RU2327347C1 - Nutrient medium for cultivation of larva wohlfahrtia magnifica - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: wohlfahrtia magnifica of all development stages. The nutrient medium contains hydrolyzate of the cattle muscular tissue, defibrinated blood of sheep, yeast extract, glucose, agar-agar, penicillin, streptomycin, and Hanks solution.

EFFECT: output of the fully developed larva increases; the larva produce is getting cheaper.

4 ex

Description

FIELD OF THE INVENTION

The invention relates to veterinary medicine, mainly to veterinary parasitology, and can be used for artificial breeding of Wohlfahrtia magnifica larvae of all developmental stages used for laboratory, in particular toxicological, studies necessary to develop new effective means and methods of combating wolfarthiosis.

State of the art

Known nutrient medium for the cultivation of larvae of gastric gadfly, containing a five percent fish hydrolyzate, Hanks solution and horse blood serum (Somov S.L., Somova N.M., Krasyukova L.S. Method for cultivating larvae of gastric gadfly. A.S. No. 1001900 A01K 67/00, 1983).

However, this method of cultivation and a nutrient medium cannot be used for the cultivation of larvae of the tungsten fly, since these parasites differ in the method of parasitization and developmental cycles.

The closest in technical essence and the achieved positive effect to the proposed nutrient medium and adopted by the authors for the prototype is a nutrient medium for growing larvae of tungsten fly, which is used as a dry serum of cattle, which is placed in a gauze bag, wetted with distilled water and riboflavin, which fits on pieces of cattle skin with larvae of flies.

However, the method does not provide continuous cultivation of all phases of the development of insects, and the number of pupae obtained depends on a complex of abiotic factors and is characterized by the breeding of inferior insects from 35 to 40% (Ternova V.I. Problems of Veterinary Sanitation, Transactions of VNIIVS, M, 1962, T. XXI p. 84-87).

Disclosure of invention

The task of the invention is the creation of an artificial nutrient medium that provides a sufficiently high yield of fully formed larvae of Wohlfahrtia magnifica.

The technical result, which can be obtained using the present invention, boils down to the possibility of obtaining in laboratory conditions, regardless of the time of year, fully formed larvae of the wolfarth fly of various developmental stages for conducting studies aimed at combating wolfarthiosis, and the cost of obtaining larvae.

The technical result is achieved using a nutrient medium for the cultivation of Wohlfahrtia magnifica larvae containing a hydrolyzed cattle muscle tissue, defibrinated sheep blood, glucose, agar-agar, yeast extract, streptomycin, penicillin and Hanks solution in the following ratio of components (g):

The muscle tissue hydrolyzate of cattle (FGM) - 30.5-31.5

Defibrinated sheep blood - 10.0-11.0

Glucose - 5.0-6.0

Agar-agar - 1.0-1.5

Yeast extract - 0.3-0.4

Streptomycin - 0.05-0.06

Penicillin - 0.05-0.06

Hanks solution - 53.1-49.48

The essence of obtaining a nutrient medium:

In 53.1-49.48 g of Hanks solution, 1-1.5 g of agar-agar is added and placed in a water bath at a temperature of 50-52 ° C until the agar-agar is completely dissolved with periodic stirring. In a warm agar-agar solution, 30.5-31.5 g of cattle muscle tissue hydrolyzate, 5-6 g of glucose, 0.3-0.4 g of yeast extract, 10-11 g of defibrated blood of sheep are added, 0.05 -0.06 g of streptomycin and 0.05-0.06 g of penicillin.

Then the medium is poured into Petri dishes. Cultivation is carried out in a thermostat at a temperature of 38-40 ° C and a humidity of 70-75%. The duration of cultivation is 5-7 days. The yield of larvae is 79-85%. The yield of flies from larvae obtained on an artificial nutrient medium is 68-72%.

The nutrient medium cannot be stored and is prepared immediately before the start of cultivation.

The implementation of the invention

Examples of specific performance of obtaining a nutrient medium for the cultivation of larvae of Wohlfahrtia magnifica.

Example 1

The ratio of the components of the medium is (g):

Cattle muscle tissue hydrolyzate - 20.0

Defibrinated Sheep Blood - 5.0

Glucose - 3.0

Agar Agar - 0.5

Yeast extract - 0.2

Streptomycin - 0.1

Penicillin - 0.1

Hanks Solution - 71.1

In 71.1 g of Hanks's solution, 0.5 g of agar-agar is added and placed in a water bath at a temperature of 50-52 ° C until the agar-agar is completely dissolved with periodic stirring. In a warm agar-agar solution, 2 g of cattle muscle tissue hydrolyzate, 3 g of glucose, 0.2 g of yeast extract, 5 g of sheep defibrinated blood, 0.1 g of streptomycin and 0.1 g of penicillin are added.

Then the medium is poured into Petri dishes. Cultivation is carried out in a thermostat at a temperature of 38-40 ° C and humidity - 70-75%. The duration of cultivation is 5-7 days.

When cultivating tungsten fly larvae on a medium with such a composition, the larval yield was 41–43%, and the yield of flies was 23–25%.

Example 2 is carried out analogously to example 1, but the ratio of the components of the medium is (g):

Cattle Muscle Tissue Hydrolyzate - 30.5

Defibrinated Sheep Blood - 10.0

Glucose - 5.0

Agar Agar - 1.0

Yeast extract - 0.3

Streptomycin - 0.05

Penicillin - 0.05

Hanks solution - 53.1

When cultivating tungsten fly larvae on a medium with such a composition, the larval yield was 79-85%, and the yield of flies was 68-72%.

Example 3 is carried out analogously to example 1, but the ratio of the components of the medium is (g):

Cattle muscle tissue hydrolyzate - 31.5

Defibrinated Sheep Blood - 11.0

Glucose - 6.0

Agar Agar 1.5

Yeast Extract - 0.4

Streptomycin - 0.06

Penicillin - 0.06

Hanks solution - 49.48

When cultivating tungsten fly larvae on a medium with such a composition, the larval yield was 79-85%, and the yield of flies was 68-72%.

Example 4 is carried out analogously to example 1, but the ratio of the components of the medium is (g):

Cattle muscle tissue hydrolyzate - 35.0

Defibrinated Sheep Blood - 16.0

Glucose - 15.0

Agarose - 3.0

Yeast Extract - 1.0

Streptomycin - 0.5

Penicillin - 0.5

Hanks Solution - 29.0

When cultivating tungsten fly larvae on a medium with such a composition, the larva yield was 61-65%, and the yield of flies was 65-67%. The decrease in the yield of larvae is explained by the excess content of nutrients in the medium and the violation of their ratio.

Thus, examples 2 and 3 are the most optimal, because the yield of larvae is the highest and is 79-85%, and the brood of flies is 68-72%.

Tests of the invention showed that the proposed nutrient medium allows to obtain a sufficiently high yield of Wohlfahrtia magnifica larvae in laboratory conditions without the use of laboratory animals, which will allow more efficient studies aimed at developing advanced methods and methods of combating wolfarthiosis.

The invention in comparison with the prototype and other known technical solutions has the following advantages:

- the possibility of obtaining in laboratory conditions fully formed larvae of the tungsten fly;

- the cost of obtaining a nutrient medium;

- high yield of larvae;

- laboratory studies regardless of the time of year.

Muscle tissue hydrolyzate preparation

Cattle meat is cleaned of bones, fat and films, minced in a meat grinder. Stuffing is placed in a wide bottle and filled with 4 times the amount of distilled water (per 1 kg of stuffing - 4 l of water). For each liter of the mixture add 30 ml of hydrochloric acid (specific gravity 1.19). The contents of the bottle are mixed well, the bottle is closed with a stopper and put into an autoclave. Hydrolysis is carried out at 2 atm for 3 hours. Then the hydrolyzate is defended and filtered. The Kjeldahl total nitrogen content is 800-1000 mg /%, amine nitrogen is 500-600 mg /%, the degree of hydrolysis is 0.60.

Preparation of Yeast Extract

250 g of pressed baking yeast are suspended until a homogeneous mass is formed in 1 liter of distilled water. The suspension is boiled for 4-5 minutes, stirring continuously. The microbial mass is precipitated by centrifugation at 3-4 thousand rpm for 30 minutes. The supernatant is drained and sterilized through a Seitz filter. The extract is stored in a dark colored container at 4-5 ° C and used for 8-10 days from the date of manufacture.

The nutrient medium cannot be stored and is prepared immediately before the start of cultivation.

Claims (1)

Nutrient medium for cultivation of Wohlfahrtia magnifica larvae containing cattle muscle tissue hydrolyzate, defibrinated sheep blood, glucose, agar-agar, yeast extract, penicillin, streptomycin and Hanks solution in the following ratio of components, g: Cattle Muscle Hydrolyzate 30.5-31.5 Defibrinated Sheep Blood 10.0-11.0 Glucose 5.0-6.0 Agar agar 1.0-1.5 Yeast extract 0.3-0.4 Streptomycin 0.05-0.06 Penicillin 0.05-0.06 Hanks Solution 53.1-49.48