1.1 Pest problem is one of the major constraints for achieving higher production in agriculture crops. India loses about 30% of its crops due to pests and diseases each year. The damage due to these is estimated to be Rs.60,000 crores annually. The use of pesticides in crop protection has certainly contributed for minimising yield losses. The pesticides, which are needed to be applied carefully, only when the threshold limits of the pest population is exceeded. However, quite often the indiscriminate and unscientific use of pesticides has led to many problems, such as pests developing resistance, resurgence of once minor pest into a major problem besides environmental and food safety hazards.

1.2. The problem of insect-pest is acute in case of all the crops and especially so in case of commercial crops. The use of insecticides and pesticides have increased manifolds during the past 3 – 4 decades with the introduction of intensive cropping. The average consumption of pesticides in India is about 570 gms per ha. as compared to developed countries like Japan, Thailand and Germany where the consumption rate is 11 kg, 17 kg and 3 kg per ha, respectively. Though the average quantum of pesticides usage in India is low, the damage caused due to their indiscriminate usage and poor quality maintenance is alarming. Interms of value, much of the pesticide application is accounted for by a few crops. For example, cotton, paddy and vegetable crops account for 80% of the value of pesticides applied in India.

1.3. Pesticides or chemicals are meant to control harmful pests such as insects, nematodes, diseases, weeds etc. However, excessive use of pesticides not only leave residues in soil, water and air but also have adverse effects on the non target organisms such as pollinators, parasitoids, predators and wild animals. This has adversely affected the ecological balance resulting in pest resurgence, development of resistance in the pest species and environmental pollution. Development of pest resurgence and resistance has resulted in high cost of production and low income especially to cotton farmers in AP, Maharashtra.

1.4. In view of the several disadvantages associated with the unscientific use of pesticides in agriculture, there is an urgent need for minimising the use of chemical pesticides in the management of insect pests. Growing public concern over potential health hazards of synthetic pesticides and also steep increase in cost of cultivation/low profit making by farmers has led to the exploration of eco-friendly pest management tactics such as Integrated Pest Management (IPM). IPM aims at suppressing the pest species by combining more than one method of pest control in a harmonious way with least emphasis on the use of insecticides. In simple terms “IPM is the right combination of cultural, biological and chemical measures which provides the most effective, environmentally sound and socially acceptable methods of managing diseases, pests and weeds”. The major components of IPM are prevention, observation and intervention. The IPM seems to be the only answer to counter some of the major pests of crops, which have become unmanageable in recent years. The success of IPM largely depends upon conservation of naturally occuring bio control agents.

2.1. In nature every ecosystem exists in a balance. Growth and multiplication of each organism depends on the food-chain, its predetors, parasites, etc. In biological control system, these interrelations are exploited. The natural enemy of a pest, disease or weed is selected, its biology is studied for mass multiplication and utilize the same to check the target pest. They are also specific in their action and perish once their feed (i.e. the pest) is exhausted. Thus they are based on natural principles, do not leave any residue, safe and economical.

2.2. Among the alternatives, biological control of pests is one of the important means for checking pest problems in almost all agro-ecological situations. Bio pesticides are living organisms which can intervene the life cycle of insect pests in such a way that the crop damage is minimized. The agents employed as biopesticides, include parasites, predetors and disease causing fungi, bacteria and viruses, which are the natural enemies of pests. Further, they complement and supplement other methods of pest control. Utilisation of naturally occurring parasites, predators and pathogens for pest control is a classical biological control. On the other hand, these bio agents can be conserved, preserved and multiplied under Laboratory condition for field release. Once these bio-agents are introduced in the field to build their population considerably, they are capable of bringing down the targeted pest’ population below economic threshold level (ETL). However, the crux lies in their mass production and application at the appropriate time.

3. Major advantages of bio pesticides

Bio-pesticides are preferred over chemical pesticides for the following reasons:

• no harmful residues;
• target specific and safe to beneficial organisms like pollinators, predetors, parasites etc.;
• growth of natural enemies of pests is not affected, thus reducing the pesticide application;
• environmental friendly;
• cost effective;
• important component of IPM as 1st line and 2nd line of defence, chemicals being the last resort.

4.1. Last decade has witnessed a tremendous breakthrough in this aspect, especially on standardization of production techniques of Trichoderma, Gliocladium, Paecilomyces, Pseudomonas, Trichogramma, NPV and Bacillus to use them against many insect pests and diseases.

4.2. There are a number of instances where bio control agents have been successfully employed in India. Some examples of these are given below :

1. Growth of lantana weed was controlled by using the bug Telonemia scrupulosa
2. Sugarcane pyrilla has been successfully controlled in a number of States by the introduction of its natural enemy Epiricania melanoleuca and Tetrastictus pyrillae.
3. Trichogramma, which feeds on the eggs of sugarcane borers, has been used against the borers in the states of Tamil Nadu, Rajasthan, UP, Bihar and Haryana.
4. Similarly Trichogramma, Bracon, Chelonus and Chrysopa spp. are being used for the control of cotton bollworms. Trichogramma has also been used against rice stem borer and leaf folder.
5. The sugarcane scale insect has been controlled with the help of predatory coccinellid beetles in UP, West Bengal, Gujarat and Karnataka.

4.3. The popularity of biopesticides has increased in recent years, as extensive and systematic research has greatly enhanced their effectiveness. Also, techniques for the mass production, storage, transport and application of biopesticides have been improved in recent years.

5. Scope for Commercial Production of Biopesticides

Though there are about 140 biopesticide production units existing in the country as on today, they are able to meet the demand of only less than 1% of cropped area. There exists a wide gap, which can only be bridged by setting up of more and more units for production of biopesticides. This requires large scale investment and private participation.

Some of the local small scale industries have already started production and marketing of Trichoderma viride (against few fungal diseases) and Trichogramma (against sugarcane early shoot borer). There is a scope to enhance production and use of biological control agents in the days to come as the demand is on the increase every year.

6. Location of Biopesticide Units

In order to achieve optimum results, care needs to be taken to set up biopesticide facilities in areas which have appropriate climatic conditions. The production of Biopesticides requires controlled climatic conditions. Temperature control is less costly in locations where there is no extreme conditions. Besides the climatic conditions, the proximity of the location to the market is also important. However, care must be taken that the production facilities are set up at least a quarter of a mile away from farming areas, so as to prevent the contamination of production facilities by insecticides from the farming areas.

• The primary objective of biopesticide projects is to establish the bankability of mass multiplication of various bioagents discussed in the models
• To serve as guidelines for extending financial assistance to entrepreneurs who may be interested in setting up biopesticide units
• To promote setting up of more bio-control production units
• To disseminate widely the technology

 9. Basic requirements for establishment of Biopesticide units

Based on the field visits to bio-control production units and in line with the technology and objective of biopesticides production, various facilities required for the successful implementation of such projects are indicated below:

9.1 Land

Land is required for construction of culture and rearing rooms, processing room, laboratory, office etc. In the present models, we have assumed only rented buildings, hence no land cost has been considered except for poly house.

9.2 Building and civil works

Biopesticides production involves rearing of insects. Hence, the basic infrastructure to be created includes only the civil structures built in such a way as to provide environmental conditions suitable for rearing of insects. The production unit has to be located away from industrial unit to avoid pollution problems

9.3 Plant and Machinery

There is no requirement of heavy plant and machinery. Racks, trays and other facilities are required for rearing insects. Apart from this centrifuge, mixers and some fabricated equipments for insect collection and rearing are required. For production of Trichoderma fermentors, laminar flow apparatus etc. are required. All the machinery required are locally manufactured.

9.4 Raw material

For rearing of insects special diet is required which comprises of pulses, vitamins, antibiotics etc. For production of Trichoderma molasses-yeast medium, is required. All these materials are available locally.

9.5 Water

The water requirement is mainly for feed preparation, washing, cleeaning, drinking etc.. Water quality should be tested to establish the suitability.

9.6 Power

Power supply is essential for bio-pesticide units. Electricity charges under recurring cost are considered in the models.

9.7 Manpower

Production of bio-pesticides required skilled manpower. There is need for a number of labourers at each stage of production. The project is labour intensive. The manpower requirement is as under:

Technical Aspects of Biopesticides

Bio pesticides are living organisms which can intervene the life cycle of insect pests in such a way that the crop damage is minimized. The agents employed as biopesticides are parasites, predetors, fungi, bacteria and viruses which are natural enemies of pests. These bio agents can be conserved, preserved and multiplied under laboratory condition for field release.

2. Major types of bio-agents available for commercial production

There are different types of bio-agents which can be commercially mass produced for large scale distribution among the farmers for control of insect pests. They are:

Field efficacy trials have been conducted by State Agricultural Universities and ICAR Research Institutes/Stations to know the extent of pest control by application of biopesticides. The percentage of pest control achieved for selected bio-control agents is as under:

• Speed/Mobility to prevent pathogen to develop resistant structures.
  • Longevity, enough to protect plant during its vulnerable period, whatever that may be
  • Environmental tolerance, to sustain activity under different soil and climatic conditions.
  • Mode of Action, varies from pathogen to pathogen, physical contact, chemical nature of killing component.

While using natural enemies, it is important to have fast growing biocontrol organism in the fields which can eventually make the conditions unfavourable for the pathogens proliferation

5. Technical consultancy for setting up of mass production of bioagents

Setting up of unit for mass production of bio agents especially those which are based on fungi, bacteria and virus is highly technical in nature. The skill required to handle the mass production process is also higher. done by larvae is avoided. It offers a lower cost but more effective plant protection option in comparison to insecticides. Two species i.e., T. chilonis and T. japonicum are predominantly used in India.

1.2 Trichogramma are dark coloured tiny wasps and the female wasp lays 20-40 eggs into the host’s eggs. The entire cycle is completed within 8-12 days. The tiny adult wasps search for the host (pest) eggs in the field and lay their eggs into the eggs of the pests. The parasitised host’s eggs turn uniformly black in 3-4 days. The Trichogramma eggs on hatching, feed the embronic contents of host’s egg, completes its development and adult comes out of the host egg by chewing a circular hole. A single Trichogramma, while multiplying itself, can thus destroy over 100 eggs of the pest.

2. Major equipment needed

Equipments like semi-automatic corcera rearing cages, trays, iron racks, hot air oven, air conditioner, UV chamber, incubator, moth breeding tins, grinder, mating chambers, parasitization jars, refrigerator, wire mesh, netlon etc. are required for mass rearing of corcera and Trichogramma production.

3. Steps involved in production

3.1 Identification of host

The Trichogramma of multiplication starts with identification of a suitable host species, with the following characteristics :

• easily available.
• easy to culture with the locally available material.
• should yield maximum host egg/larvae/pupae per unit cost.

In India Corcera cephalonica, a stored grain pest has been used for mass multiplication of targetted species.

3.2 Rearing of host insect

The host rearing containers are made of materials which are non-toxic, cheap and optimum sized to permit mating and host searching and amenable to easy cleaning. Most commonly used cages are wooden cages, which are now replaced with semi automatic corcyra rearing cages. The nuclear culture, i.e. eggs of Corcyra cephalonica are introduced in rearing cages. In the model use of semiautomatic rearing cages of 30 no. is considered.

3.3 Preparation of feed material

Corcyra feed may be prepared from bold white sorghum grains without any insecticide residues. This can be tested by taking a sample of 100 g from each bag. The crushed sample is fed to 20 number of 1st/2nd instar Corcyra larvae for 2-3 days. Based on the mortality of the larvae, suitability of grains may be decided. The requisite quantum of sorghum is milled to make 3-4 pieces of each grain. Sorghum grains are heat sterilised in oven at 1000C for 30 minutes and the grains are sprayed with 0.1% formalin. This treatment helps in preventing the growth of moulds as well as to increase the grain moisture to the optimum (15-16%), which was lost due to heat sterilisation. Then grains are air dried.

3.4 Corcyra charging

In each rearing cage, 7.5 kg of sorghum grains are filled and charged with 0.5cc eggs (1cc = 20,000 eggs) of mother culture. Yeast, groundnut kernel and streptomycin is added to enhance egg laying capacity of the adult moths and for enriching the diet.

3.5 Collection of moths

After about 40 days of charging, moths start emerging and the emergence continues for two months. 10 to 75 moths emerge daily with the peak emergence being between 65th and 75th day.

Collect the moths daily and transfer to the specially designed oviposition cages for egg laying. Roughly 2000-3000 pairs of moth can be placed in one chamber. Moth emergence reduces after 100 days of initial infestation and cages are released for cleaning.

3.6 Collection of eggs

Eggs are collected by means of manual suction and are placed in tubes and counted with measuring cylinder. Approximately one cc of eggs of Corcyra counts about 20,000 at the fresh harvest. After that due to shrinkage of eggs the count may be increased. The present model assumes 20,000 eggs per one cc for calculation purpose. The final output of Corcyra eggs from one cage has been assumed at 7.5 cc.

 4. Production of Trichocards

The demand for Trichocards will start from the onset of kharif season and extends to rabi season. The summer season vegetables offer an extra demand.

4.1 Egg preparation

The eggs of Corcyra thus collected are cleaned to make it free from insect scales etc. They are sieved thrice and then poured on a plain paper. By slowly tapping eggs come downward stick on to gummed card. Thus, the cleaned eggs are spread on the gummed cards (15 cm x 10 cm) with the help of screen. These eggs of Corcyra are exposed to UV rays of 15 watt UV tube for 45 minutes to prevent hatching. While UV exposure, egg card should be kept about 12-15 cm away from tube.

4.2 Introduction of Trichogramma

After the sterilisation the egg cards are placed in plastic bottles and are introduced with nucleus culture of Trichogramma species of egg or pupal stage. The ratio of host egg and parasite adult should be maintained at 1:5.

4.3 What is a tricho card ?

The parasitisation of Trichogramma spp., in laboratory condition on one cc eggs of Corcyra cephalonica, which are uniformly spread and pasted on a card measuring 15 cm x 10 cm is called as Tricho card. The card has 12 demarcations (stamps). About 12,000 Trichogramma adults emerge out from this card in 7-8 days after parasitisation. To delay the emergence of Trichogramma, these cards can be stored in refrigerator at 5-100C for 10-15 days. On removing the cards to room temperature, the parasitoids emerge normally. Trichocards have a shelf life of 2-3 days. However, these can be stored in a refrigerator for a period of 1 month without any spoilage.

Production technology in flow-chart form is presented in exhibit-I.

5. Dosage

5.1 For controlling sugarcane early shoot borer : Start releasing 6,000 parasites per week per acre area, for a period of 5 weeks, starting from 4th week of planting i.e., as soon as the adult male moths of early shoot borer are noticed in the field. Totally 30,000 parasites are to be released per acre. More parasites may also be released depending upon the crop and pest density.

5.2 In cotton- The Trichocards are released in the field at 45 days after sowing @ 5 cards / ha (one lakh eggs). In total three release are necessary.

6. How to use ‘Tricho card’

The cards are to be used before the emergence of the adult parasite. Cut or tear each Tricho card into small pieces and distribute them all over the field. The pieces may be stapled to sugarcane leaf at 7-8 m distance. Care is to be taken to release the parasites either in morning or evening i.e., during cool hours, in windward direction and there should not be any pesticide spray. Before releasing the parasite, the infected shoots are to be cut to ground level and buried inside the soil so as to avoid secondary infestation.

Advantages of using Tricho cards

1.       Less cost, more effective.

2.       field application (releases) is very simple as compared to other methods.

3.       Records show higher yield in sugarcane (about 4-5 tonnes), as secondary infestation is avoided while using Tricho cards.

4.       Cost of pest control is very nominal.

5.       Added to all these, environmental pollution is avoided.

   Precautions

The following precautions are required to be taken while using Trichocards :

• Trichocards should be packed in such a way that the parasitised surface is on the inner side.
• Emergence date should be specified on cards for the guidance of the users.
• Trichocards should be stapled on the inner-side of the leaf to avoid direct sunlight.
• Card should be stapled in morning hours and just before emergence to avoid predation.
• Farmers should refrain from using pesticides in the field where Trichogramma are released. If need arises selective / safer pesticides can be used and it is to be ensured that pesticides are used 15 days before or after release of Trichogramma

Technology for mass production of Trichoderma fungi

1.1 Crop losses due to soil borne plant pathogens worldwide are Pythium spp., Furarium oxysporum, sclerotium rolfsii, Rhizoctonia solani and Phytophthora spp. These fungi pathogens generally cause wilt disease in many crops. Trichoderma, a fungi, which grow saprophytically in soils have proved as an effective biocontrol agent of wilt diseases.

1.2 Trichoderma spp. are commonly found in almost any soil and other natural habitats consisting of organic matter such as decaying bark, plant material, etc. They grow trophically towards hyphae of other pathogenic fungi, coil them and degrade their cell walls. This process is called “mycoparasitism”, which limits the growth and activity of plant pathogenic fungi. In addition, they produce toxic metabolites which protect the seeds from soil borne pathogenic fungi, by forming a protective coating on them.

1.3 Trichoderma spp. are saprophytic fungi that grow best in neutral and acid soils and thrive well in moist conditions.

1.4 The important species available for mass production are Trichoderma viride and Trichoderma harzianum

1.5 Equipments required: Equipments like fermentor, rotary mixer, auto packer, rotary shaker, laminar flow, water distillation unit, refrigerator, haemo cytometer etc. are required for the production of Trichoderma fungi.

2. Major steps in production process

Inoculation | Fermenter run | Harvesting | Blending | Drying and Packing

3. Outlines for production of Trichoderma

• The pure mother culture of Trichoderma fungi is being maintained in Agri. Universities, IARI, some ICAR institutions (like PDBC, Bangalore) etc. The mother culture can be purchased from the identified sources. They have to be further sub-cultured and maintained purely for mass production by adopting standard techniques under the supervision of trained microbiologist / pathologist.
• The culture has to be mass multiplied in two levels namely (i) at primary level using shakers in flasks and (ii) secondary stage multiplication in fermenters. The important factor in this is the preparation of growing medium in which the culture is mass multiplied. For Trichoderma Fungi, the growing media used in the model is molasis and protein material.
• After the growing media is formulated and sterilised in fermenter, it is inoculated using the culture multiplied in the flasks.
• The molasis based culture media is continuously aerated by passing sterile air from compressors. After about 3-4 days fermentation period, the culture will be ready for packing in a carrier material.
• While the inoculated culture is gathering ready in the fermenters, the carrier material is sterilised in autoclaves and kept ready for mixing the culture. Talk powder is reported to be the commonly used carrier material for Trichoderma Fungi.
• The cultured (fungi) and sterilised carriers are mixed mechanically in a blender and the material is packed using semi automatic packing and sealing machine.

Talc based formulations of the fungal antagonists are applied at the rate of 4gm per kg of seed for controlling soilborne plant diseases. Mix the powder with sufficient quantity of water to make slurry for treating seed before sowing.

                 5. Advantages of Trichoderma applicatiom

• Ecofriendly
• Can be used along with organic manure
• Trichoderma spp. are also known to suppress plant parasitic nematodes (roo-knot nematodes).
• Lower cost and longer efficacy than fungicides
• Does not lead to development of resistance in plant pathogens
• No physotoxic effects
• Minimises losses and cost of production and increases yield & profit.
• Promoter plant growth

                 6. Application

               6.1 Soil application

Trichoderma spp. suppress the activity of soil borne fungal pathogens, especially Rhizoctania solani and Pythium spp. and protect transplanted seedlings by colonizing their roots.

               6.2 Seed treatment

Seed treatment is an alternative approach to introduce Trichoderma spp. into the soil. This method requires smaller amounts of biological material than soil treatment. Unlike chemical fungicides, Trichoderma spp. provide long term protection without any adverse side effects.