Baculovirus: A viral bioagent for biological control

Major Guide
Dr. R.N. Pandey
Speaker
Prashant C. Arade
Minor Guide
Ph.D. 4th Sem.
Dr. P. K.Borad
 Outline
Introduction

State of Taxonomy and Biology of Baculoviruses

Baculovirus Production Technology

Field stability and persistence

Use of Baculoviruses for Pest Control

Genetically Modified Baculoviruses to Control Insects

Final Considerations and Further Prospects on Use

of Baculoviruses as Biopesticides
Introduction
 There are at least 12 viral families associated with insects and other
arthropods (Erlandson, 2008).

 The Baculoviridae is the most commonly investigated with regard to its

development as a microbial insecticide due to its favorable characteristics
such as safety to the environment, humans, other vertebrates, plants, and
natural enemies of pests (particularly predators and parasitoids).

 The baculoviruses are ideal control agents to be used in integrated pest

management (IPM) programs in agriculture, forests, and pastures.

 Baculoviruses are a large and diverse group of viruses pathogenic to

arthropods, primarily insects from the orders Lepidoptera,
Hymenoptera, and Diptera.

 They are valuable natural control agents, but their utility in many
agricultural applications has been limited by their slow speed of kill and
narrow host specificity.

 Baculoviruses represent a promising alternative to the use of synthetic

chemical insecticides.
 Table 1: Characteristics of viruses found in insect
Nucleic Virus Inclusion body Subgroup and
Virus
acid particle shape common names
Ascoviridae dsDNA Allantoids None -
Polyhedral NPV
Baculoviridae dsDNA Bacilliform Cigar-shaped
GV
capsules
Calciviridae ssRNA Isometric None -
Iridoviridae ssDNA Isometric None Iridescent
Nodaviridae ssRNA Isometric None -
Parvoviridae ssDNA Isometric None -
Picornaviridae ssDNA Ovoid None -
Ovoid or
Poxviridae dsDNA Spheroid Entomopox viruses
brick shaped
Cytoplasmic
Reoviridae dsDNA Isometric Polyhedral
polyhedrosis
Rhabdoviridae ssRNA Helical None -
 Order Species

Approximately 60 percent of 1200 known Lepidoptera 455

insect viruses belongs to family
baculiviridae Hymenoptera 31

Dipteral 27

Baculovirus infection described in 700 Coleoptera 5

species of invertebrates
Neuropteran 2

Trichoptera 1

Thysanoptera 1

Sphanoptera 1
Table 2: Current use of Baculoviruses as biological insecticides

Crop Insect pest Virus Used

Apple, pear, walnut and Codling moth Codling moth granulosis
plum virus
Cabbage, tomatoes, Cabbage moth, American Cabbage army worm
cotton, (and see pests in bollworm,diamondback nuclear polyhedrosis virus
next column) moth, patato tuber moth
Cotton, corn, tomatoes Spodoptera littoralis Spodoptera littoralis
nuclear polyhedrosis virus
Cotton and vegetables Helicoverpa zea, and Helicoverpa zea nuclear
Heliothis virescens polyhedrosis virus
Vegetable crops, Beet armyworm (Spodoptera Spodoptera exigua nuclear
greenhouse flowers exigua) polyhedrosis virus
Vegetables Celery looper (Anagrapha Anagrapha falcifera
falcifera) nuclear polyhedrosis virus
Alfalfa and other crops Alfala looper (Autographa Autographa californica
californica) nuclear polyhedrosis virus
Taxonomy and Biology of Baculoviruses
 More than 700 baculoviruses have
been isolated from invertebrates.
 They possess circular, covalently
closed, double-stranded DNA
genomes ranging from 80 to 180 kbp
in length, encoding for 100–180
proteins.
 These viruses occur naturally in
insect populations and are normally
named for the initial host from which
they were isolated.
 Baculoviruses exist as two
phenotypes, i.e., occlusion-derived
virus (ODV) and budded virus (BV), Fig. Taxonomic classification of Baculovirus
which have a common nucleocapsid
structure and carry the same genetic
information.
 Phylogeny of Baculovirus

The viruses belong to the

family Baculoviridae, which
is currently subdivided on
the basis of phylogenetic
evidence and molecular
characteristics into four
genera:
Alphabaculovirus
(lepidopteran
nucleopolyhedrovirus),
Betabaculovirus
(lepidopteran granulovirus),
Gammabaculovirus
(hymenopteran
nucleopolyhedrovirus)
Deltabaculovirus
(dipteran nucleopolyhedrovirus).
 Ingestation of OBs polyhedra

Release of hundreds of ODVs in the gut.

The ODVs is dissolved by the alkaline environment.

Pass through the peritrophic membrane, attach to the microvilli causing

primary infection

Budded virus (BV) causes secondary infection

In the nucleus, where viral transcription, DNA replication, and assembly of

progeny nucleocapsids occur, resulting in the production of BV and ODV.

Nucleocapsids become occluded in a protein matrix to form OBs

OBs are released upon death

Infected insect can migrate to a higher elevation

Figure 1:
A life cycle of a baculovirus causing systemic infection. Occlusion bodies ingested by an
insect dissolve in the midgut, and ODV are released which then infect midgut epithelial cells
(A). The virion buds out of the cell in a basal direction and initiate a systemic infection (B).
Early in the systemic infection more BV are produced which spread the infection throughout
the insect (C). Late in infection occluded virions are produced, and the cell then dies
releasing the occlusion bodies (D). The virogenic stroma (VS) is indicated.
Nuclear Polyhedrosis Virus
(NPV)
 Nuclear Polyhedrosis Virus
 41 % of arthropod viruses
develop in host cell nuclei,
virion occluded singly/groups
in polyhedral inclusion bodies
 Rod shaped, double sranded,
POBs 0.2-15 µm in diameter
 Highly host specific
 Enters through injection of
plant material into insect gut
through mouth and cuticle
Fig.3: POBs of NPVs were purified by differential centrifugation and enumerated
under phase-contrast microscope at 1000 X magnification.
 A B C

Figure 4: Scanning electron micrograph (SEM) showing

(A) H. armigera nucleopolyhedrovirus (HaNPV) polyhedra
(B) S. litura nucleopolyhedrovirus (SlNPV) polyhedra
(C) Amsacta albistriga nucleopolyhedrovirus (AmalNPV) polyhedra
Figure 5: Circular map of the BusuNPV genome of Buzura suppressaria
 Symptoms of NPV infection
Diseased larvae typically crawl to the top of the plant to die.
Shortly after death, an infected larva’s body becomes flaccid
and its skin (integument) ruptures, releasing millions of PIBs
(the infectious virus particles) back into the environment.
When larval numbers are high, waves of natural infection can
develop as more larvae become infected, die, and spread the
infection, resulting in an outbreak (epizootic).
Left: A larvae of H. armigera that died of NPV on chickpea
Right: A larvae of H. armigera that died of NPV on cotton
 Granulosis Virus
 Develop either in the nucleus/ cytoplasm/
tracheal/matrix/epithelial cells of host
 Virions are occluded singly in small
inclusion bodies called capsules
 Rod shaped virion, dsDNA
 Oval occlusion bodies about 200x400nm
 They enter through ingestion, similar to
NPV
 Fat body is the major organ involved
 Diseased larvae- less active, flaccid, fragile,
wilted prone to rupture in later stages,
death in 6-20 days
Cydia pomonella
Baculovirus Production
Technology
In vivo production
In vitro production
 In vivo production

Commercial production of Baculoviruses in vivo:

Applying the virus against the host insect in the field

Producing the target insect in the laboratory on artificial

diet
 A B C

Supernatant

NPV pellet

Figure 6: Centrifugation of virus-infected larval extracts of HaNPV (A), SlNPV (B)

AmalNPV (C)
 In vitro production
Baculovirus production for agricultural pest control needs to be efficient,
with competitive costs, leading to a final product which is highly pathogenic
to the target pest.

Limitation for in vitro production:

 Successive passages of the virus in cell culture result in genetic
alterations (Krell, 1996; Rhodes,1996).
 Highly productive insect cell line and a highly productive culture
medium
 In vitro production remains an important requirement from a
commercial perspective for the use of baculoviruses as
insecticides.
 Many Polyhedra (MP) variants → plaque assay technique → MPs
maintain the wildtype features.
 Reduction of cost of cell culture medium
• 250 LE (LE-larval equivalent) HaNPV
– The effectiveness of the viral insecticide is critically dependent on
the concentration of POB, which is expressed as larval equivalent
(LE).
– A standard 1 LE stock preparation consists of 6 ×109 POBs/ml.

Use of HaNPV in field

The field application rate of 250-500 LE/ha of HaNPV has been
recommended for the control of H. armigera by different workers on
various crops.

The virus is highly specific to target insect. So it has no deleterious

effect on non-target organisms and environment.
 Field stability and persistence

Highly susceptible to damage by desiccation and by exposure to sunlight or

UV radiation

Additives like charcoal, egg albumin, molasses, and sugar

Addition of crude sugar to HaNPV spray fluid increased persistence of virus

both under natural sunlight and shade

Brightness reduces the LC50

Evening spray
 General considerations for field application of BVs
1 Method of application Foliar spray
2. Stage of pest Early instars are highly susceptible.

3. Dose of virus preparations HaNPV-250LE/ha (1LE=6×PIB),2 to 3 sprays

in early stages of pest.

4. Preparation of spray fluid NPV in required strength is mixed with good

quality soft water +0.1%
teepol or Trition-x-100.

5. Time of application Preferably in evening hours.

6. Frequency of application 2-4 application in case of NPV/weekly.
7. Application equipment High volume applications are more effective
than low volume.

8. Integration of chemicals NPV is compatible with most of pesticides.

9. Use of adjuvants Skimmed milk, juggary, teepol, sandovit
Table 3: Development of Cryptophlebia leucotreta on maize meal diet with different
concentrations of casein, ascorbic acid and wheat germ added.

Additive* Moths Duration of Contamination

produced/jar** ± development from
SE egg to start of
(5 jars/treatment) pupation (days)

None 33.8bc ± 4.8 28 None in any of

Rhizopus sp. 74.2d ± 5.7 12 the diets.
Ascorbic acid 0.2 g 19.0ab ± 2.8 29
Ascorbic acid 0.4 g 3.8a ± 2.1 30
Casein 0.36 g 42.8bcd ± 10.7 22
Casein 0.73 g 50.2bcd ± 6.1 21
Wheat germ 2 g 69.6cd ± 8.6 20
Wheat germ 4 g 70.6cd ± 5.9 18
*Nipagin (0.3 g) was added to each jar (to prevent fungal contamination) except to the standard Rhizopus
sp. inoculated diet.
**Values in the same column followed by the same letter are not significantly different (P > 0.05;
Bonferroni LSD multiple range test).

Moore , 2002 Grahamstown

Table 4 : Percent mortality and LT50 values of SpltNPV-infected Spodoptera litura
larvae after treatment with different SDS concentrations.

Mortality (%) LT50 (days)

Treatment
(Mean±SE)* (95% C.I)**

Water 0a 0
Untreated NPV 96.00±1.63b 3.51 (3.29-3.72)
0.005% SDS 95.00±1.66b 3.45 (3.24-3.66)
0.01% SDS 92.00±2.91b 3.71 (3.50-3.92)
0.10% SDS 72.00±4.67c 4.14 (3.93-4.35)
0.20% SDS 68.00±5.33c 4.34 (4.13-4.54)
*Means with the same letters indicate no significant difference at P<0.05 (Tukey’s test).
**Estimates of LT50 (days) calculated from means of 10 replicates with 10 larvae per treatment. No mortality was recorded
from negative control (water).

Nazli-Huda et al. 2012 Malaysia

Table 5: Amount of obtained virus material in correlation to population density of the
satin moth (Leucoma salicis) in greenhouse conditions
No. of dead larvae Total
No. of (in %) body Weight
No. of
examined mass of of virus PIBs/mg
larvae/ Total Larval stage
larvae dead powder powder
1L [WW]
[Ni] larvae [in mg]
[Nz] L4 L5 L6
[in g]

2450 567 1153 730

6.0 1000 x 3 ** 557.4 c 36.6 b 5.1x108a
(81.6) (18.9) (38.4) (24.3)

4387 2164 1488 735

9.5 1600 x 3 ** 424.1 b 28.7 b 4.8 x 108a
(91.4) (45.1) (31.0) (15.3)

429 273 129 27

18.6 186 x 3 * 37.2 a 2.4 a 4.8 x 108a
(76.9) (48.9) (23.1) (4.8)

680 306 199 175

28 280 x 3 * 53.3 a 3.3 a 4.9 x 108a
(80.9) (36.4) (23.7) (20.8)

Insect rearing conditions: * glass vessels (10 l); ** cages (168 l). Examined stage L4
Numbers followed by the different letter differ statistically
WW] = [Nz]/[Ni] x [PIBc]/[PIBi] where [PIBi] = 9 x 109

Ziemnicka, 2007 Poland

Table 6: Effect of UV protectants with and without PIBs of SlNPV on Spodoptera litura

Treatments PIB *Mean per cent mortality

Cupric Ammonium Nitrate (UE) + 97.78a (82.99)**
Cupric Ammonium Nitrate (E) + 94.43b (76.52)
Tinopal (UE) + 95.56ab (78.00)
Tinopal (E) + 84.44c (66.80)
Cupric sulphate (UE) + 95.56ab (78.00)
Cupric sulphate (E) + 95.55ab (80.02)
PIB alone (UE) + 97.78a (82.99)
PIB alone (E) + 4.44d (12.00)
Cupric Ammonium Nitrate alone - 0.00e (0.00)
Tinopal alone - 0.00e (0.00)
Cupric sulphate alone - 0.00e (0.00)
Control - 0.00e (0.00)
S.Em. % 2.88
CD@ 5% 4.94
UE- Unexposed; E- Exposed
*Values are mean of three replications. **Figures in the parentheses are arcsine transformed values.
Mean value with different alphabets differ significantly

Arivudainambi et al., 2000 Annamalai Nagar

Table 7: Field efficacy of HaNPV isolates against H. armigera on chickpea (cv. Shoba)

Number of larvae per 10 plant Pod damage

Reduction Reduction
(%)
Treatment* Pre-treatment Pooled over control over
(Pooled
count mean** (%) (%)
mean)**
CBE I 9.33 5.17a 61.88 9.72ab 54.52
NEG 10.33 5.50ab 59.42 9.26a 56.67
BAN I 9.67 6.50cd 52.03 11.11c 48.01
HYD 10.00 6.33bcd 53.28 10.80bc 49.44
MAH I 9.67 7.11de 47.53 12.48d 41.59
RHI 10.33 7.56e 44.26 13.22d 38.11
RAJ 9.67 9.33f 31.14 17.98e 15.86
Endosulfan 35 EC
9.00 6.00abc 55.75 10.48bc 50.96
(700 g/ha)
Untreated control 10.33 13.56g – 21.37f –
*NPV was applied @ 1.5 × 1012 POB/ha in teepol 0.1%; crude sugar @ 2.5 kg/ha was used as adjuvant
**Pooled mean after six rounds of spray
In a column means followed by similar letters are not statistically different by DMRT (P < 0.05). ANOVA statistics
– number of larvae F = 46.25, df = 40, P < 0.001; pod damage F = 89.30, df = 24, P < 0.001

Jeyarani et al., 2010 TNAU, Tamil Nadu

Table 8: Field efficacy of NPV-S alone and in combination with insecticides on the
infestation of Spodoptera litura on cabbage crop.
Treatment Pre- Per cent reduction in larval population
treatment after
population 1st spray 2nd Spray
(%) 7 DAS 14 DAS 7 DAS 14 DAS
NPV-S (250 LE/ha) 2.30 25.61 40.39 60.06 72.28
NPV-S (500 LE/ha) 2.27 42.80 61.98 78.26 87.24
Endosulfan (1250 ml/ha) 2.33 53.57 68.13 85.78 93.65
Neemarin (700 ml/ha) 2.26 23.14 37.65 60.78 69.13
NPV-S (250 LE/ha)+ Endosulfan 2.27 27.03 43.80 88.14 95.35
(625 ml/ha)
NPV-S (500 LE/ha)+ Endosulfan 2.13 45.56 59.63 93.85 98.25
(625 ml/ha)
NPV-S (250 LE/ha)+ Neemarin 1.99 30.19 45.00 66.34 79.34
(700 ml/ha)
NPV-S (500 LE/ha)+ Neemarin 2.23 43.12 60.03 81.71 90.53
(700 ml/ha)
Untreated check 2.40 3.41 8.39 11.67 13.45
SE.M. ± - 2.67 1.77 2.45 1.78
P≤ 0.05 NS 8.00 58.30 7.34 5.33
P≤ 0.01 NS 11.01 7.30 10.11 7.34
Kumari and Singh, 2000 Jaipur
Table 9: Pot culture evaluation of different adjuvants on cabbage
Treatments Pretreatme #Mean percent mortality
nt Under shade (DAS) Under sun (DAS)
5th 7th 10th 5th 7th 10th
Jiggery 5% + Blue 1% + 20 45.00 65.00 78.75 35.00 48.75 58.70
1X106 OB’s/ml, spltNPV- (42.12)a (53.76)a (62.66)a (36.25)ab (44.28)ab (50.06)a
Glycerol 10%
Soyaflour 10% + 1x106 20 33.75 47.50 63.75 31.25 42.50 48.70
OB’s/ml, spltNPV-glycerol (35.48)b (43.56)c (53.01)b (33.98)b (40.66)b (44.28)b
10%
1x106 OB’s/ml, spltNPV- 20 18.75 27.50 31.25 11.25 20.00 26.20
Glycerol 10% (25.54)c (31.61)d (33.94)c (19.24)c (26.48)c (30.75)c
Indoxacarb 14.5 SC, Avant® 20 45.00 55.00 71.25 41.25 52.50 63.75
1ml/l (42.12)a (47.90)b (56.83)b (39.94)a (46.44)a (53.02)a
Control 20 0.00 0.00 0.00 0.00 0.00 0.00
(0.00)d (0.00)e (0.00)d (0.00)d (0.00)d (0.00)d
F test * * * * * *
SEm ± 1.253 1.418 1.335 1.381 1.486 1.299
CD at 1% NS 5.226 5.109 5.562 5.757 6.192 5.415

# Mean of 30 larvae/treatment/replication, DAS: Days after sowing

$ Figures in the parentheses are Angular transformed values

Bhutia et al., 2012 UAS, GKVK, Bangalore

Table 10: Efficacy of selected adjuvants in protecting sunlight inactivation on SlNPV
after 16 hrs of exposure to sunlight

Mortality (%)*
Treatment LT50 Slope
(Mean±SE)**

Unexposed virus 100.00±0a 9.10 (8.57–9.63) 8.27±1.35

Exposed virus 0d - -

Virus+Tinopal 100.00±0a 7.81 (7.30–8.27) 8.01±1.24

Virus+riboflavin 29.17±1.60b - -

Virus+brown sugar 24.17±1.60c - -

Virus+palm oil 0d - -

Control 0d - -
*Means with the same letters indicate no significant difference at Pb0.05 (Tukey's test).
** Estimates of LT50 (days) calculated from means of four replicates with 30 larvae/
replicate. No mortality was recorded from all controls.

Sajap et al. 2009 Malaysia

Table 11: Combination effect of NPV with thiamethoxam against 5-day old
larvae.
Treatment Concentration of Concentration of % Mean of
insecticide NPV mortality
(ppm) (PIB/ml)
NPV alone 0 2.2 x 106 48.3 a
NPV + Thiamethoxam 30 2.2 x 106 65.0 b
NPV + Thiamethoxam 50 2.2 x 106 70.0 bc
NPV + Thiamethoxam 100 2.2 x 106 73.3 bc
NPV + Thiamethoxam 150 2.2 x 106 83.3 c
F 38.18**
CV (%) 14.50
** Significant at 1% and the values with same alphabet is not significantly different.

Trang and Chaudhari, 2002 Vietnam

Table 12: Combination effect of NPV with diflubenzuron against 5-day old larvae.

Treatment Concentration of Concentration of % Mean of

insecticide NPV mortality
(ppm) (PIB/ml)
NPV alone 0 2.2 x 106 43.3 a
NPV + Diflubenzuron 1 2.2 x 106 65.0 b
NPV + Diflubenzuron 3 2.2 x 106 66.7 b
NPV + Diflubenzuron 5 2.2 x 106 70.0 b
NPV + Diflubenzuron 7 2.2 x 106 73.3 b
NPV + Diflubenzuron 10 2.2 x 106 93.3 c
F 8.93**

CV (%) 13.5

** Significant at 1% and the values with same alphabet is not significantly different.

Trang and Chaudhari, 2002 Vietnam

Table 13: Combination effect of NPV with imidacloprid against 5-day old larvae.

Treatment Concentration of Concentration of % Mean of

insecticide (ppm) NPV(PIB/ml) mortality

NPV alone 0 2.2 x 106 46.6 a

NPV + Imidacloprid 1 2.2 x 106 73.3 b
NPV + Imidacloprid 3 2.2 x 106 80.0 bc
NPV + Imidacloprid 5 2.2 x 106 81.7 cd
NPV + Imidacloprid 7 2.2 x 106 88.3 d
NPV + Imidacloprid 10 2.2 x 106 48.3 a
NPV + Imidacloprid 20 2.2 x 106 45.0 a
F 127.5**

CV (%) 7.4

** Significant at 1% and the values with same alphabet is not significantly different.

Trang and Chaudhari, 2002 Vietnam

Table 14 : Bio – efficacy of Nuclear Polyhedrosis Virus (NPV) on protection of cotton
Bolls damaged.
Observation of Bolls damaged (%) *
Treatments Replications Pre-
Spray 1 Spray 2 Spray 3 Spray 4
Treatment
R1 8.24±1.43a 6.42±1.33g 5.68±1.99i 2.76±1.39f 2.68±1.30f
T1 R2 9.51±1.55h 6.86±1.39h 4.74±1.38g 2.18±1.39c 2.42±1.73d
R3 9.46±1.88g 7.13±2.48i 5.27±1.77h 2.35±1.55e 2.54±1.54e
R1 8.61±1.31b 2.14±1.20a 2.92±1.23f 3.17±1.99h 3.24±1.77g
T2 R2 10.41±1.92l 2.65±1.28d 2.74±1.22d 3.04±1.82g 3.75±1.42i
R3 9.23±1.44d 2.71±1.22e 2.84±1.37e 3.22±1.37i 3.69±1.26h
R1 9.31±1.33f 2.28±1.25c 1.95±1.22a 1.95±1.20a 1.84±1.27b
T3 R2 8.64±1.95c 2.19±1.39b 2.17±1.27b 2.17±1.35b 1.98±1.37c
R3 10.27±1.66j 3.18±1.28f 2.23±1.38c 2.23±1.40d 1.23±1.66a
R1 9.28±2.48e 9.32±2.46j 10.16±2.28j 11.72±2.48l 12.15±2.46j
T4 R2 10.32±2.64k 9.87±2.49k 10.28±2.83k 11.48±2.65k 11.40±2.48l
R3 39.54±1.99i 10.11±2.84l 10.71±2.75l 11.37±2.99j 12.24±3.34k
T1 = NPV 500 LE / ha; T2 = Cypermethrin 25% EC. 25% + Quinalphos 25% EC . 5%
T3 = NPV 500 LE / ha + Cypermethrin 25% EC. 25% + Quinalphos 25% EC .5%;
T4 = Control (without NPV) Each value mean ± S.D represents mean of five values.
Values in a column with a different superscript alphabet are *significantly different at P < 0.05(MANOVA; LSD -Tukey’s Test).
*Percentage of boll damaged observed for pre and post treatment of Nuclear Polyhedrosis Virus (NPV) after 24h of exposure period.
Pugalenthi et al., 2013 Pattanam (TN)
Table 15: Bio – efficacy of Nuclear Polyhedrosis Virus (NPV) on protection of cotton
Bolls damaged.
Observation of Bolls damaged (%) *
Treatment Replications Pre-
Spray 1 Spray 2 Spray 3 Spray 4
Treatment
R1 9.15±1.23c 7.31±1.27i 4.38±1.22h 3.86±1.82i 2.96±1.23h
T1 R2 8.76±1.78a 6.83±1.92h 4.16±1.36g 3.45±1.77f 2.32±1.37e
R3 9.24±1.82d 6.18±1.82g 4.42±1.30i 3.78±1.28g 2.71±1.38g
R1 8.95±1.84b 4.15±1.29f 3.25±1.85e 3.17±1.33d 2.18±1.93d
T2 R2 10.36±2.54k 3.73±1.93d 3.67±1.86f 3.43±1.54e 2.34±1.51f
R3 9.87±1.22i 3.54±1.26b 3.18±1.22d 3.84±1.93h 3.07±1.94i
R1 9.53±1.43e 3.68±1.24c 2.43±1.73c 2.14±1.33b 1.15±1.34b
T3 R2 9.71±1.83g 3.49±1.22a 2.15±1.82a 1.95±1.84a 1.15±1.34b
R3 9.84±1.66h 3.82±1.86e 2.36±1.27b 2.27±1.77c 1.28±1.83c
R1 10.42±2.73l 11.63±2.57l 12.47±2.99k 11.81±2.83k 12.56±2.94k
T4 R2 9.67±1.9 2f 10.48±2.49k 12.18±2.74j 12.17±2.94l 12.84±2.74l
R3 9.89±1.23j 10.17±2.69j 11.74±2.88l 11.64±2.59j 11.52±2.55j
T1 = NPV 500 LE / ha; T2 = Cypermethrin 25% EC. 25% + Quinalphos 25% EC . 5%
T3 = NPV 500 LE / ha + Cypermethrin 25% EC. 25% + Quinalphos 25% EC .5%;
T4 = Control (without NPV) Each value mean ± S.D represents mean of five values.
Values in a column with a different superscript alphabet are *significantly different at P < 0.05(MANOVA; LSD -Tukey’s Test).
*Percentage of boll damaged observed for pre and post treatment of Nuclear Polyhedrosis Virus (NPV) after 24h of exposure period.
Pugalenthi et al., 2013 Pattanam (TN)
 Figure 7: Time mortality curves for Spodoptera litura larvae
treated with NPV at different age.

Trang and Chaudhari, 2002 Vietnam

Table 16: Probit analysis for dose- mortality response of H. armigera to
commercial formulations of Nuclear Polyhedrosis Virus

Source of HaNPV formulation LC50 Fiducial Regression Chi

POBs/ml Limits equation Square
POBs/ml (n=5)

Pest Control India, Pvt Limited, 9.5X105 4.70 –15.03 Y= -3.094+0.817X 1.71
Bangalore (PCI)
Project Directorate of Biological 11.42X106 6.08 – 21.60 Y= -4.497+0.595X 0.26
Control, Hebbal, Bangalore (PDBC)
Margo Bio Pesticides, Bangalore 9.5X105 4.80 – 19.41 Y= -3.031+0.49X 1.44
(MBPS)
Biopest Management Services, 15.6X105 7.50 – 33.75 Y= -4.063+0.528X 1.09
Bangalore
ZARS, Gulbarga (ZARS-G) 3.12X104 2.35 – 11.17 Y= -1.711+0.387X 1.15

Srinivasa et al. , 2008 UAS, Bangalore

Table 17: Comparative susceptibility of neonate larvae of C. pomonella and three
other tortricid fruit pests to the nucleopolyhedro virus of A. falcifera

Occlusion bodies/mm2 (95% confidence limit)

Species/dasage
LC50 LC95
1.77x104 1.67x104
Cydia pomonella (1.19x103-2.52x103) (9.02x103-5.53x104)
4.30x103 3.19x104
Pandemis pyrusana (3.53x103-5.32x103) (2.14x104-5.64x104)
7.63x103 8.45x104
Choristoneura rosaceana (4.47x103-1.78x104) (2.97x104-1.35x106 )
2.06x103 9.55x103
Grapholitha molesta (1.30x103-2.91x103) (5.4x103-5.15x104)

Lacey, et al., 2002 USDA

Table 18: Control of bollworm Helicoverpa armigera in cotton
Treatments Concentration Surviving larvae (meanSEM)/25 plants a % damage
per ha (meanSEM) a;c
8 August b 11 August 15 August
HaSNPV-WT 2.1X1012 PIBs 20.8±0.6a 13.8±0.5b 9.8±0.4c 11.2±0.8b
HaSNPV-AaIT 2.1X1012 PIBs 19.0±1.0a 4.8±0.5a 2.8±0.5a 7.6±0.7a
HaSNPV-AaIT 1.5X1012 PIBs 22.5±2.1a 12.0±1.7b 6.8±0.9b 12.5±0.5bc

HaSNPV-AaIT 9.0X1011 PIBs 19.3±1.5a 11.8±0.7b 8.3±0.5bc 14.3±0.6c

B. thuringiensis 4.8X1010 IUd 19.0±1.5a 12.3±0.9b 8.0±0.6bc 10.7±0.7b

WP
Controle – 20.8±1.5a 19.5±1.3c 18.5±1.3d 17.5±0.6d

a Different letters indicate significant differences between treatments (p ¼ 0:05).

b Applications were made on August 8. Larvae were counted before spray.
c Number of squares, flowers, and bolls on 25 plants in each plot which were damaged or undamaged due to feeding by bollworm was
counted on
August 15. % damage was calculated as counts of damaged organs/(counts of undamaged organs+counts of damaged organs).
d IU, International Units.
e Control plots were treated with formulation only.

Jiayu & Hubei, 2000 Japan

 Genetically Enhanced Viral Pesticides
Two broad strategies have been pursued in laboratories worldwide
to achieve this goal:
1. Interference with host physiology and
2. Introduction of an insect-specific toxin
 The first strategy involves introducing genes coding for some
insect hormones or enzymes into the baculovirus genome.
 Maeda (1989) was the first to introduce a diuretic hormone gene
into Bombyx mori baculovirus genome to cause insects to lose
water.
 The recombinant baculovirus reduced survival time of
Trichoplusia ni larvae by more than 20% in comparison to larvae
infected with a control virus.
 LD50 values were significantly lower for the recombinant virus in
comparison to the wild-type virus (from 4.4- to 21-fold lower).
 One of the recombinants expressing cathepsin L under
baculovirus promoter of p6.9 gene generated a 51% faster speed
of kill in comparison to the wild-type virus.
 STRATEGIES TO COUNTERACT LIMITATIONS OF BACULOVIRUSES

Mixture of In vivo sequential

Substances that
baculoviruses with passage of Genetic engineering of
enhance baculovirus
reduced dosages of baculoviruses on host baculoviruses
activity in host insects
insecticides and non-host insects

PROBLEMS THAT LIMIT EXPANSION OF BACULOVIRUS USE

 Host-specificity of baculoviruses
 Large-scale production of baculoviruses
 Lack of low-cost, serum-free media and suitable cell lines and virus
strains
 Need to scale up production of highly active virus with high titers per
ml of medium
 Slow speed in stopping pest damage and killing the hosts
 Are baculoviruses safe?

Yes.
 Only found in insect (mainly lepidopteran species)
 Narrow host range, high selectivity
 No production of metabolites or toxins
 Cause no hazards to human health
 Adopted for wide-scale use
Commercial formulations used in India & Shrilanka
Conclusion
It is very likely that the growing awareness of the
benefits of the environment-friendly pesticides will result
in the reevaluation of the prospects for biological
protection with baculovirus preparations. However, until
today, baculovirus insecticides have not met their full
potential to control pest insects worldwide. The expansion
of baculovirus use, in the following years, would depend on
new developments in the areas of recombinant
baculoviruses and in the in vitro commercial production of
these agents. Future development of baculovirus pesticides
will probably depend on the attitude towards genetically
modified organisms.
Future Prospects
 There should be modification of proteins and peptides
for enhanced stability in vivo may improve all of the
recombinant baculoviruses especially those that show
minimal increases in speed of kill.
 The improvements will be at the level of diagnostics of
infection, development of the in vitro cultures and
changes in the formulations of the biopesticide.
 There should be improved efficiency of production,
better performance in the field and an increased
demand for environmentally safe, host specific
microbial pest control products may lead to greater
degree of commercialization and use of baculoviruses
in pest control agents.
Thank you