Nutrient medium for culturing plague microorganism vaccine strain

FIELD: biotechnology, microbiology.

SUBSTANCE: invention relates, in particular, to preparing nutrient media used for culturing the plague microorganism vaccine strain and can be used in medicinal microbiology. The nutrient medium for culturing the plague microorganism vaccine strain comprises additionally as a stimulating additive sodium sulfite and as a nutrient base - soybean fruits enzymatic hydrolyzate in the following ratio of components, g/l: microbiological agar, 11.0-13.0; soybean fruits enzymatic hydrolyzate, 250.0-350.0; sodium chloride, 4.5-5.5; sodium hydrogen phosphate, 3.5-4.5; sodium sulfite, 0.0003-0.0005; distilled water, the balance. Invention provides enhancing the growth property of nutrient medium.

EFFECT: valuable properties of medium.

3 ex

 

The invention relates to biotechnology and, in particular, to obtain nutrient media for cultivation of the plague microbe.

Known nutrient medium of the following composition, g/l: agar-agar to 20.0; peptone - 10,0; sodium chloride - 5,0; distilled water to 1 l; 20% sodium hydroxide to a pH of 7.2 ("Handbook of microbiological and virological methods", Birger MO, 1972, 49). The disadvantage of this environment is its low productivity.

Closest to the proposed invention relates nutrient medium for cultivation of microorganisms including, g/l: agar-agar - 20,0; gidrolat beef containing amine nitrogen of 0.12%; sodium chloride - 5,0; 20% sodium hydroxide - 0,002; distilled water to 1 l ("Handbook of microbiological and virological methods", Birger MO, 1972, p.53). The disadvantage of this environment is its high cost.

The aim of the invention is to obtain high quality cheap a nutrient medium for cultivation of vaccine strains of the plague microbe. The essence of the invention lies in the fact that the nutrient medium contains a nutrient basis of enzymatic hydrolysate of soy beans and stimulating additive - sodium sanitarily, in the following ratio of ingredients, g/l: microbiological agar - 11,0-13,0 enzymatic guy is raisat soybeans - 250,0-350,0 sodium chloride - 4,5-5,5 sodium phosphate 2-substituted - 3,5-4,5 sodium sanitarily - 0,0003-0,0005 distilled water - the rest. As a source of raw materials use soy, fruits of which beans contain about 40% protein, in addition, soy is rich in trace elements and vitamins. As you know (Y.P.Yupta, 1987) soybean proteins are characterized by a high content of glutamic and aspartic acids, which are closely associated with carbohydrate metabolism through the tricarboxylic acid cycle, supplying high energy communication. Soy proteins also contain a large number of such essential amino acids as lysine, leucine, arginine. There is also information about the ability of leucine, such as amino acids from protein branched chain, to initiate protein synthesis (ei Chazov, Hsian /USA/ 1989).

Preparation of nutrient media for cultivation of microorganisms is as follows. Fruits soya beans, in the amount of 0.5 kg, five-soaked in 5 l of hot water during the day, and last infusion of beans boiled for 40 minutes Then cast infusion in 10 l of the cylinder is cooled, the beans are crushed by a grinder and placed in a flask. Add the pancreas of cattle based 20,0 1 l, set pH to 8.2 20%solution of sodium hydroxide in the amount of 0,0003 add 1% chloroform. Hydrolysis are in a heat chamber at 37°in the course is the development of three days. The first 2 hours and the mixture is stirred every 15 min for 5 min, subsequently the mixture is stirred every 6 hours Daily for measuring the level of amine nitrogen, which on the third day of the hydrolysis reaches 0.6 + 0,05%.

The nutrient medium on the basis of soy enzymatic hydrolysate for the cultivation of microorganisms is prepared as follows. Enzymatic hydrolysate of soybean, diluted with distilled water to testimony amine nitrogen of 0.14% - 300 ml; sodium chloride - 5,0:, sodium phosphate 2-eamestly - 4,0; sodium sanitarily to 0.0004; 20%solution of sodium hydroxide 3 ml; agar microbiological - 12, 0mm; distilled water - the rest.

Wednesday boil to dissolve the ingredients, filtered through a cotton-gauze filter, then set the pH to 7.2±0.1 to 20%aqueous solution of sodium hydroxide, poured into vials are sterilized at 0.5 psi for 30 minutes

As an example, experienced the culture of the vaccine strain of the plague microbe was grown on plates, 2% agar of Hottinger pH to 7.2 at a temperature of 28°With during the day. Then prepared microbial suspension 1-day-old cultures of the test strains equal to 10 IU optical standard turbidity on CCA gisk named after. L.D. Lytvyn, then serial tenfold dilutions in physiological solution of 4.5 ml conveyed to the content in 1 ml 1000; 100 is C From each dilution of the suspension cultures were sown by 0.1 ml into three Petri dishes. Crops were incubated at 28°in terms of thermostat. Analysis was performed 24-48 hours

Example 1. The test strain was grown on a nutrient medium containing, g/l: microbiological agar - 11,0; hydrolyzed soy beans - 250,0; sodium chloride and 4.5; sodium phosphate 2-substituted - 3,5; sodium sanitarily - 0,0003; 20%solution of sodium hydroxide and 2.5; distilled water - the rest. With this ratio of ingredients, the number of grown colonies was 51 with a diameter of 1 mm.

Example 2. The test strain was grown on a nutrient medium containing, g/l: microbiological agar - 12, 0mm; hydrolyzed soy beans - 300,0; sodium chloride - 5,0; sodium phosphate 2-substituted - 4,0; sodium sanitarily to 0.0004; 20%solution of sodium hydroxide to 3.0; distilled water - the rest. With this ratio of ingredients, the number of grown colonies was 73 with a diameter of 1 mm.

Example 3. The test strain was grown on a nutrient medium containing, g/l: microbiological agar - 13,0; hydrolyzed soy beans - 350,0; sodium chloride - 5,5; sodium phosphate 2-substituted - 4,5; sodium sanitarily - 0,0005; 20%solution of sodium hydroxide and 3.5; distilled water - the rest. With this ratio of ingredients, the number of grown colonies was 50 di the meter up to 1 mm.

Thus, the claimed nutrient medium prepared on the basis of enzymatic hydrolysate of soy beans, can replace the nutrient medium on the basis of beef.

Nutrient media for cultivation of vaccine strain of the plague microbe containing a nutrient basis, microbiological agar, sodium chloride, sodium phosphate 2-substituted, distilled water, characterized in that it further comprises as stimulating additives sodium sanitarily, as well as nutrition - enzymatic hydrolysate of fruits soya beans in the following ingredients, g/l:

Agar microbiological 11,0-13,0

Enzymatic hydrolysate of soy beans 250,0-350,0

Sodium chloride, 4,5-5,5

Sodium phosphate 2-substituted 3,5-4,5

Sodium sanitarily 0,0003-0,0005

Distilled water the Rest



 

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