No

1. DEPARTMENT OF ENTOMOLOGYDEPARTMENT OF ENTOMOLOGY
CREDIT SEMINAR
ON
CREDIT SEMINAR
ON
RECENT ADVANCES IN MICROBIAL PESTICIDE
IN INSECT PEST MANAGEMENT
Presented By:-
RAM KUMAR
M.Sc. (Ag.)
Dr. Rajendra Prasad Central Agricultural University, Pusa, Bihar

2. Microbial pesticides
 Microbial pesticides are products based on living
micro-organisms which are pathogenic to targeted
pest
 Consist of a microorganism as the active ingredient
(e.g., bacteria, fungi, virus, protozoa, and nematode)
 The term entomopathogenic has a Greek origin
entomon, refers to insect, and pathogenic, which
denotes causing disease
 About 3000 microbes reported to cause diseases in
insects

3. MICROBIAL PESTICIDES PRODUCTS
WORLDWIDE
Woo, et al., 2010.
Others (3%)

4. Status of production of microbial pesticides
used against insect pests in India
Agent 2009-2010
HaNPV 15639 (L) 7958 (L) _ 12522
SlNPV 3438 (L) 2387 (L) _ 1673
Bacillus
thuringenesis
52 7.0 5.0 _
Beauveria bassiana 141 82 244 19478
Metarhizium
anisopliae
29 35 20 4730
Verticillium lecanii 45 30 190 10242
Paecilomyces
lilacinus
50 3.0 14 26436
P. fumosoroseus _ _ 2.0 4000
Solid formulations(T) Liquid formulations(L)
Ramanujam et al. (2014). Proc. Indian Natn. Sci. Acad., 80(2): 455-471.
2005-06 2006-07
(T/L) (T/L)

5. Advantages of microbial pesticides
• Microbial pesticides are difficult for insects
to develop resistance
• Safe to natural enemies and higher
organisms
• Biodegradable
• Cheaper, renewable and can be handled
safely
• Most are compatible with insecticides and
other agents
• Residue free

6. Limitations of microbial pesticides
Slow effect
Lack of persistence and narrow activity
Rapidly degraded by UV light
Not easily available everywhere
Poor water solubility and are generally
not systemic in nature
Poor storage characteristics

7. TYPES
MICROBIAL PESTICIDES
Bacteria FungiVirus NematodeProtozoa

8. Bacterial insecticides
•Spore forming
•Rod-shaped
• Isolated from soil samples
•Stomach poisons
•Genus Bacillus

9. CLASSIFICATION OF ENTOMOPATHOGENIC
BACTERIA
ENTOMOPATHOGENIC BACTERIA
Spore producers
Non-spore producer
e.g. Pseudomonus spp.
Obligate spore
producers
e.g. Bacillus popillae
Facultative spore
producers
Crystelliferous
e.g. Bacillus
thuringiensis
Non crystelliferous
e.g. Bacillus cereus

10. Some properties of the insecticidal toxins
from
various strains of B. thuringiensis
Strain/subsp. Protein
size
cry Target Insects
berliner 130-140 kDa CryI (Cry1Aa) Lepidoptera
kurstaki KTP, HD1 130-140 kDa CryI (Cry1Ab) Lepidoptera
entomocidus 6.01 130-140 kDa CryI (Cry1Ba) Lepidoptera
aizawai 7.29 130-140 kDa CryI (Cry1Ca) Lepidoptera
aizawai IC 1 135 kDa CryII (Cry1Da) Lepidoptera, Diptera
kurstaki HD-1 71 kDa CryII (Cry2Ab) Lepidoptera, Diptera
tenebrionis (sd) 66-73 kDa CryIII (Cry3Aa) Coleoptera
morrisoni PG14 125-145 kDa CryIV (Cry4Aa) Diptera
israelensis 68 kDa CryIV (Cry4Ba) Diptera

11. CRY Proteins
• Crystalline protein and are toxic
ingredient
• Each Bt species/strain produces a
unique set of CRY proteins
• Each CRY protein has unique activity
• CRY proteins are usually designated
with a series of numbers and letters:-
Cry1a, Cry2

12. Mode of
action

13. B. thuringienis parasporal crystal
composed of Cry1 protoxin protein.

14. Bt strain Trade name Uses
Bt var. aizawai Florback, Centari Diamondback
moth
Bt var. galleriae Certan Wax moth larvae
in honey combs
Bt var. israelensis Bactimos,
Bactis,
Thurimos,
Vectobac
Larvae of
mosqitoes and
balckflies
Bt var. kurstaki Bt, Biobit, Dipel,
Delfin, Javelin
Lepidopterous
larvae
Bt var. sandiego Diterra, M- one
plus
Beetles and
weevils
Bt var. thuringenesis Muscabac,
Thuricide
Flies,
Lepidopterous
larvae
Bt based commercially available
pesticides in India

15. Effect of Bt liquid formulations against Helicoverpa
armigera in pigeon pea
treatments
No. of H. armigera larvae / plant on pigeon pea
PDBC Bt1 @1% 0.73 0.47 0.40 3.60 2.20 2.07 1.28
PDBC Bt1 @ 2% 0.80 0.43 0.33 3.67 2.07 1.60 1.11
NBAII BtG4 @ 1% 0.67 0.40 0.33 3.53 2.47 1.53 1.18
NBAII BtG4 @ 2% 0.73 0.37 0.27 4.00 1.93 1.47 1.01
Beauveria bassiana
@ 1.5 Kg/ha
0.87 0.60 0.33 3.73 2.80 2.00 1.43
Beauveria bassiana @
2.0 Kg/ha
0.73 0.67 0.40 3.67 2.20 1.67 1.23
Bt k @ 0.2% 0.93 0.47 0.33 3.80 2.53 1.80 1.28
Chlorpyriphos 20EC @
0.25%
1.07 0.33 0.17 4.07 1.40 1.3 0.81
Untreated control 0.80 0.87 0.93 3.93 4.07 4.33 2.55
Kumar, et al., (2016). J. Appl. Biol. Biotech., 4 (1): 39-42.
I Spray II Spray
Pre count 3 DAS 7 DAS Pre count 3 DAS 7 DAS Mean

16. Effect of different Bt liquid formulations on pod, grain
damage and yield of pigeon pea
Treatments Pod damage (%) Seed damage (%) Yield (q/ha)
PDBC Bt1 @1% 7.74 5.54 13.5
PDBC Bt1 @ 2% 7.5 5.34 14.1
NBAII BtG4 @ 1% 6.29 4.77 14.2
NBAII BtG4 @ 2% 5.30 3.91 15.3
Beauveria bassiana @ 1.5 Kg/ha 8.07 5.77 12.1
Beauveria bassiana @ 2.0 Kg/ha 6.75 4.99 12.7
Bt k @ 0.2% 7.09 5.4 13.7
Chlorpyriphos 20EC @ 0.25% 5.07 3.24 16.9
Untreated control 12.94 8.78 9.2
Kumar, et al., (2016). Evaluation of liquid formulations of Bt against gram pod borer,
Helicoverpa armigera (Hubner) and spotted pod borer, Maruca vitrata (Geyer) in pigeon
pea. Journal of Applied Biology & Biotechnology, 4 (1): 39-42.

17. Virus
• Set of one or more nucleic acid template
molecules, normally encased in a protective
coat of protein or lipoprotein that is able to
organize its own replication only within suitable
host cells
NPV infected larva Gypsy moth larva killed by

18. Characteristics of Viruses found in
insectsVirusVirus NucleicNucleic
acidacid
VirusVirus
particleparticle
Inclusion bodyInclusion body
shapeshape
Subgroups andSubgroups and
common namescommon names
Ascoviridae Ds DNA Allantoid None -
Baculoviridae Ds DNA Bacilliform
Polyhedral NPV
Cigar- shaped GV
Calciviridae ss RNA Isometric None -
Iridoviridae ss DNA Isometric None Iridscent
Nodaviridae ss RNA Isometric None -
Parvoviridae ss DNA Isometric None -
Picornaviridae ss RNA Isometric None -
Polydnaviridae ssDNA Ovoid None -
Poxviridae Ds DNA Ovoid or brick
shaped
Spheroid Entomopox viruses
Reoviridae Ds DNA Isometric Polyhedral Cytoplasmic
polyhedrosis
Rabhdoviridae ss RNA Helical None -

19. • Approx. 60 per cent of 1200
known insect viruses
belongs to family
Baculoviridae
• Baculovirus infection
described in 700 species of
invertebrates
OrderOrder SpeciesSpecies
LepidopteraLepidoptera 455455
HymenopteraHymenoptera 3131
DipteraDiptera 2727
ColeopteraColeoptera 55
NeuropteraNeuroptera 22
TrichopteraTrichoptera 11
ThysanopteraThysanoptera 11
SiphanopteraSiphanoptera 11
Helicoverpa armigera infected with HaNPV

20. Mode of Action

21. Efficacy of Baculoviruses against
insect pests
Virus Dose Targeted
Pest
Mortality (%) Reference
HzNPV 100-250
LE/ha
Helicoverpa
zea
90- 96 Dhaliwal et
al., 2007
SlNPV 375 LE/ha Spodoptera
litura
95.00 Arora et al.,
2003
HaNPV 1 X 108
OB’s Helicoverpa
armigera
90.60 Snegapriya
and
Manjula,
2008
AgNPV 50 LE/ha Anticarsia
gemmatalis
80.00 Dhaliwal et
al., 2007
HaNPV - Helicoverpa
armigera
Effective
control
Hu et al.,
2003
SfNPV 2.5X 1011
OB’s
Spodoptera
frugiperda
Effective
control
Dhaliwal et
al., 2007

22. Effect of microbial insecticides on the larval population of
H. armigera in gram
Larval population per meter row
Treatment Dose/ha 1DBISP 3DAISP 7DAISP 1 DBIISP 3DAIISP 7DAIISP
Bt var Kenyae 1500g 14.00 7.00 8.66 2.66 3.00 1.00
Delfin
(53000SU/mg)
1500g 14.66 9.33 6.33 4.00 4.66 1.00
Dipel DF (2.3107
conidia/ml)
5000ml 16.33 8.00 4.66 2.66 3.66 1.00
Biolep
(1600IU/mg)
1500g 14.33 8.00 6.66 3.66 3.33 1.00
Halt(55000SU/mg 2000g 12.33 9.33 6.00 3.66 3.33 2.00
Basina 5000g 13.33 7.00 6.00 4.66 4.33 2.66
HaNPV 250 LE 13.33 8.00 8.66 3.66 4.00 1.66
Endosulfan 35 EC 1000 ml 15.00 7.66 7.66 4.00 4.33 1.33
Untreated check - 17.33 15.00 11.00 8.00 5.66 5.00
Gowda S.D.K and Yelshetty, S. (2005). Evaluation of Microbial Agents against Gram
Pod Borer, Helicoverpa armigera (Hubner). Karnataka J. Agric. Sci.,18(1): 44-46.

23. Effect of microbial insecticides on the pod an
yield characters in gramickpea
Treatment Dose per ha. Pod damage(%) Yield (q/ha)
Bt var Kenyae 1500g 13.73 9.40
Delfin (53000SU/mg) 1500g 12.38 8.52
Dipel DF (2.3107 conidia/ml) 500ml 13.23 9.18
Biolep (1600IU/mg) 1500g 12.90 8.43
Halt (55000SU/mg) 2000g 14.53 8.70
Basina 5000g 13.43 8.36
HaNPV (6×109
POB/LE) 250LE 11.63 (min.) 10.26 (max.)
Endosulfan 35 EC 35EC 13.38 8.16
Untreated check - 20.30 5.83
Gowda S.D.K and Yelshetty, S. (2005). Evaluation of Microbial Agents against Gram
Pod Borer, Helicoverpa armigera (Hubner). Karnataka J. Agric. Sci.,18(1): 44-46.

24. Metarhizium anisopliae
Beauveria bassiana infection of
Clover Worm
Beauveria bassiana infection of
worm within woody substrate
Fungi
 An entomopathogenic fungus is a fungus that act
as a parasite of insects and kills or disables them
seriously

25. Entomopathogenic fungi used for the
control of pests
FungusFungus ProductProduct TargetTarget
Verticillium lecanii Mycotol, Vertalec Whitefly, thrips and aphids
Metarhizium anisopliae Meta guard Termites
Metarhizium anisopliae Biogreen Locusts
Metarhizium anisopliae Bio-Path Cockroaches
Metarhizium anisopliae Bio-Blast Termites
Metarhizium anisopliae Cobicant Sugarcane spittle bug
Metarhizium anisopliae Bio-Cane Cane grubs
Metarhizium anisopliae Green Muscle Locusts, grasshoppers
Beauveria bassiana Conidia Coffee berry borer
Beauveria bassiana Cornguard European corn borer
Beauveria bassiana Naturalis- L Cotton pests
Beauveria bassiana Mycotrol GH Locusts, grasshoppers
Beauveria bassiana Mycotrol WH and Botanigard Whitefly, aphids, thrips
Paecilomyces fumosoroseus PFR- 97, Pae- Sin Whitefly

26. Entomopathogenic fungi in Insect ControlEntomopathogenic fungi in Insect Control
Tea Mites
Rice bugs
Helicoverpa
Beauveria infected Helicoverpa
Paecilomyces infected tea mites
Metarhizium infected rice bugs

27. Mode of action of Entomopathogenic
fungi

28. Selected metabolites of important
Entomopathogenic fungi
Pathogen Metabolite
Metarhizium anisopliae Destruxins (>27 types),
cytochalasin
Beauveria bassiana Bassianin, beauvericin,
bassianolide, tenellin
Beauveria brogniartii Oosporein
Paecilomyces fumosoroseus Beauvericin, beauverolies,
pyridine-2,6-dicarboxylic acid
Verticillium lecanii Dipcolonic acid,
hydroxycarboxylic acid,
vertilecannins, bassianolide
Tolypocladium spp. Cyclosporin, efrapeptins (5 types)
Hirustella thompsonii Hirsutellin A, hirsutellin B,
phomalatone

29. Efficacy of Fungi against pests
FungusFungus ConcentrationConcentration
(conidia/ ml)(conidia/ ml)
Pests (Crop)Pests (Crop) MortalityMortality
(%)(%)
ReferenceReference
Beauveria bassiana 1 X 108
Lygus lineeolaris 77.47 Sabbahi et al.,
2008
1 X 108
Anthonomus signatus 60.35
1 X 108
Otiorhynchus ovatus 54.50
1 X 1010
Holotrichia
consanguinea
56.67 Jat and
Choudhary,
20065 X 109
Holotrichia
consanguinea
53.33
1 X 107
Varroa destructor 100.00 Meikle et al.,
2006
Metarhizium
anisopliae
1 X 107
Varroa destructor 75.00 Meikle et al.,
2006
Nomurea rileyi 1 X 108
Helicoverpa armigera 82.10 Snegapriya and
Manjula, 2008
1 X 109
Helicoverpa armigera 70.00 Manjula and
krishnamurthy,
2005
1 X 109
Spodoptera litura 53.40 Manjula and
krishnamurthy,

30. Population density of Diamondback mothlarvae,
Plutella xylostella on cauliflower
Treatment Average number of DBM per m2
B.Bassiana
(VL1-SCL)
90.0 77.6 56.4 44.0 30.4
B. bassiana
(OM2-SDO)
96.4 80.8 55.2 32.8 22.4
M.anisopliae
(OM1-R)
84.8 75.6 50.4 32.8 25.6
M.anisopliae
(OM3-STO)
94.8 77.2 46.0 24.8 19.2
(min.)
Crymax35W
P
87.2 68.4 42.4 26.4 20.0
Atabron 5EC 80.4 71.6 56.4 41.6 32.8
3 DAT 5 DAT 7 DAT1
DAT
1 DBT
Loc, N.T and Chi, T.B. (2007). Biocontrol potential of Metarhizium anisopliae and
Beauveria bassiana against diamondback moth, Plutella xylostella. Omonrice.,15: 86-93.

31. Field efficacy of certain M. anisopliae and B. bassiana
isolates against DBM larvae, Plutella xylostella
Treatment concentration
mortality (%)
B.Bassiana
(VL1-SCL)
107
conidia/ml 38.6 51.0 64.3
B. bassiana
(OM2-SDO)
107
conidia/ml 43.2 65.2 75.3
M.anisopliae
(OM1-R)
107
conidia/ml 39.7 60.7 67.4
M.anisopliae
(OM3-STO)
107
conidia/ml 52.4 74.2 78.5(max.)
Crymax35WP 0.1% 53.5 70.0 76.1
Atabron 5EC 0.3% 30.0 44.8 53.7
CV (%) - 16.7 13.2 10.2
3 DAT 5 DAT 7 DAT
Loc, N.T and Chi, T.B. (2007). Biocontrol potential of Metarhizium anisopliae and
Beauveria bassiana against diamondback moth, Plutella xylostella. Omonrice.,15: 86-93.

32. Effect of M. anisopliae and B. bassiana on ratio of commodity
cauliflower and cauliflower yield
Treatment Ratio of
commodity
(cauliflower %)
Yield
B.Bassiana (VL1-SCL) 68.3 6.37 58.1
B. bassiana (OM2-
SDO)
85.0 6.78 68.2
M.anisopliae (OM1-R) 83.3 6.72 66.7
M.anisopliae (OM3-
STO)
91.7 6.98 73.2
Crymax35WP 93.3 7.04 74.76 (max.)
Atabron 5EC 63.3 5.97 48.1
Untreated control 13.3 4.03
t/ha Increased over control(%)
Loc, N.T and Chi, T.B. (2007). Biocontrol potential of Metarhizium anisopliae and
Beauveria bassiana against diamondback moth, Plutella xylostella. Omonrice.,15: 86-93.

33. Protozoans
• More than 100 species are pathogenic to insects
• Chronic nature of infection so limited efficiency
• Few are Highly virulent or fast acting so more
appropriate for long term control programmes
with high economic injury level
• Important Phyla of Protista are:
Sarcomastigophora, Apicomplexa, Microspora
and Ciliophora

34. Protozoans considered for control of
insect pests
Parasite Host
Nosema acridophagous, N.
cuneatum, N. locustae
Grasshoppers
N. algerae Anopheles albimanus, Culex
tritaeniorhynchus
N. fumifueranae Spruce budworm
N. heliothidis Helicoverpa zea
N. pyrausta Ostrinia nubialis
N. whitei Tribolium castaneum
N. spp. Helicoverpa armigera, Spodoptera
litura
Vairimorpha necatrix Agrotis ipsilon, Helicoverpa zea

35. Protozoan insecticides
Nosema locustae
Specific to grasshoppers and Mormon cricket
Slow acting
Most appropriate for long term management
Limited shelf life
Nolo bait

36.  Entomopathogenic nematodes are a group of
nematodes causing death to insects
 The infective juveniles of entomopathogenic
nematodes is 3rd instars
 Soil inhabiting
 Endoparasitic
 EPN’s have been found in all ecologically diverse
habitats, from cultivated fields to deserts
Entomopathogenic nematode

37. Infective juvenile
Host finding
Infection
Release of bacteria
Host killed
Development & reproduction
of nematodeInfective juvenile emergence
Mode of Action

38. Effect of entomopathogenic nematodes on flea beetle
at different temperature
LC50
Nematode species 150
C 200
C 250
C
S. feltiae 975 1044 730
S. carpocapsae _ 752 870
H. megidis 1354 1020 1148
H. bacteriophora 2492 912 778
Trdan, et al (2008). Acta Agriculturae Scandinavica Soil and Plant Science., 58:
169-175.

39. Commercial products of Entomopathogenic
nematode available in international market
Nematode species Product formulation Country
Steinernema carpocapsae Green commandos,
Soil commandos
India
Sanoplant Switzerland
Boden Nutz;linge Germany
Helix Canada
X-GNAT, Vector TL USA
S. feltiae Magent USA
Nemasys, Stealth UK
Entonem USA
S. riobrave Vector MG, Bio Vector USA
Heterorhabditis
bacteriophora
Otinem USA
H. megidis Nemasys UK
S. carpocapsae Ortho, Biosafe, Bio Vector, Exhibit USA
Vashisth, S., Chandel, Y.S and Sharma, P.K. (2013). Agri. Reviews, 34 (3) : 163-175.

40. Desirable characteristics required in microbial
pesticides
Characteristic Bacteria Fungi Viruses Protozoa Nematode
Time to kill Good Poor Poor Poor Good
Easy to apply Yes Yes Yes Yes Yes
Storage
Characteristics
Good Poor Good Poor Good
Environmental
stability
Poor Poor Poor Poor Poor
Safe to non
target organisms
Yes Yes Yes Yes Yes
Easy to produce Yes Yes Poor Poor Yes

41.  The microbial pesticide are potential bio-agents amenable for
mass production, handling and application on a large scale in
the field for management of insect pests.
 Microbial pesticides provide effective alternatives in an
eco-friendly way for the management of many insect pests.
 Maximum used microbial pesticides in world is bacteria (60%)
followed by fungi (27%), virus (10%), nematode, protozoa etc.
 Rapid degradation by UV light, Poor water solubility, poor
storage capacity and non-availability of microbial pesticide
limited their use.
 For expanded use of microbial pesticide there is need to
exploit the potential biodiversity in the country by continued
surveys.
Conclusion