SU1017726A1 - Strain vibrio cholerae eltor 673/67thy for use as test culture for detecting thymidine in culture media - Google Patents

Abstract

SHTAMM VIBRIO CHOtERAE ELTOR: B73 / 67 {Muei of living cultures of the All-Union NI of the Anti-Plague Institute Microbe used as a test culture: for determining tidshdine in nutrient media.

Description

The invention relates to the field of microbiology, and concerns the strain νΐΒτΐο cobalt e1COG 673/67 ^^ ^- which can be used for the microbiological determination of thymidine in various liquid and dense nutrient-5 media in the study of the quality of culture media for the diagnosis of cholera.

The aim of the invention is the creation of a strain of νϊότΐο with 10-1073 667/67 ^{+ b} * - used as a test culture to determine thymidine in nutrient media.

The strain was obtained by exposure to supermuntangen (nitrosoizanidine) d5 'for the delizogenized variant V.e. 673/67, followed by selection on trimethoprim. .

Stored in the collections of the All-Union Museum of Living Cultures of Pathogenic Microorganisms of the Institute "Microbe." under the number 673 / 67ΙΦ * -. ·

Strain Uggho so1egae eShrg b? 3/67 * ^'Ly harakterieuetsya following svoystvg.mi. .

Cultural and morphological features. The cells of the daily culture of the strain are slightly curved Gram-negative rods with a moderately pronounced polymorphism of magnitude 1.5-2.0 x 9.3 - 0.4 μm.

Agar Martin with the addition of 10 mg / ml thymidine (pH 7.6-7.8). After 12–16 hours, rounded flat colonies with a smooth edge, transparent in passing, 35 * tsishy, and slightly bluish in reflected light, grow.

Broth Martin and 1% peptone water with an alkaline pH value. The cells of the strain give a uniform clouding of the df with a delicate film on the surface.

Glucose-mineral synthetic media (Bhaskaran and Rowley M-9 medium) pH 7.8-8.0 with the addition of 10 μg / ml thymidine. Translucent ⁴⁵ colonies are formed with smooth smooth edges 1.5-2.0 mm in size with slightly slower growth (24 hours).

Physiological and biochemical signs. The strain possesses an active active oxidase-50> tures up to acid without gas;:;., Glucose in the medium of Hugh and Lyufson under aerobic and anaerobic conditions.

• decarboxylate lysine and do not have I ... give arginine dihydrolase. On Wednesday 55 Guissa ferments sucrose, mannose, mannitol. Arabinose and lactose do not collect _{: it} lives. It has diagnostic activity. Forms indole and does not form hydrogen sulfide. Produces acetyl-, methylcarbinol. Not homolytic. Grows on medium with the addition of 50 µg / mp

VNIIPI Order 3481/28

l

[polymyxin. Forms vibrios. Not lysogenic.

It is lysed by cholera bacteriophage Eltor. Agglutinated to titer by species cholera O-serpent and typical Ogawa serum.

Optional aerobic.

A feature of the strain is a chromosomal mutation, which determines its dependence on exogenous thymidine additives (0.1-10 μg / ml medium).

Determination of thymidine is carried out both in liquid media by a test tube method in a series of standard thymidine solutions in the range of 0.1-10 μg / ml, and in dense media by a plate method in a series of standard solutions of thymidine 0.5-50 μg / ml.

Example. The determination is carried out in a minimal salt medium M-9, Ia ^ NRS ^ (anhydrous) 6g, KH2PO4, (anhydrous) Zg, NaCi 0.5 g ΝΗήΰΙ 1 g, 20% glucose solution 20 ml, 0.1 M solution MDO3 ^ 10 ml, 0.01 M solution of £ aC1 ₂ 10 ml, distilled water up to 1000 ml, pH 7.6,

Prepare two parallel rows of tubes with consecutive tenfold dilutions of the studied nutrient medium. Bred to. 10 ^- ^ minimal environment without the addition of thymidine.

To construct a standard growth curve for the test shgamma, two parallel rows of tubes are filled with 4.5 ml of synthetic medium (4.5 ml) with the addition of thymidine in concentrations of 100, 10, 1, 0.1 and 0.01 μg / ml. Then in each tube, both in the experimental and in the standard rows, 0.5 ml of culture suspensions of the test strain are introduced. Crops are kept at 37 ° C for 24 hours, after the test, the intensity of the growth of the strain is recorded by measuring the turbidity of the medium with a nephelometer. -

Claims (1)

Indicator; the culture was preincubated in nutritious broth of pH 7.6 for 18 hours with addition of up to 10 μg / ml of thymidine and washed twice with broth buffer from the broth using Centrifugation before starting the assay. The cell concentration is adjusted to 10 ⁱⁿ cells / ml of saline. The use of the strain allows the determination of thymidine at a concentration of 0.350 μg / ml of medium. The proposed strain can be used to solve the problems of butymic mutagenesis, to identify the causes of cell death during the thymine starvation, to synchronize the division of bacteria and for other purposes.