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In Vitro rearing of trichogramma (Hymenoptera: Trichogrammatidae)
DOI: http://dx.doi.org/10.20534/ESR-16-9.10-11-13
Jumaev Rasul Axmatovich, Tashkent state agrarian University. Postgraduate student, the protection and quarantine E-mail: [email protected] E-mail: [email protected]
In Vitro rearing of trichogramma (Hymenoptera: Trichogrammatidae)
Abstract: This article reports the recent results of our experiments using artificial media devoid of insects additives for rearing T.pintoe, T.evanescens in vitro.
Key words: Artificial media, parasitoid growth factors, in vitro, hemolymph, egg yolk, inorganic salt mixture, cow milk, trichogrammatidae, predator, research tool, mass production.
Introduction
At the process of entirely rearing trichogramma parasite in the laboratory condition cultivates Sitotrogoce reallela Olive egg a lot of corn productions such as grain, maize, barley and worker force are spent. In the result of it, rearing price is increased. More than 150 thousand ton barley is emitted in exist biolaboratory our Republic in a year [1; 2; 3].
Emitting this barley, other diets cultivation happens in the account of decreasing producing ofplants or enterprises. This testifies that above mentioned process is damaging for diets industry [1; 2].
By rearing trichogramma commonly using artificial diets mediums and automatizing them gives an opportunity to solve above problem.
In China 1975-1978 years for 3 year' investigations rearing trichogramma in the diets medium, which belongs to artificial medium researches showed its good results [1; 3].
According to the first medium, first general ingredients: (Hemolymph from pupae Antheraea pernyi 43.1%, egg yolk 34.48%, inorganic salt mixture 13.79%, and pig serum 8.62% or hemolymph from pupal Philosamia cynthia ricini 31.75%, cow milk 31.75%, egg yolk23.81% and, inorganic salt mixture12.70% [2; 3].
By the method of rearing Entomophags in vitro many scientists investigated and achieved positive results
According to Gao and others' (1982) researches trichogramma generation which is reared in vitro when is used against H.armigers in the field of cotton-plant was achieved biologic effectiveness by 93% [1; 2].
The artificial rearing of trichogrammatidae started a long time ago, with the main goal to try obtaining a mean to multiply and produce parasitoids to be released in biological control strategies. But it is also a powerful tool to conduct studies on biology, physiology and behavior of entomophags, especially endoparasitoid species (Grenier, S., Plantevin, G. 1990.). Trichogramma spp 5 are widely used for biological control of pest insects. Currently, these egg parasitoids can be produced only from natural or factitious hosts or from insect hemolymph- based diets [2; 3].
Wu et al. (1982), Liu and Wu (1982), and Qin and Wu (1988) described artificial diets devoid of insect components for the in vitro rearing of Trichogramma dendrolimi Matsumura [2; 3].
Considerable work has been done on the development of artificial diets for Trichogramma spp. Many of these efforts have involved diets containing hemolymph (Guan et al., 1978; Hoffman et al., 1975 Hubei Research Group, 1979; Liu et al., 1979, 1982; Wu et al, 1982 Strand and Vinson, 1985; Xiet al, 1986, 1997a; Qin and Wu, 1988 Grenieret al., 1995). Development ofartificial diets for Trichogramma would foster the implementation of automated mass rearing systems. Some progress has been made along this line, primarily in China (U. S. Department of Agriculture, 1982; Li et al, 1988, 1995a, b)
where in vitro reared Trichogramma chilonis Ishii have been released for control of sugarcanestem borers (Liu et al., 1995b) and Trichogramma dendrolimi Matsumura has been reared for control of Heliothis armigera L. in cotton and Dendrolimus punctatus L. in pine trees [1; 3].
Materials and methods
Source of Trichogramma and In Vitro Rearing Trichogramma stock: T. evenecens, T.pintoe were collected from Tashkent province, Buka district, and reared in the laboratory on Heliothis armigera and Agretus segetum shif.
Ingredients for in vitro medium: Pupae hemolymphy (G.melonnellan or H.armigera), cow milk or 10% powdered milk solution, chicken embryo extract and Neisenheimer's mixture salt.
Preparation of the ingredients
1. Insect hemolymph collection: Alive pupae were immersed in water bath at 60 °C for 6 or 7 min to avoid blackening of the hemolymphy. After surface sterilization with alcohol and need sterile condition.
2. Chicken embryo extract collection: The Chicken embryo extract, only it should be the egg yolk and need sterile condition.
3. Milk: Fresh cow milk or 10% powdered milk solution also need sterile condition.
4. Inorganic mixture salt: Use Neisenheimer's mixture salt (NaCl7.5 g, KCl0.1 g, CaCl20.2 G, NaHC030.2 g, H2O 100 ml).
Artificial "egg-cards"
There are 2 types of artificial "egg-cards" [2; 3].
1) Tri-ring "egg-cards" 2 pieces of plastic film are used. The semispherical concaves are made on the upper plastic film. Artificial medium is poured into concaves fully (but without overflow) with a micro-syringe or micro-pipette. The bottom plastic film has no concaves. The upper and bottom plastic films are separated and stretched tightly by three plastic ring with different inner diameters, in our Cass they are: 5:5 cm, 5.4 cm, 5.2 cm respectively;
2) Bag-form "egg-cards": One half of a piece of film is fulled of concaves with medium and is covered by another half of film. Three sides of film are sealed. Convex side of capsules are exposed and between the concave side and bottom half of the same piece of film there should be space for aeration. The size of plastic film depends upon the number of semispherical concaves made on film [2; 3].
Culturea of G.melonnella or H.armigera fresh cocoons of G.melonnellan or H.armigera from Tashkent province.
Components of artificial diets for Trichogramma pintoi
The first medium: Hemolymph (A) G.melonnellan (Aj 45,5%, Neisenheimer's (Aj 15,5%, Chicken egg yolk (A3) 25,5%, milk (A4) 13,5%.
The second medium: Hemolymph (B) Heliothis armigera Hb (B1)34,5%, Neisenheimer's (B2) 12,5%, Chicken egg yolk (B3) 30,5% milk (B4) 22,5%.
Section 2. Biology
The three medium: Hemolymph (C) Agretus segetum shif (C1)42,5%, Neisenheimer's (C2) 13,5%, Chicken egg yolk (C3) 23,5%, milk (C4) 20,5%.
The four medium: Hemolymph (D) G.melonnellan (D1) 45,5%, Neisenheimer's (D2) 12,5%, Chicken egg yolk (D3) 30,5%, milk (D4) 11,5%.
The five medium: Hemolymph (E) G.melonnellan (E1)45,5%, Neisenheimer's (E2) 13,5%, Chicken egg yolk (E3) 20,5%, milk (E4) 20,5%.
The results of the research
It was showed in the Table 1 that the pupae hemolymphy of either G.melonnellan or H.armigera Hb could be used as the main component of the artificial diet for the development of T. pintoi and T. evenecens. There was no significant difference in their parasitism, survival, percentage of pupation, adult emergence and reproduc-tively when the pupae hemolymphyof G.melonnella was used instead of that of H.armigera Hb.
Developmental stages of T. evenecensby in vitro.
a b
Picture 1. a - Bag-form artificial "egg — card", b - Young scientist R. A. Jumaev is practicing his research on the theme of "Rearing Trichogramma in vitro"
Table 1. - Development of Trichogramma pintoi, reared in vitro laboratory experiences, 2013-2016 y. (+25±3°C, RH 80±5%)
Composition of medium % The amount Development degree of trichogramma generation in artificial medium by days Genders proportion 6: ?
of damage % Eggs larva period Pupa period Mature period (imago)
A
A, A2 A3 A4 66,3 1,4 5,6 4,4 3,4 2:4
45.5 15.5 25.5 13.5
B
B1 B2 B3 B4 46,7 1,4 3.3 - - -
34.5 12.5 30.5 22.5
C
C1 C2 C3 C4 34,4 1,2 4,8 2.4 - -
42.5 13.5 23.5 20.5
D
D, D2 D3 D4 74,6 1,4 5,6 4,1 3,3 2:6
45.5 12.5 30.5 11.5
E
E, E2 E3 E4 82,5 1,3 5,8 5,4 3,6 2:8
45.5 13.5 20.5 20.5
Jumaev R.A.
IAP 20150052
I
a b
Picture 2. Developmental stages of T. evenecens by in vitro a — egg, b — larvae of 2nd instar, early stage of pre pupae and female adult
Study on effect of a-lipoic acid on phospholipid composition of rat cardiac tissue under hypobaric hypoxia
It is obvious in the research, prepared all mediums of diets are harmed with trichogramma generation and put there eggs. But it is observed dying because of inconvenience medium of diets for developing parasite generation. According to diet mediums component development of trichogramma parasite was rather prolonged when G.melonnella hemolymphy and inorganic salt's quantity was more in diets medium. In fact quantity ofprotein and oil is more in the structure of hemolymphy as well as it is considered convenient for development ofparasite maggot.
References:
Thus, from above nominated artificial diet mediums, fifth diet medium (E) pupae hemolymph of G.melonnella (E^ 45,5%, inor-ganicsalts (E2) 13,5%, chicken yolk (E3) 20,5%, milk (E4) 20,5% composed medium of diets is defined as a suitable one for normal nourishment and development of trichogramma generation in order to rear trichogramma parasite.
1.
2.
3.
Кимсанбаев. Х. Х., Жумаев Р. А. Renewing and rearing technology of Bracon hebetor Say in Biolaboratory; Материалы VIII-оймеждународной научно - практической конфе-ренции молодых исследователей, г. Волгоград, - 2014. - С. 257-259. Li Li-ying, Liu Wen-hui 1997. Rearing Trichogramma spp, with artificial diets, containing hemolymph of different insects. Parasitoids and predators (insecta) of agricultural and forestry arthropod pests - 335-337. (In China).
Li Li-ying, Liu Wenhui, Chen Chaoshian, Han Shityou, Shin Jiachi, Du Hansun, Feng shuyi, 1997. In vitro rearing of Trichogramma spp and Anastatus sp in artificial "eggs" and the methods of mass production. Parasitoids and predators (insecta) of agricultural and forestry arthropod pests - 344-357. (In China).
DOI: http://dx.doi.org/10.20534/ESR-16-9.10-13-16
Mamadaliyeva Nodira Isakovna, Post-doctoral student Laboratory of Metabolomics, Acad. O. A. Sadykov Institute of Bioorganic Chemistry, Uzbekistan Academy of Sciences E-mail: [email protected] Saatov Talat Saatovich, Professor, Head of laboratory Laboratory of Metabolomics, Acad. O. A. Sadykov Institute of Bioorganic Chemistry, Uzbekistan Academy of Sciences E-mail: [email protected] Umerov Oybek Ilyasovich Junior researcher Laboratory of Metabolomics, Acad. O. A. Sadykov Institute of Bioorganic Chemistry, Uzbekistan Academy of Sciences E-mail: [email protected]
Study on effect of a-lipoic acid on phospholipid composition of rat cardiac tissue under hypobaric hypoxia
Abstract: Hypoxia is a rather frequent pathological or borderline state caused by the reduction of oxygen concentrations in the ambient environment or dysmetabolism of oxygen in an organism. The simulated chronic hypoxia similar to the one at the altitude of 7,000 m, caused changes in total sum of phospholipids in rat cardiac tissue and proportions of some phospholipids. Pharmacocorrection of the changes with berlithion containing a-lipoic acid was found to normalize levels of some rat cardiac tissue phospholipids.
Key words: cardiac tissue, hypobaric hypoxia, phospholipid.
Introduction [3, 49-50; 4, 10]. In many cases damage of integrity of membrane
Hypoxia is a rather frequent pathological or borderline state- bilayer's lipid phase accounts for fails indicating significant role oflip-caused by the reduction of oxygen concentrations in the ambient ids in pathogenesis of hypoxic conditions [5, 31-32; 6, 84]. environment or dysmetabolism of oxygen in an organism [1; 2183]. Changes in the lipid composition of cardiac tissue membranes
Hypoxia is capable of causing diseases independently and modi- under hypoxia are known to take place because of hypoenergetic fying courses of other diseases, primarily, cardio-vascular ones [2; condition, excessive lipid peroxidation [7, 140-141; 8, 413-415] 634-635]. and modification of membrane structures under the effect of cyto-
The findings from numerous recent studies aiming at elucidation toxic metabolites [9; 177]. Disorders in cardiac bioelectrical activity, of a mechanism underlying the onset and progression ofpathological onset and progression of arrhythmia and hypertension [10; 177], processes associated with hypoxia of cardio-vascular system are the atherosclerosis [11; 188], myocardial hypertrophy, ischemic heart evidence for the fact that changes in a cell membrane's structure and disease and myocardium infarctions [12; 380] are among the con-function are the significant link in the chain of the pathology's onset sequences taking place [13; 199-200].
