blocking the ability of the pathogen to produce disease. This can
happen by prevention of infection, restricting pathogen growth within the
plant, or some other mechanism of defense activation within the plant.
Induction of systemic resistance may involve activation of multiple
306 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

potential defense mechanisms, including increased activity of chitinases,

beta 1,3-glucanases, peroxidases, and other pathogenesis related (PR)
proteins, and accumulation of antimicrobial compounds such as phyto-
alexins and formation of protective biopolymers such as lignin, callose,
and hydroxyproline-rich glycoproteins (Kloepper et al., 1996). Examples
of this interaction are presented by certain rhizobacteria (such as strains
of P. fluorescens) that can induce systemic resistance to a number of dif-
ferent pathogens (Zhou and Paulitz, 1994; Liu et al., 1995). In cucumber,
induction of resistance by a single rhizobacterial strain provided protec-
tion against several different pathogens, including fungi, bacteria, and
viruses as well as reduction in insect feeding (Kloepper et al., 1996).
Protection of crops by induced resistance has also been demonstrated in
the field for some crops (Tuzum et al., 1992; Wei et al., 1996). Another
example of induced systemic resistance (ISR) is that incited by certain
nonpathogenic or avirulent strains of F. oxysporum, which are able to
parasitically colonize roots without causing disease and induce resistance
to Fusarial wilt in several crops (Leeman et al., 1995; Larkin et al., 1996).
Because there is generally an induction period of one to several days
required after exposure of the plant to the antagonist before ISR occurs,
this mechanism is most effective when the antagonist is applied prior
to exposure to the pathogen, such as with a seed or seedling treatment
operations. Although the mechanism of induction is unknown, a role for
salicylic acid has been confirmed in some ISR reactions.
Treatment of plants with exogenous salicylic acid induced PR protein
synthesis and enhanced resistance to a number of pathogens. In addi-
tion, endogenous salicylic acid levels rise specifically during resistance
response in plants (Chen et al., 1995).

21.6 DELIVERY OF BIOCONTROL PRODUCTS UNDER FIELD

CONDITIONS

One of the areas of biocontrol products research has the delivery system.
It is rather necessary to have an efficient economically viable mode of
application of biocontrol products in seed, soil, and seedling under field
condition.
Fungal Diseases of Medicinal and Aromatic Plants 307

21.6.1 SEED TREATMENT

Seed coating with biocontrol products has emerged as a feasible way of

delivering the antagonist for the management of plant diseases. For sow-
ing purpose, a powdered product of antagonist is used @ 5–8 gm powder/
kg seed, based on seed size and formulations of antagonist (Mukhopad-
hyay et al., 1992; Mukherjee and Mukhopadhyay, 1995; Vidhyasekaran
and Muthamilan, 1995). Trichoderma hamatum, T. harzianum, T. virens,
and T. viride are effective seed protectants against Pythium spp. and Rhi-
zoctonia solani (Mukherjee and Mukhopadhyay, 1995). A large number
of seed, seedling, root, stem, foliar, and panicle diseases have been sup-
pressed by seed treatment with biocontrol products.

21.6.2 SEEDLING TREATMENT

Treatment of planting materials with beneficial microorganisms is becom-

ing increasingly important. Seedling roots can be treated with spore or cell
suspension of antagonists. This method is generally used for vegetable
crops, rice, etc., where transplanting is practiced. Seedlings of root should be
dipped in water suspension of the powdered product of antagonist @ 10 g/L
of water for 30 minutes in shady conditions. Root dipping in antagonist sus-
pension not only reduces disease severity but also enhances seedling growth
in rice, tomato, brinjal, chili, and capsicum (Mishra and Sinha, 2000).

21.6.3 SOIL TREATMENT

Numerous attempts have been made to control several soil-borne diseases

by incorporating natural substrates colonized by a powdered product of
antagonists into field (Vidhyasekaran and Muthamilan, 1995; Sen, 2000).
For the treatment of one hectare area, 2.5 to 3 kg powdered products of
antagonist can be mixed in 70–100 kg vermicompost or well-rotted farm
yard manure (FYM) or well-rotted press-mud in about 7–10 days, and
when the substrate is fully colonized, it should be incorporated into the
soil before sowing.
308 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

21.7 CONCLUSION

Biocontrol may function through competition, antibiosis, parasitism,

induced résistance, or a combination of these mechanism. These various
mechanisms may control disease by the reduction or inhibition of the patho-
gen inoculums, protection of the infection court (prevention of pathogen
infection), or by limiting disease development after pathogen infection.
Each organism, mechanism, and activity has different traits, conditions,
and requirements associated with it that are essential to biocontrol activity.
Other areas of research that may be critical for further development and
improvement of biological control systems are the development and use of
multiple antagonists having different mechanism of action, incorporating
the influence of host plant on microbial communities to enhance biocon-
trol organism’s activity, better integration of biocontrol with other disease
control strategies, and improved formulations and delivery systems to pro-
vide the optimal starting point.

ACKNOWLEDGMENT

The authors are thankful to the authorities of Sardar Vallabhbhai Patel

University of Agriculture and Technology, Meerut, as well as to Mizoram
University, Aizawl, for providing various facilities during the research.

KEYWORDS

•• bio-control
•• biological management
•• formulation
•• fungal
•• medicinal plants
•• microbial
Fungal Diseases of Medicinal and Aromatic Plants 309

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 PART III

HORTICULTURE FOR
HEALTH AND NUTRITION
CHAPTER 22

BROAD SPECTRUM OF INDIAN

PEPPERMINT OIL AGAINST
DISEASE-CAUSING HUMAN
BACTERIAL PATHOGENS
AWADHESH KUMAR,1 AMRITESH C. SHUKLA,1
and ANUPAM DIKSHIT2
1
Department of Horticulture, Aromatic and Medicinal Plants,
Mizoram University, Aizawl–796004, India,
E-mail: [email protected]
Biological Product Lab, Department of Botany,
2

University of Allahabad, Allahabad–211004, India

CONTENTS

Abstract.................................................................................................. 315
22.1 Introduction................................................................................. 316
22.2 Materials and Methods................................................................ 318
22.3 Results and Discussion............................................................... 323
22.4 Conclusions................................................................................. 331
Acknowledgments ................................................................................. 331
Keywords............................................................................................... 332
References.............................................................................................. 332

ABSTRACT

The fresh leaves of Mentha piperita L. (Lamiaceae) were collected, and

after proper shade drying, the essential oil was extracted through hydro-
316 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

distillation using Clevenger’s apparatus. The gas chromatography-mass

spectroscopy (GC-MS) analysis of the oil showed that menthol (37.20%)
and menthone (22.52%) were the major active constituents. In the pres-
ent work, its antibacterial efficacy was screened with the disc diffusion
method, where the maximum zone of inhibition was found against Sal-
monella typhi (17 ± 0.1 mm). Further, the in vitro efficacy of oil was
validated against bacterial pathogens such as Escherichia coli (ATCC
25922), Proteus vulgaris (MTCC 1771), Salmonella typhi (MTCC 733),
Klebsiella pneumoniae (MTCC 109), and Pseudomonas aeruginosa
(MTCC 103) by using the broth microdilution method (CLSI). The mini-
mum inhibitory concentration (MIC) was recorded as 0.83 mg/mL, 1.19
mg/mL, 0.30 mg/mL, 0.61 mg/mL, and 1.09 mg/mL, respectively. Fur-
thermore, the oil showed toxicity against heavy inoculum density, quick
killing activity, broad antimicrobial spectrum, thermostability, and long
shelf-life. The in vivo investigations of its active constituents will be
needed to perform in order to confirm the mechanism of action for curing
human diseases.

22.1 INTRODUCTION

Water and plants are the two main sources on the Earth to continue life.
They are directly linked to each other. Ancient data revealed that humans
as well as animals live properly settled near the banks of rivers, lakes and
other water resources, because they got drinking water easily. Wherever
the scarcity of any of them occurs, life becomes difficult. But as the
population grew rapidly, the fresh drinking water became polluted and
several fatal bacterial, viral, protozoans, nematodal, and fungal diseases
emerged. This is due to rapid pace of urbanization, which increased the
demand of infrastructure for better livelihood (Kumar, 2011). Presently,
bacterial pathogenic diseases are emerging from unsafe drinking water.
Further, consumption of this contaminated water can cause several ill
effects in human beings such as the very common bacteria like Esche-
richia coli, Vibrio cholerae, Salmonella typhi, Salmonella typhimurium,
Shigella dysenteriae, Staphylococcus aureus, Pseudomonas aeruginosa,
Proteus vulgaris, Klebsiella pneumoniae, etc., are responsible for a
variety of diseases like cholera, typhoid, dysentery, bacillary dysentery,
Broad Spectrum of Indian Peppermint Oil 317

vomiting, urinary tract infection, and other gastrointestinal symptoms

(Kumar et al., 2012). Every day, even though the pharmaceutical indus-
tries are producing a number of new antimicrobial drugs, the microor-
ganisms have developed resistance against these drugs. It is because
the bacteria have the genetic ability to transmit and acquire resistance
to drugs used as therapeutic agents (Nascimento et al., 2000). On the
contrary, the revival of interest in plant-derived drugs is mainly due to
the current widespread belief that “green medicines” are safe and more
dependable than the costly synthetic drugs that have adverse side-effects
(Nair and Chanda, 2007). India, one of the developing countries, has
a gift of nature that the very rich botanical wealth and a large num-
ber of diverse types of medicinal plants grow wildly in every corner of
country. Hence, different forms of botanicals such as essential oils (EOs)
and plant extracts have been used to cure specific ailments from ancient
times (Bhattacharjee, 1998; Kumar et al., 2012).
According to the World Health Organization (WHO) report, about 80%
of the world’s population relies on traditional medicine, and the major-
ity of the traditional therapies involves the use of plant extracts or their
active constituents (WHO, 1993). Further, in another report, many species
of mint have already been exploited by our forefathers, and peppermint
itself has been used for 250 years ago (Hornok, 1992). The genus Mentha
of family Lamiaceae is composed of 19 geographically widespread spe-
cies and 13 named hybrids (Chambers and Hummer, 1994). M. piperita
commonly known as peppermint is a nonnative herbaceous and perennial
plant; the plant height reaches up to 80–100 cm (40 inches) with four-sided
stem. The leaves are stalked opposite and toothed. The flowering time
is from August to October; flowers are hermaphrodite (have both male
and female organs) and pollinated by insects. The flowers are irregular in
shape; they are pinkish or purplish (Kirtikar and Basu, 1972). Leaves con-
tains about 0.5–6% volatile oil commonly known as peppermint oil which
is composed of 50–78% active constituents dominated by monoterpenes,
mainly menthol, menthone and their derivatives, e.g., isomenthone, neom-
enthone, acetylmenthol, pulegone, and menthofuran. Majority of them are
widely in use as conventional medicine for antispasmodic, carminative,
refrigerant, stimulant and diuretic and antiseptic purposes (Edris et al.,
2003); their essential oils are using in chewing gums, alcoholic beverages,
318 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

cosmetics, perfumes, toothpastes and mouthwashes (Baytop, 1984). It is

one of the major constituent. Menthol is used in medicine for stomach dis-
orders and in ointments for headache, whereas infusion of leaves is used
in indigestion and rheumatic pains (Nair and Chanda, 2007). Besides, the
plants are also in use in salads, spice, and mint herbage for wool dyeing
(Leung and Foster, 2003).
Its active constituent menthol is used in medicine for stomach dis-
orders and in ointments for headache. The infusions of leaves are used
in indigestion and rheumatic pains. Mentha piperita oil, particularly as
an inhalant, relieves nausea and respiratory problems and aids diges-
tion. Digestive, carminative, respiratory, anti-inflammatory, cooling
and muscle relaxant are peppermint oil properties. The Indian Materia
Medica also proved that the various infusions of leaves of M. piperita
are used in cases of vomiting, gastric colic, cholera, diarrhea, flatulence,
weak digestion, hiccup, and palpitation of the heart (Nadkarni, 1976).
Since the plant M. piperita is well known in India, it was selected for
detailed in vitro antibacterial activity against Escherichia coli, Proteus
vulgaris, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Salmo-
nella typhi, the bacterial pathogens causing severe illnesses/diseases in
human beings.

22.2 MATERIALS AND METHODS

22.2.1 THE ESSENTIAL OIL

The plant material (fresh leaves) was collected from the Pratapgarh district
of Uttar Pradesh, India. The essential oil was extracted from the leaves of
Mentha piperita through the hydrodistillation method using Clevenger’s
apparatus (Clevenger, 1928). The extraction of oil was carried out continu-
ously on a heating mantle at the temperature of 30°C–50°C until no further
oil was extracted. The excess water content of the oil sample was removed
through anhydrous sodium sulfate (Na2SO4), and after filtration, it was
stored in a dark tightly closed bottle at +4°C for further investigations.
The yield of the obtained essential oil was about 0.63% based on the dry
weight of plant leaves.
Broad Spectrum of Indian Peppermint Oil 319

22.2.2 GAS CHROMATOGRAPHY (GC)

The gas chromatography (GC) analysis of the oil was carried out by Per-
kin-Elmer Auto XLGC and a Nucon gas chromatograph model 5765, both
equipped with an FID using two different stationary phases, PE-5 (60 m
× 0.25 mm; 0.25 µm film coating) and BP-20 (coated with a Carbowax
20 M, 30 m × 0.32 mm × 0.25 µm film thickness), fused silica columns,
respectively. Hydrogen was the carrier gas at 1.0 mL/min. The column
temperature programming was from 70–250°C at 3°C/min (for PE-5) and
from 70°C to 230°C at 4°C/min (for BP-20). The injector and detector
temperatures were 200 and 230°C on BP-20 and 220 and 300°C on PE-5
column, respectively. The injection volume of the sample was 0.02 μL
neat and the split ratio was 1:30.

22.2.3 GAS CHROMATOGRAPHY/MASS SPECTROMETRY

(GC/MS) ANALYSIS

The gas chromatography-mass spectrometry (GC–MS) analysis of the oil

was performed on a Perkin-Elmer Turbomass Quadrupole mass spectrom-
eter fitted with an Equity-5 (Perkin-Elmer) fused silica capillary column
(60 m × 0.32 mm; 0.25 µm film coating). The column temperature was pro-
grammed 70°C, initial hold time of 2 min, to 250°C at 3°C/min with a final
hold time of 3 min; the analysis was carried out using helium as the carrier
gas at a flow rate of 1 mL/min. The injector and source temperatures were
250°C. The injection volume was 0.06 μL neat with a split ratio 1:30. The
MS readings were taken at 70 eV with an EI source with mass range of m/z
40–400. The separated components were identified tentatively by match-
ing with EI-MS results of National Institute of Standards and Technology
(NIST and WILEY), 8th edition and NBS mass spectral library data. The
quantitative determination was carried out based on peak area integration.

22.2.4 PATHOGENIC BACTERIAL CULTURES

The specific bacterial cultures Escherichia coli (ATCC 25922), Proteus

vulgaris (MTCC 1771), Pseudomonas aeruginosa (MTCC 103), Kleb-
320 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

siella pneumoniae (MTCC 109), and Salmonella typhi (MTCC 733)

procured from the Microbial Type Culture Collection (MTCC), Chan-
digarh, India, and the American Type Culture Collection (ATCC), Ban-
galore (local center in India), were used in this study. The fresh culture
of each test organisms was periodically maintained on Muller Hinton
agar (MHA) slants and kept at 35 ± 2°C in a BOD incubator for further
analysis.

22.2.5 IN VITRO ANTIBACTERIAL INVESTIGATION

22.2.5.1 Preparation and Standardization of Inoculum

Standardized inoculum of each bacterium, i.e., 1×107 cells CFU/mL (col-

ony forming units) with 0.5 McFarland standard was taken for antimicro-
bial assay.

22.2.5.2 Disc Diffusion Method (Zone of Inhibition)

For screening of antibacterial properties of essential oil of M. piperita,

the disc diffusion method was used (Bauer et al., 1966). Sterilized Petri
plates were pre-seeded with 15 mL of Muller Hinton Agar medium and 0.1
mL of bacterial suspension containing 1 × 107 CFU mL−1 inoculum was
uniformly spread over the media to form a lawn of the culture. The stock
solution of various concentrations (100 mg, 50 mg, and 25 mg) of 50%
essential oil was prepared in dimethyl sulfoxide solvent (DMSO). Sterile
Whatman paper discs (10 mm diameter) were soaked in these concentra-
tions in such a manner so that ultimate amount in each disc was 100 μL, 50
μL, and 25 μL, respectively. After 5 minutes of proper soaking, the discs
were place over the bacterial lawn and incubated for 24 h at 35 ± 2°C. In
set of controls, ampicillin 10 µg (Hi-Media disc) was used as negative
control, while discs soaked in sterile distilled water were placed on the
lawn as a positive control. Further, the inhibited area called as “zone of
inhibition” of growth was measured in mm (millimeter) and compared
with the standard reference antibiotics. The zone of inhibition was calcu-
lated using the following formula:
Broad Spectrum of Indian Peppermint Oil 321

T −D
W=
2

where: W – diameter of clear zone of inhibition; T – total diameter of

including disc and clear zone; D – diameter of the test Disc.
All measurements were calculated in mm.

22.2.5.3 Evaluation of Antibacterial Activity MIC (Minimum

Inhibitory Concentration)

Further, the actual determination of antibacterial activity of the essential

oil was investigated using broth micro-dilution method CLSI (NCCLS,
2003). Out of various stock solutions, 50 mg/mL was taken for measuring
the minimum inhibitory concentration (MIC). The experiment was con-
ducted in sterile 96-well microtiter plates with lids (SPL) using ELISA
reader (Spectramax Plus384, Molecular Devices Corporation, USA). The
experiment was started by filling the column 4 to 11 with 180 μL MHB
media and 20 μL of essential oil, so that final volume 200 μL can be made.
Further, serial dilutions were made from column 4 to 11 and excess broth
(100 μL) was discarded from column 11. Now, in each well (from column
4 to 11), 100 μL of inoculum was added. In this way, column 1 (which con-
tains 200 μL media along with antibiotic and inoculum) served as negative
control; column 2 (contains 200 μL sterile MHB media) served as media
control; column 3 (contains 190 μL MHB media with 10 μL essential oil)
served as drug control; and column 12 (which contains 200 μL inoculum
but no essential oil) served as a positive control. Furthermore, mixtures
of the wells were mixed thoroughly, and the microplates were incubated
at 35 ± 2°C for 18–24 h. After the incubation, microbial growth inhibi-
tion was evaluated by measuring absorbance at 492 nm. Each experiment
was repeated in triplicate. The MIC was defined as the lowest concentra-
tion of essential oil, showing no visible bacterial growth after incubation
time. However, bactericidal concentration (MBC) was the lowest inhibi-
tory concentration at which the replicates failed to grow, even after re-
inoculating on or in the fresh culture media, i.e., MHB/MHA/NA (Kumar
et al., 2011, 2012).
322 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

22.2.5.4 Effect of Inoculum Density

The effect of inoculum density on MBC of the essential oil against the
test pathogens was determined using the method of Shukla et al. (2013).
Inoculum of each bacterium was prepared at CFU/mL of 1 × 103, 1 ×
105, 1×107, and 1 × 109. The culture plates containing essential oil at their
respective MBCs were prepared for the treatment set. However, in case of
control set, sterilized water was used instead of the essential oil. Further,
observations were recorded after the incubation period of 18–24 h.

22.2.5.5 Minimum Killing Time (MKT)

The MKT of the essential oil of M. piperita at the respective MICs and
MBCs against the test pathogens were also determined using the microti-
ter plate assay. All the wells were filled with 100 μL culture media (MHB).
Again, 80 μL more media was added to the wells of the 4th column. The
plant essential oil (20 μL) was added to the wells of the 4th column and
serially diluted up to the 11th column using a multichannel micropipette.
Then, 100 μL of inoculum was added to all the wells, except the 1st, 2nd,
and 3rd columns. The wells corresponding to the MICs, and MBCs were
marked. Prior to incubation, the loop was touched over the wells contain-
ing MICs, and MBCs essential oil and drag over the MHA/NA plates,
gently. The microtiter plate and the MHA/NA plates were kept for incuba-
tion. Further, after 5 min, 30 min, 1 h, 6 h, 12 h, 18 h, and 24 h, the same
process was repeated. The MHA/NA plates were kept for incubation at
35 ± 2°C up to 18–24 h. Observations were made after the incubation
period (Shukla et al., 2013).

22.2.5.6 Broad Antimicrobial Spectrum

To test the range of spectrum of the oil of M. piperita, the minimum bac-
tericidal concentration was subjected for investigation against 10 other
bacterial pathogens, namely Bacillus cereus (MTCC 430), Bacillus sub-
tilis (MTCC 441), E. coli (MTCC 723), Vibrio cholerae (MTCC 3906),
Salmonella enterica (MTCC 3858), Lactobacillus acidophilus (MTCC
Broad Spectrum of Indian Peppermint Oil 323

447), Staphylococcus aureus (MTCC 96), Shigella flexneri (MTCC 1457),

Yersinia enterocolitica (MTCC 859),,and Vibrio vulnificus (MTCC 1145),
which were available in the laboratory (Kumar et al., 2013; Shukla et al.,
2013).

22.2.5.7 Thermostability and Shelf-Life

Effect of some physical factors, viz. temperature (40°C, 60°C, and 80°C)
and time of storage on efficacy of the oil, at MBC, was also determined
(Kumar et al., 2013; Shukla et al., 2013). Samples of the oil (1.0 mL) in
small vials were exposed at 40°C, 60°C, and 80°C in hot water bath for
30 min separately; and their efficacy was observed against their respec-
tive bactericidal concentration. Moreover, the essential oil of M. piperita
showed the efficacy, even after 3 years, against the same test pathogens.

22.2.5.8 Statistical Analysis

The data of zone of inhibition of essential oil obtained against different

bacterial pathogens were statistically analyzed with three-way classifica-
tion and tested by F-test where significance was evaluated at 5% level.
Microsoft excel was used to analyze the standard deviation/error, etc.

22.3 RESULTS AND DISCUSSION

22.3.1 CHEMICAL COMPOSITION OF ESSENTIAL OIL

The present investigation was performed to determine the various active

constituents and antibacterial activity of M. piperita, a traditional medici-
nal plant in India, against some common infectious bacterial pathogens.
The percentage of yield of obtained essential oil from the leaves was
0.63%, though the plant has strong aromatic flavor. Its GC/MS analysis
showed that the essential oil has 56 constituents, where menthol (37.20%)
along with menthol stereoisomer’s (+ neomenthol and + isomenthol) and
menthone (22.52%) were the major constituents including the isomen-
324 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

thone (4.70%), 1,8-cineole (4.68%), and methyl acetate (4.18% and Pule-
gone (3.70%) and Limonene (0.8%) as the minor constituents. The other
monoterpenes such as β-caryophyllene, piperitone, pinene, eugenol, car-
vone, linalool, α-phellendrene, ρ-menthane cadinene, and dipentene were
observed in trace amounts.

22.3.2 ANTIBACTERIAL ACTIVITY

Later the antibacterial screening was made by using disc diffusion tech-
nique, and it was found that the essential oil possessed high antibacterial
activity. The higher concentration 100 mg/mL exhibited the maximum
zone of inhibition followed by 50 mg/mL and 25 mg/mL conc. Among
all the tested bacterial pathogens, the maximum zone of inhibition was
recorded against the S. typhi (17 ± 0.1 mm); however, the least susceptible
was P. vulgaris (8 ± 0.12 mm). From the zone of inhibition results, it was
found that the efficacy of peppermint oil increased with increasing con-
centration. On the other hand, E. coli (13 ± 0.2 mm) and K. pneumoniae
(12 ± 0.32 mm) showed more or less very similar inhibition with respect
to various concentrations. Further, the standard ampicillin (antibiotic) was
compared against the essential oil; it was showed the largest zone of inhi-
bition (31 ± 1.24). There was no growth of bacteria in the positive control
(Table 22.1). The same values are also indicated in the form of graph in
Figure 22.1.

TABLE 22.1 Zone of Inhibition of Peppermint Oil Against Bacterial Pathogens

Extract Con- Bacterial Pathogens/Diameter of Zone of Inhibition (mm±S.E.)
centration E. coil P. vulgaris S. typhi K. pneumoniae P. aeruginosa
(µg/mL)
25 3±2.3 1±1.2 6±0.7 2±0.22 1±0.14
50 8±0.1 3±0.5 13±0.31 7±0.02 4±0.56
100 13±0.2 8±0.12 17±0.1 12±0.32 10±1.2
Standard 26±0.72 27±2.5 37±0.1 27±3.6 31±1.24
(Ampicillin)
Control (Only NG NG NG NG NG
extract)
mm – millimeter; S.E. – Standard Error; NG – No growth.
Broad Spectrum of Indian Peppermint Oil 325

FIGURE 22.1 (See color insert.) Effect of essential oil showing zone of inhibition
against bacterial pathogens.

22.3.3 DETERMINATION OF MINIMUM INHIBITORY

CONCENTRATION (MIC) AND MINIMUM BACTERICIDAL
CONCENTRATION (MBC)

The plant essential oils of M. piperita results showed a wide variation in

the antibacterial properties. The validation of inhibitory action, which was
recorded in zones of inhibition at higher concentration in the disc diffu-
sion method, was done through the broth microdilution method. Among
all the tested bacterial pathogens such as Escherichia coli (ATCC 25922),
Proteus vulgaris (MTCC 1771), Pseudomonas aeruginosa (MTCC 103),
Klebsiella pneumoniae (MTCC 109), and Salmonella typhi (MTCC 733),
the most potent inhibitory activity of peppermint oil was found for S. typhi
with the MIC of 0.30 mg/mL as shown in Table 22.2. It was least effec-
tive for P. vulgaris; however, for E. coli and K. pneumoniae, the efficacy
was almost similar, which validated the observation of the disc diffusion
method. The antibacterial activity was found to progressively increase
with increasing concentration of essential oil. The lowest effective dose of
the oil was observed at 0.3125 mg/mL. Based on these various concentra-
tions of the essential oil, the minimum inhibitory concentration (MIC) of
the oil was recorded as 0.61 mg/mL, 1.19 mg/mL, 0.30 mg/mL, 0.69 mg/
mL, and 1.09 mg/mL, respectively (Table 22.2). Besides this, IC50 (50%
326 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

inhibitory concentration) values of the oil against the test pathogens was
also recorded as 0.28 against Escherichia coli; 0.54 against Proteus vul-
garis; 0.14 against Salmonella typhi; 0.31 against Klebsiella pneumoniae,
and 0.58 against Pseudomonas aeruginosa (Table 22.2). Further, their
average percent (%) growth inhibition was also recorded in Figure 22.2.

22.3.4 BROAD-SPECTRUM PROPERTIES OF ESSENTIAL OIL

Furthermore, the oil’s toxicity was also determined against heavy inocu-
lum density as well as other different gram-negative and gram-positive

TABLE 22.2 Antimicrobial Activity of Essential Oil of M. piperita (Peppermint Oil)

S. No. Name of Water Borne Antimicrobial Activity
Bacterial Pathogens IC50 MIC MBC
1. Escherichia coli 0.28 0.61 0.625
2. Proteus vulgaris 0.54 1.19 1.25
3. Salmonella typhi 0.14 0.30 0.325
4. Klebsiella pneumoniae 0.31 0.69 0.625
5. Pseudomonas aeruginosa 0.58 1.09 1.25
IC50 = 50% inhibitory conc.; MIC = minimum inhibitory concentration.

FIGURE 22.2 (See color insert.) Average percent growth inhibition of peppermint oil
against bacterial pathogens.
Broad Spectrum of Indian Peppermint Oil 327

bacterial strains. The peppermint oil showed quick killing activity, ther-
mostability, and long shelf-life (Tables 22.3 and 22.4).
The active principles of the medicinal plants are divided chemically
into a number of groups, among which are alkaloids, volatile essential oils,
phenols and phenolic glycosides, resins, oleosins, steroids, tannins, and
terpenes (Habtmaiam et al., 1993). Kazemi et al. (2012) reported that the
main components in M. piperita oil were menthol, limonene, 1,8-cineole,
sabinene, menthyl acetate, and menthone and in M. spicata oil were car-
vone, menthol, limonene, and menthone, which proved the finding of pres-
ent study. Antibacterial and antifungal activity of these oils and their
components were assayed against a variety of human pathogenic bacteria
and fungi. In every traditional system of medicine, the peppermint and its
oil have been used as an antispasmodic, aromatic, and antiseptic and in the

TABLE 22.3 Detailed In Vitro Investigation of the Peppermint Oil against the Tested
Bacterial Pathogens
Properties Tested Bacterial Pathogens
studied E. coil P. vulgaris S. typhi K. pneumoniae P. aeruginosa
Minimum Inhibitory Concentration
MBC 0.625 mg/ 1.25 mg/ 0.3125 mg/ 0.625 mg/mL 1.25 mg/mL
mL mL mL
Minimum Killing Time (MKT)
Pure oil 35 sec 55 sec 20 sec 40 sec 1 min
MBC 4:00h 5:00h 3:25h 4:25h 5:30h
Inoculum density (CFU/mL)
1×103– No Growth No Growth No Growth No Growth No Growth
1×105
1×107– No Growth No Growth No Growth No Growth No Growth
1×109
Thermostability (°C)
50°C– Effective Effective Effective Effective Effective
100°C
Effect of Storage (in months)
12 months +++ +++ +++ +++ +++
24 months +++ +++ +++ +++ +++
36 months +++ ++ +++ ++ ++
328 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

TABLE 22.4 Toxicological Spectrum of Peppermint Oil against Other Common

Bacterial Pathogens
S. No. Names of Water Lethal concentration Hyper lethal
Borne Bacterial (1.25 mg/mL) concentration (3.2 mg/mL)
Pathogens
1. Bacillus cereus 100C 100C
2. Bacillus subtilis 100 C
100C
3. E.coli (MTCC 723) 100S 100C
4. Vibrio cholerae 100 S
100C
5. Salmonella enterica 100C 100C
6. Lactobacillus aci- 100S 100C
dophilus
7. Staphylococcus aureus 100C 100C
8. Shigella flexneri 100 S
100C
9. Yersinia enterocolitica 100S 100S
10 Vibrio vulnificus 100 S
100C
c – cidal concentration; s – static concentration.

treatment of cancers, colds, cramps, indigestion, nausea, sore throat, and

toothaches (Briggs, 1993). In the in vitro experiment, it was seen that the
peppermint oil possesses antibacterial activity; moreover, its different
commercial preparations exhibit various activities (Lis-Balchin et al.,
1997). Therefore, the plants are important source of potentially useful
structures for the development of new chemotherapeutic agents. Many
reports are available on the antiviral, antibacterial, antifungal, anthelmin-
tic, antimolluscal and anti-inflammatory properties of the plants (Stepa-
novic et al., 2003). The MIC values observed in the present study showed
the correlation with the studies of Fabio et al. (2007), where the MIC val-
ues of peppermint oil was reported to range between 1.25 μL/mL (E. coli)
and 3.125 μL/mL (K. pneumoniae); the present study results are also in
accordance with finding of Sartoratto et al. (2004), who reported that
gram-positive organisms were inhibited by peppermint oil at very low
concentrations. In another study, it was seen that C. albicans was the most
sensitive microorganism (MIC=0.312 mg/mL) to peppermint oil, followed
by S. epidermidis (MIC=0.625 mg/mL), B. cereus, S. typhimurium, E.
aerogenes, S. aureus, and E. coli (MIC = 1.25 mg/mL). However, P. aeru-
Broad Spectrum of Indian Peppermint Oil 329

ginosa with an MIC value of 5 mg/mL was more resistant that others
(Iscan et al., 2002). So, in comparison with experiment of Iscan et al.
(2002), the efficacy of oil was found to be more potent in the present study
as mentioned in Table 22.3. The peppermint oil is non-toxic and non-irri-
tant in low dilutions, but sensitization may be a problem due to the men-
thol content, and due to this, it can cause irritation to the skin and mucus
membranes and should be kept well away from the eyes. It should also be
avoided during pregnancy and should not be used in children below 7
years (German Commission E Monographs (Phytotherapy), 1990). Some
reports on peppermint oil also suggested that, in any form, the oil is not
recommended for those with hiatal hernia, gallbladder disease, or while
pregnant or nursing. Its overdose symptoms are slow breathing, rapid
breathing, abdominal pain, diarrhea, nausea, vomiting, blood in urine, no
urine production, convulsions, depression, dizziness, twitching, uncon-
sciousness, uncoordinated movement, and flushing (http://www.drugs.
com/enc/peppermint-oil-overdose). Although the essential oils have vari-
able composition of components, some of which are actually toxic to
human, insects and plants if ingested in large quantities; even though, a
large number of active principle responsible for such activities are used in
the development of drugs for the therapeutic use in human being (Gubaron,
2000; Talebi et al., 2006). But the difference between antibacterial activi-
ties of essential oils may be related to the concentration and nature of
contents, the respective composition, the functional groups, the structural
configuration of the essential oil against bacterial pathogens, their possible
synergistic interaction, and ecological/plant growth factors (Chang et al.,
2001). Besides, seasonal variation, especially harvest time, also affects
biological activities of oils (Kizil, 2010; Mimica-Dukic et al., 2003). The
hydrophobicity, an important characteristic of essential oils which enables
them to associate in the lipids of the bacterial cell membrane and mito-
chondria, disturbing the structures and rendering them more permeable
leading to leakage of ions and other cell contents which decreased the
intracellular ATP pool of bacterial or fungal cell and also increased extra-
cellular ATP, indicating disruptive action on the cytoplasmic membrane
(Kumar et al., 2012; Sikkema et al., 1994). The differences in MIC values
of bacteria may be related to differential susceptibility of the bacterial cell
wall, which is the functional barrier, to minor differences present in the
330 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

outer membrane in the cell wall composition (Zhao et al., 2001). Accord-
ing to Jeyakumar et al. (2011), the diffusion of oil was found to be less as
compared with the broth microdilution method; this is due to the insuffi-
cient penetration of oil in the agar medium, and hence, concentration of oil
was required to inhibit the growth of pathogenic bacteria in the agar well/
disc diffusion method. Moreover, the phylogenetic variations also play an
important role in the differences between the MIC of the oil (Kumar et al.,
2011, 2012). A previous study showed the toxic effect of garlic oil was
attributed to sulfur and allicin components, which had ability to react with
SH groups of enzymes and change their properties (Mitchell, 1980). Lit-
erature also revealed that gram-negative bacteria are more resistant to the
essential oils present in plants (Kumar et al., 2011; Smith-Palmer et al.,
1998). The region behind that the cell wall of gram-negative bacteria
essentially contains lipopolysaccharides (LPS), which creates a barrier to
accumulate the oil on their cell surfaces. Further, in another study, the
active constituent of the oil, menthol, has bactericidal effects and a spicy
odor (Hornok, 1992). The antimicrobial activity of peppermint oil is due
to the presence of terpenoides menthol, 1-8 cineole, methyl acetate, men-
thofuran, isomenthone, limonene, β-pinene, germacerene-d, trans-sabi-
nene hydrate, and pulegone (Sartoratto, 2004). Furthermore, some essential
oils with the same common name may be derived from different plant
species. Secondly, the method used to assess antimicrobial activity, strains,
and the choice of the test organisms varies between different studies. Men-
thol has also been reported to have antimicrobial activity (Iscan et al.,
2002). This may be because of phenolic nature of the menthol, and com-
ponents that are phenolic in nature generally decreased the intracellular
ATP pool of bacterial cell and also increased extracellular ATP, indicating
disruptive action on the cytoplasmic membrane (Helander et al., 1998;
Kumar et al., 2011, 2012). The interaction of peppermint oil and menthol
with the antibiotics was studied on the same bacterial strain with the
checkerboard method, and the antiplasmid activity of peppermint oil and
its main constituent menthol was observed; this implies that menthol-con-
taining substances are potential agents that could eliminate the resistance
plasmids of bacteria. The compound preferentially kills the plasmid-con-
taining bacteria due to their increased sensitivity to menthol (Shrivastava,
2009). This was hypothesized through some research data that indicated
Broad Spectrum of Indian Peppermint Oil 331

irritation and toxicity caused by eugenol, menthol, and thymol; hence, the
cytotoxicity study of these compounds suggested that gum irritation may
be related to membrane lysis and surface activity and that tissue penetra-
tion may be related at least partly to membrane affinity and lipid solubility
(Manabe et al., 1987). On the other hand, the experiment clearly picturized
that, the cells appeared oblong; edges become abnormal, triangular, elon­
gated and incapable of formation of septum in dividing. Cell wall dis-
rupted and exhibited thickened in some parts and breakdown in other
which leading to leakage of cytoplas­mic materials. Therefore, these varia-
tions in the activity between different organisms were observed, and hence,
the oil was highly effective against S. typhi but was least effective against
P. vulgaris; however, for E. coli and K. pneumoniae, the bactericidal effi-
cacy was almost similar at the same concentration (0.625 mg/mL).

22.4 CONCLUSIONS

The use of essential oils of herbs and spices are playing more important
role in food and other beverages because of their specific taste. Likewise,
the medicinal plant Mentha piperita was chosen for the present work, as
this plant is traditionally known and has continuously been in practice
of use as a therapeutic agent against a variety of diseases. The system-
atic studies conducted on peppermint oil indicated the immense potential
against the infectious bacterial pathogens. Moreover, the present finding
provides enough experimental data like heavy inoculum density, quick
killing activity, broad antimicrobial spectrum, thermo stability, long-shelf
life, etc., through both the techniques. Further in vivo experiments can
help to explore the actual mechanism of action of peppermint oil. There-
fore, it is recommended to check the toxicity at lower doses for medicinal
formulation in the favor of human welfare.

ACKNOWLEDGMENTS

The author is thankful to the authorities of the Mizoram University, Aizawl,

as well as the Biological Product Laboratory, Department of Botany, Uni-
versity of Allahabad, for providing the research facilities.
332 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

KEYWORDS

•• antibacterial
•• CLSI
•• GC-MS
•• hydrodistillation
•• thermostability

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CHAPTER 23

UNDEREXPLOITED VEGETABLES IN
NORTH EASTERN INDIA: A GATEWAY
TO FOOD SECURITY
B. LALRAMHLIMI,1 AKOIJAM RANJITA DEVI,2 and KHAIDEM NIRJA3
1
Department of Vegetable Crops, 2Department of Spices and
Plantation Crops, Faculty of Horticulture, 3Department of
Agricultural Extension, Faculty of Agriculture, Bidhan Chandra
Krishi Viswavidyalaya, Mohanpur–741252, Nadia, West Bengal,
India, E-mail: [email protected]

CONTENTS

Abstract.................................................................................................. 335
23.1 Introduction................................................................................. 336
23.2 Importance for Promotion of Underutilized Vegetables............. 338
23.3 Potential Contribution of Underexploited Crops........................ 340
23.4 The Depletion of Bio-Diversity of Underutilized Vegetables:
A Major Concern......................................................................... 348
23.5 Conclusion and Recommendation.............................................. 348
Keywords............................................................................................... 349
References.............................................................................................. 349

ABSTRACT

Northeastern states of India are very diverse climatically, agro-ecologi-

cally, and ethnically. About 50% of wild edible plants wealth is located in
336 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

this region, which is also one of the richest reservoirs of different underex-
ploited vegetable crop species (Arora, 1997). Underexploited vegetables
play a crucial role in poor people’s livelihood and may have a signifi-
cant potential for commercialization in this region. Moreover, they also
possess several desired medicinal properties. Some underexploited veg-
etables in northeastern regions are rice bean (Vigna umbellate) and tree
bean (Parkia javanica Merr. Syn. P. Roxburghii), Manipur Loosestrife/
Kengoi (Lysimachia obovata) in Manipur, Chingit/Indian Pepper (Zan-
thoxylum rhetsa) and Anhling/Black nightshade (Solanum americanum)
in Mizoram, Indian pennywort (Centella asiatica), Zanthoxylum armatum
in Arunachal Pradesh, Chichiri (Monochoria hastate) in Tripura, and East
Indian Glory Bower (Clerodendrum colebrookianum) in Sikkim. Most of
them are very rich sources of vitamins, minerals, and other nutrients such
as carbohydrates, proteins, fats and phytochemicals that have anticancer
and anti-inflammatory properties, which confer many health benefits.
They have the potential to contribute to food security, nutrition, health,
income generation, and environmental services as they are adapted to mar-
ginal soil and diverse climatic condition. The high nutritional qualities
indicate that the cultivation and consumption of these crops may be help-
ful in overcoming the nutritional deficiencies predominant in many rural
areas of this region and boost the socioeconomic conditions. Owing to
various human activities, there is depletion of this biodiversity. The topic
has been taken up for harnessing diversity of these vegetables that have
enormous potential with much diversity for exploitation, their conserva-
tion, and food security.

23.1 INTRODUCTION

The northeastern (NE) region of India comprising eight states, namely

Arunachal Pradesh, Assam, Manipur, Meghalaya, Mizoram, Nagaland,
Tripura, and Sikkim, has vast physiographical variations, which have been
represented in six agro-climatic zones. The NE region is one of the richest
reservoir of genetic variability and diversity of different vegetable crops
(Asati and Yadav, 2004). The northeast India is a part of the Himalaya and
the Indo-Burma biodiversity hotspots in the world (Moa et al., 2008). Many
local vegetables that have the potential to contribute to food security are
Underexploited Vegetables in North Eastern India 337

produced in this area, and their knowledge on distribution of their genetic

diversity and use patterns is still largely limited. It is estimated that half
of India’s floristic diversity of higher plants of about 7150 species out of
a total of 15000 species is concentrated in this region. About 50% of wild
edible plant wealth is located in this region, which is also one of the rich-
est reservoirs of different underexploited vegetable crop species (Arora,
1997). With increasing population and consequent shortage of food grains
and vegetables, the collection and utilization of various types of unutilized
crops are considered very essential. The high nutritional qualities indicate
that the cultivation and consumption of these crops may be helpful in over-
coming the nutritional deficiencies predominant in many rural areas of
the country and boost the socioeconomic condition of the society (Chitta
Ranjan et al., 2013). The causes of malnutrition and hunger in our country
are not the scarcity of food but an inability to access the available food due
to poverty and negligence of important local vegetables. Underexploited
vegetables play a crucial role in poor people’s livelihood in the rural areas
as improved varieties of different vegetables do not reach these regions
and contribute significantly in the food of rural masses of the people;
these vegetables have a significant potential for commercialization in this
region. Moreover, they also possess several desirable medicinal properties
to curb diseases and malnutrition as they are rich sources of vitamins, min-
erals, and other nutrients such as carbohydrates, proteins, and fats. These
crops require proper studies to know their values and unknown potentials
as a means of improving livelihoods, especially in support of the rural poor
vegetable farmers of these regions. They have an unimaginable potential
to contribute to food security, nutrition, health, income generation, and
environmental services as they are adapted to marginal soil and adverse
climatic condition. However, the availability of most of these wild crops is
now depleting rapidly owing to various factors such as “Jhum/shifting cul-
tivation,” forest fire, construction of industries, felling of trees for timber
and other socio-economic anthropogenic activities in the area. Therefore,
it is extremely necessary to tap their potentials and conserve these regions
wealth and work out the ways for commercial-scale propagation of these
plant species. The lack of improved varieties, information on unutilized
crops and unavailability of capital for farmers makes the commercial-scale
production difficult. The farmers’ security is as important as food security
338 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

as they are the sole runner in the food production. Chief Minister’s Uzha-
var Patthukappu Thittam 2011, has been implemented in Tamil Nadu for
the security and welfare of the farmers.

23.2 IMPORTANCE FOR PROMOTION OF UNDERUTILIZED

VEGETABLES

The country’s population is increasing at the rate of 1.548%, and it is pro-

jected that it will be around 1256 million up to 2015 and 1331 million in
2020 (Pandey et al., 2014). The food requirement increases day by day.
Although India is the second largest producer of vegetables in the world,
the available vegetables are not sufficient to meet the requirement of the
people especially in rural areas. A large mass of people depend on local
vegetables for their livelihood to meet their daily vegetable requirement to
sustain life. Some underexploited tuber crops in the NE regions, like Colo-
casia, Alocasia, Yams, Cassava, and Sweet potato contributes to carbohy-
drate requirement of many people in this region. The state of Nagaland is
a remote tribal state in NE India and is very rich in floristic diversity. In
Nagaland, the angiosperm is represented by over 2500 species belong-
ing to 963 genera and 186 families (Deb and Imchen, 2008). Apart from
their use as source of food, some are important due to their medicinal
properties, vegetables, fibers, construction materials, dyes, etc., (Chitta
Ranjan et al., 2013). These plants/parts contribute significantly in the food
of rural masses of Nagaland. Fern (Diplazium esculentum) is grown wild
in this region and is eaten by the local tribals. Indian pennywort (Centella
asiatica) is an important unutilized leafy vegetable that has innumerable
medicinal properties. Colocasia esculenta/Taro is a tuber crop grown in
tropical areas as a vegetable food for its edible corm and secondarily as
a leaf vegetable. The local tribals of Arunachal Pradesh grow a vegetable
having red tomato-like fruits that are slightly bitter in taste and belong to
the genus Solanum (Rai et al., 2004). Bamboo shoot (Bambusa tulda) is
used to make fermented bamboo shoot which is eaten and cooked with
other vegetables and meat. Amaranth (Amaranthus caudatus, A. viridis,
A. lividus, A. retroflexus, and A. Spinosus) which has high vitamins and
mineral content is eaten as boiled stuff. Drumstick (Moringa oleifera) is a
Underexploited Vegetables in North Eastern India 339

highly nutritious plant whose whole parts are edible. In Manipur, another
kind of brinjal, having round fruit and intermediate in appearance between
tomato and brinjal, is grown. In the hilly areas, tree tomato (Cyphomandra
betacca), a perennial shrub producing red tomato-like vegetables, is also
grown. Tree tomato is consumed as delicious chutney when raw or after
roasting and peeling off the skin (Rai et al., 2004). Euryale ferox grown
in ponds and other water bodies of Madhubani, Bihar, is considered one
of the most viable sources of income of the local people (Pandey et al.,
2014). It is also an important source of income of local people in Manipur.
Bathua/Jimilsag (Chenopodium album) is eaten as boil leafy vegetable
and highly nutritious. Rice bean (Vigna umbellate) and Manipur Loose-
strife /Kengoi (Lysimachia obovata) are also very nutritious and important
vegetable. Tree bean (Parkia roxburghii G. Don.) is one of the most com-
mon of multipurpose tree species in the Manipur and Mizoram (Kumar
et al., 2002). In Mizoram, Chingit/Indian Pepper (Zanthoxylum rhetsa)
and Anhling/Black nightshade (Solanum americanum) are important local
leafy vegetable with many unknown medicinal properties. Sword bean
(Canavalia ensiformis) and Jack bean (Canavalia gladiata) are eaten as
salad and cooked with other vegetables. Bitter brinjal (Solanum torvum)
is sold in the market of Mizoram and is eaten as vegetable (Asati and
Yadav, 2004). Climbing wattle/Khanghu/Biswal/Agla bel (Acacia pen-
nata), which has a stinky leaf, is a delicious leafy vegetable in its season
and can be eaten as fried or boiled. A wide diversity of Rosella (Hibiscus
sabdariffa) is present in this region, which is eaten as boiled stuff. Tooth-
ache plant/Paracress (Spilanthes acmella) is marketed by local tribals as
a vegetable. Snake gourd (Trichosanthes cucumerina) is also an impor-
tant crop for this region. One of the interesting species of Vigna namely
V. vexillata is grown by the tribals of Tripura. It is a legume cum tuber
crop with much variation in edible tubers (Arora and Pandey, 1996). Chi-
chiri (Monochoria hastate) in Tripura is also an important underexploited
aquatic vegetable whose whole part is edible. The wild species Cucumis
hardwickii, the likely progenitor of cultivated cucumber, is found growing
in natural habitats in the foothills of Himalayas and NE region, particu-
larly Meghalaya (Asati and Yadav, 2004). Sophlong (Moghania vestita)
is sold in the market and eaten as raw with salt. Chow-Chow (Sechium
edule), a native of tropical America, is a very popular vegetable in the
340 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

region. Commonly called as squash, a wide diversity of this vegetable

grows abundantly without much care and attention. Chow-Chow produces
large starchy edible roots in addition to fruits (Rai et al., 2004). In Sikkim
and many parts of NE regions, East Indian Glory Bower (Clerodendrum
colebrookianum) is grown wild without proper care but is an important
crop in this region. Rhubarb (Rheum nobile) native of Himalayas, Sikkim,
Nepal is an important vegetable occurring in the alpine zone at 4000–4800
m altitude is also a rare and important crop in India (Anonymous d., 2015).
In Assam and many other regions, spine gourd (Momordica dioica) is a
very important underexploited vegetable. Kakrol (Momordica cochinchi-
nesis) and kartoli (M. dioica) are widely spread in Assam, the Garo hills
of Meghalaya (Ram et al., 2002). Many more underexploited vegetables
are in these regions that require deeper studies to know their importance,
their potentials to contribute to food security, and their immediate need for
promotions and improvements.

23.3 POTENTIAL CONTRIBUTION OF UNDEREXPLOITED

CROPS

23.3.1 FOOD SECURITY

Food security exists when all people, at all times, have physical and
economic access to sufficient, safe and nutritious food that meets their
dietary needs and food preferences for an active and healthy life (World
Food Summit, 1996). Later definitions added demand and access issues
to the definition. The four pillars of food security, namely food avail-
ability, food access, food utilization, and food stability, were stated later
(World Food Summit, 2009). Food availability decreases each year as
human population increases and the cultivated area decreases, collec-
tion and utilization of various types of unutilized crops are very much
important. Most of them are very rich sources of vitamins, minerals and
other nutrients such as carbohydrates, proteins and fats (Rai et al., 2005).
In our country, where problem of malnutrition is prevailing in general
and micronutrient malnutrition in particular, addressing the household
nutritional security is indispensable. A recent study indicates that intake
of micronutrients in daily diet is far from satisfactory and largely less
Underexploited Vegetables in North Eastern India 341

than 50% recommended dietary allowance (RDA) is consumed by over

70% of Indian population (Pandey et al., 2014). Northeast states com-
prising eight states, namely Arunachal Pradesh, Assam, Manipur, Megha-
laya, Mizoram, Nagaland, Sikkim, and Tripura, has a total population of
75,587,982 (Census of India, 2011). Vitamin A, iron, and zinc deficiency
when combined constitute the second largest risk factor in the global bur-
den of diseases; 330,000 child deaths occur every year in India due to
vitamin A deficiency; 22,000 people, mainly pregnant women, die every
year in India from severe anemia; 6.6 million children are born men-
tally impaired every year in India due to iodine deficiency; intellectual
capacity is reduced by 15% across India due to iodine deficiency; and
200,000 babies are born every year with neural tube defects in India due
to folic acid deficiency (Pandey et al., 2014). In northeast India, these
vitamins and minerals deficiencies are very prevalent. Considering the
above, it has become the need of the hour for local governments along
with researchers, farmers, and nutritionists to search for cheap, reliable,
and safe plant-based resources to overcome these deficiencies. Exploring
underexploited vegetables could be of high significance for food security,
meeting nutritional requirements, and agricultural development as well as
an efficient means for crop alternates and thus can effectively contribute
to overall improvement of a nation’s economy.

23.3.2 NUTRITION

The RDA per capita per day recommended by the Indian Council of Medi-
cal Research (ICMR) for adult males includes 475 g cereals, 125 g green
leafy vegetables, 80 g pulses, 100 g roots and tubers, 30 g fruits, and 75
g other vegetables. The adult females should have the same quantities of
vegetables, roots and tubers and fruits as recommended for the adult males
but less quantities of cereals (350 g) and pulses (70 g). Human nutrient
requirements vary with sex, age, weight, height, and physical activity. The
balanced diet should contain adequate energy source (calories) and nutri-
ents like proteins, carbohydrates, fats, vitamins, minerals, and essential
amino acids. The minimum amount of calories and nutrients required per
capita per day for adult male and adult female, respectively, are listed in
the following table.
342 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

Nutrients and calories Adult male Adult female

Calories 2800 kcal 2200 kcal
Protein 55 g 45 g
Calcium 400–500 mg 400–500 mg
Iron 20 mg 30 mg
Riboflavin 1.5 mg 1.2 mg
Carotene 3000 µg 3000 µg
Thiamine 1.4 mg 1.1 mg
Ascorbic acid 50 mg 50 mg
Folic acid 100 µg 100 µg
Vitamin B12 1.0 µg 1.0 µg

Underutilized vegetables have immense potential for contribution to

a particular pocket’s of food production because they are well adapted
to existing as well as adverse environmental conditions and are gener-
ally resistant to pests and pathogens. They are a cheap source of car-
bohydrates, proteins, fats, vitamins, and minerals. They are available
locally and moreover have many nutritional and medicinal benefits. To
supply the daily requirements of nutrients and minerals, these underuti-
lized crops are an important option. According to Pandey et al. (2014),
the free and total folic acid content of Amaranthus is 41.0 μg and 149.0
μg per 100 g edible portion, respectively. Colocasia also contains 16.0
μg free and 94.0 μg total folic acid per 100 g edible portion, and snake
gourd contains 7.5 μg free and 15.5 μg total folic acid per 100 g edible
portion. Cluster bean (guar) is also a rich source of folic acid with 50.0
μg free and 144.0 μg total per 100 g edible portion. Gupta et al. (2005)
have given the mineral contents and trace element contents of different
underutilized vegetables. Trianthema portulacastrum has mineral con-
tents of ash (2.29 g), calcium (52.0 mg), phosphorus (22.0 mg), potas-
sium (317.0 mg), sodium (16.0 mg), and magnesium (153.0 mg) per
100 g of edible portion. Centella asiatica has a high potassium content
of 345.0 mg, ash (2.06 g), calcium (174.0 mg), phosphorus (17.0 mg),
sodium (107.8 mg), and magnesium (87.0 mg). Celosia argentea has
high potassium content of 476.0 mg, ash (2.65 g), calcium (188.0 mg),
Underexploited Vegetables in North Eastern India 343

phosphorus (35.0 mg), sodium (240.6 mg), and magnesium (233.0 mg).
Boerhaavia diffusa has ash (2.91 g), calcium (330.0 mg), phosphorus
(27.0 mg), potassium (381.0 mg), and magnesium (167.0 mg). Digera
arvensis has a high calcium content of 506.0 mg, ash (3.54 g), phospho-
rus (63.0 mg), potassium (604.0 mg), and magnesium (232 mg). The
trace element contents of underutilized leafy greens such as Trianthema
portulacastrum are iron (4.16 mg), zinc (0.46 mg), copper (0.12 mg),
chromium (0.20 mg), and manganese (0.43 mg) per 100 g of edible por-
tion. Celosia argentea also contains iron (13.15 mg), zinc (0.49 mg),
copper (0.15 mg), chromium (0.153 mg), and manganese (0.27 mg) per
100 g of edible portion. Boerhaavia diffusa contains iron (7.83 mg), zinc
(0.44 mg), copper (0.22 mg), and chromium (0.040 mg) per 100 g of
edible portion. Centella asiatica has high iron content of 14.86 mg, zinc
(0.97 mg), copper (0.24 mg), and chromium (0.046 mg) per 100 g of
edible portion. These crops showed the mineral and nutrient contents of
different underutilized crops, which are higher than most of the crops
that are commercially grown. Winged bean has excellent nutritional
qualities and is particularly very rich in protein (Rao and Dora, 2002).
Jack bean seed has been promoted in developing nations as a potential
source of affordable and abundant protein. It has 29.0% protein content
(Adebowale and Lawal, 2004) (Table 23.1).

TABLE 23.1 Calories and Protein from Some Important Underexploited Vegetables
Vegetables Calories (kcal per Protein (grams/ Reference
100g fresh weight) 100grams fresh
weight)
Grain amaranth 371 14 Anonymous b. (2017)
Taro (cooked) 142 0.52 Anonymous a. (2012)
Lotus root 66 1.58 Anonymous c. (2017)
Rice bean 327 20.9 Rajerison, (2006)
Tree bean 426 18.8 Longvah, & Deost-
(mature pod) hale, (1998).
Fox nut 360 9.7 Shankar, M. (2010)
Sword bean 59 3 Ekanayake, Jansz, &
Nair, (2000).
344 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

23.3.3 MEDICINAL PROPERTIES OF SOME IMPORTANT

UNDERUTILIZED VEGETABLES

Besides being the source of food, these underexploited vegetables possess

several desirable medicinal properties. They are a rich source of phyto-
chemicals that have anti-cancer and anti-inflammatory properties which
confer many health benefits. Fern (Diplazium esculentum) is used in the
treatment of diabetes mellitus in an Aboriginal community in Lohit district
of the eastern zone of Arunachal Pradesh and Himalaya. The extracts of
Diplazium esculentum show anti-inflammatory activity and are also good
for constipation (Hui et al., 2012). Indian pennywort (Centella asiatica)
may help protect the brain from damage due to toxic metal exposure of
aluminum. It may help protect against skin cancer caused by exposure to
ultraviolet radiation. An extract of Centella asiatica, Asiaticoside, may
help prevent liver cancer. Oxalis corniculata is known to cure dysentery,
diarrhea, and skin diseases (Raghavendra et al., 2006). Taro (Colocasia
esculenta, Colocasia affinis) leaf is eaten for fever and respiratory disor-
der. It has a medicinal use for leprosy and tuberculosis, earache, alopecia
or hair loss and being styptic, it stops the flow of blood; this action helps
to arrest arterial disorder (Anonymous a., 2012). The fruit of Solanum kur-
zii, one of the Solanum spp, is used as an anti-allergic agent by the Mao
Naga tribe of Manipur. The fruit is crushed and applied to the allergic area
of the body. This is very effective for any types of allergies. The seed is
also edible and used by the tribals of the region as vegetables. The fruit is
used for cough and worm infestation (Mao et al., 2009). Solanum myria-
canthum is a perennial shrub that is used in the folk medicine of Tangkhul
Naga tribe of India for treating intestinal worms (Yadav and Tangpu,
2012). Solanum nigrum is considered good for cooling hot inflammation,
ringworms, ulcers, testicular swellings, gout and ear pain. The Arabs used
the bruised fresh leaves to alleviate pain and reduce inflammation. Sola-
num torvum leaf juice is taken orally to reduce body pain (Muthu et al.,
2006). According to Asati and Yadav (2004), there are around 12 culti-
vated species of Solanum in NE India, viz, Solanum macrocarpon (intro-
duced in the NE region), Solanum mammosum (possibly introduced,
ornamental with high solasodine percentage), Solanum xanthcarpum (used
Underexploited Vegetables in North Eastern India 345

as vegetable and medicinal purpose), Solanum indicum (domesticated,

used as vegetable and medicine), Solanum khasianum (wild and cultivated
for solasodine alkaloid), Solanum torvum (wild, sold in the market in
Mizoram), Solanum berbisetum (ripe fruits are eaten), Solanum ferox
(wild, leaves are used medicinally), Solanum spirale (wild but domesti-
cated for medicinal use in Arunachal Pradesh), Solanum sisymbrifolium
(native of Africa, wildly grown in Meghalaya), Solanum kurzii (endemic
in Garo hills, Meghalaya), and Solanum gilo (introduced in the NE region
as a vegetable). For bamboo shoot (Bambusa tulda), the fermented bam-
boo shoot juice has preservative property similar to vinegar, and meat,
fish, or vegetables are cooked with it have longer shelf-life. Methanol
extracts of Amaranthus caudatus, Amaranthus spinosus, and Amaranthus
viridis showed significant anti-diabetic and anti-cholesterolemic activity,
which provides the scientific proof for their traditional claims (Girija et
al., 2011). High magnesium content in Amaranthus cruentus (2.53 mg/100
g), T. triangulare (2.22 mg/100 g), Celosia (1.41 mg/100 g), and G. latifo-
lium (1.32 mg/100 g) may explain their blood pressure lowering properties
(Mensah et al., 2008). Amaranthus cruentus is used as tapeworm expellant
and causes relief of respiratory disease (Mensah et al., 2008). Amaranthus
spinosus stem and leaf are useful to treat dysentery. Amaranthus virides
stem and leaf are useful against small pox. Red amaranth is used for the
treatment of skin problems, diarrhea, sores, aching, and bleeding gums.
Fox nut/ Thangjing is recommended for the treatment of diseases of respi-
ratory, circulatory, digestive, excretory, and reproductive systems (Shan-
kar et al., 2010). Bathua/Jimilsag (Chenopodium album) in Ayurveda is
used to treat diseases of blood, heart, spleen, and eye; in biliousness condi-
tions, cough, abdominal pain, pulmonary obstruction; and in nervous ail-
ments (Sikawar et al., 2013). Chingit/Indian Pepper (Zanthoxylum rhetsa)
fruit and stem bark are aromatic, stimulant, astringent, stomachic and
digestive; they are prescribed in urinary diseases, dyspepsia, diarrhea and
with honey for rheumatism. Fruits are appetizer; useful for treating chol-
era, asthma, bronchitis, heart troubles, piles and toothache, and relief from
hiccup. The carpels yield an essential oil, which is given for treating chol-
era. The seed oil is antiseptic and disinfectant and is applied on inflamma-
tory dermatosis. The seed oil is used in dry eczema and dandruff of children
in Jointiapur of Sylhet. The root barks have cholinergic, hypoglycaemic
346 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

and spasmolytic activity. Most of the secondary metabolites present in the

plant possess antimicrobial and cytotoxic properties. In addition, the fruits,
seeds, stem bark and heart wood contain furoquinoline and indolequinazo-
line alkaloids and terpenoids (Ghani, 2003). Zanthoxylum acanthopodium
seeds and bark are used in the treatment of dyspepsia, fever, and cholera.
Zanthoxylum armatum exhibits spasmolytic effects, mediated possibly
through Ca(++) antagonist mechanism, which provides pharmacological
base for its medicinal use in the gastrointestinal, respiratory, and cardio-
vascular disorders (Gilani et al., 2010). Zanthoxylum oxyphyllum fruit is
used for stomach disorder. In China, Anhling/American black nightshade
(S. americanum) tea from the whole plant is used to treat cancer of the
cervix. Extracts from S. americanum were found to have selective antivi-
ral activity against the herpes simplex type-1 virus (HSV-1). Methanol
extracts of S. americanum have high antimicrobial activity against Esch-
erichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Asper-
gillus niger. In Nigeria, jack bean seed is used as an antibiotic and
antiseptic (Olowokudejo et al., 2008). There is also pharmaceutical inter-
est in the use of C. ensiformis as a source for the anticancer agents trigo-
nelline and canavanine (Morris, 1999). The bark of Climbing wattle/
Khanghu/Biswal/Agla bel (Acacia pennata) is used as antiasthmatic and
antibilious. The leaf is used as stomachic, styptic (for bleeding gum), and
antiseptic (for scalding of urine), and a decoction of young leaves is taken
for headache, fever, and body pain (Khare, 2008). In folk medicine, the
calyx extracts of Rosella (Hibiscus sabdariffa) are used for the treatment
of several complaints, including high blood pressure, liver diseases, and
fever. In healthy men, consumption of H. sabdariffa showed significant
decreases in the urinary concentrations of creatinine, uric acid, citrate, tar-
trate, calcium, sodium, potassium and phosphate, but not oxalate (Ali et
al., 2005). Toothache plant/Paracress (Spilanthes acmella) is used for
toothache in local areas. Spilanthes paniculata young stem and leaf are
taken for deworming. Fresh flower is used for relieving toothache. Zombi
pea (Vigna vexillate), is grown by the tribals of Tripura. The whole plant
part is used for medicine. It is effective for joint disorders, arthritis, and
swellings in joints. As a hemostatic, it checks hemorrhaging and thus pro-
longs life in individuals suffering from internal bleeding while building
their strength with its nutritive action. In Rice bean (Vigna umbellate),
Underexploited Vegetables in North Eastern India 347

Catechin-7-O-glucoside can be found in the seed. In vitro, this compound

has an antioxidant activity leading to a cytoprotective effect. Catechin-7-
O-β-d-glucopyranoside scavenges free radicals and protects human B
lymphoma BJAB cells on H2O2-mediated oxidative stress (Baek Jin-A et
al., 2011). Loosestrife /Kengoi (Lysimachia obovata) is a herb used as
traditional herbal medicine for the treatment of diabetes (Devi et al., 2011).
The green portion of the fruit Tree bean (Parkia roxburghii) is mixed with
little amount of water and applied to wounds and scabies. The fruit of
young shoot is eaten for curing diarrhea, dysentery, and food poisoning
(Bhardwaj and Gakhar, 2005). It has a very nutritious pod. Sophlong
(Moghania vestita) is an anthelmintic (Mali and Mehta, 2008). East Indian
Glory Bower (Clerodendrum inerme) leaf is grounded in water, and the
juice is taken orally to treat fever (Muthu et al., 2006) and boiled leaves
are taken to get relief from high blood pressure in local areas. Rhubarb
(Rheum nobile) root is used as an anticholesterol, antiseptic, antispas-
modic, antitumor, astringent, demulcent, diuretic, laxative, purgative, sto-
machic and tonic. Rhubarb roots contain anthraquinones which have a
purgative effect and anti-cancer properties (Qing Huang et al., 2006).
Spine gourd (Momordica dioica) is a protein-rich vegetable. Fruits are
used in ulcers, piles, sores, and obstruction of the liver and spleen. It pos-
sesses several medicinal properties and is said to be good for those suffer-
ing from cough, bile, and other digestive problems. The seeds are used for
chest problems and simulate urinary discharge (Ram et al., 2004). Mor-
inga/Drumstick leaf is taken as food, and it reduces body heat and to treat
indigestion and eye diseases. Flower is taken as food, and it gives cools
eyes and increases sperm production in men (Muthu et al., 2006).

23.3.4 INCOME GENERATION

These underexploited vegetables play an important role that constitutes

the daily vegetable requirement of the people. They are locally available
wild in different parts of these regions. The tribals collect and sell these
vegetables to earn income that could support their daily needs and to earn
small amount of income. These crops require proper studies to know their
values and unknown potentials as a means of improving livelihoods, espe-
cially in support of the rural poor vegetable farmers of these regions. Thus,
348 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

if this hidden wealth of novel leaves, fruits and its medicinal compound
are explored without further delay, these NE states, which are a rich source
of genetic biodiversity, will be in a position to occupy a sizeable share in
the national and international market for vegetables and herbal medicine.

23.4 THE DEPLETION OF BIO-DIVERSITY OF UNDERUTILIZED

VEGETABLES: A MAJOR CONCERN

According to a study of Food and Agricultural Organization (FAO), crop

genetic resources are being wiped out at the rate of 1.2% per annum. Trop-
ical forests are falling at a rate of just under 1% per annum or 29 hectares
per minute. From 1980 to 1990, this is equivalent to an area the size of
Ecuador and Peru combined (Shand, 2000). However, the availability of
most of these wild crops are now depleting rapidly owing to various factors
such as “Jhum/shifting cultivation,” forest fire, construction of industries,
felling of trees for timber, and other socio-economic anthropogenic activi-
ties in the area. Therefore, it is a need of the hour to tap their potentials and
conserve these regions’ wealth and devise the ways for commercial-scale
propagation of these plant species. If these underutilized plants are not
conserved, research studies and commercialization of these crops are far
beyond the reach of the researchers and the industries.

23.5 CONCLUSION AND RECOMMENDATION

Besides their commercial utility by the local tribes as food, these under-
exploited vegetables have immeasurable potential to contribute to food
security and have desirable medicinal properties, which require further
studies to exploit their potential. The possible reasons for the low utiliza-
tion of underutilized vegetables in spite of their recognized importance are
the lack of seeds, lack of information about their performance and input
requirements, lack of information on how they can fit into production sys-
tems, and nonviability of indigenous vegetable production like the major
cultivated species of vegetables such as tomato, pepper, eggplant, cauli-
flowers, cabbage, etc., whose improvement and seed production are taken
care by the private sector as well as government institutions, while the
Underexploited Vegetables in North Eastern India 349

underutilized vegetables are a neglected lot (Pandey et al., 2014). How-

ever, there is still limited research on these underutilized crops and com-
mercialization is still impossible due to negligence. Therefore, there is an
immediate need to tap their potentials and conserve these regions’ wealth,
to harnessing the diversity of these crops, and to popularize and use them
as crop alternatives as they are important sources of nutrients with innu-
merable medicinal properties.

KEYWORDS

•• food security
•• north-eastern India
•• phytochemicals
•• underexploited vegetables

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CHAPTER 24

ESSENTIAL OIL OF THYMUS

SATUREJOIDES COSS. IN THE HIGH
ATLAS OF MOROCCO: FROM
TRADITIONAL MEDICINE TO
COMMUNITY NATURAL PRODUCT
DEVELOPMENT
BERNADETTE MONTANARI1* and AMRITESH C. SHUKLA2
1
Department of Geography, University of Urbana Champaign,
USA (Social Dimensions of Environmental Policy (SDEP)),
Tel: +12173774102, E-mail: [email protected]
Department of Horticulture, Aromatic and Medicinal Plants,
2

Mizoram University, Aizawl–796004, India

CONTENTS

Abstract.................................................................................................. 354
24.1 Introduction................................................................................. 354
24.2 Materials and Methods................................................................ 358
24.3 Results and Discussion............................................................... 360
24.4 Results of the Essential Oil Distillate of Thyme from Megz...... 361
24.5 Conclusion.................................................................................. 364
Acknowledgments.................................................................................. 365
Keywords............................................................................................... 365
References.............................................................................................. 366
354 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

ABSTRACT

This article provides an insight into the phytochemistry of the essential

oil of Thymus saturejoides Coss. (thyme) and its potential for natural
product development in Megz, a village of the Agoundis valley, in the
High Atlas mountains of Morocco. The aromatic plant is widely har-
vested by the local population as an important source of income and
used in traditional Berber medicine. To assess the commercial viability
of the plant, the essential oil was distilled, analyzed, and compared with
thyme oil distillates from Asni Moulay Brahim situated at 55 km, and a
recent study of thyme samples collected in the lower part of the Agoun-
dis valley. The gas chromatography–mass spectrometry (GC-MS) analy-
sis of the thyme oil from Megz revealed borneol (32.89%), carvacrol
(18.05%), and thymol (0.46%) as the active constituents. These have
already been reported for their antimicrobial, antiseptic, antioxidant, and
other pharmaceutical properties. We suggest that the added value of the
essential oil of thyme from Megz may therefore be destined for the aro-
matherapy market, and for more specific indications in the pharmaceuti-
cal and sanitary industries.

24.1 INTRODUCTION

People have been resorting to traditional medicine since ancient times often
in the lack of conventional medicine and proximity to medical facilities. An
estimated 70,000 plant species have medicinal value and are employed in
traditional medicine worldwide (Lange, 2006; Schippmann, et al., 2002).
Medicinal and aromatic plants (MAPs) are also gathered to generate income
and to enhance livelihood (Ticktin, 2004). While many have condemned
the practices of wild harvesting as unsustainable, because of the enormous
pressure exerted at local, national, and environmental levels, and which
may result in species extinction (Ticktin, 2004; Sheldon, et al., 1997),
wild harvesting of medicinal plants does, nonetheless, allow isolated rural
populations to maintain vital subsistence (Robins, 2000; Schipmann, et al.,
2005; Ticktin, et al., 2002). Medicinal plants used in the traditional systems
of medicine hold the potential to add value to the socioeconomic welfare of
communities and to contribute to conservation strategies (Hamilton, 2004).
Essential Oil of Thymus Saturejoides Coss. 355

Morocco is the second most diverse country of species in the Mediter-

ranean basin in terms of biological resources after Turkey. The country
offers a rich flora and high endemism, 41 ecosystems, and 7000 vegetal
species of which 4500 are vascular plants. Six hundred species, which
have been reported as endemic, are listed as having medicinal and aro-
matic uses and harvested from the wild or cultivated (Vasisht, Kumar,
2004). Within the Mediterranean region, Morocco stands as the ninth larg-
est exporter of MAPs on the global scale (Lange, 2004; Ozhatay, et al.,
1997). These are used mainly commercially in the pharmaceutical, cos-
metic, culinary, and food industries (USAID, 2006) The country’s export
of pharmaceutical plants alone has increased from 5510 metric tons (MT)
to 12133 MT between 1993 and 2007, which represents more than US$ 34
million of MAPs and an estimated US$ 18 million worth for essential oils
(CFC, 2012). The main species harvested and processed in various parts of
the country are R. officinalis L., (rosemary) (1 million ha of the plant for
the production of rosemary essential oil, with an estimated annual yield of
60 MT), thyme, lavender, and Artemisia species, M. pulegium L., (penny-
royal), O. vulgare L., (oregano), and C. sativum L., (coriander). Prices for
essential oils vary widely from US$ 2 to US$ 10 for 10 mL and potentially
can reach US$ 200 to 600 per kg, especially for specialized production
used in the food, cosmetic, and pharmaceutical industries.
The present study was conducted between 2007 and 2009 in Megz, a vil-
lage of the Agoundis valley in the High Atlas mountains of Morocco (Figure
24.1). As one of the seven valleys originating from the Toubkal, the Agoun-

FIGURE 24.1 (See color insert.) Map of Morocco and the study area.
356 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

dis valley benefits from streams descending in altitude that carry snowmelt
water across various altitudinal zones and ecosystems. Temperatures may
vary throughout the year from 5°C to 6°C during the winter season and
over 30°C in the months of July and August. Rain fall also varies consid-
erably according to the seasons and may range from a low of 0.92 mm in
July to a high 67.36 mm in December and 70.13 mm in January. Snow fall
usually occurs from October until May, more or less abundant at 1200 m;
however, it becomes substantial at 2000 m (Benaboubou, 2004). Owing to
its topographic and geographic position, Agoundis is one of the narrowest
and enclave valleys of the High Atlas, enclosed between abrupt forested
slopes, offering very little cultivable space. The duality of this spatial struc-
ture produces noticeable differences in the landscape and in the availability
of resources. The strong declivity of the slopes favors the streaming and ero-
sion of the ground, thus necessitating the construction of terraces. Because
of the altitude ranges, local families have traditionally diversified livelihood
strategies according to the seasons. Millennia of human modification have
shaped the typicality and diversity of these landscapes to control erosion
and to promote agriculture. The Agoundis valley has, therefore, access to a
remarkable human-shaped landscape (Gerbati, 2004).
Although the environment is biologically rich, especially in MAPs, the
natural resources of the region are overall declining owing to overharvest-
ing in the face of the increasing demand for phytoaromatic products and the
needs of a growing population. In the mountains during the summer months,
local people harvest the aromatic plants thyme, Salvia aucheri Bentham var.
canescens Boiss and Heldr (sage), and Lavandula dentata L. (lavender) for
both herbal medicine and for trade (Montanari, 2004), the most lucrative
being thyme. These plants are not only one of the few sources of income,
but their utilization and collection also represent an important aspect of the
transmission of herbal plant knowledge within the community. The plants
are then traded down the valley via several middlemen to urban markets
in Marrakech and beyond. The trade follows two commodity chains, one
official (legal) and one unofficial (illegal) (Montanari, 2013). This income,
which varies in terms of the amount of plant material collected, represents
a significant contribution to the household economy.
Although this huge demand for plant products for domestic and com-
mercial use puts enormous pressure at the local and regional levels, these
figures suggest that the essential oil sector has potential for adding value
Essential Oil of Thymus Saturejoides Coss. 357

to otherwise fragile and marginal landscapes and to provide employment,

especially in isolated rural communities. Some communities have indeed
managed to overcome these obstacles and to achieve positive outcomes
in the plant trade. This is the case, for example for P. Africana Hook.f.,
Kalkman (red stinkwood) in Cameroon where local communities signed
agreements with external companies to ensure sustainable revenues and
practices (Ndam and Marcelin, 2004). Similarly, in Madagascar, middle-
men buy dried Centella asiatica L., Urban (gotu kola) from harvesters and
are responsible for packaging (Rasoanaivo, 2009). Exporters in Namibia
pay a percentage to the harvesters for good harvesting practices of Harp-
agophytum procumbens Burch., (devil’s claw) (Cole, 2009; Tonye Mahop,
2009), as is the case for the minor millets in the Kolli Hills of Tamil Nadu,
India (Gruère, et al., 2007). These cases have in common the full and
active integration of local people in either self-help groups or small-scale
enterprises, and agreements signed directly between external companies
and the communities. Such arrangements have potential for positive finan-
cial outcomes at all levels, i.e., village, local, and national.
Thyme (Figure 24.2) for instance is a promising source of antibacterial
and anti-inflammatory products according to the European Pharmacopeia
(WHO, 2002). The essential oils, which are mainly found in the flower-
ing stems of the plant (Hmamouchi, 2001), are rich in borneol (with high
antimicrobial activity), flavonoids (derived from apigenol and luteolol),

FIGURE 24.2 T. saturejoides Coss.

358 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

phenolic acids (particularly cafeic and rosmarinic acids), tannins, resins,

and other chemical compounds responsible for the majority of these phar-
macological effects (Garcia-Martin, et al., 1974; Grieve, 1974; Guenther,
1955). This article therefore seeks to demonstrate that the essential oil of
thyme could significantly increase the potential of a natural product devel-
opment on a commercial scale operated by the local people in the valley,
with the aim of returning benefits to the local population.

24.2 MATERIALS AND METHODS

24.2.1 STUDY AREA

The study was conducted between 2007 and 2009 in Megz, a village of
the Agoundis valley in the High Atlas mountans of Morocco. The village
is situated at an altitude of 1300 m, 8 km from the main rural commune
of Ijoukak, 100 km from Marrakech in Al Haouz province, close to the
Toubkal National Park, and has a rich biodiversity.

24.2.2 PLANT DATA COLLECTION WITH THE INHABITANTS

OF MEGZ

The data regarding the use of thyme were randomly collected with a total
of 60 female informants aged between 20 and 65 years and a total of 50
male informants aged between 20 and 67 years. This coverage allowed a
wide age spectrum of individuals involved (or with a history of involve-
ment) of herbal knowledge and collection, but excluding minors. All
interviews were conducted using an interpreter assisting with the trans-
lation of the Berber language back to French. The interviews sought to
identify the ethnomedicinal uses of the plant, the used parts for treating
specific ailments as well as the locations where the plant was collected.
Plant information/collections were made in various locations, i.e., with
the women while they were collecting vegetables, weeding the gardens, or
doing laundry down by the river. The men were accompanied while they
were working in the gardens, ploughing the land, when they went to the
mountains to collect the aromatic plants and firewood, or while they were
Essential Oil of Thymus Saturejoides Coss. 359

building a dam in the river. The identification of the collected plant thyme
(voucher specimen n° BML07) was compared with botanical plates from
“Medicinal plants of North Africa” (Boulos, 1983), and their therapeutic
properties were confirmed with the available literatures, viz., “La phar-
macopéemarocainetraditionnelle,” “Les plantesmédicinales du Maroc”
(Bellakhdar, 1997; Sijelmassi, 2003). A plant taxonomist at the Laboratory
of Vegetal Ecology, University Cadi Ayyad, Marrakech, assisted with the
identification of plant voucher specimens. These were then deposited at
the Laboratory of Vegetal Ecology at the Faculty of Sciences, University
Cadi Ayyad in Marrakech.

24.2.3 THYME ESSENTIAL OIL DISTILLATE

Flowering tops of thyme were randomly collected from four different loca-
tions on Tanammirt, Tissilu, Wankrim, and Wijdane Mountains around
Megz in the Agoundis valley. However, for the purpose of this study, only
the sample collected from Wijdane Mountain was analyzed. The fresh aer-
ial parts of the plant, when just coming into flower (discarding the lower
portion of the stem, together with any yellow or brown leaves), were used
for steam distillation and for estimation of the bioactive constituents. The
fresh aerial parts (1 kg) were steam distilled for 24 h in a 20-liter distilla-
tion flask fitted with an oil estimator. Light amber colored oil (2.73 mL, w/
dry weight) was obtained. This was done at the Faculty of Sciences, Cadi
Ayyad University in Marrakech.
Staff at the “Laboratoire de Biotechnologies végétalesappliquées aux
plantesaromatiques et médicinales” at Jean Monnet University in St Eti-
enne, France, performed the gas chromatography (GC) and mass spec-
trometry (MS) analyses. The essential oil sample was diluted 1/50 with
hexane and injected with a 1:200 split. The GC analysis of the oil was per-
formed on a Perkin-Elmer GC 8500, using a fused silica capillary column
(25 m × 0.55 mm, film thickness 0.245 µm), coated with dimethyl siloxane
(BP-1). The oven temperature was programmed from 60°C to 220°C at
5°C/min, then held isothermally at 220°C; detector temperature, 300°C;
carrier gas-nitrogen at an inlet pressure of psi; split, 1: 80. GC-MS data
were obtained on a Shimadzu QP-2000 Mass Spectrometer instrument at
360 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

70 eV and 250°C. GC column: Ulbon HR-1 (equivalent to OV –1), fused

silica capillary column (0.25 mm × 50 m, film thickness 0.25 µm). The
initial temperature was 100°C for 7 mm, and then heated at 5°C/min to
250°C. The carrier gas helium was used at a flow rate of 2 mL/min. The
results of the GC-MS analysis were then compared with studies of thyme
distillate conducted by Jafaari et al. (2007) and by Ghalbane et al. (2011)
in the lower part of the Agoundis valley. Moreover, data were analyzed
using statistical method of randomized block design (Gomez and Gomez,
1983) in Table 24.2.

24.3 RESULTS AND DISCUSSION

24.3.1 HERBAL MEDICINE USE

The Agoundis valley is botanically rich; many species are endemic, while
others are cultivated in gardens. In the absence of conventional medicine,
the villagers resort to medicinal plants that are widely used in ethno-med-
icine, and most families have harvested supplies of plants stored in their
houses. It is women who usually prepare and administer plant mixtures
that are consumed on a daily basis in tea, coffee, infusion and other more
complex herbal preparations for treating and bringing relief from com-
mon ailments (Montanari, 2014). Thyme is used profusely in herbal medi-
cine. It is taken regularly, more or less on a daily basis, as a fresh herbal
tea infusion during the harvest season or outside the harvesting season in
the dried herb form. Because thyme contains antioxidant, anti-infectious,
antispasmodic, anti-microbial, and anti-inflammatory properties (Jafaari,
et al., 2007), the dried herb is used to relieve common ailments in tradi-
tional folk medicine throughout Morocco (Bellakhdar, 1997). In Megz,
women take regularly the powdered herb for painful menses, to relieve
gastric disorders (stomachache, bile complaints, indigestion, and intestinal
trouble). It is also administered for respiratory disorders such as colds,
coughs, chills, and headache. People, however, have cautioned that thyme
should not be taken over long periods of time as it will damage teeth and
gums (Montanari, 2014). It has another important use as a preservative in
the confection of Smen. Smen is the term applied to preserved butter that
is prepared with the addition of salt and thyme. The addition of the leaves
Essential Oil of Thymus Saturejoides Coss. 361

and stems allows the mixture to be preserved for a year due to its antimi-
crobial properties (Banqour, 1985; Gutierrez, et al., 2008).

24.4 RESULTS OF THE ESSENTIAL OIL DISTILLATE OF THYME

FROM MEGZ

According to the literatures, the chemical composition of thyme oil has

also been studied by various researchers (Table 24.1).
It is carvacrol and thymol that gives thyme its antioxidant properties, a
phenolic structure present in various concentrations of thyme essential oil
(El Bouzidi, et al., 2013). Studies conducted on the essential oil of thyme
reveal important radical scavenger actions and their potential antibacterial,
antifungal, antiviral, and antioxidant properties (Ghalbane, et al., 2011;
Alaoui, et al., 2012). However, the borneol contained in the essential oil
of thyme contributes to its antimicrobial and anti-inflammatory proper-
ties, indicated in respiratory viral or bacterial chronic infections arthritis,
rheumatism, deep physical and sexual asthenia, candida infections, cysti-
tis, leucorrhoea, and externally for acne, infected wounds, dermatitis, and
other skin problems (Laboratoire de Combe d’As, 2014; Zhiri and Bau-
doux, 2008). Further research on the aqueous extract of thyme has shown
that its analgesic action is more potent than acetyl salicylic acid (ASA),
acting through an opioid-mediated mechanism (Elhabazi, et al., 2008).
From the studies, carvacrol, thymol and borneol stand as the major
constituents of interest of thyme from these three mountain locations and
are typically of the carvacrol-thymol chemotype. However, based on the
observations of Tables 24.1 and 24.2, the carvacrol from Megz is lower
than in the thyme species samples collected from Asni Moulay Brahim in
the High Atlas (35.90%), and it is higher than the thyme species samples
collected in the lower part of the Agoundis valley (9.1%). On the other
hand, the thymol content (0.46%) is lower than that in both samples col-
lected by Jafaari et al. (2007) and the lower part of the Agoundis valley by
Ghalbane et al. (2011), that is, 0.94% and 0.1%, respectively. However,
the borneol content of thyme collected from Megz (32.89%) is predomi-
nantly higher than that of the other thyme analyzed from the two locations
(Table 24.2). This phytochemical variation shows that thyme species and
TABLE 24.1 Chemical Composition of Thyme Oil: An Overview 362
Authors Chemical Constituents Remarks
Rovesti, 1971 camphene-pinene (0.15%), ρ-cymene (15–50%), linalool (3– Reported as additions in the chemical composition
13%), linalyl acetate (0–6%), bornol(2–8%), carvacrol (0–20%), from various Italian essential oils of thyme
thymol (5–60%).
Garcia-Mar- α-thujene (0.5%), β-pinene (4.6–4.7%), myrcene (0.4–0.9%), Reported as additions in the chemical composition
tin, et al., α-phellandrene(0.1–0.2%), limonene + 1,8-cineole 35.7–44.4%, of Spanish essential oils of thyme
1974 γ-terpinene 0.3%, trans-linalool oxide 0.5%, cis-linalool oxide
1.0–1.1%, camphor 11.6–16.3%, β-terpinelol 0.6–0.9%, α-ter-
pineol + borneol + bornyl acetate 7.8- 8.9%, α-terpinyl acetate
0.7–1.4%, geranyl acetate 0–0.5%, geraniol 0.1–0.2%.
Richards, et a-terpinene 1.0%, trans–sabinene hydrate 0.3%, methyl carvacrol Reported as additions in the chemical composition
al., 1975 3.0%, terpinen-4-ol 0.3% and caryophylln 0.9%. of French essential oils of thyme
Lawrence, sabinene trace, verbenene trace, 1-octen-3-ol 0.1%, methyl Reported as additions in the chemical composition
1984 thymol 1.5%, verbenone 0.2%, α-muurolene 0.3%, ρ-cymen-8-ol of Spanish essential oils of thyme
0.1%.
Jafaari, et al., camphene 0.58%, β-pinene trace, α-terpinene 0.37%, p-cymene Reported as additions in the chemical composition
2007 2.17%, g-terpinene 0.37%, linalool 30.03%, camphor 0.48%, of essential oil of thyme from AsniMoulayBrahim
borneol 30.03%, bornyl acetate 1.73%, thymol 0.94%, carvacrol
35.90%, α-copaene trace, caryophyllene 0.16%.
Ghalbane, et α-Tricyclene 0.1%, Tricyclene 0.1%, α-Thujene 0.1%, α-Pinene Reported as additions in the chemical composition
al., 2011 1.6%, Thuja-2,4(10)-diene trace, Camphene 4.0%, Verbenene of essential oils of thyme from the lower part of the
trace, Octen-3-ol trace, Sabinene trace, β-Pinene 0.5%, Myrcene Agoundis valley
0.2%, 3-δ-carene trace, α-terpinene 0.1%, p-cymene 5.1%,
Limonene 0.5%, γ-Terpinene 0.1%, Dehydro-p-cymene trace,
(E)-Sabinene hydrate 0.2%, Linalol 5.8%, Thujone trace,
Sustainable Horticulture Volume 2: Food, Health, and Nutrition
TABLE 24.1 (continued)

Authors Chemical Constituents Remarks

p-Menth-2-en-1-ol 0.1%, Camphor 1.0%, Pinocarveol0.3%,
(E)-Verbenol 0.2%, Isoborneol 0.1%, Borneol 29.5%, Terpin-
en-4-ol 2.1%, Dihydrocarvone 1 1..0%, α-Terpineol 6.5%, (Z)-di-
hydrocarvone trace, carveol trace, bornylformate 1.1%, Methyl
thymol 0.6%, Thymol 0.1%, Bornyl acetate 5.5%, Carvacrol
9.1%, α-Cubébène 0.1%, Copaène 0.9%, β-Bourbonene 0.4%,
β-Patchoulene 0.1%, α-Gurjunene 0.4%, β-caryophyllene 8.2%,
β-Cubebene 0.2%, α-Panasisen trace, Aromadendrene trace,
α-Humulene 0.4%, allo-Aromadendrene 0.3%, γ-Muurolene
0.3%, β-Gurjunene 0.2%, Ledene 0.3%, γ-Cadinene 1.0%, δ-Ca-
dinene 0.7%, β-Ionone 0.1%, Guaiazulene 0.3%, Caryophyllene
oxide 2.5%, Isoaromadendreneepoxyde 0.2%, GeranylLinalol
Essential Oil of Thymus Saturejoides Coss.

0.2%.
Present study gtricyclene 0.27%, a-thujene 0.30%, a-pinene 5.62%, camphene Reported as additions in the chemical composition
11.00%, β-pinene 0.71%, myrcene 0.23%, a-terpinene 0.31%, of essential oil of thyme from Megz
p-cymene 4.15%, limonene 0.41%, g-terpinene 1.74%, linalool
3.65%, camphor 0.31%, borneol 32.89%, a-terpineol 7.07%,
methyl carvacrol 0.82%, bornyl acetate 2.39%, thymol 0.46%,
carvacrol 18.05%, a-copaene 0.23%, caryophyllene 5.13%, a-hu-
mulene 0.09%, g-cadinene 0.17%, d-cadinene 0.46%, caryophyl-
lene oxide 0.15%.
363
364 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

TABLE 24.2 Comparison of the Three Main Compounds of Interest Found in Thyme
Essential Oil in the Three Different Locations of the High Atlas
Average of T. saturejoides, T. saturejoides, T. saturejoidesCoss.,
bioactive Coss., (Megz, High Coss., (Asni, High (Agoundis valley,
compounds Atlas) Atlas) High Atlas)
Area% Area% Area%
Borneol 32.89 30.03 29.5
Carvacrol 18.05 35.90 9.1
Thymol 0.46 0.94 0.1

their chemical composition may vary according to soil variation, vegeta-

tive cycle, seasonal variations, and extreme climatic conditions that may
affect their chemotyped identity (Thompson, et al., 2013). Therefore, in
concordance with Ghalbane et al. (2011) and Jafaari et al. (2007), these
results suggest that the essential oil of thyme from these three locations
has powerful antibacterial and antioxidant activities and can make an
important contribution to aromatherapy, natural preservation of food, and
as antibacterial ingredients for food and pharmaceutical industry.

24.5 CONCLUSION

Molecules from natural products will in the future continue to play a pre-
ponderant role as active substances in the discovery and validation of
new drugs. Of approximately 420,000 species, less than 5% have gone
through screening for one or several biological actions and the vast major-
ity of antibacterials or 78% of new chemicals are derived from natural
product molecules. With the advance in technology, especially the use
of high-performance liquid chromatography (HPLC), and spectrometry
for the rapid characterization of extracts and molecules, phytochemical
analysis,has a major impact in research on natural products. Natural com-
pounds obtained from indigenous drugs therefore hold the potential to
be utilized not only in the discovery of new drugs but also to benefit
local communities from which the substances are extracted. The study
presented here has shown that thyme species and their chemical compo-
sitions are largely altered by climate condition, soil variation, vegetative
Essential Oil of Thymus Saturejoides Coss. 365

cycle, and seasonal variation. However, the phytochemical analysis of

thyme collected from Megz as well as the significantly varied bioactive
compound, viz., borneol, carvacrol, and thymol contents of thyme col-
lected from the other two agro-climatic locations are worthy of attention.
The quality of the phytochemical content of thyme from Megz therefore
contributes to the European markets for aromatherapy use. While the aim
is not to distill the essential oil of thyme from Megz on an industrial scale,
it is rather to target a niche market production, an emblem of community-
produced essential oil. This would necessitate contractual agreements
with fair trade companies to ensure that ethical practices are respected to
return benefits to the local population.

ACKNOWLEDGMENTS

The authors are thankful to the communities of Megz (Agoundis val-

ley, High Atlas of Morocco) for sharing their valuable information, the
funding organizations (The Gen Foundation, FfWG (Funds for Women
Graduates), Radcliffe-Brown & Firth Trust, Funds (Royal Anthropology
Institute), John Ray Trust) who have permitted fieldwork, and the authori-
ties of Mizoram University, Aizawl (India), for providing various types of
support during the compilation of the paper.

KEYWORDS

•• aromatic plant
•• community development
•• essential oil
•• morocco
•• natural product
•• thyme
366 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

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(2013). Evolution of a genetic polymorphism with climate change in a Mediterra-
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its for non-timber products species in Mexico. Conservation Biology, 16, 691–705.
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CHAPTER 25

POTENT NUTRIMENTAL AND

ETHNOMEDICINAL HORTICULTURAL
FLORA FROM NORTH CENTRAL
TERAI FORESTS OF U.P., INDIA
S. C. TRIPATHI and T. P. MALL
Postgraduate Department of Botany, Kisan PG College,
Bahraich, U.P., India, Tel: +91-5252-235113, 9450259294,
E-mail: [email protected]

CONTENTS

Abstract.................................................................................................. 369
25.1 Introduction................................................................................. 370
25.2 Materials and Methods................................................................ 372
25.3 Result and Discussion................................................................. 373
25.4 Conclusion.................................................................................. 375
Acknowledgments.................................................................................. 376
Keywords............................................................................................... 376
References.............................................................................................. 376

ABSTRACT

Bahraich, Uttar Pradesh, India, is blessed with diversified flora of more

than 1200 plant species, and has Tharu tribals inside as well as around the
forests. The tribals have strong belief in magicotherapeutic properties of
plants for treatment of their ailments. The vegetation of the area is mainly
370 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

characterized by large member of herbaceous plants growing on variety of

habitats along with scattered occurrence of many indigenous and exotic
species of trees and shrubs in open areas or cultivated in gardens and along
road sides. The North Central Terai belt in which Bahraich is situated is
next to North East and Western Ghats, which represents one of the 18 hot
spots of the world mega biodiversity. Despite this richness, the wealth of
traditional knowledge is being lost, as the traditional culture is gradually
disappearing; hence, we have taken this project to document and describe
the potent, nutrimental, ethnomedicinal, and horticultural flora of the stud-
ied area.

25.1 INTRODUCTION

Bahraich district is one of the Terai districts of eastern Uttar Pradesh, situ-
ated in Upper Gangetic Plane. It lies between 27°43ʹ and 28°51ʹ North
Latitude and 81°8ʹ, and 82°10ʹ, East longitude, with a total area of about
6944 sq km. Botanically, the area is very interesting. In north, the Himala-
yas rise as a virtual wall beyond the snow line. Above the alluvial plain lies
the Terai strip, a seasonally marshy zone of sand and clay soils.
This north Terai region has higher rain fall than the plains, and the
downward rushing rivers of the Himalayas slow down and spread out in
the flatter Terai zone, depositing fertile silt and reproductive means during
the monsoon season and receding in the dry season. Terai, as a result, has
higher water level and is characterized by moist sub-tropical condition and
a luxuriant turnover of green vegetation all the year around.
The study area is blessed with several floras by nature; it is referred
as natural paradise, and it is very rich in ethnic and floristic diversity. The
Tharu tribes are endowed with vast knowledge of medicinal plant and
have strong belief in magicotheropeutic properties of plants for the treat-
ment of various ailments. The district is having good population of tribals,
mostly Tharus residing in villages, viz., Phakeerpuri, Amba, Balai gaon,
and Ramapur of Mihinpurwa block; their knowledge regarding plants has
descended from one generation to another as a domestic practice (Brah-
man, 2000). Due to the vast area of natural forests, the Bahraich is also
known as City of Forests.
Potent Nutrimental and Ethnomedicinal Horticultural Flora 371

The land surface is a level tract sloping gently from North West to
South East. A remarkable feature fills landscape is the total absence of
any hill or hillocks. The soil is composed of Gangetic alluvium. Because
much of the ground is liable to inundation, the particles deposited are very
fine. Bahraich enjoys monsoon type of climate, very much influenced by
the Himalaya being nearer to the region. The climate is markedly periodic
and is divided into three seasons, i.e., rainy, winter, and summer season.
The general temperature range between 3°C and 43°C. The general veg-
etation of the area is tropical deciduous type. However, some of the trees
are evergreen and semi-evergreen. The forests are only restricted to north-
ern portion of the district bordering up to foothills of Nepal. The middle
and southern part of the area are under the influence of human and their
domestic animals. Thus, the vegetation of this area is being damaged by
intense grazing, fire, and cutting down of plants for fodder, fuel, and for
various developmental projects. A vast area is also under cultivation. The
vegetation of these areas is mainly characterized by a large number of her-
baceous plants growing on variety of habitat along with scattered occur-
rence of many indigenous and exotic species of trees and shrubs in open
areas or cultivated in gardens and along road sides.
Plants have a significant contribution toward the wealth of a country.
During recent years, the exploration of our plant wealth and its economic
utilization have rightly been given due importance. The contribution on the
economic aspects of our plants are scattered over numerous literatures. The
revision of the information based on modern collection and field observa-
tion has been advocated by Rao (1958). Gupta (1967) emphasized that the
information we already possess on the economic aspect of plants should
be revised thoroughly based on personal enquiries and experimentations.
India presents colorful mosaic of about 563 tribal communities that have
acquired considerable knowledge on the use of plants for their livelihood,
healthcare and other purposes through their long association with the for-
ests, inheritance, practices, and experiences. Plants with medicinal proper-
ties enjoyed the highest reputation in the indigenous system of medicines
all over the world. India has one of the oldest, richest, and most diverse
cultural traditions called folk tradition associated with the use of medicinal
plants. Traditional folk medicine is the application of indigenous beliefs,
knowledge, skills, and cultural practices concerned with human health.
372 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

The ethnic people have provided several miraculous plants of medicinal

value for modern civilization. Both the ayurvedic and Siddha system of
medicine originated more than 300 years ago and are prevalent in North
and South India (Lgnacimuthu et al., 2006). The traditional definition of
medicinal plant is given in Ashtasane Hrdays 2006 AD Sutra sihana ch
9, Vrse 10 as “There is nothing in this universe which is non-medicinal
which cannot be used for many purposes and by many modes” (Shanker
et al., 2000). India represents one of the 12 mega biodiversity centers of
the world, and has two of the world’s 18 biodiversity hot spots. North East
and Western Ghats ranks first followed by the north central forests of Terai
region. This Terai belt, well blessed and inhabited by tribal community
inside the forest as well as around the forest area, is a natural paradise for
ethnobotanical, mycological, plant pathological as well as work related
with wildlife alone or interdisciplinary work. The World Health Organiza-
tion (WHO) has also recognized the role of traditional system of medi-
cine and considers it as a part of strategy to provide healthcare to masses.
Folk medicine is gaining importance. Much of this wealth of knowledge is
being lost as traditional culture is gradually disappearing (Hamilton, 1995).
Tribal people throughout the world have developed their own culture,
customs, cults, religious rites, myths, folk tales and songs, foods, medici-
nal practices, etc. Numerous wild as well as cultivated plants play a very
important and vital role among these cultures, and this interrelationship
has evolved over generations of experience and practices (Maheswari,
1983).

25.2 MATERIALS AND METHODS

The survey of the Bahraich districti was carried out during 2010–2014. Rap-
port was established with local elderly persons and the vaids (Ayurvedic
physicians), Hakims of the locality as well as Tharu tribes of the surveyed
area. Inquiries were made on the plant material used for curing different
ailments. Elderly men and women folk were interviewed by the question-
naire method, which resulted in heterogeneity of information. Participa-
tion in their feasts, festivals, and other social events, etc., was of great use
in collecting information on flora and their use. The plant was collected in
Potent Nutrimental and Ethnomedicinal Horticultural Flora 373

flowering and fruiting stage so as to facilitate identification. It was identi-

fied by authentic literatures and floras, viz Hooker (1872–1997); Sharma
et al. (1993a,b); Dubey (2004); Saini et al. (2010); and Saini (2006). The
herbarium of flora was prepared according to the method described by
Jain and Rao (1976) and Rao (1958) and deposited in Herbarium of the
Department for the record and reference.

25.3 RESULT AND DISCUSSION

25.3.1 ENUMERATION

The study area has eight floras of family Apocynaceae (dicot) and three
floras of family Liliaceae m(Monocot).

25.3.2 FAMILY APOCYANACEAE

1. Carissa carendas—A horticultural plant used in cure of anemia,

acid reflux, analgesic for headache and migrane anorexia, analge-
sic for Aphthous ulcers, antihelminthic, and antihistamine. It con-
tains alkaloids and is sour in taste due to presence of vitamin C. It is
antiscorbutic. Parts used: leaves, fruits, roots, and dried stem bark.
Center of origin is India.
2. Carrisa edulis—Roots contain an active ingredient, carissin, that
is proved useful in the treatment of cancer; the twigs contain que-
brachytol and cardioglycosides that are useful as an antihelmin-
thic against tapeworm. The boiled leaves and poultice is applied
to relieve toothache. Root bark is mixed with spices and used as
an enema for lumbago and other pain. Root scrapings are used
for glandular inflammation; ground up roots is used as a remedy
for venereal diseases, to restore vitality and to treat gastric ulcers,
cause abortion, and as an expectorant. An infusion of roots along
with other medicinal plants is used for treating chest pains, and a
root decoction is also used for treating malaria. Carrisa edulis is
an attractive ornamental tree and is suitable for planting in amenity
374 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

areas; the abundant branching naturet and the presence of spines

make the plant suitable for planting as a protective hedge.
3. Catharanthus roseus—Flower extracts have wound healing activ-
ity, it is used to treat diabetes, it has anticancer and antimicrobial
properties.
4. Holarrheria antidysentrica—Stem, root bark and seeds are used,
it is primarily used for the treatment of dysentery, bleeding dis-
order menorrhea, hemorrhoids, edema, tumors, abscesses aches,
branchites, colic disorders diarrhea, spinitis and as a vermifuge,
laxative and astringent.
5. Nerium indicum—The whole plant is used; it’s constituents are ole-
andrin, tanin, nerrin, phytosterine and L—stroph­nathinrosaginin,
nerlin, volatile oil, fixed oil, neriodorine and nerriodorein. Leaves
& flowers are thought to have action as cardiotonic, diaphoretic,
diuretic, emetic, expectorant and used to treat malaria and in termi-
nation of embryo. Tincture or decoction is used to reduce swelling
and scabes, Root powder is used as an external remedy for hemor-
rhoids and ulcers around genitals. It also has anticancer properties.
6. Rauwolfia serpentina—It is used in high blood pressure, nervous
disorders insomnia, and mental disorder such as agitated psychosis
and in insanity. The juice of the root is given internally to treat
snake bite. The decoction of leaf is invariably used in typhoid,
malaria, and other fevers. It is anti-inflammatory, antidiurectic, and
anticholinergic. It is one of significant medicinal herb used by the
people as indigenous medicine.
7. Rauwolfia tetraphylla—Roots are sedative tonic febrifuge; it is a
valuable remedy in high blood pressure and is used in insomnia,
mental disorders, and painful ailments of the bowels, hypochon-
dria and irritating condition of the central nervous system. Roots
are used in anxiety, excitement, schizophrenia, and epilepsy. Root
extracts are also used a valuable remedy in diarrhea, dysentery,
cholera, colic, and fever, Root paste along with orange peel is
used to treat fever by the tribal of Mihinpurwa Block. The root
or leaf juice is used against piles and as a remedy for sterility in
women.
Potent Nutrimental and Ethnomedicinal Horticultural Flora 375

8. Thevetia peruviana—Bark or leaf decoction is taken to loosen the

bowels as an emetic and is said to be an effective cure in intermit-
tent fevers. In Senegal, water in which leaves and bark are macer-
ated is taken to cure amenorrhea.

25.3.3 FAMILY – LILIACEAE

1. Aloe barbadensis—It is used to treat wounds, heals skin infections,

cure chopping, decrease hair loss, and eliminate hemorrhoids. It
is also used to cure eczema, sun burns, amputation, stump ulcers,
lacerations, colds, tuberculosis, gonorrhea, asthma, dysentery, and
headache. It is also used as insect repellent and as a laxative.
2. Asparagus racemosus—The roots are used following a regime of
processing and drying. It is used as uterine tonic and as a galac-
togogue (to improve production of breast milk); it is also used in
hyperacidity and as a best general health tonic.
3. Glorisa superba—It is also known as flame lily and has a wide
variety of uses within traditional medicine. It contains alkaloid
colchicines, which has been used effectively to treat gout, intesti-
nal worms, infertility, wounds and other skin problems. The roots
and leaves are used as an antidote for snake bite, as a laxative,
and to induce abortion. It has proven useful in the treatment of
chronic ulcers, arthritis, cholera, colic, and kidney problems. Glory
lily extract is useful to rectify many respiratory disorders. The sap
from the leaf tip is used for pimples and skin eruptions. Glorisa
paste can be applied for crump inflammation like wounds, piles,
and skin-related problems. Its powder is also helpful in relieving
from menstrual disturbances.

25.4 CONCLUSION

The rural and tribal repository of the studied area contains many medi-
cines for the treatment of various ailments. It is hoped that this effort will
376 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

not only provide additional support to the earlier findings but also provide
clues for new materials having traditional potentiality for the benefits of
mankind.

ACKNOWLEDGMENTS

The authors are thankful to all the local knowledge holders who helped in
one way or the other. Thanks are also due to The Principal, Kisan P.G. Col-
lege, Bahraich, for his permission to conduct this project and for facilities
and to The Chief Wildlife Warden, Uttar Pradesh Government, Lucknow,
for due permission and facilities. Thanks to Prof. S. K. Singh, Retired
Professor and Head, Department of Botany, DDU University of Gorakh-
pur and Late Dr. D.C. Saini, Scientist Grade F, Birbal Sahni Paleobotany
Research Institute, Lucknow, for identification of certain plants and for
their admirable help and encouragements.

KEYWORDS

•• ethnomedicinal
•• horticultural
•• mega biodiversity
•• nutrimental
•• potent
•• tribals

REFERENCES

Brahman, M., (2000). Some Ethnomedicinal plants of Akola and Sanganer talukes of
Ahmadnagar. J. Indian Bot. Soc., 81, 213–215.
Dubey, N. K., (2004). Flora of BHU Campus, Printed and Published by BHU Publication
Cell, 1–180.
Gupta, R., (1967). Seasonal Flowers of the Indian Summer Resorts, Mussoorie Hills, New
Delhi.
Potent Nutrimental and Ethnomedicinal Horticultural Flora 377

Hamilton, A., (1995). The people and plants initiative, In: Ethnobotany: Methods and Man-
ual, by Martin, G. J., WWW International, Chapman and Hall, London, pp. 10–11.
Hooker, J. D., (1872–1897). The Flora of British India, 1–7, (London).
Ignacimuthu, S., Ayyangar, M., & Shanker, S. K., (2006). Ehhnobolanical investigations
among tribes in Madurai district of Tamil Nadu (India). J. Ethnobiol Ethnomedi, 2,
25.
Jain, S. K., & Rao, R. R., (1976). Hand Book of Field and Herbarium Methods, Today and
Tomorrow Printers and Publishers, New Delhi, 33–58.
Maheswari, J. K., (1983). Developments in Ethnobotany. J. Econ. Tax. Bot., 4(1), 1–5.
Rao, R. S., (1958). History and importance of Indian herbaria. J. Ind. Bot. Soc., 3, 152–159.
Saini, D. C., (2006). Flora of Bahraich District, Uttar Pradesh, J. Eco Taxon Bot., 29(V),
843–886.
Shanker, D., Ved, D. K., & Geeta, V. G. A., (2000). Green Pharmacy Indian Health Tradi-
tions, The Hindu Special issue with the Sunday Magazine, 1–2.
Sharma, B. D., & Samjappa, M., (1993). With assistance from Bal Krishnan, N. P., Ed.
Flora of India, vol. 3, BSI. Calcutta Deep Printers New Delhi.
CHAPTER 26

ELEMENTAL DETERMINATION OF
TWO MEDICINAL PLANTS OF
MIZORAM USING EDXRF
R. LAWMZUALI,1 K. BIRLA SINGH,2 and N. MOHONDAS SINGH1
Department of Chemistry, School of Physical Sciences,
1

Mizoram University, Aizawl–796004, India

Department of Zoology, Pachhunga University College,
2

Aizawl–796001, India, E-mail: [email protected]

CONTENTS

Abstract.................................................................................................. 379
26.1 Introduction................................................................................. 380
26.2 Methodology............................................................................... 381
26.3 Result and Discussion................................................................. 382
26.4 Conclusion.................................................................................. 385
Acknowledgment................................................................................... 386
Keywords............................................................................................... 386
References.............................................................................................. 386

ABSTRACT

Medicinal plants have been used by humans for centuries in folklore medi-
cine. Medicinal plants are also incorporated into the historical medicine
of virtually all human cultures. The present work discusses the elemental
determination of elements in two medicinal plants Solanum nigrum linn
380 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

and Spilanthes acmella by using energy dispersive X-ray fluorescence

(ED-XRF). S. nigrum linn belongs to the Solanaceae family and is used
in the treatment of cardiac, skin disease, and inflammation of kidney, etc.
S. acmella belongs to the family Compositae and is called the toothache
plant and it is also used as an anti-inflammatory and analgesic. Twelve
elements were determined in this study.

26.1 INTRODUCTION

Mizoram is a state rich in plant life, and traditional medicines procured

from plants are employed for the treatment of numerous ailments since
time memorial. Such knowledge has been handed down through genera-
tions, and although modern medicines have seen a huge breakthrough,
these traditional medicines are still being used to a certain degree. Among
these, two of the most commonly used medicinal plants are Solanum
nigrum linn and Spilanthes acmella (Figure 26.1).
Solanum nigrum linn commonly known as Black Nightshade is a dicot
weed in the Solanaceae family. It is an annual branched herb of up to 90
cm high, with dull dark green leaves, juicy, ovate or lanceolate, and tooth-
less to slightly toothed on the margins. Flowers are small and white with

FIGURE 26.1 Medicinal plants: (a) Solanum nigrum linn and (b) Spilanthes acmella.
Elemental Determination of Two Medicinal Plants of Mizoram 381

a short pedicellate and five widely spread petals. Fruits are small, black
when ripe. The herb is antiseptic, antidysenteric, and antidiuretic and is
used in the treatment of cardiac, skin disease, psoriasis, herpesvirus infec-
tion, and inflammation of the kidney. The root bark is laxative and is use-
ful in the treatment of ulcers on the neck, burning of throat, inflammation
of the liver, and chronic fever. Berries are bitter and pungent useful for
treating heart disease, piles, and dysentery.
Spilanthes acmella is an indigenous herb belonging to the family Com-
positae. The plant has yellow/red gumdrop-shaped flowers. The leaves are
arranged opposite to one another and are 2.5 cm to 5 cm long. It is an
important medicinal plant and is commonly known as Akkalkara plant
with rich source of therapeutic constituents. It is called the toothache plant
because by chewing the leaves or flowers, it produces a numbing effect on
the tongue and gums. The flower heads of S. acmella can be used to relieve
toothache and also has anti-inflammatory and analgesic effects.
However, complete data regarding the elemental composition of these
plants are not known. The World Health Organization (WHO) has esti-
mated that 80% of the population of developing countries relies on plant-
based traditional medicines to maintain their primary healthcare needs.
High treatment cost and side effects along with drug resistance are the
major problems associated with synthetic drugs. Because traditional medi-
cine is not only easily accessible but also affordable, there is an increased
emphasis on the the use of plants to treat human diseases. Therefore, the
global markets are turning to plants as a potential and realistic source of
ingredients for healthcare products.

26.2 METHODOLOGY

26.2.1 SAMPLE PREPARATION

The two plants were thoroughly washed with triple distilled water to elim-
inate contamination due to dust and environmental pollution, air-dried,
and then oven dried at 60°C. After drying, they were grounded into fine
powder using mortar and pestle. The powdered samples were then formed
into pellets.
382 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

26.2.2 ANALYSIS USING ED-XRF SPECTROMETER

The elemental analysis of plant samples was carried out using a Xen-
ematrix Ex-3600 energy dispersive X-ray fluorescence (ED-XRF) spec-
trometer, which consists of an oil-cooled Rh anode X-ray tube (maximum
voltage 50 kV, current 1 mA). The measurements were carried out in
vacuum using different filters (between the source and sample) for the
optimum detection of elements. For example, for P, S, Cl, K, and Ca, no
filter was used, and a voltage of 6 kV and current of 200 mA were used,
and samples were run for 200 sec. A 0.05-mm-thick Ti filter was used
in front of the source for Mn, Fe, Cu, and Zn, with an applied voltage
of 14 kV and a current 900 mA, and samples were run for 400 sec. For
higher Z elements such as Se, Br, Rb and Sr, Fe filter of 0.05 mm thick-
ness was used at a voltage of 23 kV and 200 mA current, and samples
were run for 600 sec. The X-rays were detected using a liquid nitrogen-
cooled 12.5 mm2 Si (Li) semiconductor detector (resolution 150 eV at 5.9
keV). The X-ray fluorescence spectra were quantitatively analyzed by the
software integrated with the system. This software uses the fundamental
parameter method approach, which combines a theoretical basis of X-ray
emission and absorption with experimental measurements for unknown
sample analyses. Here, all matrix corrections, etc., are taken into account.
The experimental results were subjected to statistical analysis using Excel
2007 and SPSS package v.17.0. Values are presented as standard error of
mean (SEM).

26.3 RESULT AND DISCUSSION

The elemental determination of the two medicinal plants, viz,. S. nigrum

linn and S. acmella using ED-XRF yielded 12 different elements that are
given in Table 26.1.
From the result, we can see that both contain high concentration of
potassium (K), calcium (Ca), phosphorus (P), and sulfur (S) (Figure 26.2).
K is one of the essential elements of human diet and play an important
role in vital cellular mechanisms. As a cofactor, it catalyzes the conversion
of ADP to ATP, and K deficiency can create resistance in fat and muscle
Elemental Determination of Two Medicinal Plants of Mizoram 383

TABLE 26.1 Elemental Concentration (mg/L) of the Selected Medicinal Plants of

Mizoram. Values are Mean ± SEM of 6 Observations Each
Solanum nigrum linn Spilanthes acmella
Calcium (Ca) 16659.7±329.5 12542.9±346.2
Potassium (K) 42180.9±789.3 46224.4±690.6
Phosphorus (P) 9816.7±125.1 12310.1±312.8
Sulphur (S) 7577.9±333.1 5608.2±116.1
Iron (Fe) 577.2±110.6 736.1±90.8
Manganese (Mn) 89.4±6.5 140±3.9
Zinc (Zn) 59.1±9.9 152.5±8.9
Copper (Cu) 11.3±0.1 10.2±0.7
Selenium (Se) 0.23 13.9±0.1
Strontium (Sr) 91.5±5.2 58.1±2.4
Rubidium (Rb) 20.7±0.7 16.2±3.0
Bromine (Br) 15.3±1.4 110.4±2.2

FIGURE 26.2 (See color insert.) EDXRF spectra of major elements in Solanum nigrum
linn and Spilanthes acmella.

cells, etc., from insulin, increase serum triglycerides, may lower HDL and
blood supply to the vital organs, and increase the chances of stroke. Ca
is also an important element that plays a pivotal role in the physiology
and biochemistry of the cells in humans. It has an important role in signal
384 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

transduction pathways, where it acts as a secondary messenger, in neu-

rotransmitter release from neurons, contraction of all muscle cell types,
and fertilization. P is an element that makes up 1% of a person’s total body
weight and is present in every cell of the body. The main function of P in
the human body is in the formation of bones and teeth. As a macroelement
of the body, the role of S is to act as an integral part of many important
compounds found in all body cells which are indispensable for life.
The presence of trace elements like iron (Fe), manganese (Mn), zinc
(Zn), copper (Cu), selenium (Se), strontium (Sr), rubidium (Rb), and bro-
mine (Br) in notable concentrations explained to a certain degree their
medicinal properties (Figures 26.3 and 26.4).
Trace element plays an important role in human health because they
participate in biological functions that contribute to growth and good
health. Fe is a necessary nutrient element and is a core component of RBC.
It is needed for healthy immune system and for energy production. Mn is
one of the important essential elements required in carbohydrate metabo-
lism as well as an antioxidant in SOD enzymes. As a contaminant, no
maximum permissible limit (MPL) has been fixed for Mn in vegetables.
The upper tolerable limit of Mn for humans is 2–11 mg/day.
Zn is an important trace element involved in numerous aspects of cel-
lular metabolism and required for the catalytic activity of more than 200
enzymes. Zn also plays an important role in immune function, wound
healing, protein synthesis, DNA synthesis, and cell division. Cu is known

FIGURE 26.3 (See color insert.) ED-XRF spectra of minor elements in Solanum nigrum
linn and Spilanthes acmella.
Elemental Determination of Two Medicinal Plants of Mizoram 385

FIGURE 26.4 (See color insert.) ED-XRF spectra of earth elements in Solanum nigrum
linn and Spilanthes acmella.

to play an important role in human metabolism, largely because it allows

many critical enzymes to function properly. As an antioxidant, Cu scav-
enges or neutralize free radicals and may reduce or help prevent some of
the damage they cause. The concentration of Cu in plants varied much
with dependent nearby factors like proximity industries and use of fertil-
izers and Cu-based fungicides. Se is an element which behaves as both
an antioxidant and anti-inflammatory agent. As a contaminant, there is no
limit for Se. Rb ions are utilized by human body in a manner similar toK
ions, being actively taken up by plant and animal cells.

26.4 CONCLUSION

The chemical characterization of the two medicinal plants, viz., Solanum

nigrum linn and Spilanthes acmella by ED-XRF reveals the presence of
variable amounts of different types of major elements (Ca, P, K, and S),
minor elements (Zn, Fe, Cu, Mn, and Se), and earth elements (Sr, Rb,
and Br).
386 Sustainable Horticulture Volume 2: Food, Health, and Nutrition

ACKNOWLEDGMENT

The authors are thankful to the UGC-DAE Consortium for Scientific

Research, Kolkata Center, for aiding in the research work.

KEYWORDS

•• EDXRF
•• elements
•• medicinal plants

REFERENCES

Acharya, E., & Pokhrel, B., (2006). Ethno-medicinal, Plants used by Bantar of Bhaudaha,
Morang, Nepal. Our Nature, 4, 96–103.
Chakraborty, A., et al., (2004). Preliminary studies on anti-inflammatory and analgesic
activities of Spilanthes acmella in experimental animal models. Indian. J. Pharma-
cology, 36(3), 148–150.
Chopra, R. N., Nayara, S. L., & Chopra, I. C., (1956). Glossary of Indian Medicinal Plants,
Council of Scientific & Industrial Research, New Delhi, 168–169.
Cooper, M. R., & Johnson, A. W., (1984). Poisonous Plants in Britain and Other Effects on
Animals and Man. Ministry of Agriculture, Fisheries Food, 161, 219–220.
Gokhle, V. G., et al., (1945). Chemical investigation of Spilanthes acmella (Murr.). J.
Chemical Society, 1, 250–252.
Hsiu-chen, et al., (2010). Chemical composition of Solanum nigrum Linn extract and
induction of autophagy by leaf extract and its major flavonoids, Journal of Agricul-
tural and food Chemistry, 58(15), 8699–8708.
Kritikar, K. R., & Basu, B. D., (1935). Indian Medicinal Plants. M/S Bishen Singh Mahen-
drapal, New Delhi, India, pp. 267–268.
Pronob Gagoi, & Islam, M., (2012). Phytochemical screening of Solanum nigrum L and
S. myriacanthus dunal from districts of Upper Assam, India, IOSR Journal of Phar-
macy, 2(3), 455–459.
Zakaria, Z. A., Gopalan, H. K., Zainal, H., et al., (2006). Antinociceptive, anti-inflamma-
tory and antipyretic effects of Solanum nigrum chloroform extract in animal models.
Yakugaku Zasshi, 126, 1171–1178.
INDEX

1,8-cineole, 195, 200, 201, 205, 206, 209, Africa, 70, 195, 200, 201, 223, 293, 345,
324, 327, 362 359
2,2-diphenyl-1-picrylhydrazyl (DPPH), 91, Agoundis valley, 354–356, 358–362, 364,
98, 176, 179, 180, 182, 183 365
radical scavenging activity, 176, 179, Agricultural cash crops, 152, 164
180, 182, 183 Agriculture, 3, 175, 176, 218, 222, 223,
3,3 thiobis-1-propene, 202 225, 237, 238, 240, 301, 308
5-log reduction, 6–8 Agriotes obscurus, 204
8-hydroxyquinoline citrate (8-HQC), Agro-forestry, 57, 67
104–110 Alcohols, 201, 203
Alexivirus, 232
α Aliphatic compounds, 203
Alkaloids, 195, 237, 327, 346, 373
α-cubébène, 363
Allamanda catharitica, 245
α-gurjunene, 363
Allium sativum, 202, 251–253, 255–257
α-thujene, 201, 362 Allylpropyl disulfide, 202
Almond, 56
β Aloe barbadensis, 375
β-caryophyllene, 200, 201, 324, 363 Aloysia citriodora, 208
β-dihydropseudoionone, 199 Alphonso, 39, 40, 42–52
β-gurjunene, 363 Alternaria, 251–258, 286, 297
β-pinene, 195, 199, 201, 330, 362, 363 sesame, 257
solani, 251–258
γ Aluminum
γ-cadinene, 363 chloride colorimetric method, 176
foil, 106
γ-muurolene, 363
sulfate, 104, 105
A Amaranthus
caudatus, 338, 345
Absorption, 142, 146, 148, 176, 182, 382 cruentus, 345
Acanthoscelides obtectus, 194 lividus, 338
Acaricidal, 202, 237, 242, 248 retroflexus, 338
Acetic acid, 72, 282 spinosus, 345
Acetogenins, 236 viridis, 345, 338
Acetyl salicylic acid (ASA), 361 Amaryllidaceae, 202
Actinomycetes, 263, 281, 295, 299 Amino acids, 55, 56, 303, 305, 341
Advanced Center of Plant Virology Aminooxy acetic acid (AOA), 104–106,
(ACPV), 231–233 108–110, 112
Adverse effects, 35 Amomum dealbatum, 163, 164
Aegle marmelos, 257 Amorphophallus paeonifolius, 163
388 Index

Anastrepha Aromatic plant, 194, 201, 354, 356, 358, 365

ludens, 244 Arthropods, 193, 205, 236
lutescens, 244, 245 Artocarpus
Annona camansi, 55, 56
diversifolia, 244, 245 heterophyllus, 162
muricate, 236, 243–245 Ascorbic acid, 6, 9, 47, 49, 70–72, 80, 82,
squamosa, 236, 237, 242–244, 248 83, 176, 177, 179, 182
Annonaceae, 236, 237, 248 equivalent (AAE), 179, 181, 182
Anoglissus acumulata, 165 Ash, 62, 66
Anopheles Asimia, 236
stephensi, 244 Asimina triloba, 236
subpictus, 243 Asparagus, 141, 375
Anthocyanin, 70, 72, 74, 76, 80, 83, 84 Aspergillus
Anthracnose, 87–89, 94, 99, 279, 280, 288, flavus, 286
294, 296 niger, 297, 301, 302, 346
Anti-inflammatory, 15, 177, 195, 197, 198, parasiticus, 33
318, 328, 336, 344, 357, 360, 361, 374, Asteraceae, 193, 194
380, 381, 385 Astringency, 72
Anti-pathogenicity, 89 Ataulfo mango, 93
Anti-rheumatic, 195 Atmospheric pressure, 61
Antibacterial, 316, 318, 320, 321, Atranorin, 287
323–325, 328, 329, 332, 357, 361, 364 Atropa belladonna, 297
Antifeedant, 192 Attractant, 192
Antifungal, 34–36, 89, 95, 200, 251, 256, Average number of spores, 284
280–283, 285–288, 305, 327, 328, 361 Azadirachta
lichen extracts, 283 aegypti, 243, 244
Antimicrobial indica, 244, 251, 253, 255–257
activity, 27, 36, 109, 254, 281, 330, 346,
357 B
properties, 257, 361, 374
Antioxidant, 9, 15, 40, 70–72, 76, 80, 88, Bacillary dysentery, 316
90, 91, 97–99, 175–184, 197, 347, 354, Bacillus thuringiensis, 261, 263
360, 361, 364, 384, 385 Bacteria, 4, 52, 231, 232, 281, 293, 295,
Antirrhinum, 104–114 303, 304, 306, 316, 317, 324, 327, 329,
Antisorbutic, 71 330
Antiviral, 71, 328, 346, 361 Bactericides, 107, 225
Aonla, 69–76, 79–85 Badnavirus, 232
grape, 80, 82 Baking, 125, 135
litchi, 80, 82 Bamboo, 152, 157, 158, 163, 164, 169,
Apiaceae, 193 172, 338, 345
Apis mellifera, 209 flowering, 164
Application method, 222 pole, 152, 158, 159
Araucaria, 140, 141, 143–145, 147, 148 Bambusa
cunninghamii, 140–147 tulda, 158, 338, 345
Aroma, 40, 41, 51, 71, 79, 80, 122, 124, vulgaris, 158
133, 176 Banana, 298
therapy market, 354 Bankura district, 237
Index 389

Batrafine, 35 production, 64
Beauveria bassiana, 261 uses, 65
Begomovirus, 232 frozen seeds, 59
Benzaldehyde nutritional value of boiled seeds, 62
camphor oil, 208 seeds flour, 59
propionic acid, 208 seeds in brine, 59
Benzyl adenine (BA), 104–110, 112, 113 uses, 61
Beverages, 3, 65, 69–72, 74–76, 79–82, Broad
85, 195, 236, 317, 331 antimicrobial spectrum, 316, 322, 331
ready-to-serve (RTS), 69 spectrum properties of essential oil, 326
Bibliometric analysis, 190 Bromelain, 177
Bio-agents, 87 Bromeliaceae family, 176
Bioassay, 196, 239, 248, 283 Bromine, 383
Biochemical analysis, 71 Broom
Biochemistry, 183, 383 grass, 157–159, 161, 162, 169, 172
Biocontrol, 89, 286, 308 stick, 152
medicinal plants disease management, Bruchidae, 193, 194
295 Bruchus chinensis, 193
products delivery under field conditions, Bud drop, 107
306 Building materials, 152
seed treatment, 307 Burning, 35, 241, 381
seedling treatment, 307
soil treatment, 307 C
Biodiversity conservation, 152
Bio-efficacy, 261, 263, 264 Cabbage
Biological fresh yield, 274
control, 222, 292, 294 head borer, 261–263, 265, 276
management, 219, 223, 308 Cabinet dryer, 14, 18
Biopesticides, 220, 222, 224, 226, 227, Calcium, 56, 62, 63, 66, 176, 262, 282,
299, 300 342, 343, 346, 382, 383
Bischofia javanica, 165 Callicarpa arborez, 165
Black Nightshade, 380 Camellia sinensis, 236
Black rot, 175–177, 183, 184 Canavalia ensiformis, 339
Boric acid, 140, 142, 143, 145, 148 Cancer, 4, 177, 220, 293, 344, 346, 347, 373
Bostrichidae, 194 Cantharanthus roseus, 296
Botrytis cineraria, 89 Capsaicin, 15
Bouquets, 140 Capsicum, 279, 307
Brazil nut, 56 Carbamate pesticides, 191
Breadfruit, 56, 57, 65, 117–125, 127, Carbohydrates, 55, 56, 62, 63, 66, 303,
129–136 336, 337, 340–342
Breadnut, 55–67 Cardamom, 14
brine, 63 Cardiotonic, 71, 374
flour, 56, 60 Cardiovascular diseases, 4, 202
trees, 57 Caribbean islands, 123
frozen, 63 Carica papaya, 298
flour, 64 Carissa carendas, 373
nutritional value, 65 Carlavirus, 232
390 Index

Carotenoids, 47, 90, 97, 99, 129 Cinnamic acid, 193, 197
Carrisa edulis, 373 Cinnamomum, 197, 198
Carvacrol, 193, 197, 354, 361–363, 365 Cinnamon, 197, 198, 200, 203, 206, 207
Carveol, 193, 363 Circumference, 57, 60, 121, 123, 124
Carvones, 193 Citral, 193, 199, 202
Cashew, 56 Citronella, 198, 203
Cassia angustifolia, 296 Citronella oil, 192, 198
Catechin equivalent (CE), 178, 180, 181 Citronellal, 193, 198–201, 204
Catharanthus roseus, 257, 374 Citronellyl
Cavitation, 7 acetate, 201
Celiac patients, 55, 57, 66, 67 formate, 201
Cell Citrus
death, 130 aurantium L., 194
scrapper, 90 sinensis, 194
structures, 146 Cladosporium, 33, 118, 130
Celosia argentea, 342, 343 cladosporioides, 33
Centella asiatica, 336, 338, 342–344, 357 Cleanliness, 131
Central Biological Control Stations Clerodendrum
(CBCS), 222 colebrookianum, 167, 336, 340
Central Mechanical Engineering Research inerme, 347
Institute, 14, 16 viscosum Ventenat, 246
Central Plant Protection Stations (CPPS), Clevenger’s apparatus, 29, 316, 318
222 Cling filmed fruits, 118, 127
Central Surveillance Stations (CSS), 222 Clip-on
Centrifuge tubes, 105 barquettes, 132, 133, 135
Chaetomium, 297, 305 packaging, 132, 133
Characteristics of fruit and seed, 60 Closterovirus, 232
Chaunsa mango, 99 Clove, 199, 200, 203, 206, 207, 209
Chavyanprash, 71 Coat protein (CP), 231
Chemical composition of essential oil, 323 Coccinellids, 225
Chenopodium ambrosioides, 206 Cold stored bread fruit slices, 130
Chestnut, 61 Coleus forskohlli, 297
Chili, 15, 16, 279, 280, 307 Colletotrichum, 33, 88, 90, 92, 118, 130,
Chlorophyll, 129, 145, 236, 293 279, 280, 283, 288, 297
Chlorophytum borivilianum, 296 acutatum, 280
Cholera, 316, 318, 345, 346, 374, 375 capsici, 33, 279, 280, 283, 285–288
Choristoneura rosaceana, 197, 202 coccodes, 280
Chow-Chow (Sechium edule), 339 dematium, 280
Chronic human diseases, 3 falcatum, 33
Chrysanthemum gloeosporioides, 88, 90–92, 99, 280
ciner ariifolium, 298 Colocasia
indicum, 242, 243 affinis, 344
Chrysomelidae, 194 esculenta, 338, 344
Chrysoperla, 225 Color, 4–8, 15, 39–41, 49–51, 60, 61,
Chutney, 40, 339 64, 70, 71, 74, 75, 79, 80, 82–85, 92,
Cinchona ledgeriana, 297 93, 95–98, 110, 111, 113, 118, 119,
Cinnamaldehyde, 193, 197, 198 121–130, 132, 135, 136, 146, 148, 155,
Index 391

158, 168–171, 256, 282, 285, 286, 325, Dapchhuah village, 156, 162, 165, 167
326, 355, 383–385 Datura stramonium, 297
Community Days after treatment (DAT), 239, 245, 262,
development, 169, 365 264
hall, 156, 169 Days to ripening, 127
information center, 169 Decay organisms, 130
randomized block design (CRBD), 107, Deeringothamnus, 236
142, 255 longispathus, 158, 163, 164
Conical flask, 90 Dehydrated, 40–45, 47, 49–52, 139, 140,
Consumers, 4, 8, 9, 61, 131, 192, 210, 252 149, 262
Conyza bonariensis, 245 Dehydration, 40, 139, 146
Cooling system, 9 Dehydroascorbic acid, 72
Copper, 62, 66, 383 Deltamethrin, 261–263, 265, 272, 274
Cosmetics, 196, 318 Dendrocalamus
Cost-benefit management, 226 giganteus, 158
Cost-effective, 28, 222 hookeri, 158
Critical difference (CD), 48, 50, 91, 255, longispathus, 158, 161, 163
257, 264, 267, 269, 271, 273, 275 Dermatophtes, 36
Crop research station (CRS), 118, 120, Dermestidae, 194
125–128, 131, 132 Determination of antioxidant properties,
Crop yields, 190, 191, 217 178
Crude fiber, 62, 66, 176 Di-2-propenyl trisulfide, 202
Cucujidae, 194 Diabetes, 4, 70, 220, 344, 347, 374
Cucumber mosaic virus, 231 Diabrotica virgifera virgifera, 200
Cucumovirus, 232 Diagnosis, 219, 226, 230, 233
Culex Diagnostic methods, 230
quinquefasciatus, 244 Diallyl
disulfide, 202
tritaeniorhynchus, 243
thiosulfinate, 202
Curculionidae, 192, 194
trisulfide, 202
Curcuma
Diarrhea, 35, 318, 329, 344–347, 374
angustifolia, 27, 29, 30, 34
Dichlorodiphenyltrichloroethane (DDT),
aromatica, 27, 29, 30, 34
191
domestica, 27–36
Diflubenzuron, 261–263, 265, 272, 274
longa, 14, 298
Digestive agent, 177
zedoaria, 27, 29, 30, 34
Digitalis purpurea, 297
Curcumin, 15
Dimethyl sulfoxide solvent (DMSO), 320
Curvularia lunata, 33
Dimorphotheca, 104–114
Custard apple, 237, 243, 248 Dioscoria dettoidea, 297
Custard apple seed (CASE), 237, 242–248 Dipentene, 199, 324
Cyclocurcumin, 15 Diphenyl-1-picrylhydrazyl (DPPH), 88, 91
Cymbopogon, 198 Diplazium esculentum, 338, 344
Cyphomandra betacca, 339 Direct hand plucking, 164
Disc diffusion method, 316, 320, 325, 330
D Disease
Dactrine, 34–36 incidence, 126
Dal, 65 management, 226, 294
392 Index

Dried treatments, 111

fruit, 40 yeasts, 92, 93, 95–98
mango, 40 yield parameters, 264
materials, 140, 141 Electric field strength, 9
Drosophila melanogaster, 200 Electromagnetic spectrum, 8
Dryer, 14, 16, 17, 19–21, 23, 24 Elements, 105, 176, 300, 379, 380,
Drying 382–386
chamber, 19 Embedding, 140–142, 145, 149
process, 16, 39 Emblica officinalis, 69, 70, 79, 80, 162
rates, 21 Energy dispersive, 380, 382
Dual culture technique, 90 Entry point activities (EPA), 169, 171, 206
Duncan’s multiple range test (DMRT), social benefits, 169
264, 267, 269, 271, 273, 275 Environmental services, 336, 337
Dysentery, 316, 344, 345, 347, 374, 375, Enzymatic browning, 41, 47, 49, 51, 132
381 Enzyme-linked immunosorbent assay
Dyspepsia, 345, 346 (ELISA), 230, 233, 321
Enzymes, 4, 5, 7, 176, 304, 305, 330, 384,
E 385
Ephestia
E-pest surveillance, 218, 221, 224 cautella, 201
Early blight, 251–256, 258 ceratoniae, 201
Eastern Himalayan, 281 kuehniella, 201
Eco-friendly, 139–141, 149, 218, 221, 222, Epidermal cells, 130
224, 239, 240, 261–265, 276 Epidermophyton floccosum, 28–32, 34, 36
pesticides, 261–265, 276 Epiphytic plants, 281
Economic Equipment development, 16
injury level (EIL), 219 Escherichia coli, 5, 52, 231, 232, 316, 318,
threshold (ETL), 219, 221 319, 322, 324–326, 328, 331, 346
Ecotoxicity, 202 Essential oil (EO), 190–211, 315, 317,
Ectomyelois ceratoniae, 201 318, 320–325, 327, 329–331, 345,
Edible portion, 342, 343 354–359, 361–365
Edoes, 61 Ethnomedicinal, 358, 370, 376
EDXRF, 379, 383, 386 Eucalyptus, 146, 200, 251, 255–257
Effect of chamadulonsis, 201, 251–253, 256, 257
carotenoid contents, 49 cinerea, 146
different osmotic treatments, 45, 50 dundasii, 201
drying air temperature, 21 oil, 200
eco-friendly pesticides, 264–266, 268, rudis, 201
270 Eugenol, 199, 200, 204, 324, 331
larval population, 264 acetate, 200
inoculum density, 30, 322 Eukaryotic, 293
moisture content on thermal conduc- Euphorbiaceae, 80
tivity, 22 European Union, 191, 206
osmotic treatments, 47 Eurya acuminatea, 163, 164
plant extracts, 256 Euryale ferox, 339
sensory quality, 42 Evaluation of breadnut, 58
some physical factors, 31 Evapo-transpiration, 92, 93
Index 393

Evernia prunastri, 287 utilization, 340

Everniastrum, 280, 282, 285, 287, 288 Forest product, 151, 152, 154, 172
cirrhatum, 280, 285, 287 Formation of slime, 5
Evernic acid, 287 Formulation, 203, 299, 300, 308, 331
Experimental station (ES), 118, 120, 125, Fresh cut, 119, 132, 136, 167
127, 128, 131, 132, 134 Fruit
bars, 40
F juice, 3–9, 71
Farm juice processing, 5
bio-security, 217 non-thermal processing, 6
yard manure, 295, 307 thermal processing, 5
Farnesol, 199 pulp, 90, 123, 126
Fat, 62, 66 transpiration, 92
Fenchone, 193 Fuelwood, 152, 153, 157, 165, 166, 169,
Ferric–tripyridyltriazine complex, 90 172
Filtration, 18, 283, 318 Fumigant, 192–194, 196–201, 204, 208,
Firmness, 59, 88, 92, 93, 99, 118, 119, 209, 211
123, 124, 126, 129, 136, 178 Fungal, 16, 29, 36, 95, 126, 219, 252, 254,
Flakes, 40 257, 263, 284, 287, 292, 294, 295, 299,
Flash pasteurization, 6 304, 305, 308, 316, 329
Flavonoid, 8, 176, 180, 181, 183, 184 growth inhibition (FGI), 29
Flavonoids, 176–178, 180, 181, 357 Fungicide, 89, 130, 203, 225, 251–256,
Flavoparmelia caperata, 287 258, 280, 284, 286, 287, 294, 295, 385
Flavor, 4–6, 14, 39–41, 51, 59–62, 64, Fungitoxic spectrum, 32
79–82, 84, 118, 119, 129, 136, 176, 190, Fusarium, 33, 295–298, 302, 303
209, 323 oxysporum, 33, 296, 298, 302, 303
Flexibility, 141, 225
Flies, 192, 193 G
Floriculture Galangal, 14
industry, 139 Gallic acid equivalent (GAE), 178, 180
trade, 104 Gamma-aminobutyric acid, 194
Flow rate, 9, 319, 360 Gandul, 177
Flower drop, 107, 109 Garcinia lanceifolia, 162
Foliage, 142, 145, 148, 149, 244 Garlic, 195, 202, 203, 206, 223, 243, 252,
Folin-Ciocalteu 257, 330
assay, 178 oil, 202, 206
method, 176 Gas
Folklore medicine, 379 chromatography (GC), 195, 199, 316,
Food 319, 323, 332, 354, 359, 360
access, 340 GC-MS, 195, 199, 316, 332, 354,
availability, 340 359, 360
FAO, 176, 348 production, 5
industry, 6, 8, 15 Gastrointestinal symptoms, 317
requirements, 152 Geraniaceae, 201
security, 55, 120, 152, 336, 337, 340, Geraniol, 193, 198, 199, 201, 202, 204,
341, 348, 349 206, 362
394 Index

Geranium, 201–203 Heterodermia leucomela, 287

Geranyl High
acetate, 201, 362 density polyethylene (HDPE), 60, 128
acetone, 193 temperature short time (HTST), 6
Germination, 257, 280, 283, 284, 287, 288, voltage power supply, 8
299, 303, 305 Highest score, 50–52
Ginger, 14–25 Holarrheria antidysentrica, 374
Gingerols, 15 Homalomena aromatica, 163, 168
Gliocladium, 296, 297, 302, 304, 305 Honestly significant difference (HSD), 179
Global population, 190 Horticultural, 171, 190, 191, 205, 211,
Glorisa superba, 375 219, 223, 229, 230, 233, 280, 282, 287,
Gluten, 56, 57, 62, 66, 67 370, 373, 376
Glycerin, 140–142, 146, 148 Horticulture, 40, 156, 227, 233
Glycoprotein, 293 Humidity, 16, 89, 106, 118, 135, 148, 238,
Gmelina arborea, 166 253
Good agricultural practices (GAP), 226 Hydrocarbons, 203
Gooseberry, 70, 80 Hydrodistillation, 318, 332
Government of India (GoI), 222, 241, 226 Hypoglycemic activity, 71
Grape, 79–85, 89, 210 Hypogymnia physodes, 287
Green
medicines, 317 I
pesticide, 207, 209, 211 Ilarvirus, 232
revolution, 191 Immunodiagnosis, 230–233
Gregarious, 262 Immunoglobulin G (IgG), 232
Grinder, 14, 16, 19, 20, 24, 237, 254 Immunological, 229, 233
Grinding, 25, 60 In vitro antibacterial investigation, 29, 320
Groundnut bud necrosis virus (GBNV), In vitro screening, 254
231, 232 Income generation, 25, 120, 336, 337
Growth Incubator, 178, 320
inhibition, 90, 92, 254, 321, 326 Indian Agricultural Research Institute
regulatory activities, 192 (IARI), 104, 105, 223, 231–233
Indian Council of Agricultural Research
H (ICAR), 220, 223
Harpagophytum procumbens, 357 Indian Council of Medical Research, 341
Harvest, 57, 89, 105, 106, 118, 119, 123, Induced systemic resistance (ISR), 306
130, 135, 156, 158, 162, 164–166, 171, Industrial agriculture, 191
177, 178, 329, 356, 360 Information communication technology
Harvesting, 118, 120, 130, 135, 152, 154, (ICT), 225
157, 158, 161, 162, 164–169, 171, 195, Infrastructure, 84, 156, 224, 316
354, 357, 360 Initial
Healthy snack, 57, 63, 67 fresh weight (IFW), 106
Hedonic scale, 40, 82 volume of medium, 284
Helminthosporium maydis, 33 Inoculation, 253–255
Herbaceous plants, 104, 370, 371 Insecticidal, 192–195, 199–201, 236, 272
Herbal drug, 36 Insecticides, 203, 205–209, 225, 236, 262
Herbicides, 203 Integrated
Index 395

decision support system (IDSS), 218, Litchi, 79–82, 84, 85

224 Litchi chinensis, 80
health management, 219 Litsea monopetala, 166, 167
pest management (IPM), 192, 210, Livelihood, 24, 151, 152, 172, 218, 262,
217–227, 248 316, 336–338, 354, 356, 371
Integrated, 192, 208, 210, 218, 224, 227, Localized heating, 7
248, 292, 294, 299, 382 Low-density polyethylene, 128
Intensive agriculture (IA), 220, 227 LPG connection, 165, 169
Interactions, 292, 304 Lupin, 104–110, 112, 114
Intrinsic properties, 4 Lycopersicon esculentum, 298
Iron, 62, 66, 303, 383 Lysimachia obovata, 336, 339, 347
Irritation, 35, 329, 331
M
J
Macluravirus, 232
Jam, 40, 71 Macrophomina, 295, 296, 298
Jamun, 69–76 Magnesium, 56, 62, 66, 129, 176, 262,
Jhum cultivation, 154 342, 343, 345
Josapine–N36 pineapple hybrids, 177 Magnolia grandiflora, 146
Juice, 4, 6–8, 70–72, 76, 80–82, 84, Malaysian Agricultural Research and
243–345, 347, 374 Development Institute (MARDI), 177
Juice blends, 76 Management, 28, 89, 95, 119, 153, 205,
Juniperus chinensis, 140–142, 145, 146, 208, 217–222, 224–227, 230, 236, 248,
148 280, 281, 287, 292, 307
Mancozeb, 280, 284–286
K Mandrivirus, 232
Kalisena, 223, 301 Manganese, 62, 66, 176, 343, 383, 384
Klebsiella pneumoniae, 316, 318, 325, 326 Mango, 39–52, 87–89, 92, 93, 95–99, 223
Mangifera indica L., 40, 88
L Marketability, 4, 135
Laboratory-based techniques, 230 Maspine, 177
Lamiaceae, 190, 192–195, 315, 317 Mass
Land use pattern, 154 spectrometry (MS), 195, 199, 316, 319,
Larkspur, 104–110, 112–114 323, 332, 354, 359, 360
Lateral flow assay (LFA), 230 transfer rates, 41
Lauraceae, 190, 195, 197 Mathematical models, 21
Lavandula dentata, 356 Mauritius, 55, 57, 62, 63, 66, 119, 120,
Laxative, 71, 347, 374, 375, 381 126, 136
Lemongrass, 198, 199, 203 Maximum
Lichenic acids, 287 permissible limit (MPL), 384
Lichens, 280–282 residue limit (MRLs), 226
Lima oil, 194 Mechanisms, 6, 190, 281, 292, 306, 308,
Limonene, 193, 194, 196, 327, 330, 362, 382
363 Medicinal
Linalool, 193, 196, 197, 199–202, 324, aromatic plants (MAPs), 354–356
362, 363 plants, 28, 36, 69, 152, 157, 158, 160,
Lipopolysaccharides (LPS), 330 166, 167, 211, 292–295, 299, 308,
396 Index

317, 327, 354, 359, 360, 371, 373, killing time (MKT), 28, 32, 322, 327
379, 380, 382, 385, 386 Mint, 196, 203, 206, 207, 210, 298, 317,
Mediterranean region/area, 194, 195, 197, 318
355 Mizoram, 14–16, 23, 24, 36, 152, 157,
Mega biodiversity, 370, 372, 376 166, 219, 282, 287, 308, 331, 336, 339,
Melaleuca alternifolia, 206 341, 345, 365, 380, 383
Melocanna baccifera, 158, 163–165 Modified
Mentha × Piperita, 196 cytometer technique, 283
Mentha spore germination inhibition technique
citrata, 193 (MSGIT), 280, 283, 288
piperita, 315, 317, 318, 320, 322, 323, Moisture, 18, 21–23, 40, 43–45, 51, 52,
325–327, 331 62, 64, 66, 140–143, 145–149, 176, 221
Menthol, 193, 196, 206, 209, 316–318, content, 44
323, 327, 329–331 loss, 40, 52, 140, 142, 143, 145–148
Menthone, 193, 196, 316, 317, 323, 327 ratio (MR), 21, 22
Mesua ferrea, 165 Molecular diagnostic kits, 229
Methodology, 154, 381 Momordica
Methyl cochinchinesis, 340
1-propenyl disulfide, 202 dioica, 340, 347
2-propenyl trisulfide, 202 Monochoria hastate, 336, 339
heptenone, 199 Monoclonal antibody (MAb), 232
Metric tons (MT), 41, 176, 355 Monoterpenes, 190, 196, 199, 203, 317,
Miconazole nitrate, 34 324
Microbial, 4–7, 9, 41, 57, 70, 72, 82, 90, Moringa oleifera, 338
91, 105, 107, 130, 132, 135, 263, 264, Morocco, 365
272, 292, 294, 300, 303, 308, 321, 360 Morris, 7, 9, 175–177, 179–184, 346
biocontrol product formulations, 295 Morris pineapple, 182, 183
inactivation efficiency, 8 Mosquitoes, 192, 193, 207
quality in osmo-dried mango slices, 52
Mouthwashes, 196, 318
Microorganisms, 4– 6, 8, 89, 91, 105, 107,
Multipurpose tree species, 171, 339
135, 199, 295, 298, 300, 307, 317
Musa sapientum, 298
Microscopic field, 283, 284, 293
Mushroom, 163, 164
Microsporum
Mycelia growth, 251, 257
auddouinii, 32, 33
Mycelial discs, 31
gypseum, 28, 29, 32, 34, 36
Mycoplasma, 281
Microwave oven, 140–143, 145, 146, 148
Myrtaceae, 70, 190, 193–195, 199
Mikania micrantha, 167, 223
Myzus persicae, 193
Minerals, 39, 55, 56, 63, 66, 176, 262, 336,
337, 340–342
Minimum
N
bactericidal concentration (MBC), National Botanical Research Institute
321–323, 325–327 (NBRI), 282, 287
cidal concentration (MCC), 30–32 National Research Centre for Integrated
effective concentration (MEC), 29, 31, 34 Pest Management (NCIPM), 222–224
fungicidal concentration, 28, 31, 32 Natural
inhibitory concentration (MIC), 28, 36, fruit juices, 4
316, 321, 325, 326, 328–330 product, 207, 236, 281, 354, 358, 364, 365
Index 397

Nausea, 35, 318, 328, 329 Organochlorine, 220

Nectars, 40 pesticides, 191
Neem, 224, 244, 257, 261, 263 Organoleptic, 4–6, 49, 52, 82
seed kernel extract (NSKE), 261–263, Organophosphate pesticides, 191
265, 272, 274 Origanum vulgare, 208
Nematicide, 15 Oryzaephilus surinamensis, 197, 201
Nematodes, 218, 281, 293 Osmia cornifrons, 209
Neonicotinoids, 191 Osmo-dehydrated mango slices, 47, 52
Neral, 199 Osmosis, 40–43
Nerium indicum, 374 Osmotic
Nerol, 199 concentration, 41
Neurohormonal influences, 193 dehydration, 39–43, 46, 47, 51, 52
Nezara viridula, 208 technique, 41
Nizaral, 34–36 pre-treatment, 42
Non-timber forest products (NTFPs), Ovicidal activities, 193, 246
151–154, 156, 158, 159, 165, 167–171 Oviposition
market flow, 167 deterrent, 192
Nongovernmental organization (NGO), 14, inhibition, 193
16, 24, 166 Oxalis corniculata, 344
Nonthermal processing, 4, 5, 9 Oxidation, 72, 80, 82, 132, 304
North American genera, 236 Oxygen, 4, 72, 133, 195
North-east (NE), 219, 223, 336, 338–340,
344, 345, 348, 349 P
North-Eastern Hill (NEH) region, 223 Pale yellow flesh, 124, 125
North-eastern India, 349 Pamplemousses, 57, 118, 120, 121, 125,
Nutrient retention, 41 127, 128, 131, 132, 134
Nutrimental, 370, 376 Panax ginseng, 297
Nutrition, 57, 63, 67, 336, 337, 341 Papaver somniferum, 296
Nutritional Papaya, 298
qualities, 4, 6, 336, 337, 343 ringspot virus (PRSV), 231, 232
value, 3, 4, 6, 60, 62, 67 Paracress (Spilanthes acmella), 339, 346
Parkia
O javanica, 336
Oatmeal, 65 roxburghii, 339, 347
Ocimum basilicum, 192 Parmelia sulcata, 287
Octopamine receptors, 193 Parmotrema, 280, 282, 285–288
Odor, 5, 124, 330 grayanum, 286
Oligonychus coffeae, 236, 238, 243, 245, hysterophorus, 245
246, 248 praesorediosum, 286
One-way ANOVA, 179 reticulatum, 280, 282, 285–287
Orange, 6–9, 206, 285, 374 tinctorum, 286
Organic Partial dehydration, 39, 41
farming, 191, 299 Participatory rural appraisal (PRA), 152,
food production, 205, 210 154
Organisms, 5, 219, 225, 263, 281, Pasteurization, 6
292–294, 299, 300, 320, 328, 330, 331 Patchouli, 192
398 Index

Pathogenesis related (PR), 306 Plant

Pathogenic bacterial cultures, 319 bioregulators, 104, 112, 114
P-cymene, 197, 201, 362, 363 diseases, 226, 280, 281, 295, 299, 307
Pediculus humanus capitis, 244 forecasting, 226
Pelargonium, 201 extracts, 243, 251, 252, 254, 256–258,
Penetrometer, 90, 123, 126 317
Penicillin G, 283 protection, 211, 218–221, 225
Penicillium virus, 229–231, 233
implicatum, 33 Planttago ovata, 296
italicum, 286 Poaceae, 190, 195, 198
Peppermint, 195, 196, 203, 205, 257, 317, Pogostenmon heyneanus, 192
318, 324–331 Policy makers, 225
Percentage Pollinators, 208, 209, 225
disease index (PDI), 253, 255 Polyclonal antibody (PAb), 232
flower opening, 107 Polyethylene, 60, 127, 128
weight loss, 125 Polygonal segments, 118, 124, 135
Persistent toxicity (PT), 237, 239, Polymerase chain reaction (PCR), 230,
241–244, 246, 248 233
Pesticides, 191, 192, 207, 210, 264–266, Polymers, 93
268, 270, 272, 273 Polypeptides, 195, 305
Pests, 190–196, 201, 206, 207, 210, Polyphenol, 4, 8, 72, 130, 176, 178–180,
218–225, 227, 236, 262, 264, 342 183, 184
Petal wilting, 105, 107 oxidase, 133
Petri plates, 254, 320 Polyphenolic compounds, 15
Petroleum ether, 90 Polyphenols, 80, 176, 180, 182, 195
PH, 4, 6, 49, 59, 63, 91, 129, 133, 221 Polysaccharides, 293
Pharmaceuticals, 28, 196 Post treatment observation, 239, 240
Phenethyl alcohol, 201 Postharvest, 52, 99, 146
Phenolic compounds, 177, 199 disease, 87, 89, 175, 177
Phenolics, 15, 74, 80, 82, 130 fungi Rhizopus, 135
Philippines, 70 incidence, 118, 130, 135
Phosphomolybdenum method, 179 laboratory, 123, 125, 131
Phosphorus, 56, 62, 63, 66, 176, 262, 342, processing package, 14
343, 382, 383 Posttreatment observation, 241
Phytic acid, 64 Potassium, 56, 62, 63, 66, 81, 176, 262,
Phytochemicals, 3, 336, 344, 349 282, 342, 343, 346, 382, 383
Phytonutrients, 4 Potato dextrose broth, 283
Phytopathogenic fungi, 279, 281, 282, Potent, 242, 325, 329, 361, 370, 376
286, 287 Potexvirus, 232
Phytopathogens, 280 Potyvirus, 232
Phytophthora, 287, 295 Power ultrasound, 7
Phytotoxicity, 210, 237, 240, 241, 246, Preservation, 4–9, 39, 41, 52, 141, 142,
247, 257 209, 364
Pickle, 71 Pressure, 7, 31, 34, 46, 59, 61, 64, 71, 167,
Pigment level, 146 169, 171, 176, 283, 345–347, 354, 356,
Pineapple, 6, 46, 175–184 359, 374
Piperitone, 200, 324 Pre-treatments, 43
Index 399

Probit R
analysis, 241, 242
Radical scavenging activity, 179, 182, 183
mortality curve, 238
Randomized block design (RBD), 262,
Processing, 4–9, 14–17, 23, 24, 41, 57–59,
263, 360
67, 130, 131, 154, 157, 163, 167, 375
Rauvolfia
technology, 14, 23
serpentine, 28, 374
Productivity, 41, 158, 218, 221, 224, 227,
tetraphylla, 374
262, 280 Ready-to-serve (RTS), 69–72, 74–76
Profitability, 227 Recombinant, 231–233
Protection, 3, 118, 123, 135, 153, 190, Recombinant
192, 194, 202, 205, 207, 211, 217, 218, antibody, 232
222–226, 306, 308 antigen, 231
Protein, 55, 56, 62, 63, 65, 66, 229–231, Recommended dietary allowance (RDA),
262, 305, 306, 343, 347, 384 341
Proteolytic enzyme, 177 Red spider mite (RSM), 236–238, 242,
Proteus vulgaris, 316, 318, 319, 325, 326 243, 246, 248
Protolchesterinic acid, 287 Redox potential, 4
Provitamin A, 40 Reducing sugar, 62, 66
Proximate analysis, 56 Refractometer, 71, 82, 90
Pseudaletia unipuncta, 195 Regeneration, 153
Pseudomonas Relative fresh weight (RFW), 106, 107,
aeruginosa, 316, 318, 319, 325, 326, 109
346 Repellent, 192, 194, 201, 375
fluorescens, 223, 304 Rheum nobile, 340, 347
Puccinia spegazzinii, 223 Rhizoctonia solani, 295–297, 304, 305,
Pulegone, 193, 317, 324, 330 307
Pulse Rhizome (Z. officinale Roscoe), 14–17,
generator, 8 168
width, 9 Rhyzopertha dominica, 196, 201
Pulsed electric field (PEF), 7–9 Ringworm infection, 27, 28
Pyrethroids, 191, 193 Roasting, 57, 125, 339
Pyrethrum, 298 Rodents, 218
Pythium, 287, 295–298, 302, 304, 307 Rosella (Hibiscus sabdariffa), 339, 346
aphanidermatum, 305 Rosemary, 195, 196, 204, 205, 207, 355
ultimum, 287, 296, 304 Rosmarinus officinalis, 195, 204
Rotary drum washer, 14, 16, 18, 23
Q Royal Horticultural Society, 123, 126
Quality, 4–9, 14, 16, 41, 42, 51, 52, 56, 59, Rural areas, 40, 153, 336–338
62, 64, 69, 71, 72, 80, 84, 88, 89, 99, Rutaceae, 193
104, 109, 119, 122, 123, 127, 128, 131,
132, 136, 139–141, 146, 195, 222–226, S
231, 280, 365 Sabouraud dextrose agar, 29
Quantity, 29, 41, 84, 218, 222, 223, 231, Saccharomyces
241, 284 cerevisiae, 88–90, 92, 94, 95, 97–99
Quercus pachyphylla, 165 pombe, 89
400 Index

Safety, 4–6, 9, 88, 94, 131, 135, 152, 217, gilo, 345
226, 294 khasianum, 345
Salmonella kurzii, 345
typhi, 316, 318, 320, 325, 326 macrocarpon, 344
typhimurium, 316 myriacanthum, 344
Salsola bayosma, 252 nigrum linn, 379, 380
Salvia aucheri Bentham, 356 sisymbrifolium, 345
Sand media, 146, 148 spirale, 345
Sanitation, 130, 131 torvum, 163, 339, 344, 345
Sarawak, 175–177, 179–184 xanthcarpum, 344
pineapple, 181–183 Solid gain (SG), 40, 42, 46, 47, 52
Sartorius electronic balance, 123 Sophlong (Moghania vestita), 339, 347
Schima wallichi, 165 Southern Asia, 194
Schinus molle, 208 Spasmolytic applications, 195
Schizostachylum Spectrophotometrically, 82, 91
dullooa, 158 Spiders, 225
fuchsiamum, 158 Spilanthes
Schizzophylum commune, 164 acmella, 339, 346, 380, 381, 383–385
Sclerotium rolfsii, 295–297, 305 paniculata, 346
Senescence, 105, 107, 109, 113 Spine gourd, 340
Sensory Spinosad, 261, 262, 272, 274
appraisal, 126, 129, 133 Spodoptera, 193, 204, 261, 262, 266, 268,
evaluation, 71, 80, 84, 126 270, 273, 276
qualities, 81, 85 litura, 193, 204, 261, 262, 266, 268, 276
quality, 42, 123, 131, 136 Spoilage, 4–6, 41, 88–91, 93–95, 99
score, 40, 50–52, 70 Stability, 7, 227, 331, 340
Shelf-life, 4, 6, 59, 60, 63, 70, 118–120, Standard deviation (SD), 179, 323
125, 127, 128, 131–133, 135, 136, 299, Standard error of mean (SEM), 382
300, 316, 327, 331, 345 Staphylococcus aureus, 316, 323, 328, 346
Shigella dysenteriae, 316 Staple, 56, 57, 63, 67, 119
Silica gel, 140–143, 145, 148 Statistical analysis, 91, 179, 255, 285, 323,
Silver oak (Grevillea robusta), 141 382
Single chain variable fragment, 232 Stegobium paniceum, 192
Sitophilus oryzae, 192–194, 198 Stem
Skin disease, 380, 381 bending, 105, 107
Slicing, 14, 16–18, 23–25, 131, 132, 163 elongation, 104, 109, 110
Snake gourd, 342 rotting, 107
Trichosanthes cucumerina, 339 Sterilized chopping board, 131
Socioeconomic survey, 154 Storability, 51, 89, 99, 135
Sodium, 62, 66, 132 Storage, 7, 14, 28, 40, 41, 47, 52, 63, 70, 72,
chloride, 90 74, 75, 80–85, 88–90, 92–99, 118–120,
metabisulfite, 119, 131–133, 136 126–130, 132–136, 199, 202, 323
Solanum, 163, 252, 336, 338, 339, 344, trial, 132
345, 380, 383–385 Strawberry, 6
americanum, 336, 339 Streptomycin sulfate, 283
berbisetum, 345 Striking colors, 103
ferox, 345 Sucrose, 72, 104–108, 113, 136
Index 401

Sugar, 41–43, 46, 49, 51, 52, 70–72, 81, saturejoides, 354, 357, 364
95, 97, 105, 124, 146, 176, 209 vulgaris, 197, 204, 208
Sun drying, 14, 15 Thysanolaena maxima, 158
Sustainability, 227 Ticks, 192, 207
Sweet William, 104–107, 109–114 Timeliness, 222
Sword bean, 339, 343 Titratable acidity (TA), 70, 82, 88, 90, 95,
Synergistic effect, 203, 204 96, 99
Syrup, 40, 42–44, 46, 47, 49, 51, 52, 209 Tobamovirus, 232
Syzygium Tomato, 92, 95, 204, 224, 232, 251–258,
aromaticum, 199 298, 307, 338, 339, 348
cumini, 69, 70, 76 Solanum lycopersicon L., 252
Tospovirus, 232
T Total
folic acid, 342
Tamarix aphylla, 252
inoculums density (TID), 284
Tannins, 64, 80, 327, 358
soluble solids (TSS), 44, 70, 71, 82, 88,
Tea, 198, 206, 236–240, 243, 246, 248,
90, 95–97, 99
346, 360
Totapuri, 39, 40, 42–52
Temperature, 6, 7, 9, 15, 21, 22, 31, 39–43,
slices, 39, 40, 43, 44, 46–52
52, 76, 89, 106, 127, 128, 131, 132, 135, Toxic effect, 199, 202, 330
145, 178, 221, 237, 238, 282, 283, 318, Toxicity, 35, 190–205, 207–209, 211, 237,
319, 323, 359, 360, 371 239–244, 246, 248, 293, 316, 326, 331
Tenaderm, 34–36 Training, 25, 222
Tenebrionidae, 192, 194 Transformation, 82, 241, 264, 293
Terpineol, 193, 197, 199–201, 204, 363 Transportation, 166
Testa, 56, 58–61, 63, 64 Treatment
Testosterone, 35 chamber, 8
Tetraanychus kanzawai, 244 temperature, 9
Tetranychus urticae, 194, 196, 204, 243, Tremellomycetes fuciformis, 164
244 Trialeurodes vaporariorum, 196
Texture, 40, 41, 50, 51, 56, 60, 61, 118, Tribals, 338, 339, 344, 346, 347, 369, 370,
120, 123, 124, 129, 133, 135, 145, 146 376
Thermal Tribolium castaneum, 192, 196, 208
conductivity, 21–23 Trichoderma, 220, 223, 224, 296–298,
pasteurization, 4, 9 300–303, 305, 307
processing, 6, 7, 9 Trichophyton
treatment, 4–6 mentagrophytes, 33
Thermostability, 316, 327, 332 rubrum, 28–32, 34, 36
Thevetia peruviana, 375 tonsurans, 33
Thielaviopsis paradoxa, 177 violaceum, 33
Thin layer chromatography (TLC), 282 Tridax procumbens, 242, 243
Thrips tabaci, 195, 196 Triplicates, 29, 90, 91, 284
Thuja orientalis, 140–148 Trissolcus basalis, 208
Thujone, 193 Trypsin inhibitors, 64
Thyme, 195, 197, 204, 207, 210, 354–356, Turbidity, 5
358–365 Turmeric, 14–18, 20, 23–25, 280, 298
Thymus C. longa, 15
402 Index

Typhoid, 316, 374 W

Tyramine, 193
Water
Tyrophagus putrescentiae, 202
activity (AW), 4, 6, 41, 47
U dehydrated sample, 47
loss (WL), 40–42, 46, 47, 52, 93, 128,
Ultrasonic processing, 7 130
Ultrasound, 7 Weight
Ultraviolet light irradiation, 7 loss, 19, 40, 47, 90, 118, 125, 127, 128,
Ultraviolet-C (UV-C), 8 133, 134, 136
Underexploited vegetables, 336, 340, 341,
reduction (WR), 40, 42, 47, 52
344, 347–349
during osmosis, 47
Unpasteurized juices, 4, 5
West Indies, 62, 63, 70
Unripe fruit, 119, 125
White sand, 140, 143
Urinary
Wild
diseases, 345
foods, 152, 157, 163, 164
tract infection, 317
Usnic acid, 287 fruits, 162, 172
Ustilago maydis, 287 Winter annuals, 103–105, 113, 114
flowers, 103–105, 113
V World Health Organization (WHO), 28,
220, 293, 317, 357, 372, 381
Vase
decoration, 140, 141 X
life, 104, 105, 107, 109, 110, 112–114
solution uptake rate (VSUR), 104, 106 X-rays, 382
Vegetable, 41, 55–59, 61, 63, 67, 124, 164, emission, 382
169, 223, 224, 262, 299, 307, 336–340, fluorescence, 380, 382
345, 347, 348 Xylem vessels, 105, 107
Verbenol, 193, 199
Verbenone, 193, 195, 362 Y
Vernier caliper, 107 Yeast, 4, 52, 89, 90, 92–99, 232
Victorian age, 140 potato dextrose agar (YPDA), 89, 90
Vigna Yield of,
umbellate, 336, 339, 346 osmotically dehydrated mango slice, 46
vexillate, 346 Young Mizo Association (YMA), 166
Village Forest Development Committee
(VFDC), 152, 154, 156, 157, 165 Z
Vitamin C, 40, 49, 71, 80, 176, 373
Vitis vinifera, 80 Zanthoxylum
Volatile acanthopodium, 346
monoterpenes, 192 armatum, 336, 346
organic compounds (VOCs), 95 oxyphyllum, 346
Volume of, rhetsa, 336, 339, 345
liquid medium (VLM), 284 Zinc, 62, 66, 383
microscopic field (VMF), 283, 284 Zingiber officinale, 14
Volumetric flask, 90 Zingiberaceae, 14, 27, 29
Vomiting, 35, 317, 318, 329 Zone of inhibition, 324