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The technology of rearing Braconidae in Vitro in biolaboratory
Section 1. Biology
DOI: http://dx.doi.org/10.20534/ESR-17-3.4-3-5
Jumaev Rasul Axmatovich, Pisa University, Italy Postgraduate (PhD) student E-mail: [email protected]/[email protected] Kimsanboev Xojimurod Xamrakulovich, Tashkent state agrarian University.
Professor, the protection and quarantine. Uzbekistan.
Adilov Makhsud Mirvasitovich, Tashkent state agrarian University Professor, the viticulture and horticulture Rustamov Anvar Axmatovich, Tashkent state agrarian University Postgraduate (PhD) student. Uzbekistan
The technology of rearing Braconidae in Vitro in biolaboratory
Abstract: In the article researches were given about creating artificial medium of diets in order to rear Bracon and choosing effective medium of diets. In our researches medium of diets which consists of some components was created and they were harmed with Bracon generations. According to it, generation of Bracon's nutrition and developing three main types of medium were produced in order to rear Bracon parasite that was composed wax moths (G. melonnella) and (H. armigera) hemolymph, egg yolk, natural milk.
Keywords: Bracon, entomophagous, vermin, unripe cotton bell's caterpillar, wax moths, in vitro, artificial medium, hemo-lymph, egg yolk, genders proportion, inhabitation, result.
Introduction
Nowadays the safety of foodstuff is considered as one of the world global problems. More than 60 countries are suffering from this problem in the world. Increasing population of our country and exceeding demand for quality food, cultivating agricultural products and searching new sources of rearing is becoming problem [1; 2; 6].
That's why our motherland is paying attention widely to provide people with the safety of foodstuff in order to cultivate ecological healthy food and protect environment [1; 2; 3].
It is required much financial expenses to rear entomophages in laboratories in order to cultivate agricultural products and protect pests. In our Republic exist bio laboratories separate more than 140150 thousand tons barley, maize, wheat to culture entomophages. It means that it causes great loss for food industry [1; 2; 3].
To carry artificial mediums in rear entomophages is considered as the best way and it notes the duties of directions such as produce them, save and select effective sorts [1; 2].
In rear parasite entomophages, using artificial mediums widely and automatizing them gives much opportunity to solve above mentioned problems [3; 4; 6].
It was investigated scientifically by learning inhabitation of Bracon parasite entomophages. In this article creating artificial diets medium for growing Bracon parasite entomophagous in bio laboratories, choosing productive diet medium and learning inhabitation of Bracon parasite entomophag, which reared in artificial way, was investigated scientifically [1; 4; 5; 6].
Bracon is reared widely in biolaboratories as an effective parasite which is against for rodent pests of agricultural plants. It was widely
spread in Uzbekistan and other countries of the world. By developing biological methods of learning insects for struggling against pests in the field of researching by these organisms is being payed attention much more. However, in spite of this few researches were done in the field of developing these organisms artificially. That's why we chose this branch [1; 3; 4; 5].
The materials and methods of research
Preparing of artificial mediums. The basic ingredient of medium is prepared insect hemolymph and in addition it is added chicken yolk, solution of cow milk (10 gr/100 ml water). In each 100 ml mixture, 40-60 thousand i.u of gentamisin is added. The mixture of diets are shown at the 6th table.
It is defined there is albumen, oil, water when it is checked ingredients of master caterpillar's type that belong to Bracon in nature. Ingredients of caterpillar resemble each other. That's why it is taken as a base hemolymph of wax moths' caterpillar to rear properly in laboratory. Besides, Heliothis armigera Hb, Agrotis segetum Sciss and cabbage white moths' pupal liquid is used for research.
Then it is dried with filter paper after taking water. When it dries, by cutting a small part of the tail or pressing to disjoin liquid, it is pulled with special syringe.
Natural milk mixture is added to hemolymph. In this case 1 gr of dry milk is dissolved at 10 ml distilled water and chicken yolk is added diet medium and put at ultraviolet lamp. It is rotated for 5 minutes at 2000 second speed in centrifuge. As a result, above separated additional oil and foam is distinguished. Diet medium should be kept in the clean, without microbe room and at 20 °C cool [1; 3; 5].
Section 1. Biology
After medium has been ready, it is placed in special artificial caterpillars which are cleaned with 75% ethyl spirit made of politilen.
The following laboratory tools and instruments are necessary to produce artificial caterpillar:
Particular thermostat, test-tube 50-PX, politilen material 4mm, spirit 75%, ultraviolet lamp, petri plate, antiseptic pipette, centrifuge 2500, medical syringe 5 ml are necessity [1; 3; 4; 6].
Produce depicted artificial caterpillar:
A small part (1x1) of parafilm is extended 2 x 4 cm, absorbed in 75% alcohol for 15 minutes. Then it is dried with sterile printed paper, folded as a sack and fixed in order to stick both sides. 0,5 ml of mediums is placed by pipette into each parafilm box. When parafilm box is fulled with mediums, it yields such depicted maggot. 15-20 small holes are opened with sterile entomologic needle for properly prepared artificial caterpillars. All the processes of the preparation are required to carry out in sterile room [1; 3; 4; 5].
Diet mediums which are ready are put in test-tube (50-PX), a day ago flown and nourished Bracon's (Bracon hebetor Say) female genders are flown in 1 : 5 proportion.
In order to separate 100 mostly effective diet mediums,3 kinds of diet mediums are prepared, they are investigated repeatedly 100 times. Their ingredients are following; [1; 3; 4; 6].
First: (A) Hemolymph wax moths (G.melonnella) (Aj) 40,04%, chicken yolk (a3) 30,03%, milk (A4) 29,03%.
Second: (B) Hemolymph wax moths (G.melonnella) (Bj) 45,01%, chicken yolk (B3) 28,02%, milk (B4) 26,07%.
Third: (C) Hemolymph unripe cotton caterpillar (H.armigera Hb) (Cj) 50,04%, chicken yolk (C3) 25,03%, milk (C4) 24,03%.
In order to prevent diet mediums from microorganisms, gentamisin solution is added for each prepared medium 100 ml. i.u 40 thousand - 60 thousand i.u quantity.
Results of the research
In each experiment 200 artificial caterpillar are formed from diet mediums of above structure and each of them is experienced 100 times (1 - picture).
We chose a type of Bracon to be harmed with Bracons. Bracon hebetor Say, this type is tolerant, durable in extreme condition. It is practiced by means of choosing convenient state in developing each type to be harmed diet mediums with Bracon and in this condition it is put in thermostat.
Damaging the type of Bracon with diet mediums at + 28±1 °C temperature, at 68±3% moisture.
Figures 1. Artificial caterpillar and developing the generation of Bracon
To compare Bracon generation of all diet mediums with The indexes of specifying Bracon generation's developing artifi-
(H. armigera Hb) caterpillars, cotton-plant caterpillars are also dam- cially made caterpillars in different expenditure limit. aged with brac Bracon on. (1 - table).
Table 1. - (Experiments of the laboratory, 2014-2016 years) (+ 29 ± 2 °C, RH 78 ± 3%)
Types of diet mediums and their ingredients' expenditure limit% The amount of damage% with parasite for artificial caterpillar. Development degree of Bracon in diet mediums by days Genders proportion <$: ?
W Larva Pupa Undone developing Imago All the numbers of generation days
A 86,5 1,8±0,2 5,4±0,3 4,6±0,4 - 1,5±0,9 13,3±0,8 1:6
A1 A2 A3
40,04 30,03 29,03
B 61,3 1,3±0,5 3.3±0,9 4,8±0,9 1,8±0,2 - 12,2±0,5 1:1
B1 B2 B3
45,01 28,02 26,07
C 73,4 1,7±0,5 4,8±0,2 4.5±0,3 - 2,3±0,2 12±0,3 1:4
C1 C2 C3
50,04 25,03 24,03
To define the biological active matter-flavonoids in medicinal plants
According to it, the first diet medium (A) organizes damaging degree with Bracon 86,6% of all prepared diet medium, It takes about 13,3 days from eggs to imago (mature) period. Imagoes live 5,8 days. It takes 1,8 days to birth larvae from eggs,5,4 days for larva period,4,6 days for pupa period.
Genders of Bracons which went out from medium are male: female proportion will be 1:6.
In next diet medium (B) damaging degree of Bracon mediums will be 61,3%. It takes 1,3 days to go out larvae from eggs, in diet medium caterpillars inhabit 3,3 days and transform to pupa. Pupa period continues 4,8 days, imagoes develop a little and in pupa period they die in spite of observing their flying. In next mediums (C) Bracon damage diet mediums till 73,4%. It takes 1,7 days to appear larvae from layed eggs and they begin to consume nutrition.
Larvae develop for 4,8 days in this diet mediums and pupa period includes 4,6 days. Genders of Bracons flown from medium are male: females proportion is 1 : 4.
Conclusion
It is defined during research that prepared all diet mediums are detrimented with Bracon generation, they layed their eggs there. But some of diet mediums die because of inconvenience for developing parasite generation.
According to mediums' structure the portion of only wax moths hemolymph (A) is 40,4 and in this diet medium Bracon parasites progress continuingly. The reason of it, there is average albumen and oil in the ingredient of hemolymph. They are convenient for developing parasite larva.
Thus, in order to develop and consume of Bracon generation well from above mentioned artificial diet mediums first medium (A) hemolymph wax moths (G.melonella) (Ax) 40,04%, chicken yolk (A3) 30,03%, milk (A4) 29,03% is confirmed as the best diet medium to rear Bracon parasite.
References:
1. Kimsanboev Х. Х., Jumaev R. А. Renewing and rearing technology of Bracon hebetor Say in Biolaboratory; Материалы VIII-оймеждународной научно-практической конференции молодых исследователей, - г. Волгоград, - 2014. - 257-259.
2. Kimsanboyev Khujamurot. Renewing technology of Bracon hebetor Say in biolaboratory - 50. - Tashkent, - 2003.
3. Jumaev R. А., Kimsanboev Х. Х. Using the efficacy of parasite entomophags rared in artificial medium for cotton-plant's agrobiosynos at controlling the number of rodent pests./^ишлок; хужалигини инновацион ривожлантришда олий ва уртамахсус, касб-хунар таълим муассасалари ёш олимларининг рули. - Тошкент, - 2016. - 211.
4. Li Li-ying et al - 1988, The artificial diets ofEocanthecona furcellata (Wolff). Bulletin of Biologic control - 4 (1): 45 (In china) References.
5. Huang Xin - fei - 1986. Study on the use of Habrobracon hebetor. Bulletin of Biological Control - 2 (2): 70-75 (In Chinese).
6. Consoli F. L., Parra J. R. P., - 1999. - In vitro rearing of parasitoids: constraints and perspectives. - Trends in Entomology, - 2: 20-30.
DOI: http://dx.doi.org/10.20534/ESR-17-3.4-5-7
Khujanazarov Uktam Eshtemirovich, Tashkent State Pedagogical University, Docent, the faculty of Natural sciences E-mail: [email protected] Mirkhamidova Parida, Tashkent State Pedagogical University, Doctor of biology, professor, the faculty of Natural sciences E-mail: [email protected] Valikhanova Akida Kamoliddin qizi, Tashkent State Pedagogical University, Bachelor student, the faculty of Natural sciences E-mail: [email protected] Nizamova Aziza Rakhmatulla qizi, Tashkent State Pedagogical University, Bachelor student, the faculty of Natural sciences E-mail: [email protected] Riskitillaeva Nikhola Abduvoxid qizi, Tashkent State Pedagogical University, Bachelor student, the faculty of Natural sciences named after Nizami, Tashkent city, Uzbekistan E-mail: [email protected]
To define the biological active matter-flavonoids in medicinal plants
Abstract: This paper summarizes the general flavonoids quantity, having antioxidant peculiarities where is situated in the southern-western Zarafshan ranges in Kashkadarya basin in medicinal plants such as Origanum tytthanthum, Ziziphora
