DEPARTMENT OF ENTOMOLOGYALLAHABAD SCHOOL OF AGRICULTURE

SAM HIGGINBOTTOM INSTITUTE OF AGRICULTURE TECHNOLOGY & SCIENCES

[Formerly-Allahabad Agricultural Institute](Deemed-to-be-University)

ALLAHABAD- 211007, U.P., INDIA 

PRESENTATION ON

MASS MULTIPLICATION OF CORCYRA CEPHALONIA ”

BY,RAKESH KR.

MEENA

MASS MULTIPLICATION OF CORCYRA CEPHALONIA

PRODUCTION OF HOSTThe 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:

1). 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.

2). 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.

3). 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).

4). 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.

5). 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 steps1. 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.

STORAGE

PRECAUTION1)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 10Maize 75,000 06Tomato 50,000 06Paddy 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.

MASS PRODUCTION TECHNIIQUE OF VIRUSES

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 viruses4.Purification5.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.

Mass culture of host insects

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.

MASS PRODUCTION OF VIRUS

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 QuantityKabuli gram flour 105 gSorbic acid 1gMethyl parahydroxybenzoate 2 gAscorbic acid 3.25 gYeast tablets 10 gWater 780 gAgar-Agar 12.75 gFormalin(10%) 2mlVitamin 2 capsulesMultivitaplex 2 capsulesStreptomycin 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.

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