Summer project viva voce presentation for the partial
Fulfilment for
the Degree of Master of Science in Microbiology
“Screening, Isolation, and characterization of pesticide
Degrading microorganisms”
resented by
Nanditha H.S Under the Guidance of
ost graduate student Dr. SUMANA k
3L11260 Associate professor
Department of Microbiology Department of Microbiology
SS Academy of Higher education and research JSS Academy of Higher education and
Mysuru - 570015 research
Mysuru - 570015
ABSTRACT
A pesticide is a material or combination of materials designed to prevent, eliminate,
repel, or reduce the harm the bug has created an insect, plant pathogen, weed,
bacteria, bird, etc. that competes with humans for food, damages property, or
spreads disease can be considered a pesticide chemicals, biological agents, Oti
microbes’ disinfectants, and more can all be considered pesticide. Both humans and
animals can be poisoned by chemical pesticides. The aim of this study is
screening isolation and testing of pesticide degrading bacteria. it helps
for the agriculture field and the bioremediation. Soil was collected from the
different area of rice field in Mysore and Madhugiri. 10 colonies where isolated and
subjected for biochemical test to which 100% were positive for catalase test, 40%
were positive for indole test 50% were positive for methyl red test, 100% were
positive for citrate test, 0% were vages-proskauer test, and 30% were positive for
gelatin test, the colonies were also subjected for cultural and morphological
characterization on nutrient agar, cetramide agar and MSM media. And performing
a gram staining respectively 90% of the selected colonies were gram negative rod
shaped 10 % of the them were gram positive rod shaped on that 10 isolated
colonies subjected for qualitative analysis are studied on that isolated colonies are
showed pesticide degrading quality for that their antibiotic sensitivity test by disc
diffusion method was studied. Thus, this pesticide degrading microorganism species
can be used for bioremediation of contaminated soil.
CONTENTS
Introduction
Literature review
Aim and objectives
Materials and method
Result
Discussion
References
Introduction
In agriculture, the use of chemical pesticides has become a vital
tool for pest management
A pesticide is a material or combination of materials used to
prevent, control, and eradicate any pests, such as rodents,
insects, weeds, birds, nematodes
Pesticides harm and even kill wild mammals and birds
microorganisms are a group of bacteria, fungi, and other
microbes capable of breaking down pesticides into less harmful
or non-toxic substances.
They play a vital role in bioremediation, helping reduce pesticide
residues in the environment and mitigating their negative impacts
on ecosystems and human health. Here's an overview of key aspects
By entering into the food chain and interfering with both male and
female reproductive systems, pesticides also endanger human
health by resulting in infertility
Pesticide-degrading bacteria offer a sustainable and eco-
friendly solution to these challenges by:
1. Detoxifying Pesticides: Converting harmful compounds
into harmless byproducts.
2. Preventing Bioaccumulation: Reducing the persistence of
pesticides in the environment.
3. Restoring Soil Health: Improving soil microbial diversity
and fertility.
Types of pesticide
Literature review
Pesticides, including insecticides, herbicides, and fungicides, are
widely used in agriculture and industry to control pests and
diseases. However, their persistence in the environment and
potential to harm ecosystems and human health has led to
concerns.
Microorganisms degrade pesticides through various mechanisms,
including hydrolysis, oxidation, reduction, and dechlorination.
Pseudomonas spp., Bacillus spp., Ralstonia spp., and Arthrobacter
spp. are commonly studied for their ability to degrade
organophosphates, carbamates, and other pesticide groups.
Fungi such as Trichoderma spp. and Penicillium spp. have been
identified as potential degraders of fungicides and other pesticides.
These fungi can produce enzymes like laccases, peroxidases, and
hydrolases.
Various enzymes, such as organophosphate hydrolases,
carboxylesterases, and laccases, are involved in the microbial
breakdown of pesticides.
Environmental factors like temperature, pH, moisture, and nutrient
availability can influence the efficiency of pesticide degradation.
Bioremediation involves using microorganisms to clean up pesticide-
contaminated environments. Studies have shown the potential of
using both indigenous and introduced microbial communities to
degrade pesticide residues in contaminated soils, water, and
sediments.
Bioremediation strategies include bioaugmentation (adding
pesticide-degrading microbes) and biostimulation (altering
environmental conditions to promote the growth of indigenous
microbes).
Future research may focus on improving the efficiency of
biodegradation through genetic engineering of microorganisms,
optimization of degradation pathways, and exploring novel microbial
communities that can degrade a broader range of pesticides.
Aim and objectives
ISOLATION OF PESTICIDE DEGRADING
BACTERIA
DENTIFICATION OF BACTERIA
SCREENING OF PESTICIDE DEGRADING
BACTERA
QUALITATIVE ANALYSIS OF PESTICIDE
DEGRADING BACTERIA
ANTIBIOTIC SENSITIVITY TEST BY DISC
DIFFUSION METHOD
Materials and method
SAMPLE COLLECTION
Soil sample were collected from rice
field
BACTERIAL ISOLATION AND PLATING
Culture enrichment in MSM ,serial
dilution
Spread plate method used on NA Media
MORPHOLOGICAL IDENTIGICATION
Colony characterization
Gram staining
BIOCHEMICAL TEST
Catalase test Citrate utilization test
Indole test Vages-proskauer test
Methyl red test Gelatin test
PRIMARY SCREENING FOR PESTICIDE
DEGRADATION
Incubate the culture in minimal salt media
ANTIBIOTIC SENSITIVITY TEST BY DISC DIFFUSION
METOD
Ampicillin,methicilln,tetracyclin,gentamicin,
Rifampicin.
Result
1.Isolation bacteria 2.Gram staining bacteria
colonies Subculture coloniesG(+)ve rod shaped G(+)ve rod shaped G(-)ve rod shaped
Bacter color margi elevat opacit Gram shape
ial n ion y staini
isolat ng 90%
es 80%
Bi1 white Irregul flat opaqu + rod 70%
ar e 60%
Bi2 white circular raised opaqu - Rod, 50% Gram postive
e chain 40% Gram negative
30%
Bi3 white circular raised opaqu + rod
20%
e 10%
Bi4 white circular convex opaqu - rod 0%
e 1
Bi5 white circular flat opaqu - rod
e
Bi6 white circular raised opaqu + rod
e
Bi7 white circular raised opaqu - rod
e
3.Biochemical test
Graphical result of biochemical test
100%
80%
60%
40%
Catalase test Indole test Methyl red test
20% Series1
Series2
0%
st st st st st st
te te te te te te
se le d
tio
n Vp at
in
ala ndo re a el
t I hy ili
z G
ca et
M ut
e
Citrate utilization at
VP test Gelatin test i tr
test C
.Primary screening for pesticide degradation
Discoloration found in test tube Turbidity found in test tube
Qualitative analysis of isolated pesticide degrading bacteria
Bacterial isolates Clear zone (mm)
1 32mm
2 20mm
3 35mm
4 10mm
Clear zone of isolated pesticide degrading bacteria.
ntibiotic sensitivity test by disc diffusion method of isolated pesticide degrading bacteria
Anti Bi1 Bi2 Bi3 Bi4
bioti
c Bi5
Amp - - - + -
icilli
n
Met - - - - -
hicill
in
Tetr - ++ - + +
acyc
line
stre - +++ - +++ -
pto
myci
Antibiotic sensitivity test for pesticide degrading microorganismn
Gent - + - +++ +++
amic
in
Discussion
Microbial degradation of pesticides is the process by which, in the presence of
favorable environment condition
Environment microorganism break down pesticide molecules into small
molecules of organic or inorganic compounds by using the residual pesticide in
the environment as nutrients such as, carbon, nitrogen, and phosphors source
However few tests for pesticide degrading bacteria were done, the test done
were, biochemical test for genus level identification of the isolated bacteria such
as catalase test, indole test, methyl red test citrate test vages-proskauer test,
gelatin test. In- vitro screening on that 10 isolated colonies 5 were showed zone
of inhibition on pesticide agar
Four types of bacteria were identified up to genus level according to Bergey`s
manual after performing bio chemical test the four bacteria Pseudomonas
putida, Bacillus cereus, Pseudomonas griseus, Streptomyces griseus
Some microorganisms are highly specific to certain pesticides, while others
exhibit broad-spectrum degradation capabilities. For example, Pseudomonas
spp. and Bacillus spp. are widely studied for their ability to degrade
organophosphates and carbamates, respectively
The application of pesticide-degrading microorganisms in bioremediation offers
a sustainable solution to pesticide contamination.
Advancements in biotechnology and genetic engineering can enhance the
degradation capabilities of microorganisms. For example, genetically modified
strains could be developed to target specific pesticides or operate efficiently
under extreme conditions.
References
1. Singh BK, Kuhad RC, Singh A, Lal R, Tripathi KK. Biochemical and Molecular Basis of
Pesticide Degradation by Microorganisms. Vol. 19, Critical Reviews in Biotechnology.
1999.
2. Mustapha MU, Halimoon NB, Lutfi W, Johari W, Yunus M, Shukor A, et al. SCIENCE &
TECHNOLOGY An Overview on Biodegradation of Carbamate Pesticides by Soil
Bacteria CORE View metadata, citation and similar papers at core.ac.uk provided by
Universiti Putra Malaysia Institutional Repository. Pertanika J Sci & Technol.
2019;27(2):547–63.
3. Mohammed AI. Isolation of Pesticide Degrading Microorganisms from soil. Available
from: http://www.soeagra.com/abr.html
4. Shannon MJ, Unterman R. Evaluating bioremediation: distinguishing fact from
fiction. Annu Rev Microbiol. 1993; 47:715–38.
5. Hussaini SZ, Shaker M, Asset Iqbal M. Advances in Bioresearch O OR RI IG GI IN NA
AL L A AR RT TI IC CL LE E Isolation of Bacterial for Degradation of Selected Pesticides.
Adv Biores. 2013;4(3):82–5.
6. Ahmad S, Ahmad HW, Bhatt P. Microbial adaptation and impact into the pesticide’s
degradation. Arch Microbiol. 2022 May 1;204(5).
THANK YOU
