TRICHOGRAMMA.pptx

Production technology of bio agents

1. Trichogramma Production Technology for Bio-agents and Bio-fertilizers ENT-422 10(0+10) Submitted To:- Dr. Kanchan G. Padwal Assistant Professor(Entomology) Institute of Agricultural Sciences,BHU

2. What is Biocontrol ? Biological control  or  biocontrol  is a method of  controlling pests  such as  insects ,  mites ,  weeds  and  plant diseases   using other organisms . There are three basic strategies for biological pest control: Classical (importation) – Where a natural enemy of a pest is introduced in the hope of achieving control . Inundative (augmentation) – In which a large population of natural enemies are administered for quick pest control. Inoculative (conservation) – In which measures are taken to maintain natural enemies through regular re-establishment .

3. TRICHOGRAMMA Trichogramma is a genus of minute polyphagous wasps that are endoparasitoids of insect eggs. Trichogramma is genera from the family Trichogrammatidae , with over 200 species worldwide. Trichogramma wasps are small and very uniform in structure, which causes difficulty in identifying the separate species. As females are all relatively similar, taxonomists rely upon examination of males to tell the different species apart, using features of their antennae and genitalia . Trichogramma spp. have been used for control of lepidopteran pests for many years. They can be considered the Drosophila of the parasitoid world, as they have been used for inundative releases Play important role in pest suppression programme by destroying the early stage of the pest.

4. More than 150 species of Trichogrammatids are distributed throughout the world parasitizing eggs of over 200 insect species. In India, 26 Trichogrammatids are recorded; out of which, 1. Trichogramma chilonis (Ishii) 2. T. japonicum (Ashmead) and 3. T. acheae 4. T. brasiliensis are of significant importance. Worldwide over 32 million ha area under agricultural crops and forest are treated annually with different Trichogramma species in 19 countries. Mostly in China and Republic of Soviet Union (Li, 1994). Their levels of parasitism sometimes reach to 100 percent depending upon the environmental conditions.

5. CLASSIFICATION Domain: Eukaryota Kingdom: Metazoa Phylum: Arthropoda Subphylum: Uniramia Class: Insecta Order: Hymenoptera Family: Trichogrammatidae Genus: Trichogramma

6. Biology and Life Cycle The development of all Trichogramma spp. is very similar . The female wasp inserts its ovipositor through the chorion and deposits its eggs within the egg of the host. It avoids super parasitism or multiple-parasitism under natural conditions. During the 3rd instar, dark melanin granules are deposited on the inner surface of the egg chorion, causing the host egg to turn black .

7. How does Trichogramma actually Parasitise on other host egg Immediately after they hatch, female Trichogramma wasps begin their search for the egg of a 'target species’. Once host egg is found – through pheromones and visual cues.The wasp drills a hole into the egg and deposits two to three eggs of her own. How many of their own eggs to lay, is calculated by timing walks across moth egg surfaces. In a mere 24 hours, the wasps eggs hatch within the host egg and the larvae consume the yolk and embryo. The larvae then undergo three instars before a pupae stage.

8. They emerging about nine days after they were initially laid within the host egg (Knutson 1998). Trichogramma larvae eat out the insides of pest eggs, pupate, and cutan exit hole in moth egg shells for winged adults to squeeze through. Males emerge first, wait for females, and immediately mate. The life cycle from egg to adult is completed in 7 to 10 days (longer in cool weather). Short life cycle -30 generations per season, and rapid population increase. Hence, early season releases produce large populations positioned to fight pest invasions.

9. Trichogramma wasps are used worldwide as highly effective biocontrol agents. Female wasps lay their eggs inside the eggs of a host insect, usually eggs the pest lays on plants, adult wasps emerge7-10 days later. Pest eggs parasitized by trichogramma wasps turn black. No pest larvae will emerge.

10. Corcyra Rearing and mass production Step 1 – Preparation of medium : Heat sterilized broken grains is taken at the rate of 2.5 kg per plastic basin to which groundnut kernel powder is added @ 100 gm /basin and yeast powder @ 5g/basin. To prevent bacterial infection in the food medium Streptomycin sulphate @ 0.05 % spray is given at 10—20 ml/basin using a hand-operated sprayer or atomizer.

11. Step 2 – Inoculation of Corcyra eggs : Nucleus culture of Corcyra eggs were added @ 0.5 CC / basin containing 2.5 kg of grain medium. After uniform mixing of the contents of the basin it was covered with khadi cloth and secured by twine. The young Corcyra larva that hatch out from the egg in 3-4 days feeds on the medium by constructing webs . The adult Corcyra starts emerging from the medium from 30-35 days onwards and continues to emerge up to 90 days after inoculation of the eggs due to staggered development of larvae in the medium.

12. Step 3 – Collection of moths for mating : The emerged adult Corcyra moth rest on the inner surface of the cloth cover. They are to be collected in the morning hours using glass specimen tubes (15x 2.5 cm) or a specially designed modified vacuum aspirator . The moth collection is effectively done by keeping the basin inside a mosquito net so that the moth cannot escape. The adult moths are transferred to a specially designed mating drum made of G.I sheets with wire mesh at the bottom. Adult moths are provided with honey solution (50 %) added with vitamin E (1 capsule per 20 ml of 50% honey) to boost the vigor of the adult and to get higher quantity of healthy eggs. The adult food is given by dipping cotton swab and allowed to hang inside the drum with a thread.

13. Step 4 – Collection of Corcyra eggs : Corcyra eggs are loosely laid and they are collected through the wire mesh at the bottom on a receiving container with funnel setup on an enamel tray. Eggs are to be collected daily for 4 days from each drum. On the fifth day it is to be vacated and cleaned. The eggs are cleaned and separated from the moth’s scales.

14. Step 5 – Cleaning of eggs : The eggs thus obtained are further cleaned with the help of plastic sieves of different meshes. One CC of Corcyra eggs contains approximately 18,000 eggs . About 100 pairs of Corcyra moth (50 % female) produce 1.5 cc of eggs during their egg laying period of 4 days . From each Corcyra rearing basin an average of 2,500 moths emerge.

15. Precautions while rearing Corcyra Bacterial disease some time attacks Corcyra culture. To control this, streptomycin sulphate is added to the crushed sorghum @0.23gm/kg and mixed thoroughly. Occasionally the mite Pyemotes ventricosus (Newport) contaminate the culture and affect egg laying and larval development of Corcyra. in case of infestation the racks, cages, boxes etc. should be disinfected with formalin and placed in the sun for six hours. In case of severe infestation, acaricide dicofol ( Kelthane ) can be applied. For this,muslin sheets are dipped in a 0.05% solution of dicofol and air dried for couple of hours. Control of Bracon hebetor infestation: A plastic tub filled with water near racks containing Corcyra boxes and keeps a table lamp with 60 or100 W bulb. The light of the table lamp should face the surface of water

16. The trap should be operated daily and side by side the culture should also be replaced with fresh stock. The boxes should be sterilized by spraying 1% formaldehyde or sodium hypochlorite solution and sun dried for 6 hours. Care should be taken to remove host/parasitoid cocoons from boxes. The feed material jowar should also be sterilized. Mix all the ingredients in separate room and charge with Corcyra eggs. Close the lid and these sterilized boxes should not be open for 40 days and kept in separate room. By this method, Bracon can be eliminated from the culture. In case of labs having separate Corcyra rearing rooms, on one corner disinfect all boxes one by one and keep them in one room. All the Bracons in previous room can be killed by spraying low concentration of any pesticide. All the windows and doors can be kept open for 15 days. .

17. Thereafter, sprinkle whole room with 1% formaldehyde (40%) solution. After about 10 days bring back whole culture to same room. Thereafter, regularly keep light-water trap to monitor Bracon . If we found discoloured larvae and white cocoons, then they are collected and immersed in 0.5% formaldehyde to kill the parasitoids. Provide windows with fine iron mesh, wire mesh and double door to avoid any entry of parasitoids Scales of laboratory host C. cephalonica may cause the respiratory problems; it is advised to wear a mask while rearing C. cephalonica . Trichocard should be properly sealed before keeping in refrigerator to avoid any type of accident.

18. Trichocard preparation Materials required Corcyra eggs, Nucleus culture of Trichogramma , Polythene bags, Rubber bands, Scissors, Gum, Brush, Mesh sieve, Tricho cards, 50% honey solution, Stapler, Refrigerator/fridge and UV lamp/LED light.

19. Preparation of Trichocard Step 1 – Preparation of “ Tricho cards ”: The Corcyra eggs collected from the mating drum are cleaned and finally taken in glass petri dishes (15-20 cm dia ). The eggs were sterilized by exposing to UV light (15 W) in a closed chamber for ½ hr. duration so that the embryo may be killed without damaging the egg contents. The UV sterilized eggs were sprinkled on drawing board cards (30 x 20 cm) smeared with thin layer of diluted gum , at 6.0 CC of egg/ card. The card was already drawn with lines to separate 30 rectangles (7 x 2 cm) to accommodate 6.0 CC eggs .

20. Step 2 – Inoculation of parasitoids : The cards pasted with Corcyra eggs were dried under fan and taken into polythene bags (45x 30 cm) containing nucleus parasitized cards at 6: 1 (fresh eggs to parasitized eggs) for exposure. The parasitoids emerging from the nucleus parasitoid eggs start parasitizing the fresh Corcyra eggs. After 2 days exposure the cards were kept for another 1-2 days , during which the parasitized cards were then stored in refrigeration at 10 C for 21 days. The parasitoids under normal room temperature emerge in 7 days where as they emerge in 2 days after removing from storage for field use.

21. Field release : Trichocards can be stapled inside plastic tea cup and the plastic tea cup can be tied to the leaf or shoot of the crop in an inverted manner to safeguard the eggs from adverse climatic conditions and hyperparasitism . The small rectangular cards were used in the field @ 5 cards /acre for sugar cane and @ 12 cards / acre for cotton. The dose being 1.0 CC and 2.5 CC /acre respectively . The cards were tied in different spots avoiding border rows. For sugarcane 6 releases are to be made at fortnight intervals commencing from 3- 3 ½ months onwards. For cotton crop the parasitoids are released soon after noticing the adult moths of bollworms flying in the field or trapped in pheromone trap or light traps. Two or three releases at fortnightly intervals are needed for cotton.

22. Precautionary measure 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. 5. If need arises uses selective/safer insecticides. 6. Ensure that insecticides are used 15 days after or before Trichogramma release.

23. DOSAGE

24. Maintenance of Trichogramma quality : The following practices may be followed to maintain the quality of Trichogramma : Production colonies should be periodically replaced with individuals from a stock culture maintained on the natural or target host. Producers should also periodically assess the per cent host egg parasitization , adult emergence and the sex ratio of emerged adults to be sure they are within acceptable standards.

25. Reference https://en.m.wikipedia.org/wiki/Trichogramma https://www.sciencedirect.com/topics/agricultural-and-biological-sciences/trichogramma https://agritech.tnau.ac.in/crop_protection/crop_prot_bio_mass_parasitoids7.html https://agritech.tnau.ac.in/crop_protection/crop_prot_bio_mass_parasitoids1.html http://www.vsisugar.com/india/agriculture http://www.slideshare.net/ http://www.insectary.com/

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