mysemonbiopesticide-140212080729-phpapp01-140911071156-phpapp01.pdf

9/11/2014 Division of Agricultural Chemicals 1
Deepak yadav & Rohit kumar
m.sc-iii
University of allahabad
Guided by-PRF. ANUPAM DIXIT

Agrochemicals for food & nutritional security:
BIOPESTICIDES IN IPM
2
Division of Agricultural Chemicals

Per capita land availability
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Problem of food security
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4

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5

GREEN REVOLUTION
RICE WHEAT PULSES ALL FOOD GRAINS
104.2
98.8
83.6
77.4
243.3
239.3
Press Information Bureau, 27-10-2008
Production and Demand of Food grains in 2011-2012 (million tonnes)
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Bacteria Fungi
Viruses
Nematodes
Attack to Crops
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Food plants of the world are damaged by more than 10,000 species of insects, 30,000
species of weeds, 100,000 diseases (caused by fungi, viruses, bacteria and other microbes)
and 1000 species of nematodes (Hall, 1995; Dhaliwal et al., 2007)
Insects
Weeds

Estimation of crop losses caused by insect pests
to major agricultural crops in India
Dhaliwal et al., 2010
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9

Role of Pesticides
Crop production without
pesticide is unimaginable
To ensure better production at harvest against
unpredictable losses caused by plant diseases & pests
 To improve both quality & quantity of food
 To decrease the extent of vector born & other
diseases in humans & animals
“Complete ban on agrochemicals use in agriculture might
result in 50% reduction in global food production and 4 to 5
times increase in food prices”
Nobel Laureate Norman Borlaug
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Risk Associated With Chemical Pesticides
Toxicity to plants
Toxicity to mammals
Toxicity to aquatic creatures
Toxicity to beneficial organisms
High persistence of residues
• Indiscriminate use leads to the Three sad R’s :
Resistance, Resurgence and Residues
• Elimination of Natural enemies of pests
• Upsetting the ecological balance
• Environmental degradation/Pollution
• Enters food chain and lead to Bio-Accumulation
and Bio-Magnification
As a result of The misuse and overuse of pesticides crop
losses have consistently shown an increasing trend (Dhaliwal
and Koul, 2010)
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New form of pesticide
Low residual toxicity
Environmentally safe
Host specific in action

Active ingredient- Living organisms
1st Biopesticide discovered in the year 1835
Biopesticides are used to control pests, pathogens, and weeds by a variety
of means
Microbial biopesticides may include a pathogen or parasite that infects the
target
Alternatively, they might act as competitors or inducers of plant host
resistance

Bio means involving life or living
organisms
Pesticide includes substance or mixture
of substances intended for preventing,
destroying or controlling any pest
Biopesticide refers introduction of any
living organism such as microorganism
including bacteria , fungi , nematodes
viruses, protozoa and parasitoids and
predators that controls pests by biological non-toxic means
e.g. Trichoderma sp., Bacillus thuringiensis, Beauveria etc.
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All the living organisms, which are cultivated in the laboratory on large
scale & used and exploited experimentally for the control of harmful
organisms are called biopesticides

Global biopesticides & synthetic pesticides market, 2003-2010
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Locked Horns:
Synthetic pesticides Vs. Bio-pesticides
(Source : agriculture Today. Nov. 2005)
Factors Synthetic Pesticides Bio-pesticides
Cost effectiveness Cheap but increased
spraying cost
Costlier but reduced
number of applications
Persistence and residual
effect
High Low
Knockdown effect Immediate Delayed
Handling and Bulkiness Easy but danger and
Hazardous
Bulky : Carrier based
Easy : Liquid formulation
Pest resurgence More Less
Effect on Beneficial flora More harmful Less harmful
Target specificity Mostly broad spectrum Mostly host specific
Nature of control Curative Preventive
Shelf life More Less
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The market share of bio-pesticide is only 2% as compared to synthetic pesticide

Woo et al., 2010
MICROBIAL PESTICIDE
Active ingredient : Microorganism (Fungi, bacteria, virus, nematode etc.)
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List of registered microbial products by CIB
Name of microbes Type
Bacillus sp. Bacteria
Trichoderma sp. Fungi
Pseudomonas fluorescens Bacteria
Gliocladium sp. Fungi
Beauveria bassiana Fungi
Verticillium lecanii Fungi
Metarhizium anisopliae Fungi
Nomuraea rileyi Fungi
Nuclear Polyhedrosis Viruses Virus
Granulosis Viruses Virus
Courtesy: http://www.cibrc.nic.in
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MICROBIAL PESTICIDE

Characteristics
Storable
Economical
Easy to produce
Safe & acceptable
Convenient to apply
Virulent against target pest
Advantages
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High degree of specificity
Compatible with chemical pesticides
Easy to apply & aid growth through out
No adverse effect on non-target organisms
Absence of residue build-up in the environment
Relatively cheaper by 50% as compared to chemical pesticides
(Narayanasamy, 1995)

Bio-pesticides
Entomopathogenic Fungi
Fungal Antagonists
Bacterial Antagonists
Entomopathogenic Bacteria
Parasites & Predators
Moore & Prior, 1993

Entomopathogenic Fungi
Entomopathogenic fungi are fungi that can act as parasites of insects and
kill or seriously disable them
Mode of Action
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Entomopathogenic fungi in insect control

Beauveria
Beauveria bassiana most common
Habitat: Foliage
Insect Host: White flies, beetles & caterpillars (including Helicoverpa sp.)
Dose: 2 treatments made at 15-day intervals with 1.5 kg/ha concentrated product of
B. bassiana (3.0 × 109 conidia)
Treatment:
i) Foliar spray: 400-500 g in ½ bigha (5g/L of water)
ii) Soil drench: 250-500 g/3 bigha
Health impact: It causes granulosis disease in human ear
Grasshoppers killed by B. bassiana
Beauveria bassiana
Cultures of B. bassiana
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Metarhizium
Metarhizium anisopliae var. anisopliae & var. major
Habitat: Foliage
Insect host: Frog hoppers, beetles
Dose: Aerial treatment at 50 l/ha with 6 × 1011 to 1.2 × 1012 conidia/l of water
Conidia
Different cultures of M. anisopliae
Cockroach killed by
M. anisopliae
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Verticillium
Verticillium (Cephalosporium) lecanii
Habitat: Glasshouse foliage
Insect host: Aphids, whiteflies & scales
Dose: 41 × 107 active spores/g either undiluted or as a 10% concentration (diluted
with talc or water)
Whitefly scale infected
with V. lecanii
Cultures of Verticillium lecanii
Conidia
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Fungal Antagonists
 Principal fungi: Gliocladium virens & Trichoderma sp.
Trichoderma sp. mainly T. harzianum & T. viride
 Habitat: Soil
 Effective against: damping-off & wilt
Parasitize Rhizoctonia & Sclerotium
Inhibit growth of Pythium, Phytophthora & Fusarium
T. harzianum T. viride
Disease: T. harzianum causes green mold in cultivated button mushrooms & T.
viride causes green mold rot of onion
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Mode of action
 Direct parasitism or lysis (lytic enzymes like chitinase, cellulase & glucanase) & death
of the pathogen
 Direct toxic effects on the pathogen by antibiotic substances released by the
antagonist
Mycoparasitism by a Trichoderma
strain on the plant pathogen Pythium
 Competition with pathogen for food
 Indirect toxic effects on the pathogen by volatile substances released by the
metabolic activities of the antagonist
Cultures of Trichoderma harzianum
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The aim of investigations was to confirm the effect of Trichoderma
harzianum on Rhizoctonia solani and make a possibility for its usage in
tobacco production
T. harzianum was applied before and after sowing including a fungicide Top
M (0.1%)
At additional treatment with Trichoderma after use of fungicide, had a
better result than fungicide alone
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The influence of T. harzianum on intensity of disease attack
Artificial inoculation
Natural inoculation
The best results have shown by a variant with T. harzianum applied on a soil before
sowing and further application at certain intervals any time in a growing season of
tobacco seedlings
Additional treatment with T. harzianum after a fungicide Top M is advantageous to
the situation with a disease, so, it may be applied with this fungicide treatment

Bacterial Antagonists
• Pseudomonas sp. are gram negative, aerobic, rods that are inhabitants of wide
range of soil, water & plant surfaces
• P. fluorescens recognized by fluorescent pigment called ‘pyoverdines’
• Bio-control abilities of strains depend on aggressive root colonization, induction
of systemic resistance in the plant & production of diffusible or volatile
antifungal antibiotics
• Antibiotics with bio-control properties include – phenazines, hydrogen cyanide,
2,4-diacetylphloroglucinol, pyoluteorin, pyrrolnitrin, lipopeptides etc.
Phenazin
2,4-diacetylphloroglucinol
pyoluteorin
pyrrolnitrin
Lipopeptide
Hydrogen cyanide
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Mode of Action
Control of diseases
• Different strains of P. fluorescens extensively used in bioremediation of
various organic compounds & bio-controls of pathogens in agriculture
• P. fluorescens found effective in controlling fungal pathogens such as
wilt/root rot, Fusarium oxysporum f. sp. Cubense, Pythium sp., R. solani, R.
oryzae, S. rolfsii & bacterial pathogens like Xanthomonas citri & P.
solanacearum in field tests
• Bacterial preparations widely used in organic spice cultivation of southern
India
Theories include -
• Induction of systemic resistance – resist attack by true pathogen
• Competition with other (pathogenic) soil microbes, e.g. siderophores
• Production of compounds (antibiotics) antagonistic to other soil microbes
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Entomopathogenic Bacteria
• Bacillus thuringiensis (Bt), a Gram-positive, motile, rod shaped bacterium
produces a parasporal crystal composed of one or more proteins
• The strains of Bt characterized so far affect members of 3 insect orders:
Lepidoptera (butterflies and moths), Diptera (mosquitoes & biting flies), and
Coleoptera (beetles)
• EPA registered Bt products include
B.t. israelensis (Diptera)—frequently used for mosquitoes
B.t. kurstaki (Lepidoptera)—frequently used for gypsy moth, spruce budworm,
and many vegetable pests
B.t. sandiego and tenebrionis (Coleoptera)—frequently used for leaf beetle,
Colorado potato beetle
B.t. kurstaki is the most commonly used Bt formulation
Bacillus thuringiensis
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Mode of Action
Bacillus thuringiensis strains
produce crystalline proteins
(called δ-endotoxins)
Caterpillar consumes the Bt spore
(diagram 1) & crystalline toxin-
treated leaf
The Bt crystalline toxin (diamond shapes in
diagram 2) binds to gut wall receptors, and
the caterpillar stops feeding
Within hours, the gut wall breaks down,
allowing spores (oval tube shapes) and normal
gut bacteria (circular shapes) to enter body
cavity, where the toxin dissolves
The caterpillar dies in 24 to 48 hours from septicemia, as spores and gut
bacteria proliferate in its blood (diagram 3)
Treatments:
Dose:
i) 100 – 150 g/ bigha for field crops.
ii) 150-200 g /bigha for orchards.
Method: The powder is first mixed with small quantity of
water to prepare a uniform suspension. Then the required
quantity of water is added and thoroughly mixed before spray.
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Laboratory assays were done to evaluate the effect of Bacillus thuringiensis,
neem seed kernel extract (Azadirachta indica), Vitex negundo leaf
extract, & applied separately or together, on nutritional indices of the
rice leaf-folder Cnaphalocrocis medinalis
Bt biopesticide & other 2 botanical pesticide suppressed feeding and larval
growth and low concentrations affected the larval performance
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(Nathan et al. ,2005)
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The combined effect of these resulted in a considerable decrease in
nutritional indices indicating strong deterrence

• Bt is considered to be “practically nontoxic” to humans and other
vertebrates
• It can cause a “very slight irritation” if inhaled & can cause eye irritation
• Bt is not carcinogenic, mutagenic, or teratogenic
• Bt does not persist in the brains, lungs, or digestive systems of animals,
including humans
• Bt has been found in fecal samples of exposed greenhouse workers, no
gastrointestinal symptoms were associated with its presence
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Human Health & Safety

• Bt appears to be a normal component in the feces of vegetable-
consuming animals, where it apparently causes no problem
• Like the active bacterial ingredient, the inert ingredients in Bt
formulations have also been studied and modified for safety
• Granular and microcapsule formulations reduce the inhalation hazard
• Volatile agents associated with some Bt formulations do not appear to
constitute a significant health hazard.
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Human Health & Safety…

Environmental Impacts
• No danger has been found to aquatic communities accidentally
exposed to Bt or to non-target organisms including beneficial insects,
amphibians, fish, and mammals
• Few reports of Bt lethality upon non-target organisms, such as leaf-
feeding caterpillars
• Clay soils may bind the bacterial toxin, increasing its environmental
persistence and possible toxicity to non-target species
• Newer formulations employ preservatives, like sorbitol, that are safer
than the xylene used decades ago
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Phytonematode management through
bacteria
Bacteria Genus/species Target nematode Mode of action References
Parasitic
bacteria
Pasteuria penetrans,
P. thornei
Phytonematodes Parasitism Bekal et al.(2001),
Bird et al. (2003)
Opportunistic
bacteria
Brevibacillus
laterosporus,
Bacillus nematocida
Free living &
Phytonematodes
Parasitism Niu et al. (2006),
Tian et al. (2007)
Rhizobacteria Bacillus sp.,
Pseudomonas sp.
Meloidogyne sp.,
Heterodera sp.
Interfering with
recognition,
production of
toxin, nutrient
competition, plant
growth promotion
Marleny et al.
(2008),
Meyer (2003)
Crystal
forming
bacteria
Bacillus thuringiensis
(Cry 5,6,12,13,14,21)
Trichostrongylus
colubriformis,
Caenorhabditis
elegans
Cry proteins cause
damage to the
intestines of
nematodes
Kotze et al.(2005),
Wei et al. (2003)
Endophytic
bacteria
Root knot
nematode,
Cyst nematode
Rhizo-bacterial &
endophytic
bacterial mode of
action
Sturz et al. (2004),
Compant et al.
(2005)
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Nuclear polyhedrosis virus (NPV)
A) NPV (Helicoverpa): It is highly effective on H. armigera, pest of
cotton,gram, pea, pigeon pea, tomato, cabbage, ground nut, millets,
oilseeds & roses
A) NPV (Spodoptera): It is highly effective against S. litura caterpillar, pest
of cotton, gram, pigeon pea, cabbage, tomato, chillies & oilseeds
Treatments: Dose: 250 – 500 LE/ha
Method:
i) Shake the bottle properly and prepare a solution @ 1 ml/litre of water
ii) Spray the solution 2-3 times at 10-15 days interval
iii) Spray preferably in the evening and on young larval stages or on sighting of
egg laying
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Enhancing food security by the local production of
microbial bio-pesticides against insect crop pests:
African armyworms as a case study
2 types of application studied
A) Aerial spray of SpexNPV
B) Ground spray of SpexNPV & OP pesticide Diazinon
separately
(Wilson et al., 2008)
SpexNPV = Spodoptera exempta Nucleo polyhedrovirus
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Commercial bio-pesticides for the control of
plant pathogens
Microorganisms Trade Name Pathogens/ Diseases
Bacteriophages of
Xanthomonas sp. and
Pseudomonas syringae pv.
Tomato
Agriphage™ Bacterial spot in pepper &
tomatoes & bacterial speck in
tomatoes
Pseudomonas syringae strain
ESC 10
Bio-Save® 10LP3 Ice inducing bacteria &
biological decay
Pantoea agglomerans strain
E325
Bloomtime,
Biological™ 3
Fire blight( Erwinia
amylovora)
Bactericides
9/11/2014 45

Microorganisms Trade Name Pathogens/ Diseases
Streptomyces
lydicus WYEC 108
Actinovate®AG,
Actinovate®SP
Soiborne pathogens: Pythium sp.,
Rhizoctonia sp., Phytophthora sp.,
Fusarium sp.
Foliar pathogens: Alternaria sp.,
Peronospora sp.
Bacillus subtilis
GB03
Kodiak® Concentrate Rhizoctonia, Fusarium, Alternaria,
Aspergillus /
Phoma. root rot, damping off,
crown rot
Trichoderma
harzianum
Rifai strain KRL-
AG2
T-22™HC, Plant Shield®,
T-22™. Planter Box,
Serenade® MAX™
Fusarium, Pythium & Rhizoctonia/
Root rot, powdery mildew
Bacillus pumilus
QST 2808
Ballad® Plus Cercospora sp./ Rust, powdery
mildew,, and brown spot
Fungicides
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Types of bio-
control agents
Names of bio-
control agents
Target species
PARASITOIDS Trichogramma chilonis Brinjal shoot and fruit
borer, shoot borers of
cotton, sugarcane, rice
T. brasiliensis and T.
pretiosum (egg
parasitoids)
tomato fruit borer
PREDATORS Cryptolaemus
montrouzieri
(Austrtralian ladybird
beetle)
several species of mealy
bugs and soft scales
Chrysoparla sp. (green
lacewing bug)
aphids, white flies
Parasitoids & Predators

Few examples of bio-control
Muscodor albus strain QST 20799 acts as bio-fumigant & controls
bacteria and soil borne pest by releasing volatile toxin
Aspergillus flavus strain AF36 can act as bio-fungicide for cotton. Unlike
other strains it will not produce carcinogenic ‘Aflatoxin’
Pasteuria sp. acts as bio-nematicide & controls microscopic worms &
other nematodes that feed on plant roots
Cydia pomonella granulosis virus acts as bio-insecticide & controls
codling moth in fruits like apples & pears
Phytophthora palmivora acts as bio-herbicide & controls milkweed
(Asclepias sp.) in citrus orchards

Path Ahead
More studies needed to determine the environmental effects on the fate
of bio-agents
New technologies such as micro encapsulation of bio-control agents may
be of high priority in enhancing their potential
Integration of bio-pesticides with botanical pesticides has a lot of
potential in pest management
Integration of bio-pesticides with chemical pesticides as part of Bio-
intensive Integrated Pest Management (BIPM)
49
9/11/2014 Division of Agricultural Chemicals

Conclusion
Microbials such as bacteria, fungi, viruses are the major bio-pesticides being
studied mostly to develop alternatives to chemicals
The no. & growth rate of bio-pesticide showing an increasing marketing trend
in past few decades
Bio-pesticides are host specific & bio-degradable resulting in least persistency
of residual toxicity
Bio-pesticides саn mаkе vital contributions tο IPM & can greatly reduce
conventional pesticides, while crop yield remains high
Bio-pesticides having lesser health hazard provides an important alternative
in the search for an environmentally sound and equitable solution to the
problem of food security
9/11/2014 50

“Life is not living, but being in health.”
- Latin poet Martial

9/11/2014 52

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