1. MASS PRODUCTION OF BIOAGENTS & BIOPESTICIDES Balram Solunke balram2424@gmail.co

2. CONTENT: Mass production of.... Trichoderma viride Corcyra cephalonica cryptolaemus montrouzieri Trichogramma chilonis Zygogramma bicolarata Nuclear polyhydrosis virus of Helicoverpa armigera Nuclear polyhydrosis virus of Spodoptera litura 3. Mass production of Trichoderma viride Materials : 1)Potato 2)Dextrose 3)Glass wares 4)Spirit lamp 5)Pure culture of fungus 6)Laminar air flow 7)Ethyl alcohol 8)Cotton plug, rubber band,alluminium foil etc. 4. Inoculation: 5. MIXING OF CULTURE 6. mix well with talcum powder for final product of Trichoderma viride 7. mix well with talcum powder for final product of Trichoderma viride 8. Packing: 9. Prepare POB broth media Cooking of 200gm potato for 1 lit media than extract taken & add water &mix 20gm dextrose Than make final volume 1 lit Procedure 10. Fill media in 1 lit conical flask & plugged with aluminum foil than tied with rubber band Sterilize media & requiered glasswares like test tube petriplates & inoculation bottles for 15 min in 1210c in autoclave Kept these for cooling 11. Sterile LAF with ethyl alcohol than transfer glass wares & media in LAF Pour media in bottle , Petridis Inoculate pure culture of trchoderma viridae with the help of inoculating needle using spirit lamp to avoid contamination 12. Collect the fungus growth & crush these culture with the help of blander mix these with the help of blander After that mix well with talcum powder for final product of Trichoderma viridae Drying of product at room temp for 1 day 13. Mix CMC (carbon methyl cellulose) as sticky agent in dried powdery form Packing product in polythene bags & labeling 14. Mass production of Corcyra cephalonica Materials : 1) One rearing room of 15x12x8 cm size with a.c installation 2) Wooden tray of 45x30x15 cm size 3) Coarsely ground grains of sorghum/bajra 4) Yeast 5) Eggs of corcera 15. Procedure sorghum / bajra Grains should be milled coarsely to make 3-4 pieces of each grains Heat/sterilized in an oven at 1000 c for 30 min 16. Spray 0.1% formalin solution uniformly & mix well Dry the grain with fan air Pour 5 kg coarsely ground grains of jawar or bajra in each wooden box & add 10gm yeast powder + 200gm g.nut 17. Obtain pure culture of corcyra cephalonica Mix 1cc 18000 to 20000 egg in each box Cover box for 30 days approximately 18. Maintain temperature of rearing room at 300c After 20-40 days on alternate day collect adults Put 1000 adults oviposition cage 19. Collect the deposited eggs, clean from scales & debris Store for 3-4 days in refrigerator at 100c it requiered,these eggs treat with UV rays for 45 min. 20. Mass production of cryptolaemus montrouzieri a predator on mealy bug 21. REARING OF MEALY BUG ON POTATO : 22. Fill wooden/plastic trays with sandy silt soil up to 2 to 3 cm deapth for planting Place 10-15 potato tubers & cover with moist soil Fill such tray frequently 1) REARING OF MEALY BUG ON POTATO : 23. Watering such trays Put ovisacks of mealy bugs over potato sprouts With in 20-25 days mass culture of mealy bugs could be obtained such trays Maintain the temperature in rearing room between 20 to 300c 24. REARING OF MEALY BUG ON RED PUMPKINS : 25. 1) REARING OF MEALY BUG ON RED PUMPKINS : Medium sized with ridges & furrows & grooves with small stack & pumpkin should be selected for the mealy bug rearing Clean with water to remove dust particles 26. The wound if any pumpkin plug with paraffin wax Than tied the whole pumpkine with thread for griftting the mealy bug on it Treat 0.1% bavistine solution (1gm/lit) to prevent its rotting during rearing process 27. Collected mealy bug release on pumpkin Cages placed on working tables/racks in tearing room Keep the pumpkin in wooden cage 28. 1) REARING OF PREDATOR cryptolaemus montrouzieri : When mealy bugs are 8-10 days old a stock of 15-20females of cryptolaemus montrouzieri at temperature of 20-300c The released female feed on mealy bugs eggs , numph & adults 29. On hatching larvae feed on mealy bug Female deposit eggs on pumpkins Put fully developed larvae with mealy bugs on as food plastic jar & provide paper strips for its pupation The emerging beetles collect in plastic bowl & feed with honey agar or mealy bugs 30. REARING OF PREDATOR cryptolaemus montrouzieri 31. MASS PRODUCTION TECHNIQUES OF TRICHOGRAMMA CHILONIS 32. Procure eggs of corcyra cephalonica Paste 1cc (18000 to 20000) egg on thinly smeared gum arabic on egg card with the help of suitable strain Expose the eggs to UV rays for 45 min On backside of the trichocards written down the mentioned information 33. Transfer 6 days old daily parasitoid egg card with 1:6 ratio (parasite : host) for 24 hrs. After 4 days of parasitization brush out host larvae of day hatched Such parasitized eggs cards could be stored for about 20 to 30 days at 100c temperature in refrigerator 34. PREPARATION OF TRICHOCARD : 35. MASS PRODUCTION TECHNIQUES OF Zygogramma bicolarata 36. Take parthanium leaves in transparent plastic container Release 10 pairs of beetles female& male on such leaves Egg laying observed in leaves replaced & fresh bouquets are provided 37. these process repeated for month After few days eggs collect in plastic small bottle & kept leaves of parthanium in bottle Emergence of larvae & feed on leaves & pupa in soil 38. 15-20 small plants transplanted in aluminum cage these cages yield about 100-125 adults 39. MASS PRODUCTION TECHNIQUES OF NUCLEAR POLYHYDROSIS VIRUS OF HELICOVERPA ARMIGERA 40. FIELD COLLECTION OF HELICOVERPA ARMIGERA FROM TUR FIELD : 41. PREPARATION OF ARTIFICIAL DIET : 42. MATING OF HELICOVERPA ARMIGERA MOTH IN GLASS CHMBER : 43. PUPATION BOX : 44. Select late third instar or early 4th instar larvae of HELICOVERPA ARMIGERA for virus inoculation Pre- strave these larvae for 6-8 hrs. before virus inoculation Introduce piece of the virus contaminated diet in each container with pre-starved larvae 45. The larvae begin to show symptoms of virus infections after about 4-5 days Collect the dead diseased larvae in distilled water Replace uneaten part of diet after 2-3 days & clean the containers 46. Pullet staved at the bottom may be discard & centrifuge at 2500 rpm for 10 min Supernatant is discard & pellet mix in water 10 times in purified virus Filter this suspension through double layers of muslin cloth & centrifuge for 3 to 5 min at 500 rpm 47. MASS PRODUCTION TECHNIQUES OF NUCLEAR POLYHYDROSIS VIRUS OF SPODOPTERA LITURA 48. Collection of SPODOPTERA LITURA from cabbage field : 49. Prepare NPV suspension in water for leaf treatment Pluck castor leaves with the petiole & dip in the virus solution Dry treated leaf under shade Starve larvae for 4 to 5 hrs before feeding 50. Keep the news paper at the bottom of bucket & provide the trated leaf for feeding cover mouth of bucket with cloth Everyday remove the excreta of larvae Provide untrated leaf for feeding 51. Larvae will start dying after 4-5 days Collect the dead larvae in distilled water & grind the larvae thoroughly stir the solution & filter it by cloth Collect filtrate store in amber coloured bottles in colol place 52. Thank You