WOet

Shimaa Ral.
LD JOURNAL OF PHAJournal
World RMAC ofYPharmacy
AND PH ARPharmaceutical
and MACEUTICSciences
AL SCIENCES
SJIF Impact Factor 5.210

Volume 4, Issue 05, XXX-XXX. Research Article ISSN 2278 – 4357

ANTIPROTOZOAL AND ANTIMICROBIAL ACTIVITY OF

SELECTED MEDICINAL PLANTS GROWING IN UPPER EGYPT,
BENI-SUEF REGION

Shimaa Mohammed Abdel Gawad1, 2*, Mona Hafez Hetta1, Samir Anis Ross2 and
Farid Abd El- Reheim Badria3

1
Pharmacognosy Department, Faculty of Pharmacy, Beni-Suef University, 62514, Egypt.
2
National Center for Natural Products Research, and Department of Bio Molecular Sciences,
School of Pharmacy, University of Mississippi, MS 38677, USA.
3
Pharmacognosy Department, Faculty of Pharmacy, Mansoura University, 35516, Egypt.

Article Received on ABSTRACT

15 March 2015,
Alcoholic extracts of fifty six plants cultivated in Beni-Suef
Revised on 06 April 2015, Governorate (Egypt) were screened in vitro for their antiprotozoal and
Accepted on 27 April 2015
antimicrobial activities. Emblica officinalis, Quercus infectoria and
Punica granatum were the most active as antimalarial with IC50 4.92,
*Correspondence for
Author 2.51, 10.61µg/mL respectively against the chloroquine sensitive strain
Shimaa Mohammed of Plasmodium falciparum and IC50 3.1, 2, 7.4µg/mL respectively
Abdel Gawad against chloroquine resistant strain of Plasmodium falciparum. The
Pharmacognosy
extracts of Ricinus communi, Corchorus olitorius and Psidium guajava
Department, Faculty of
Pharmacy, Beni-Suef
were the most active as antileishmanial and the percentage of
University, 62514, Egypt. inhibition were 91.4%, 90.9% and 90.3% respectively. The extract of
[email protected]
Emblica officinalis, Punica granatum, Quercus infectoria, Ricinus
[email protected]
communi, Tamarix nilotica, Camellia sinensis and Curcuma aromatic
were active against Candida glabrata with IC50 <8, <8, <8, 52.25, 17.12, 45.3, 26.91 µg/mL
respectively while the extract of Emblica officinalis, Quercus infectoria galls and Curcuma
aromatica were the most active against Cryptococcus neoformans with IC50 10.8, <8, 50.6
µg/mL respectively. A good antibacterial activity against MRSA was shown by the ethanol
extracts of Spinacia oleracea, Corchorus olitorius, Cyperus alopecuroids, and Sesamum
indicum with IC50 13.5, 45.31, 18.73 and 19.32µg/mL respectively. Tannins and phenolic
acids constituents of these plants proposed to be responsible for the activity through carbonic
anhydrase inhibition.

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

KEYWORDS: Medicinal plants; Beni-Suef; Egypt; Antiprotozoal; Antimicrobial; Carbonic

anhydrase inhibitors.

INTRODUCTION
Natural products were used by humans in many countries such as Egypt, China and India as
drugs for thousands of years.[1] It was reported in a recent study conducted by the information
and decision support centre in Egypt that 23% of the Egyptian use medicinal plants as a
remedy.[2] There are more than 250,000 species of higher plants in the world among them
only a small percentage of them have been investigated for their potential value as drugs.[3]

There are several approaches for selecting plants as candidate for drug discovery such as
random selection followed by chemical or biological screening, follow up of ethnomedical
uses of plants and biological activity reports.[4,5]

There is no previous study could be traced on plants growing in Beni-Suef Governorate as

antiprotozoal and antimicrobial agents, therefore the present study focused on a systemic
evaluation of a selection of commonly growing plant species in this area.

Infectious diseases either protozoal such as malaria and leishmania or microbial infections are
considered as the major killing factors in the third world countries and the most important
causes of premature death.[6] Difficulty of controlling the sources of infection, the high cost
of treatment/prevention, poor compliance, low efficacy, poor safety and drug resistance are
the major factors that may retard the treatment of these diseases. The drug resistance has
further complicated the treatment of infectious diseases in immune-compromised AIDS and
cancer patients. Therefore, there is always need for the development of new and more
effective drugs. In this respect, natural products offer good sources for new drug discovery.[6]
Various antiparasitic drugs have been developed from natural sources, including Quinine,
Artemisinine and Atovaquone as antimalarials and Amphotericin B as antileishmanial drug.[7]
The Centers for Disease Control and Prevention[8] received information that 19 locally
transmitted malaria cases have been reported in one village in Aswan Governorate in Egypt.
These are the first malaria cases seen in Egypt since 1998.

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

MATERIALS AND METHODS

Plant material
The fifty six plants used in this study (Table 1) were collected at the period from September
to December 2010 from different regions: Beni-Suef fields and herbal markets. The plants
were kindly authenticated by Prof. Dr. Lotfy Boules, Prof. Dr. Mohammed El Gibali, (senior
botanists, Faculty of Science, Cairo University), Dr. Mahmoud Omar (lecturer at plant
taxonomy Department, Faculty of Science, Beni-Suef University) and Mrs Thérèz Labib
(botanist specialist at Orman garden, Giza, Egypt). Voucher specimens (BUPD-35: BUPD-
90) were kept at the Herbarium of the Department of Pharmacognosy, Faculty of Pharmacy,
Beni-Suef University.

Preparation of extracts
The air-dried powder of each plant material (100 g) was extracted with 75% ethanol (500 mL
×3). Each extract was filtered off and concentrated using a rotatory evaporator. Each residue
was weighed and stored in a refrigerator at −4 °C until used.

Microbial strains used

 Antimalarial activity was tested in vitro against Chloroquine sensitive (D6, Sierraleon)
and Chloroquine resistant (W2, Indo-China) strains of Plasmodium falciparum.
 Antileishmanial activity was tested against Leishmania donovani promastigotes.
 The used fungal strains were Candida albicans (ATCC 90028), Candida glabrata
(ATCC 90030), Candida krusei (ATCC 6258), and Aspergillus fumigates (ATCC 90906).
 The used bacterial strains were methicillin-resistant Staphylococcus aureus (ATCC
33591), Cryptococcus neoformans (ATTC 90113), Staphylococcus aureus (ATTC
29213), Escherichia coli (ATCC 35218), Pseudomonas aeruginosa (ATCC 27853), and
Mycobacterium intracellulare (ATCC 23068).

Standards used
For the antimalarial activity, Chloroquine and Artemisine were used as positive controls
while Pentamidine and Amphotericin B were used as positive controls in the antileishmanial
testing. Amphotericin B was used also as a positive control in the antifungal activity and
Ciprofloxacine in the antibacterial. All reference drugs were obtained from (ICN
Biomedicals, Ohio).

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

Biological Assays
Antimalarial screening[9]
Crude extracts were initially tested in duplicate against D6 Plasmodium falciparum strain as
a primary screen at 15.867 µg/mL and percent inhibition (% inhibition) was calculated
relative to negative and positive controls. The extracts which showed more than 50%
inhibition proceeded to the secondary assay. In the Secondary antimalarial assay, the samples
were dissolved to 20 mg/mL and tested at 47.6, 15.867, and 5.289 µg/mL and IC50s in µg/mL
were calculated against both D6 and W2 strains. In addition to P. falciparum strains, samples
were tested in the VERO mammalian cell line as an indicator of general cytotoxicity. The
selectivity indices (SI) – ratio of VERO IC50 to D6 or W2 IC50- were calculated. All IC50
were calculated using XLFit fit curve fitting software.

Antileishmanial screening[10, 11]

A culture of Leishmania donovani promastigotes was grown in RPMI 1640 medium
supplemented with 10% GIBCO fetal calf serum at 26°C. Growth of leishmanial
promastigotes was determined by the Alamar Blue assay (BioSource International, Camarillo,
CA). Standard fluorescence was measured by a Fluostar Galaxy plate reader (excitation
wavelength, 544 nm; emission wavelength, 590 nm). Crude extracts were initially tested in
duplicate as a primary screen at 80µg/ mL and % inhibitions were calculated.

Antifungal and antibacterial screening[9]

Crude extracts were initially tested in duplicate as a primary screen at 50µg/ mL and %
inhibitions were calculated relative to negative and positive controls. The extracts which
showed more than 50% inhibition were proceeded to a secondary assay. In the latter, the
samples were dissolved to 20mg/ mL and tested at 200, 40, 8µg/mL and then IC50 were
determined.

RESULTS
Antimalarial activity (Table 1)
Results showed full inhibition with Emblica officinalis and Quercus infectoria (100%
inhibition) followed by Punica granatum (96 % inhibition) with IC50 values of 4.92, 2.51,
10.61µg/mL respectively against chloroquine-sensitive (D6) strain of P. falciparum and IC50
values of 3.1, 2.0, 7.4 µg/mL respectively against chloroquine- resistant (W2) strain of P.
falciparum compared to Artemisine (IC50 value of 0.0269 µg/mL ; D6, 0.0165; W2) and

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

Chloroquine (IC50 of 0.0098 µg/mL ; D6, 0.187; W2) without showing any cytotoxicity to
the mammalian cells (Table 2).

Antileishmanial activity (Table1)

Ricinus communi, Corchorus olitorius and Psidium guajava showed the best activity as
antileishmanial among the selected plants and their % of inhibition were 91.4%, 90.9% and
90.3% respectively.

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

Table 1. Results of antimalarial and antileishmanial activity of selected plants collected from Beni-Suef Governorate, Egypt.

Antimalarial D6 Antileishmanial
Plant Common name Family Part used
(%Inhibition) (%Inhibition)
Alhagi graecorum Boiss Camel thorn, manna tree Papilionaceae Leaves 47 0
Amaranthus lividus L. Pigweed Amaranthaceae Herb 46 6.5
Dinosaur plant, Jericho rose, Mary’s
Anastatica hierochuntica L. flower, Palestinian tumbleweed, Cruciferae Herb 44 0
resurrection plant
Leaves ,
Artemisia Absinthium L. Absinthe wormwood, green ginger Asteraceae 52 25.1
Flowers
Aster squamatous Sprengel Aster, Starwort Compositae Herb 45 52.4
Beta vulgaris var.cicla L. Spinach beet Chenopodiaceae Leaves 32 9.6
Camellia sinensis (L.)
Green tea plant Theaceae Leaves 44 48.5
Kuntze
Cartagena ipecacuanha Brot. Cartagena Ipecacuanha, Rio ipecac Rubiaceae Root 70 84.9
Chenopodium murale L. Nettleleaf goosefoot Chenopodiaceae Herb 39 6.3
Cichorium endivia L. cultivated endive Asteraceae Leaves 44 0
Roots,
Cichorium intybus L. Chicory Asteraceae 42 0
Leaves
Cinnamomum cassia (Nees
Chinese cassia, Chinese cinnamon Lauraceae Bark 44 0
& T.Nees) Farw.
West African Cherry Orange,
Citrus reticulate Blanco Rutaceae Leaves 33 75.1
Omuboro
Conyza dioscoridis (L.) Desf Horseweed, butterweed, fleabane Compositae Herb 38 60.2
Corchorus olitorius L. Jew's mallow Tiliaceae Leaves 37 90.9
Curcuma aromatic Salisb. Curcuma Zingebracea Rhizomes 52 44.1
Cymbopogon Proximus
Halfa bar, Lemongrass Poaceae Leaves 47 6.1
Spreng.

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

Cyperus alopecuroides Leaves,

Foxtail flatsedge Cyperaceae 28 63
Rottb. Flowers
Leaves,
Cyperus Rotundus L. Nut grass, Tiririca Cyperaceae 44 0
Flowers
Daucus carota L. Wild carrot, Queen Anne's lace Apiaceae Leaves 41 0
Desmostachia bipinnata (L.) Halfa grass, big cordgrass, salt reed-
Poaceae Herb 44 0
Stapf grass
Emblica officinalis (L.) Kurz Indian gooseberry Phyllanthaceae Herb 100 64.1
Eruca sativa Mill. Brassicaceae Leaves 33 0
Ficus carica L. Common fig Moraceae Leaves 36 0
Roots and
Glycyrrhiza glabra L. Liquorice Fabaceae 49 0
rhizomes
Flowers
Hibiscus sabdariffa L. Roselle, karkadeh Malvaceae Calyx and 34 0
epi-calyx
Hyphaene thebaica (L.)
Doum palm Arecaceae Fruits 35 0
Mart.
Lawsonia inermis L. Loose strife, Henna Lythraceae Leaves 0 0
Lupinus termis L. Lupine Fabaceae Seeds 0 0
Malva parviflora L. Cheeseweed Malvaceae Leaves 50 68.3
Mentha longifolia (L.) Huds. Horsemint Labiatae Herb 47 28.4
Morus alba L. White mulberry Moraceae Leaves 29 81.3
Opuntia ficus indica (L.)
Prickly pear Cactaceae Leaves 52 0
Mill.
Origanum majorana L. Sweet marjoram Lamiaceae Herb 41 31.1
Peganum harmal L. Harmal, Syrian rue Nitrariaceae Seeds 70 83
Phaseolus vulgaris L. French bean Papilionaceae Leaves 41 79.3
Phragmites communis Leaves ,
Common reed Poaceae 0 0
(Cav.) Trin. ex Steud. Flowers
Pimpinella anisum L. Aniseed Umbelliferae Fruits 44 10.1
Psidium guajava L. Guava Myrtaceae Leaves 50 90.3

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

Fruit
Punica granatum L. Pomegranate Lythraceae 96 13.9
pericarp
Quercus infectoria gall Aleppo oak Fagaceae Galls 100 74.6
Ricinus communi L. Castor-oil plant Euphorbiaceae Leaves 40 91.4
Salix subserrata Willd. Salix Salicaceae Leaves 0 49.2
Sesamum indicum L. Sesame Pedaliaceae Leaves 0 75.9
Leaves ,
Sesbania sesban (L.) Merr. Riverhemp Leguminosae 0 0
Flowers
Sisymbrium irio L. London rocket Brassicaceae Herb 3 0
Solenostemma argel (Delile)
Argel Apocyanaceae Leaves 43 1.7
Hayne
Spinacia oleracea L. Spinach Chenopodiaceae Leaves 2 81
Fruit and
Tamarindus indica L. Tamarind Fabaceae 0 0
Seeds
Tamarix nilotica L. Nile tamarisk Tamaricaceae Herb 8 0
Thymus vulgaris L. Common thyme Lamiaceae Herb 0 0
Leaves ,
Tilia cordata Mill. Small leaved lime Tiliaceae 3 31.6
Flowers
Trifolium alexandrinum L. Egyptian clover Leguminosae Leaves 0 58.5
Withania somnifera (L.)
Ashwagandha Solanaceae Leaves 15 35
Dunal.
Zingiber officinale Roscoe Ginger Zingebracea Rhizome 38 88.2
Zizyphus Spina-christi (L.)
Christ’s Thorn Jujube Rhamnaccae Leaves 0 24.9
Desf.
Standard
Chloroquine 97 -
Amphotericin B - 98.7

www.wjpps.com Vol 4, Issue 05, 2015

 Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

Table 2. IC50 ( µg/mL) of most active plants as anti-malarial

Plasmodium
Plant Plasmodium falciparum (D6) D6 (SI) W2 (SI) VERO
falciparum (W2)
Emblica officinalis (L.)
4.92 >9.7 3.1 >15.4 No cytotoxicity
Kurz
Quercus infectoria gall 2.51 >19 2 >22.9 No cytotoxicity
Punica granatum L. 10.61 >4.5 7.4 >6.4 No cytotoxicity
Standard
Chloroquine 0.0098 >24.4 0.187 >1.3 No cytotoxicity
Artemisine 0.0269 >8.8 0.0165 >14.4 No cytotoxicity
IC50s were calculated based on 2 replicates, SI: selectivity index, D6: Chloroquine sensitive strainW2: Chloroquine resistant strain.

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

Antimicrobial activity (Table 3)

 The ethanol extract of Emblica officinalis, Punica granatum, Quercus infectoria, Ricinus
communi, Tamarix nilotica, Camellia sinensis and Curcuma aromatic were active against
Candida glabrata with IC50 values of <8, <8, <8, 52.25, 17.12, 45.3, 26.91 µg/mL
respectively when compared to Amphotericin B (IC50 value of 0.283 µg/mL) (Table 4).
 The ethanol extract of Emblica officinalis, Quercus infectoria (galls) and Curcuma
aromatica were the most active against Cryptococcus neoformans with IC50 values of 10.8,
<8, 50.6µg/mL respectively when compared to Amphotericin B (IC50 Value of 0.269
µg/mL) (Table 4).
 The ethanol extract of Spinacia oleracea, Corchorus olitorius, Cyperus alopecuroids and
Sesamum indicum were the most active against MRSA with IC50 values of 13.5, 45.31,
18.73 and 19.32µg/mL respectively when compared to Ciprofloxacin (IC50 value of
0.091µg/mL) (Table 4).
 The ethanol extract of Quercus infectoria was active against E. coli with IC50 value of
40µg/mL when compared to Ciprofloxacin (IC50 value of 0.003µg/mL) (Table 4).
 The ethanol extract of Quercus infectoria was the most active extract against P. aeruginosa
with IC50 = <8µg/mL when compared to Ciprofloxacin (IC50 = 0.092 µg/mL) (Table 4).

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

Table 3. Results of antimicrobial activity (% inhibition) of selected plants collected from Beni-Suef Governorate, Egypt
Fungi Bacteria
Plant
Ca Cg Ck Af Cn Sa MRSA Ec Pa Mi
Alhagi graecorum Boiss 0 0 0 0 58 1 0 4 0 4
Amaranthus lividus L. 0 1 0 1 59 2 0 4 0 0
Anastatica hierochuntica L. 0 8 0 5 56 0 0 17 3 0
Artemisia Absinthium L. 0 21 0 2 41 3 4 1 0 0
Aster squamatous Sprengel 0 7 0 3 17 14 9 9 0 0
Beta vulgaris var.cicla L. 0 1 0 3 7 0 0 3 0 2
Camellia sinensis (L.) Kuntze 0 95 0 12 15 16 31 30 8 0
Cartagena ipecacuanha Brot. 11 17 17 9 58 7 4 18 0 1
Chenopodium murale L. 0 0 0 14 60 6 2 11 0 0
Cichorium endivia L. 0 4 0 10 40 0 0 9 0 0
Cichorium intybus L. 0 13 1 8 42 0 0 16 0 0
Cinnamomum cassia (Nees & T.Nees)
0 7 0 5 61 0 1 6 1 0
Farw.
Citrus reticulate Blanco 0 9 0 10 29 4 10 4 0 0
Conyza dioscoridis (L.) Desf 0 10 0 8 24 0 7 14 0 0
Corchorus olitorius L. 1 15 0 10 33 34 77 0 0 0
Curcuma aromatic Salisb. 1 80 3 3 77 9 2 16 1 0
Cymbopogon Proximus Spreng. 1 11 3 9 10 7 1 10 3 0
Cyperus alopecuroides Rottb. 0 5 0 6 45 26 74 0 0 0
Cyperus Rotundus L. 11 5 1 11 57 7 2 12 1 0
Daucus carota L. 3 6 0 11 22 0 4 11 1 0
Desmostachia bipinnata (L.) Stapf 5 6 4 10 21 1 5 18 0 0
Emblica officinalis (L.) Kurz 0 100 25 12 99 24 19 42 13 0
Eruca sativa Mill. 2 3 0 10 14 0 0 23 0 0
Ficus carica L. 0 2 0 3 63 1 1 13 0 0
Glycyrrhiza glabra L. 1 7 3 5 34 21 12 21 0 0
Hibiscus sabdariffa L. 0 11 0 3 43 0 14 27 2 0

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

Hyphaene thebaica (L.) Mart. 7 6 0 8 8 4 3 18 2 0

Lawsonia inermis L. 0 38 0 8 2 7 3 4 6 0
Lupinus termis L. 0 13 6 8 33 5 2 21 0 0
Malva parviflora L. 1 4 6 12 14 14 9 16 0 0
Mentha longifolia (L.) Huds. 1 6 0 10 7 4 6 25 0 0
Morus alba L. 8 12 4 13 48 12 14 17 0 0
Opuntia ficus indica (L.) Mill. 3 1 0 8 22 3 1 8 0 0
Origanum majorana L. 0 7 0 10 1 2 4 22 0 0
Peganum harmal L. 0 0 0 6 0 0 2 12 7 14
Phaseolus vulgaris L. 0 0 0 5 8 18 7 8 4 0
Phragmites communis
0 0 0 5 6 0 1 0 2 3
(Cav.) Trin. ex Steud.
Pimpinella anisum L. 0 6 0 3 51 0 0 12 3 0
Psidium guajava L. 4 18 0 14 24 40 36 5 3 0
Punica granatum L. 0 100 3 4 56 7 29 43 32 0
Quercus infectoria gall 0 100 62 5 99 40 36 53 74 0
Ricinus communi L. 0 100 0 5 64 20 47 0 0 0
Salix subserrata Willd. 0 0 0 5 9 12 22 17 0 3
Sesamum indicum L. 0 0 0 7 2 33 71 27 1 0
Sesbania sesban (L.) Merr. 0 6 0 4 2 0 1 9 2 0
Sisymbrium irio L. 0 0 0 6 2 0 0 8 4 1
Solenostemma argel (Delile) Hayne 0 0 0 5 24 0 0 6 0 0
Spinacia oleracea L. 0 2 0 3 5 10 80 18 7 4
Tamarindus indica L. 0 0 0 2 1 0 1 4 11 0
Tamarix nilotica L. 0 99 0 7 1 0 0 28 3 0
Thymus vulgaris L. 0 16 0 10 0 0 1 22 3 0
Tilia cordata Mill. 0 10 0 9 4 14 37 16 1 1
Trifolium alexandrinum L. 0 6 0 10 3 14 19 18 4 0
Withania somnifera (L.)Dunal. 0 5 0 7 3 0 6 13 1 0
Zingiber officinale Roscoe 6 22 13 11 41 2 8 10 0 0
Zizyphus Spina-christi (L.) Desf. 0 3 0 6 4 0 0 9 2 5

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

Standard
Amphotericin B (for fungi) 100 99 100 99 100 - - - - -
Ciprofloxacin (for bacteria) - - - - - 89 96 98 97 85
Candida albicans (Ca), Candida glabrata (Cg), Candida krusei (Ck), and Aspergillus fumigates (Af) and the bacteria methicillin-resistant
Staphylococcus aureus (MRSA), Cryptococcus neoformans (Cn), Staphylococcus aureus (Sa), Escherichia coli (Ec), Pseudomonas aeruginosa
(Pa), and Mycobacterium intracellulare (Mi).

Table 4. IC50 (µg/mL) of some of the active plants in primary antimicrobial screening

Fungi Bacteria
Plant
Cg Cn MRSA Ec Pa
Camellia sinensis 45.3 - - - -
Corchorus olitorius - - 45.31 - -
Curcuma aromatic 26.91 50.6 - - -
Cyperus alopecuroids - 18.73 - -
Emblica officinalis ˂8 10.8 - - -
Punica granatum ˂8 9.8 - - -
Quercus infectoria ˂8 ˂8 - 40 <8
Ricinus communi 52.25 - - - -
Sesamum indicum - - 19.32 - -
Spinacia oleracea L. - - 13.5 - -
Tamarix nilotica 17.12 - - - -
Standards
Amphotericin B (for fungi) 0.283 0.269 - - -
Ciprofloxacin (for bacteria) - - 0.091 0.003 0.092
IC50s were calculated based on 2 replicates
Candida glabrata (Cg), Cryptococcus neoformans (Cn), methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli (Ec) and
Pseudomonas aeruginosa (Pa).

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

DISCUSSION
Based on the activity and selectivity, twelve plant extracts among the tested plants could be
considered as promising and interesting to be further elaborated through purification and
biological evaluation on an individual compound basis. Camellia sinensis, Corchorus
olitorius, Curcuma aromatica, Cyperus alopecuroids, Emblica officinalis, Psidium guajava,
Punica granatum, Quercus infectoria galls, Ricinus communi, Sesamum indicum Spinacia
oleracea, and Tamarix nilotica.

Emblica officinalis, Quercus infectoria and Punica granatum were found to be the most active
antimalarial plants. The activities of Punica granatum[12] and Emblica officinalis[13] were
previously reported but it is the first report on the activity of Quercus infectoria galls as
antimalarial plant. Ricinus communi, Corchorus olitorius and Psidium guajava were reported
to have significant inhibitory effect as antileishmanial agents. The antileishmanial activity of
Ricinus communi was previously reported[14] but it is the first report on the activity of
Corchorus olitorius and Psidium guajava as antileishmanial plants.

Reviewing the antifungal and antibacterial screening results; it could be assessed that the
activity of Quercus infectoria,[15] Ricinus communi,[16] Punica granatum,[16] and Camellia
sinensis[17] against C. glabrata were previously reported but it is the first report on the activity
of Emblica officinalis, Curcuma aromatic and Tamarix nilotica as antifungal agents against
this strain. Quercus infectoria was previously reported against Cryptococcus neoformans,[18]
but it is the first report on the activity of Emblica officinalis and Curcuma aromatic as
antifungal agents against Cryptococcus neoformans. Activity of Corchorus olitorius against
MRSA was previously reported[19] but it is the first report on the activity of Spinacia oleracea,
Cyperus alopecuroids and Sesamum indicum. The activity of Quercus infectoria against E.
coli was previously reported.[20] The activity of Quercus infectoria against P. aeruginosa was
previously reported.[21]

Emblica officinalis, Quercus infectoria galls and Punica granatum showed prominent activity
as antimicrobial and antiprotozoal against Plasmodium falciparum, Candida glabrata and
Cryptococcus neoformans.

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

On studying the reported phytochemical constituents of these plants (Table 5) we noticed that
the major active constituents in most of them are tannins and phenolic acids and their esters
and these could be related to their biological activity.

In a trial to correlate the activity to the phytochemical constituents of these plants, carbonic
anhydrase inhibition activity was studied. It has been noticed that gallic and ellagic acids and
their derivatives are common constituents in the these plants.

Carbonic anhydrase inhibitors (CAIs) had many uses such as topically acting antiglaucoma,
anticonvulsants, antiobesity and antitumor agents. Thirteen catalytically active isoforms of CA
enzyme were present and belong to alpha, beta, gamma-, delta-, and zeta families and found in
many organisms.[22] CA inhibition was studied ultimately for some pathogenic protozoa
(Plasmodium falsiparum) α-CA,[23] fungi -CA (Cryptococcus neoformans,[24] Candida
albicans,[22] Candida glabrata[25] and some Sacharomyces cerevisiae,[22] and bacteria α-, -,
and/or -CA (Helicopacter pylori, Mycobacterium tuberculosis and Brucella suis), -class
enzymes from E. coli and M. tuberculosis [26].

In addition to sulfonamides and sulfamate, novel chemotypes of CAIs as coumarins, phenols

and fullerenes were also recently reported.[22] A series of phenolic acids and phenol natural
products, such as p-hydroxybenzoic acid, p-coumaric acid, caffeic acid, ferulic acid, gallic
acid, syringic acid, quercetin, and ellagic acid showed inhibitory effects against the
metalloenzyme carbonic anhydrase against all isozymes.[27]

Ellagitannins as inhibitors against carbonic anhydrase have been previously isolated from the
pericarps of Punica granatum L.[28]

www.wjpps.com Vol 4, Issue 05, 2015

 Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

Table 5. The reported active constituents of the active antiprotozoal and antimicrobial plants
Medicinal plant Active constituents
Camellia sinensis Catechin-based flavonoids such as epigallocatechin-3-gallate (EGCG), epicatechin-3-gallate (ECG), epigallocatechin
(EGC) and epicatechin (EC).[17]
Corchorus olitorius Chlorogenic acid, 3, 5-dicaffeoylquinic acid, quercetin 3-galactoside, quercetin 3-glucoside, quercetin 3- (6-
malonylglucoside), and quercetin 3- (6 malonyl galactoside) and cardiac glycosides.[29, 30]
Curcuma aromatica Zederonecurdione, neocurdione, curcumol, tetramethyl pyrazine, 1, 2-hexadecanediol, 9-oxo-neoprocurcumenol),
neoprocurcumenol and curcumin.[31]
Cyperus alopecuroids Alopecuquinone (benzoquinone) [32].
Vicenin 2, orientin, diosmetin, quercetin 3, 3'-dimethyl ether and its 3, 4'-dimethyl ether (flavonoids).[32]
Emblica officinalis - Emblicanin A and B, punigluconin, pedunculagin (hydrolysable tannins).[33]
- Gallic acid, ellagic acid, chebulinic acid, chebulagic acid, emblicanin A, emblicanin B, punigluconin, pedunculagin,
citric acid, ellagotannin, trigallayl glucose, pectin, 1-O-galloyl- -D-glucose, 3,6-di-O-galloyl-D-glucose, chebulagic acid,
corilagin, 1,6-di-O-galloyl- -D-glucose, 3 ethylgallic acid (3 ethoxy 4,5 dihydroxy benzoic acid), and isostrictiniin.[33]
- Phyllantine and phyllantidine (alkaloids).[33]
Psidium guajava - Gallic acid, caffeic acid and ellagic acid (phenolic acids).[34]
- Guavanoic acid, guavacoumaric acid, 2 -hydroxyursolic acid, jacoumaric acid, asiatic acid and isoneriucoumaric acid
(triterpene acids).[35]
Punica granatum - Punicalagin, punicalin, gallic acid, ellagic acid and ellagic acid derivative such as ellagic acid,3, 3'-di-O-methyl,ellagic
acid,3,3',4'-tri-O-methyl,ellagicacid,3'-O-methyl-3,4-methylene (hydrolysable tannins).[36]
- Pedunculagin, punicacortein A–D, granatin A and B, punicafolin, punigluconin and corilagin (Phenolic compounds).[36]
- Quercetin-3-O-rutinoside (flavonoid).[36]
Quercus infectoria Tannic acid (gallotannic acid, the principal constituent 50-70%), gallic acid, syringic acid, ellagic acid,
hexagalloylglucose and polygalloylglucose.[11]
Ricinus communi - Ricinine (0.55%) and N-demethyl ricinine (0.016%) (Alkaloids) [37]. Quercetin, rutin.[37]
- Six flavones glycosides.[37] Gallic acid, gentisic acid, epicatechin and ellagic acid (phenolic compounds).[37]
Sesamum indicum Tannins, saponins, flavonoids, terpenes and cardiac glycosides as phytoconstituents.[38]
Spinacia oleracea L. - para-coumaric acid, ferulic acid and ortho- coumaric acid (phenolic compounds).[39]
- Querecetin; myricetin; kampeferol; apigenin; luteolin; patuletin; spinacetin (flavonoids).[39]
Tamarixnilotica Methyl ferulate 3-O-sulphate, coniferyl alcohol 4-O-sulphate , kaempferol 4′-methyl ether , tamarixetin and quercetin 3-
O-β-D-glucupyranuronide.[40]

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

CONCLUSIONS
Natural products have been the most productive source of leads for new drugs. There is a need
for developing new antiprotozoal and antimicrobial agents from natural sources to overcome
resistance and safety problems of the currently existing drugs. As a part of our research
programme of investigating some medicinal plants cultivated in Beni-Suef Governorate
(Egypt) as antiprotozoal and antimicrobial agents, twelve plant extracts among the tested
plants could be considered as promising and interesting to be further elaborated through
purification and biological evaluation on an individual compound basis; Camellia sinensis,
Corchorus olitorius, Curcuma aromatica, Cyperus alopecuroids, Emblica officinalis, Psidium
guajava, Punica granatum, Quercus infectoria galls, Ricinus communi, Sesamum indicum
Spinacia oleracea, and Tamarix nilotica. The majority of studies still presented only
preliminary screening data and there is a need for further studies on the standardization or
chemical characterization of the extracts used and more description about the mechanisms of
action.

ACKNOWLEDGMENTS
This work was supported by research grants from the ParOwn program 2012 (Project No.
0911), Ministry of Higher Education at Egypt and in part from National Center for Natural
Products Research, School of Pharmacy, University of Mississippi.
We are grateful to the Egyptian Government, AI 27094 program from the NIH NCRR
(antifungal), 58-6408-1-603 program from USDA Agricultural Research Service Specific
Cooperative Agreement (antibacterial) and 58-6408-1-603 program from USDA Agricultural
Research Service Specific Cooperative Agreement (antimalarial and antileishmanial).
Thanks go to the National Center for Natural Products Research, also thanks to Dr. Melissa
Jacob, Dr. Babu Tekwani and Dr. Shabana Khan for carrying out the antimicrobial,
antileishmanial and antimalarial testing.

CONFLICT OF INTERESTS
The authors declare no conflict of interests.

REFERENCES
1. Balandrin N F KAD and Farnsworth N R. Human Medicinal Agents from Plants;. In A. D.
B. Kinghorn, N. F., Eds. (Ed.), ACS Symposium Series 534:1993, pp. 2-12.
2. AbouZid S F and Mohamed A A. Survey on medicinal plants and spices used in Beni-
Sueif, Upper Egypt. Journal of ethnobiology and ethnomedicine, 2011; 7(18): 1-6.

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

3. Grifo F, Newman D J, Fairfield A S, Bhattacharya B, Grupenhoff J T. The Origin of

Prescription Drugs, Grifo F. and Rosenthal J. Eds., Island Press, Washington D.C: 1997, p
131.
4. Khafagi IK and Dewedar A. The efficiency of random versus ethno-directed research in
the evaluation of Sinai medicinal plants for bioactive compounds. Journal of
Ethnopharmacology, 2000; 71(3): 365-376.
5. Fabricant D S and Farnsworth N R. The value of plants used in traditional medicine for
drug discovery. Environmental health perspectives, 2001; 109: 69.
6. Singh M, Khatoon S, Singh S, Kumar V, Rawat A K, Mehrotra S. Antimicrobial screening
of ethnobotanically important stem bark of medicinal plants. Pharmacognosy Res., 2010;
2(4): 254.
7. Al-Musayeib N M, Mothana R A, Al-Massarani S, Matheeussen A, Cos P, Maes L. Study
of the in vitro antiplasmodial, antileishmanial and antitrypanosomal activities of medicinal
plants from Saudi Arabia. Molecules, 2012; 17(10): 11379-11390.
8. Centers for Disease Control and Prevention (CDC), June 26, 2014,
http://wwwnc.cdc.gov/travel/notices/watch/malaria-in-egypt#fn1
9. Bharate SB, Khan SI, Yunus NA, Chauthe SK, Jacob MR, Tekwani BL, Khan IA, Singh
IP. Antiprotozoal and antimicrobial activities of O-alkylated and formylated
acylphloroglucinols. Bioorganic and Medicinal Chemistry, 2007; 15: 87-96.
10. Ma G, Khan S I, Jacob M R, Tekwani B L, Li Z, Pasco D S, Walker L A, Khan I A.
Antimicrobial and antileishmanial activities of hypocrellins A and B. Antimicrobial agents
and chemotherapy, 2004; 48(11): 4450-4452.
11. Hamid H, Kaur G, Abdullah S T, Ali M, Athar M, Alam M S. Two New Compounds from
the Galls of Quercus infectoria with nitric oxide and superoxide inhibiting ability.
Pharmaceutical Biology, 2005; 43(4): 317-323.
12. Ferreira D, Khan S, Jacope M, Gupta S, Reddy M. Antioxidant, antimalarial and
antimicrobial activities of tannin-rich fractions, ellagitannins and phenolic acids from
Punica granatum L. Planta Med., 2007; 73(5): 461.
13. Bagavan A, Rahuman A A, Kaushik N K, Sahal D. In vitro antimalarial activity of
medicinal plant extracts against Plasmodium falciparum. Parasitology research, 2011;
108(1): 15-22.
14. Zahir A A, Rahuman A A, Pakrashi S, Ghosh D, Bagavan A, Kamaraj C, et al. Evaluation
of antileishmanial activity of South Indian medicinal plants against Leishmania donovani.
Experimental Parasitology, 2012; 132(2): 180-184.

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

15. Hassan H F. Study the Effect of Quercus infectoria Galls Extracts on Growth of Candida
albicans and Candida glabrata In Vitro Which Isolated from Vaginal Swabs. Iraqi J. Vet.
Med., 2011; 35(2): 85 – 94.
16. Jain P and Nafis G. Antifungal Activity and Phytochemical Analysis of Aqueous Extracts
of Ricinus communis and Punica granatum. Journal of Pharmacy Research, 2011-a; 4(1):
128.
17. Aladag H, Ercisli S, Yesil D Z, Gormez A, Yesil M. Antifungal activity of green tea leaves
(Camellia sinensis L.) sampled in different harvest time. Pharmacognosy Magazine, 2009;
5(20): 437.
18. Abu-Mejdad N M. Antifungal Activity of Some Plant Extracts Against Two Yeasts
Isolates In Vitro. Research journal of pharmaceutical, biological and chemical sciences,
2014; 5(2).
19. Ashidi Joseph Senu E M O, Odunbaku O A, Biliaminu S A. The in-vitro effect of
Corchorus olitorius (L.) on the antibacterial activities of five antibiotics Journal of
Microbiology, 2012; 2(2): 113-117.
20. Voravuthikunchai S P and Suwalak S. Antibacterial activities of semipurified fractions of
Quercus infectoria against enterohemorrhagic Escherichia coli O157: H7 and its
verocytotoxin production. Journal of Food Protection, 2008; 71(6): 1223-1227.
21. Darogha S N. Antibacterial activity of Quercus infectoria extracts against bacterial
isolated from wound infection. Journal of Kirkuk University – Scientific Studies, 2009;
4(1), 20-30.
22. Supuran C T. Carbonic anhydrase inhibitors. Bioorganic and Medicinal Chemistry Letters,
2010a; 20(12): 3467-3474.
23. Krungkrai J and Supuran C T. The alpha-carbonic anhydrase from the malaria parasite and
its inhibition. Current pharmaceutical design, 2008; 14(7): 631-640.
24. Mogensen E G, Janbon G, Chaloupka J, Steegborn C, Fu M S, Moyrand F d r, et al.
Cryptococcus neoformans senses CO2 through the carbonic anhydrase and the adenylyl
cyclase Cac1. Eukaryot Cell, 2006; 5(1): 103-111.
25. Supuran C T, Vullo D, Leewattanapasuk W, Mühlschlegel F A, Mastrolorenzo A, Capasso
C. Carbonic anhydrase inhibitors: Inhibition of the β-class enzyme from the pathogenic
yeast Candida glabrata with sulfonamides, sulfamates and sulfamides. Bioorg Med Chem
Lett., 2013; 23(9): 2647-2652.

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

26. Supuran C. Inhibition of bacterial carbonic anhydrases and zinc proteases: from orphan
targets to innovative new antibiotic drugs. Current medicinal chemistry, 2012; 19(6): 831-
844.
27. Supuran C T, Innocenti A, Sarıkaya B Ö, Gülçin I. Carbonic anhydrase inhibitors.
Inhibition of mammalian isoforms I-XIV with a series of natural product polyphenols and
phenolic acids. Bioorganic and Medicinal Chemistry, 2010-b; 18(6): 2159-2164.
28. Satomi H, Ueno A, Hatano T, Okuda T, Noro T. Carbonic anhydrase inhibitors from the
pericarps of Punica granatum L. Biol. Pharm. Bull., 1993; 16 (8): 787-790.
29. Azuma K, Nakayama M, Koshioka M, Ippoushi K, Yamaguchi Y, Kohata K, et al.
Phenolic Antioxidants from the Leaves of Corchorus olitorius L. Journal of agricultural
and food chemistry, 1999; 47(10): 3963-3966.
30. Khan M S Y, Bano S, Javed K, Mueed M A. A comprehensive review on the chemistry
and pharmacology of Corchorus species-A source of cardiac glycosides, triterpenoids,
ionones, flavonoids, coumarins, steroids and some other compounds. Journal of the
Chemical Society, Perkin Transactions, 2006; 65(4): 283.
31. Ahmad S, Ali M, Ansari S H, Ahmed F. Phytoconstituents from the rhizomes of Curcuma
aromatica Salisb. Journal of Saudi Chemical Society, 2011; 15(3): 287-290.
32. Nassar M I, Abdel-Razik A F, El-Khrisy E D, Dawidar A M, Bystrom A, Mabry T J. A
benzoquinone and flavonoids from Cyperus alopecuroides. Phytochemistry, 2002; 60(4):
385-387.
33. Bhandari P R and Kamdod M A. Emblica officinalis (Amla): A review of potential
therapeutic applications. International Journal of Green Pharmacy, 2012; 6(4): 257.
34. Okuda T, Hatano T, Yazaki K. Guavin B, and ellagitannin of novel type. Chemical and
pharmaceutical bulletin, 1984; 32(9): 3787-3788.
35. Oliver-Bever B E P. Medicinal plants in tropical West Africa: Cambridge University
Press, 1986.
36. Jain V, Murugananthan G, Deepak M, Viswanatha G L, Manohar D. Isolation and
Standardization of Various Phytochemical Constituents from Methanolic Extracts of Fruit
Rinds of Punica granatum. Chinese Journal of Natural Medicines, 2011-b; 9(6): 414-420.
37. Jena J and Gupta A K. Ricinus communis Linn: a phytopharmacological review.
International Journal of Pharmacy and Pharmaceutical Sciences, 2012; 4(4): 25-29.
38. Okon J E and Umoh N S. Influence of Sesamum indicum L. ethanolic leaf extract on
haematological paramaters on albino rats in akwa ibom state, Nigeria. Indian Journal of
Fundamental and Applied Life Sciences, 2013; 3(1): 106-110.

www.wjpps.com Vol 4, Issue 05, 2015

Shimaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

39. Subhash G P, Virbhadrappa S R, Vasant O K. Spinacia oleracea LINN: a

pharmacognostic and pharmacological overview. IJRAP, 2010; 1(1): 78-84.
40. Abouzid S F, Ali S A, Choudhary M I. A new ferulic acid ester and other constituents
from Tamarix nilotica leaves. Chemical and Pharmaceutical Bulletin, 2009; 57(7): 740-
742.

www.wjpps.com Vol 4, Issue 05, 2015