WELCOME

TECHNIQUES FOR

APPLICATION OF BIOCONTROL AGENTS

BY : JAYANT YADAV, CCSHAU, HISAR, HARYANA

INTRoDUCTIoN

BIOCONTROL- use of natural or modified organisms, genes, or gene products” to reduce the effects of pests and diseases.

The production of food grain should increase to 250 million tones by the year 2020 in order to meet the needs of the growing population.

The environmental pollution caused by excessive use and misuse of agrochemicals, has led to considerable changes in people’s attitudes towards the use of pesticides in agriculture. (1)Biological control is less costly and cheaper than any other methods.(2) Biocontrol agents give protection to the crop throughout the crop period.(3) They do not cause toxicity to the plants.(4) Application of biocontrol agents is safer to the environment and to the person who applies them.(5) They multiply easily in the soil and leave no residual problem.(6) It increases the crop yield .(7) Biocontrol agents are very easy to handle and apply to the target.(8) Biocontrol agent can be combined with bio-fertilizers.(9) They are easy to manufacture.

Biological control practices involve 3 major techniques ,i.e. Introduction , augmentation and Conservation.1.Introduction or classical biological control: It is deliberate introduction and establishment of natural enemies to a new locality where they did not occur or originate naturally.2.Augmentation: It is the mass rearing and releasing of natural enemies to supplement the numbers of naturally occurring natural enemies . It is 2 types: A. Inoculative release: In this case control is expected from

progeny and subsequent generation . It is done infrequently once in a year .

B. Inundative release: In this case control is expected directly from released individuals. Here NE are mass reared and released frequently.

3.conservation: It is the action taken to preserve and increase natural enemies by environmental manipulation.The microbial biocontrol agents are applied in field generally by 3 ways;i. Seed treatmentii. Soil applicationiii. Foliar application

TECHNIQUE FOR APPLICATION OF beetles.These beetle are applied into field by general biocontrol technique: Augmentation technique: The beetles are mass reared and released periodically in the field against the target pest. •Mass ProductionTo release the predator in large numbers in the field, mass culture of Cryptolaemus is a pre-requisite. Cryptolaemus is easily cultured on a large scale on the mealybugs particularly pink hibiscus mealybug and citrus mealybug. In the culture room, it is ideal to maintain 25 to 30°C. •Stage to be releasedAdults and larvae can be released in the field for the suppression of pests.•Time of releaseUsually the releases are made between 8.00 AM and 10.00 AM and 3 PM and 5 PM. •Dose of releaseA release rate of 5000 beetles/ha is recommended to control mealy bugs by Creptolaemus montrouzieri.•Likewise beetles are released against weeds and insects.

SEVEN SPOTTED BEETLE

VEDALIA BEETLE

ZYGOGRAMMA BICOLORATA

CREPTOLAEMUS MONTROUZIERI

Trichogramma Release TechniquesCorrect timing of Trichogramma release requires good coordination between the rearing facility and field staff. There can be 2 methods of field applications : 1. RELEASE ON CARDS: The Trichogramma are shipped (as pupae) inside of grain moth eggs ready to emerge as adults. They can be loose or glued to perforated cards. Moth eggs glued on cards with 100,000 Trichogramma per card (up to 120,000) .

The card can be broken into 30 squares (hangable units)with 4,000 parasites per square inch that permits even distribution in fields and orchards. The cards can be stapled under leaves.The wasps emerge in two to five days, depending on temperature, which should ideally be 80º to 90ºF. Emergence can be delayed by holding parasitized moth eggs at cooler temperatures (not less than 40 ºF) and emerging wasps are usually seen in the morning. To maximize pest fighting time, do not delay release after adult wasps emerge. Keep Trichogramma cards in the shade, out of the hot sun and application rate protocols for Trichogramma spp, can be changed, according to the height of the crop to determine the proper species to choose .

Advantages are superior percentage emergence and survival in the field. It can be done by several methods:A. Glass Jar Incubation Containers:•Any size jar can work from pint to gallon according to convenience and size of release.•Use a tight-fitting lid. •Shredded paper or thin strips of cloth in the container makes it easier to lift the live Trichogramma adults to the tree. •If growers do not use shredded paper and they do not fly out of the jar easily, wrap the jar in a jar cloth or bag so they can be attracted to the mouth of the jar. • Benefit of using a jar is ability to see the wasps & the proportion remains to be released .

2.RELEASE AS ADULTS :

B. Paper Bag Incubation Containers: •Any size paper bag can work from a lunch bag to a grocery bag, but there should not have holes in it. •Again, put some material like shredded paper, short pieces of string or other material in the bag. •If the bag and other material are white or light coloured it is easier to see the moving specks that are the wasps. •The benefit of using a paper bag is ease of handling and Trichogramma that is not walking on the paper shreds can be more likely to leave the bag than a clear jar .

GENERAL PROCEDURE FOR PREPARING MICROBIAL FORMULATIONS.

PRECAUTIONS:Users are advised to handle all microbial insecticides cautiously. Bacterial spores, as well as other microbes, become “foreign proteins” if they are inhaled or rubbed into the skin and can cause allergic reactions.

There are many microbial agent used as biocontrol agent.

•Methods used are : soil application , seed pelleting , seed treatment , foliar application etc.•The microbial insecticides are: NPV, Bt, Fungi, EPN etc.•Biocontrol agent is available as suspension or dry powder.•Treating the seed material with bio control agent is most economical against seed borne or soil borne pathogen.•sticker are also used for increasing efficiency such as :Carboxymethyl cellulose(1%) , Polysulf(0.8%) , Polymethyl alcohol(20%) for treating seed.•Dipping of roots in preparation of Pseudomonas fluorescens reduces the disease by 60-80%.

1.TECHNIQUE FOR APPLICATION OF Bt AND ITS FORMULATIONS•Bt and its toxins can be formulated into liquid concentrates, wettable powders, and ready -to-use dusts and granules or expressed in transgenic plants. •Bt formulations may be applied to foliage, soil, water environment or food storage facilities. •The best-known and most widely used Bt insecticides are formulated from Bacillus thuringiensis var. kurstaki isolates that are pathogenic and toxic only to larvae of the butterflies and moths. Many such Bt products have been registered with the United States Environmental Protection Agency. •The most common trade names for these commercial products include Dipel®, Thuricide®, Worm Attack®, Caterpillar Killer®, and SOK-Bt®. •Each Bt insecticide shows high degree of specificity. Susceptible insects must consume Bt to be poisoned, so treatments must be directed to the plant parts or other material that the target pest will eat.

•Where Bt is applied to plant surfaces or other sites exposed to sunlight, it is deactivated rapidly by direct ultraviolet radiation. •To maximize the effectiveness of Bt treatments, sprays should thoroughly cover all plant surfaces, including the undersides of leaves. •Treating in the late afternoon or evening can be helpful because the insecticide remains effective on foliage overnight before being inactivated by exposure to intense sunlight the following day. •Treating on cloudy (but not rainy) days provides a similar result. •Production processes that encapsulate Bt spores or toxins in a granular matrix (such as starch) or within killed cells of other bacteria also provide protection from ultraviolet radiation.

APPLICATION TECHNIQUE FOR ENTOMOPATHOGENIC FUNGI

•The most common method used has been the immersion of any insect stage (larva, pupa or adult) in a conidia solution .

•Application of conidia to the soil have been suggested as a method to use these entomopathogenic fungi for fruit fly control .

•Howse and Underwood (2000) proposed the use of electrostatic charged powders to spread a killing agent within a pest population.

Dry root rot of mung bean caused by Macrophomina phaseolina was reduced by seed pelleting of Trichoderma viride isolates when talc was used as carrier. Among the isolates, T viride-III supported higher plant growth, better native Rhizobium nodulation and grain yield. SEED PELLETING: •For this eight-day old culture of T viride isolates and T harzianum maintained in 250 ml Erlenmeyer flask containing 70 ml of yeast molasses broth were mixed including the mycelial mat and metabolites using a mixie.•Further, it was mixed with taIc powder @ 1:2 (v/w) and shade dried for 2 days.•Carboxymethyl cellulose (10g) was added as sticky material for 1 kg of talc powder. •The talc based Trichoderma formulation was used for treating the seeds at 4g/kg of seeds.•This talc based formulation contains 1 x 107 CFU/g of talc on serial dilution (Ramakrishnan et al., 1994). •For comparison, carbendazim was used for seed treatment at 2g/kg of seed.

•The EPN can be applied in the field by mixing with irrigation water through irrigation channel.

Technique for application of EPN

REFFERANCESarwar. M. and Salman. M, (2015). Biological insecticide Trichogramma spp. (Hymenoptera: Trichogrammatidae) strikes for caterpillar control . International Journal of Entomology Research. (1): 31-36 Toledo.J, Liedo. P, Flores.S, Campos.S.E, Villaseñor. A and Montoya,P. (2006).use of Beauveria bassiana and Metarhizium anisopliae for Fruit Fly control: A novel Approach. Fruit Flies of Economic Importance: From Basic to Applied Knowledge Proceedings of the 7th International Symposium on Fruit Flies of Economic Importance .10-15 September 2006, Salvador, Brazil.pp. 127-132Raguchander .T., Rajappan. K. and Samiappan R. (1997). Evaluating methods of application of biocontrol agent in the control of mungbean root rot. Indian Phytopath. 50 (2) : 229-234 .Ramakrishnan, G., Jeyarajan, R. and Dinakaran, D. (1994). Talc based formulation of Trichoderma viride for biocontrol of Macrophomina phaseolina. J. Biological Control 8: 41-44.Howse, P. E., and K. L. Underwood. 2000. Environmentally safe pest control using novel bioelectrastatic techniques: Initial results and prospects for area-wide usage, 295-299. In: K. H. Tan [ed.]. Area-wide control of fruit flies and other insect pests. Penerbit University Sains Malaysia. Penang, Malaysia.

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