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DEPARTMENT OF ENTOMOLOGY
ALLAHABAD SCHOOL OF AGRICULTURE
SAM HIGGINBOTTOM INSTITUTE OF AGRICULTURE TECHNOLOGY &
SCIENCES
[Formerly-Allahabad Agricultural Institute]
(Deemed-to-be-University)
ALLAHABAD- 211007, U.P., INDIA BY,
RAKESH KR.
MEENA
MASS MULTIPLICATION OF
The egg of Sitotroga cerealella are
generally used as laboratory host for
production of Trichogramma in the
USA, France, Germany etc. Whereas in
India ,rice grain moth,Corcyra
cephalonica is used as the laboratory
host.
STEPS IN CORCYRA PRODUCTION:
The quantities of sorghum/pearl
millet/maize grain, free from
insecticides, are coarsely milled and
broken into 4-5 pieces in a milling
machine. The broken grains are heat
sterilized at1000C for 1 hour to
eliminate the residual population of
stored product insect.
After sterilization, the grains are cooled
and sprayed with 0.1% formalin to prevent
the growth of molds as well as to increase
the grain humidity lost during heat
sterilization.
2 kg of broken grain(provide
carbohydrate for developing larvae) are
then of transferred to plastic or Corcyra
cages along with 100 gm of roasted
groundnut powder(supply protein and
fat for developing larvae) and 5gm
yeast(regulate larval growth).
All the above gradients are properly
mixed and Corcyra eggs are sprinkled on
top of the mixture. The cages are tightly
closed with the lid.When plastic basins are
used, they are, covered with cloth and tied
with elastic band/rubber band and special
precautions are taken to protection from
Bracon sp infestation.
A total of 200 boxes/plastic or
metallic basins are charged with
rearing medium and Corcyra Eggs
in the first instance in march.
Similar sets of 500 boxes are
charged thrice at 45 days interval
in May, June and August.
MASS PRODUCTIO TECHNIQUES
FOR TRICHOGRAMMA
In india,about 26 Trichogramma
spp. Are recorded,of which
T.chilonis,T.japonicum and
T.achaeae are mortality factors for
many crops pest.These parasitoids
attack eggs of sugarcane
borers,scirpophaga insertulas;cut
worms,agrotis.
 spp;cotton bollworms,pectiniphora
gossypirlla and Erias spp.maize stem
borer,chilo pertellus etc.
 Angoumois grain moth,sitotroga
cerealella is used as fastitious host fr
mass production of trichogrammatids in
USA,USSR and many European
countries.
 In India,rice grain moth,Corcyra
cephalonica is used as the laboratory
host.
PRODUCTION OF TRICOGRAMMA
The production of Trichogramma involves
the following steps
1. The cards of 15 X 7.5 cm size prepared to
obtain 10/12 equal pieces are used. Name
of the laboratory, releasing instructions,
date of preparation of card and expected
date of parasitoid emergence are printed
on the backside of the cards. Dilute
Acacia gum is used for pasting the eggs.
 2. The egg are pasted on cards. The
extra loose egg laying on the cards can
be collected in Petri plates by tiling and
tapping the card with a finger. The
quantity of eggs laid by Corcyra is
assessed in measuring cylinder
volumetrically; about 16,000 to 18,000.
3. The egg pasted on the cards are
sterilized by exposing the cards under
30 Watt UV tube at a distance of 35 cm
from the source for 10 minutes. UV
sterilized eggs can be stored in the
lower chamber of the refrigerator up to
5 days can be used for parasitization.
4. The host eggs can also be made
nonviable by exposing the to very low
temp. of 0-20C in the freezer chamber
of a refrigerator for 3-4 hrs adopting the
freezing method the quality of the host
eggs is affected due to shrinkage.
5. The eggs on cards are exposed to adult
Trichogramma in the ratio of 8 :1 for 24
hours at 28+20C in transparent plastic
container. In case the cards are exposed in
polythene bags the host egg to parasitized
egg ratio should be 30:1but in this method
the females are allowed to parasitize till
they die.
Two trichocards can be accommodated in
each polythene bag for parasitization.
 6. Parasitized eggs start turning black
on the 3day after parasitization and the
blacking is the life cycle of
Trichogramma is completed in 7-8 days
whereas in the case of Trichogramma it
is completed in 8-9days.
7.The eggs when turned black can be
stored in the refrigerator or can be
transported for field releases. The
emergence of adults generally takes
places after 3-6 days after blacking of
the eggs.
 Parasitized egg cards showing blackening of
eggs can be stored in a refrigerator at 12-150C
for 10-15 days, the best stage for storage is
pupal stage.
 When eggs turn black. However, during the
storage the quality of emerging adults is
affected. Prolonged storage beyond 15 day
would impair emergence as well as longevity
and fecundity of the resulting progeny.
PRECAUTION
1)Emergence date should be specified on
the cards to guide the user.
 2)The cards should be stapled on the inner
side of the leaves to avoid direct sunlight.
 3)The cards should be stapled in morning
hours and just before emergence to avoid
predation.
 4)Avoid application of insecticides in the
field where Trichogramma are released.if
need arises uses selective/safer
insecticides.
Ensure that insecticides are used 15
days after or before Trichogramma
release.
DOSES EGGS/ha. NO.OF
RELEASES
Cotton 1,50,000 06
Sugarcane 50,000 10
Maize 75,000 06
Tomato 50,000 06
Paddy 50,000 06
FIELD USE-FREQUENCY OF RELEASES
 Early shoot borer of sugarcane,chilo
infuscatalus-Trichogramma chilonis,4-6
releases@50,000/ha at 10 days interval
starting from 45th day after planting or
with appearance of the pest.
 Cotton bollworms,Helicoverpa
armigera;pectinophora gossypiella; Erias
spp.-T.Chilonis@1,50,000 eggs/ha from
45th day onwards,6weekly releases or
with the appearance of the pest.Moths
are to be monitored by pheromone
trapes.
Among viruses of the group
baculoviridae,nuclear
polyhedrosis virus is utilized for
the successful of various insect
pest.
 NPVs are obligating pathogens they
need their specific live hosts for
multiplication.so production if
viruses for use as insecticides
needs mass production of their
hosts as a first step.
Basic steps in the production of
NPVs of any insect are-
1.Mass production of/culturing of
host insects
2.Host inoculation with viruses
3.Harvesting of viruses
4.Purification
5.Storage
1. Host insects viz. Helicoverpa armigera and
Spodoptera litura can be reared either on their
natural host plants(foliage, pod, fruits etc.) or on
artificial diet.
 2. Since natural host plants can not be found
throughout the year, maintenance of host insects
on artificial diet has the advantages 0f rearing
under sterile conditions, avoiding contamination,
saving space, time and labor. Thus the use of
artificial diet for mass production of host insects
is economical and easy.
3. Mass culturing of host insects can be
started either from field collected adults
using light traps or from field collected
larvae. The male and female adults of
insects that are allowed in a oviposition jar
for mating should be provided 1% honey as
food in soaked cotton swabs.
 4. Egg laid are collected daily and inner
surface be sterilized by using 0.15% sodium
hypo chloride. Eggs so collected should be
incubated in Petri dishes.
5. Rearing of newly hatched H.armigera
larvae is done by transferring
individually(since larvae are cannibalistic)
into rearing vials containing artificial diet.
6. In case of S.litura since the eggs are laid
in masses, the larval stages are not
cannibalistic; first two instars can be reared
in groups on castor leaves.
7. Gram flour, yeast tablets, methyl
parahydroxy benzoate and ascorbic acid are
added to half the quantity of in a blender and
mixed for 2-3 minutes. Simultaneously, Agar-
Agar is boiled with the remaining quantity of
water and cooled down to 700C.
 Hot Agar- Agar liquid is added to the
blender and mixed with other ingredients.
Finally multivitaplax, vitamin E,Ascorbic acid
and formalin are added and blended for
about 2 minutes.
8. Hot liquid diet is dispended into rearing
vials(approx. 20 ml vials) and allowed for
solidification at room temp. for about 20
minutes, Single larva should be introduced
in each vial on the diet surface and closed
with cotton plug.
9. While 20% of the larval population can be
allowed to pupate(can be collected after 20
days) for the continuous maintenance of the
host insect culture, the rest can be used for
virus production.
o Infection 8 to 9 days larvae are
introduced into vials inoculating the
surface of the diet poured in plastic
cues(as against the vials used for
healthy host insect culture) with a virus.
o Dose of 1.1 X104 polyhedral inclusion
bodies(PIB) per larva. Larvae are placed
individually on virus contaminated diet
cups and capped.
Observation on larval mortality
is made daily and the dead
larvae with viral symptoms
(cuticle of larvae becomes
fragile and ruptures easily when
touched;body colour changes
to blue or bluish purple)are
harvested and placed in conical
flask.
The disesed larvae are macerated in
a mixer using sterile distilled
water.The contents are allowed as
such in the conical flask for several
days at room temperature,during
which time the polyhedra settle
down as a white layer.
Alternatively macerated contents
can be sieved through a double
layer of muslin cloth and the filtrate
is standardized for the polyhedral
inclusion body/ml and packed in
white plastic cans for field use.
Ingredients Quantity
Kabuli gram flour 105 g
Sorbic acid 1g
Methyl parahydroxybenzoate 2 g
Ascorbic acid 3.25 g
Yeast tablets 10 g
Water 780 g
Agar-Agar 12.75 g
Formalin(10%) 2ml
Vitamin 2 capsules
Multivitaplex 2 capsules
Streptomycin sulphate 25 g
FIELD DOSE
 250 LE per hactare.
 One LE=6X109
 Cotton:-H.armigera-HaNPV-450LE/HA
with 5%jaggery and 0.1%Ranipal at ETL 7
ll instar larvae/20 plants.
 Chickpea:-H.armigera-HaNPV-250LE/ha
directed on the inflorescence,one or two
well timed applications to protect seed
crop.
Pigeon pea:-H.armigera-HaNPV-250
LE/ha,3-4 sprays required.
Crusiferous vegetables:-S.litura
SINPV-250-450 LE/ha;two-three
sprays at 7-10 days interval;virus in
200-400 lit.of water +1%crude sugar.
THANK YOU

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MASS MULTIPLICATION OF Corcyra cephalonia PPT

  • 1. DEPARTMENT OF ENTOMOLOGY ALLAHABAD SCHOOL OF AGRICULTURE SAM HIGGINBOTTOM INSTITUTE OF AGRICULTURE TECHNOLOGY & SCIENCES [Formerly-Allahabad Agricultural Institute] (Deemed-to-be-University) ALLAHABAD- 211007, U.P., INDIA BY, RAKESH KR. MEENA
  • 3. The egg of Sitotroga cerealella are generally used as laboratory host for production of Trichogramma in the USA, France, Germany etc. Whereas in India ,rice grain moth,Corcyra cephalonica is used as the laboratory host.
  • 4. STEPS IN CORCYRA PRODUCTION: The quantities of sorghum/pearl millet/maize grain, free from insecticides, are coarsely milled and broken into 4-5 pieces in a milling machine. The broken grains are heat sterilized at1000C for 1 hour to eliminate the residual population of stored product insect.
  • 5. After sterilization, the grains are cooled and sprayed with 0.1% formalin to prevent the growth of molds as well as to increase the grain humidity lost during heat sterilization. 2 kg of broken grain(provide carbohydrate for developing larvae) are then of transferred to plastic or Corcyra cages along with 100 gm of roasted groundnut powder(supply protein and fat for developing larvae) and 5gm yeast(regulate larval growth).
  • 6. All the above gradients are properly mixed and Corcyra eggs are sprinkled on top of the mixture. The cages are tightly closed with the lid.When plastic basins are used, they are, covered with cloth and tied with elastic band/rubber band and special precautions are taken to protection from Bracon sp infestation.
  • 7. A total of 200 boxes/plastic or metallic basins are charged with rearing medium and Corcyra Eggs in the first instance in march. Similar sets of 500 boxes are charged thrice at 45 days interval in May, June and August.
  • 8. MASS PRODUCTIO TECHNIQUES FOR TRICHOGRAMMA In india,about 26 Trichogramma spp. Are recorded,of which T.chilonis,T.japonicum and T.achaeae are mortality factors for many crops pest.These parasitoids attack eggs of sugarcane borers,scirpophaga insertulas;cut worms,agrotis.
  • 9.  spp;cotton bollworms,pectiniphora gossypirlla and Erias spp.maize stem borer,chilo pertellus etc.  Angoumois grain moth,sitotroga cerealella is used as fastitious host fr mass production of trichogrammatids in USA,USSR and many European countries.  In India,rice grain moth,Corcyra cephalonica is used as the laboratory host.
  • 10. PRODUCTION OF TRICOGRAMMA The production of Trichogramma involves the following steps 1. The cards of 15 X 7.5 cm size prepared to obtain 10/12 equal pieces are used. Name of the laboratory, releasing instructions, date of preparation of card and expected date of parasitoid emergence are printed on the backside of the cards. Dilute Acacia gum is used for pasting the eggs.
  • 11.
  • 12.  2. The egg are pasted on cards. The extra loose egg laying on the cards can be collected in Petri plates by tiling and tapping the card with a finger. The quantity of eggs laid by Corcyra is assessed in measuring cylinder volumetrically; about 16,000 to 18,000.
  • 13. 3. The egg pasted on the cards are sterilized by exposing the cards under 30 Watt UV tube at a distance of 35 cm from the source for 10 minutes. UV sterilized eggs can be stored in the lower chamber of the refrigerator up to 5 days can be used for parasitization.
  • 14. 4. The host eggs can also be made nonviable by exposing the to very low temp. of 0-20C in the freezer chamber of a refrigerator for 3-4 hrs adopting the freezing method the quality of the host eggs is affected due to shrinkage.
  • 15. 5. The eggs on cards are exposed to adult Trichogramma in the ratio of 8 :1 for 24 hours at 28+20C in transparent plastic container. In case the cards are exposed in polythene bags the host egg to parasitized egg ratio should be 30:1but in this method the females are allowed to parasitize till they die. Two trichocards can be accommodated in each polythene bag for parasitization.
  • 16.  6. Parasitized eggs start turning black on the 3day after parasitization and the blacking is the life cycle of Trichogramma is completed in 7-8 days whereas in the case of Trichogramma it is completed in 8-9days.
  • 17. 7.The eggs when turned black can be stored in the refrigerator or can be transported for field releases. The emergence of adults generally takes places after 3-6 days after blacking of the eggs.
  • 18.  Parasitized egg cards showing blackening of eggs can be stored in a refrigerator at 12-150C for 10-15 days, the best stage for storage is pupal stage.  When eggs turn black. However, during the storage the quality of emerging adults is affected. Prolonged storage beyond 15 day would impair emergence as well as longevity and fecundity of the resulting progeny.
  • 19.
  • 20. PRECAUTION 1)Emergence date should be specified on the cards to guide the user.  2)The cards should be stapled on the inner side of the leaves to avoid direct sunlight.  3)The cards should be stapled in morning hours and just before emergence to avoid predation.  4)Avoid application of insecticides in the field where Trichogramma are released.if need arises uses selective/safer insecticides.
  • 21. Ensure that insecticides are used 15 days after or before Trichogramma release. DOSES EGGS/ha. NO.OF RELEASES Cotton 1,50,000 06 Sugarcane 50,000 10 Maize 75,000 06 Tomato 50,000 06 Paddy 50,000 06
  • 22. FIELD USE-FREQUENCY OF RELEASES  Early shoot borer of sugarcane,chilo infuscatalus-Trichogramma chilonis,4-6 releases@50,000/ha at 10 days interval starting from 45th day after planting or with appearance of the pest.  Cotton bollworms,Helicoverpa armigera;pectinophora gossypiella; Erias spp.-T.Chilonis@1,50,000 eggs/ha from 45th day onwards,6weekly releases or with the appearance of the pest.Moths are to be monitored by pheromone trapes.
  • 23. Among viruses of the group baculoviridae,nuclear polyhedrosis virus is utilized for the successful of various insect pest.  NPVs are obligating pathogens they need their specific live hosts for multiplication.so production if viruses for use as insecticides needs mass production of their hosts as a first step.
  • 24. Basic steps in the production of NPVs of any insect are- 1.Mass production of/culturing of host insects 2.Host inoculation with viruses 3.Harvesting of viruses 4.Purification 5.Storage
  • 25.
  • 26. 1. Host insects viz. Helicoverpa armigera and Spodoptera litura can be reared either on their natural host plants(foliage, pod, fruits etc.) or on artificial diet.  2. Since natural host plants can not be found throughout the year, maintenance of host insects on artificial diet has the advantages 0f rearing under sterile conditions, avoiding contamination, saving space, time and labor. Thus the use of artificial diet for mass production of host insects is economical and easy.
  • 27. 3. Mass culturing of host insects can be started either from field collected adults using light traps or from field collected larvae. The male and female adults of insects that are allowed in a oviposition jar for mating should be provided 1% honey as food in soaked cotton swabs.  4. Egg laid are collected daily and inner surface be sterilized by using 0.15% sodium hypo chloride. Eggs so collected should be incubated in Petri dishes.
  • 28. 5. Rearing of newly hatched H.armigera larvae is done by transferring individually(since larvae are cannibalistic) into rearing vials containing artificial diet. 6. In case of S.litura since the eggs are laid in masses, the larval stages are not cannibalistic; first two instars can be reared in groups on castor leaves.
  • 29. 7. Gram flour, yeast tablets, methyl parahydroxy benzoate and ascorbic acid are added to half the quantity of in a blender and mixed for 2-3 minutes. Simultaneously, Agar- Agar is boiled with the remaining quantity of water and cooled down to 700C.  Hot Agar- Agar liquid is added to the blender and mixed with other ingredients. Finally multivitaplax, vitamin E,Ascorbic acid and formalin are added and blended for about 2 minutes.
  • 30. 8. Hot liquid diet is dispended into rearing vials(approx. 20 ml vials) and allowed for solidification at room temp. for about 20 minutes, Single larva should be introduced in each vial on the diet surface and closed with cotton plug. 9. While 20% of the larval population can be allowed to pupate(can be collected after 20 days) for the continuous maintenance of the host insect culture, the rest can be used for virus production.
  • 31. o Infection 8 to 9 days larvae are introduced into vials inoculating the surface of the diet poured in plastic cues(as against the vials used for healthy host insect culture) with a virus. o Dose of 1.1 X104 polyhedral inclusion bodies(PIB) per larva. Larvae are placed individually on virus contaminated diet cups and capped.
  • 32. Observation on larval mortality is made daily and the dead larvae with viral symptoms (cuticle of larvae becomes fragile and ruptures easily when touched;body colour changes to blue or bluish purple)are harvested and placed in conical flask.
  • 33. The disesed larvae are macerated in a mixer using sterile distilled water.The contents are allowed as such in the conical flask for several days at room temperature,during which time the polyhedra settle down as a white layer.
  • 34. Alternatively macerated contents can be sieved through a double layer of muslin cloth and the filtrate is standardized for the polyhedral inclusion body/ml and packed in white plastic cans for field use.
  • 35. Ingredients Quantity Kabuli gram flour 105 g Sorbic acid 1g Methyl parahydroxybenzoate 2 g Ascorbic acid 3.25 g Yeast tablets 10 g Water 780 g Agar-Agar 12.75 g Formalin(10%) 2ml Vitamin 2 capsules Multivitaplex 2 capsules Streptomycin sulphate 25 g
  • 36. FIELD DOSE  250 LE per hactare.  One LE=6X109  Cotton:-H.armigera-HaNPV-450LE/HA with 5%jaggery and 0.1%Ranipal at ETL 7 ll instar larvae/20 plants.  Chickpea:-H.armigera-HaNPV-250LE/ha directed on the inflorescence,one or two well timed applications to protect seed crop.
  • 37. Pigeon pea:-H.armigera-HaNPV-250 LE/ha,3-4 sprays required. Crusiferous vegetables:-S.litura SINPV-250-450 LE/ha;two-three sprays at 7-10 days interval;virus in 200-400 lit.of water +1%crude sugar.