. : Coffee Board of India - PRODUCTION, STORAGE, FORMULATION AND APPLICATION OF BEAUVERIA BASSIA...

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PRODUCTION, STORAGE, FORMULATION AND APPLICATION OF BEAUVERIA BASSIANA FOR THE MANAGEMENT OF COFFEE BERRY BORER
The white muscardine fungus Beauveria bassiana is a potential bio-control agent that could be used against the berry borer very effectively. This pathogen, which kills the borer can be cultured at the estate level and sprayed on the infested plants. B. bassiana, the white muscardine fungus, is a common pathogen of a range of insects belonging to different groups. This fungus often exerts a good degree of natural biological control under humid conditions. Under conditions not so favourable for natural epizootics, widespread artificial infection can be induced by application of the fungus (as spores or conidia), resulting in good pest suppression. One of the major constraints hindering the use of this fungus as a bio-pesticide is the non availability of cost effective and genuine formulations in the market. However, this fungus can be produced by farmers with some basic facilities available on the farms. REQUIREMENTS Capital 1. 2. 3. 4. 5. 6. 7. 8. Two rat proof rooms of approximately 10 x 10 ft Work table with hood UV lamp and fluorescent lamp Pressure cooker of 20 litre capacity Stove Heat sealer Veterinary syringe 20 ml capacity Burner and lighter

Recurring 1. 2. 3. 4. Par boiled rice as per requirement Poly propylene bags of size 14 x 10 Sponge pieces rolled into small plugs Disinfectant Sodium Hypochlorite solution

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. : Coffee Board of India - PRODUCTION, STORAGE, FORMULATION AND APPLICATION OF BEAUVERIA BASSIA...

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5. 6. 7. 8. 9. 10.

Cellophane tape and insulated binding wire Pure liquid culture of B. bassiana Cotton/tissue paper rolls Surfactant (APSA 80 or ACTIVE 80) Antibiotic (chloramphenicol / streptomycin / tetracycline) Workers - 2

METHODOLOGY 1. 2. 3. 4. 5. If not properly sealed, re-seal the bottom of the polypropylene /polyethylene bags 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. Close the mouth of the bag with a sponge plug and fasten tightly with a piece of insulated binding wire. Hold the plugs of five bags together and cover with a piece of paper. Place about 15 bags in the pressure cooker 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. 6. 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. 7. 8. 9. 10. 11. 12. 13. Take out the bags and remove water adhering to the sides with blotting/tissue paper placed in a clean tray. Switch off the UV lamp, place the bags on the work table and allow further cooling. 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. 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. 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. 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. 14. Separate the spores by sieving, allow drying on the work bench for about three days.

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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. 15. Pack the dry spores in laminated aluminium foil and store in the freezer compartment of a refrigerator. Formulation and application To prepare one barrel (200L) of spray fluid with about 10 million conidia per ml (the optimum dose for field application), take the conidia extracted from one kilo of rice in a polypropylene bag. Add 100 ml of any surfactant like APSA80 or ACTIVE 80, and mix thoroughly to form a uniform paste (the conidia are not miscible with most of the ordinary wetting agents and hence they cannot be used). Add small quantities of water into the bag and mix thoroughly so that no lumps remain in the suspension. Make up to 200 litres. If the culture bags are used for immediate field application, the following method may be adopted. Crush the contents of five bags (prepared with one kg of rice) and empty into a container with a screw cap. Add 100 ml of APSA 80/ACTIVE 80 and about 2 litres of water. Close the container tightly and mix thoroughly by vigorous agitation (a churner may be used for better mixing). Filter the suspension into the barrel. Repeat mixing and filtration, three or four times, to extract maximum spores. Make up to 200 litres. Unlike chemical insecticides, B. bassiana has got very little persistence on the treated plant surface. Hence, for target pests like berry borer or shot hole borer, direct contact of the formulation with the insect cuticle is essential for effective control. To ensure proper coverage of berries or twigs, it is better to use a knapsack or rocker sprayer fitted with solid cone or adjustable nozzle of 350 to 450 cc output per minute. If suitable lances and nozzles are available, motorized sprayers can also be used. The effect of the fungus can be observed in the field in about 10 days with the development of white hyphal mass or spores on the dead insects, often projecting through the bore holes on the berries. Around 80% suppression can be achieved. Apart from environmental safety, the advantage of the fungus over insecticides is its ability to kill the stages inside the tunnels through cross contamination. Hence, field application can be taken up till berry ripening starts. June to September, when temperature is below 30o C and humidity above 70%, is the ideal period for best results. Mist as well as the microclimate in the tunnels also provides high

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humidity required for fungal development. The important precaution to be followed while handling the fungus is to avoid inhalation of spores; some people may be allergic. Training on production and use of B. bassiana is being imparted at the Central Coffee Research Institute at Balehonnur and the Regional Coffee Research Stations at Chettali in Kodagu, Chundale in Wayanad and Thandigudi in Tamil Nadu. Individual growers or groups interested in production of this fungus can source the pure culture from the Coffee Research Stations. Since there is a chance of contamination at several points of the production chain, utmost care needs to be taken to keep the conditions hygienic

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