TECHNOLOGY FOR IN VITRO REARING OF PARASITICS

Аннотация

INTRODUCTION developed rapidly too. In this paper, artificial diets As the main procedure in biological control of and simulated hosts of these parasitoids are agricultural insect, mass production of several described[1;2;6;7]. species of parasitoids has been conducted in Uzbekistan. Their factitious hosts for mass METHODS production are still the eggs or larvae of insects. For 1) Artificial diets: For Trichogrammatidae. instance, the eggs of Helicoverpa armigera and Agrotis Segetum are used for mass rearing Best 3 species Trichogramma were tested: T. Trichogramma spp. The larvae of Helicoverpa pintoe, T. evanescens, T. chilonis. The first 2 species armigera is the factitious host for mass producing are indigenous in Uzbekistan, the others were Bracon habetor Say or Bracon greeni. However, introduced from China[1;2;3;4]. searching for more convenient and cheaper This Trichogramma were collected from Tashkent methods of parasitoid’ mass rearing is still an province, Buka district, and reared in the important challenge in biological control. laboratory on Galleria mellonella. Manufacture of the simulated “host eggs” and a) Pupa hemolymph of Galleria mellonella (heated “larvae” for in vitro rearing these parasitoids is https://www.theamericanjournals.com/index.php/tajabe 12 THE USA JOURNALS THE AMERICAN JOURNAL OF AGRICULTURE AND BIOMEDICAL ENGINEERING (ISSN – 2689-1018) VOLUME 06 ISSUE12 in a water bath at 60 0C for 4 or 5 min) 45.5+1.5 %, %, chicken yolk 24+1,0.8 %, milk 10% / 21.5+0.6 chicken yolk 22.5+1,0.3 %, milk 10% / 20.5+0.5 %, %, inorganic salts 11.5+1.0 %, Penicillin 400/ml. inorganic salts 11,5+1,0.3 % (Neisenheimer’s Streptomycin 400 units/ml. The development of T. mixture salt NaCl7.5 g, KCl0.1 g, CaCl20.2 G, chilonis, reared in vitro successively for many NaHCO30.2 g, H2O 100 ml), Penicillin 400/ml. generation, is showed in Table 1[1;2;3]. Streptomycin 400 units/ml. 2) Artificial diets for Braconidae: Best 2 species b) Pupa hemolymph of Helicoverpa armigera Braconinae were tested: Bracon hebetor Say, 42+1,0.2 %, chicken yolk 25+1,0.8 %, milk 10% / Bracon greeni. This Bracon were collected from 22.5+2,0.2 %, inorganic salts 10.5 +0.5 %, Tashkent province, Buka district, and reared in the Penicillin 400/ml. Streptomycin 400 units/ml.(Fig- laboratory on Galleria mellonella and Helicoverpa 1;2). armigera[4;5;6;7]. c) Pupa hemolymph of Agretus segetum 43+2,0.2 Table 1. DEVELOPMENT OF HYMENOPTERA, REARED IN VITRO SUCCESSIVELY FOR 10 GENERATION Generation 1-5 6 7 8 9 10 % parasitism 100 100 100 100 100 100 % adult emergence 89+3 88+3 87+4 86+4 86+3 85+3 % adult with expanded 82+3 82+2 81+3 81+2 80+3 80+2 wings % pupa development 93+3 92+4 92+3 90+4 90+2 87+3 % normal adult 86+2 86+3 86+4 85+2 82+2 80+3 ♂: ♀ genders proportion 2:8 2:8 2.7 2.6 2:5 2:4 armigera Hb, Agrotis segetum Sciss and Galleria Preparing of artificial mediums. mellonella’s pupa liquid is used for research (Fig- 1. Insect hemolymph collection: Pupa were 2). immersed in water bath at 600C for 6 or 7 min to Natural milk 10% mixture is added to hemolymph avoid blackening of the hemolymphy. After surface and hicken yolk is added diet medium and put at sterilization with alcohol and need sterile ultraviolet lamp. It is rotated for 5 minutes at 2000 condition. second speed in centrifuge. Diet medium should be 2. Chicken embryo extract collection: The kept in the clean, without microbe room and at Chicken embryo extract, only it should be the egg 20°C cool [1.3.5;6;7]. yolk and need sterile condition. After medium has been ready, it is placed in special 3. Milk: Fresh cow milk or 10% powdered milk artificial caterpillars which are cleaned with 75% solution also need sterile condition. ethyl spirit made of politilen. 4. Inorganic mixture salt: Use Neisenheimer’s MANUFACTURE OF SIMULATED "ARTIFICIAL mixture salt (NaCl7.5 g, KCl0.1 g, CaCl20.2 G, EGG-CARDS AND LARVAE" NaHCO30.2 g, H2O 100 ml). 1) Artificial “egg-cards”: There are 2 types of It is defined there is albumen, oil, water when it is artificial “egg-cards”[1;2;3]. checked ingredients of master caterpillar’s type a) Tri-ring “egg-cards” 2 pieces of plastic film are that belong to Bracon in nature. Besides, Heliothis https://www.theamericanjournals.com/index.php/tajabe 13 THE USA JOURNALS THE AMERICAN JOURNAL OF AGRICULTURE AND BIOMEDICAL ENGINEERING (ISSN – 2689-1018) VOLUME 06 ISSUE12 used. The semispherical concaves are made on the b) Bag-form “egg-cards”: One half of a piece of film upper plastic film. Artificial medium is poured into is fulled of concaves with medium and is covered concaves fully (but without overflow) with a micro- by another half of film. Three sides of film are syringe or micro-pipette. The bottom plastic film sealed. Convex side of capsules are exposed and has no concaves. The upper and bottom plastic between the concave side and bottom half of the films are separated and stretched tightly by three same piece of film there should be space for plastic ring with