How Mass production is done for microbial insecticides?

Mass production of Entomopthaogenic fungi - Biopesticide

REQUIREMENTS

1. Two rat proof rooms of approximately 10 x 10 ft
2. Work table with hood
3. UV lamp and fluorescent lamp
4. Auto clave Stove
5. Heat sealer
6. Veterinary syringe 20 ml capacity
7. Burner and lighter

Materials needed

1. Par boiled rice as per requirement
2. Poly propylene bags of size 14" x 10"
3. Sponge pieces rolled into small plugs
4. Disinfectant - Sodium Hypochlorite solution
5. Cellophane tape and insulated binding wire
6. Pure liquid culture of B. bassiana
7. Cotton/tissue paper rolls
8. Surfactant (tween 80)
9. Antibiotic (chloramphenicol / streptomycin / tetracycline)
10. Workers - 2

METHODOLOGY

1. If polythene bags not properly sealed, re-seal the bottom using the heat sealer. Soak rice in water for about half an hour, drain and fill in the bags at the rate of one kg equally distributed in five bags.
2. Close the mouth of the bag with a sponge plug and fasten tightly with a piece of insulated binding wire.
3. Hold the plugs of five bags together and cover with a piece of paper.
4. Place about 15 bags in the Autoclave containing some quantity of water. Close the lid and when the steam starts issuing continuously, put the weight. After three whistles reduce the flame and allow heating for 15 minutes. After this interval, put off the stove and allow the cooker to cool.
5. While pressure cooking is going on, clean the work table using 5% sodium hypochlorite solution and switch on the UV lamp. Avoid looking at the lit lamp and direct contact of the body with UV light.
6. Take out the bags and remove water adhering to the sides with blotting/tissue paper placed in a clean tray.
7. Switch off the UV lamp, place the bags on the work table and allow further cooling.
8. Light the spirit lamp/burner and heat sterilize the syringe and allow cooling. The syringe can also be sterilized in the cooker along with the rice bags.
9. Holding the liquid culture bottle above the burner flame, remove the plug, add two drops of the surfactant and 10 drops of antibiotic and shake well.
10. Draw 15 ml of the liquid culture into the syringe and inject into the bag through the sponge plug. Care should be taken to avoid puncturing the bag. Mix well, spread the rice grains loosely and place the bags on racks.
11. Incubate for two weeks and watch for development of spores. Once profuse sporulation is seen, gently crush the lumps into individual grains, cut open the bags along the sides, spread the culture in the same bag and allow drying for four or five days.
12. Separate the spores by sieving, allow drying on the work bench for about three days. For faster drying, silica gel may be used. A good culture with one kg of rice yields about 40grams of dry conidia in two or three sieving at weekly intervals. Sieving may be done under the hood to avoid inhalation of spores.