SU1405298A1 - Strain of vibrio cholerae 042 serovar bacteria for producing virulent phage 781 serovar - Google Patents

Description

(46) 08/30/90. Bksh. No. 32

(21) 4066689 / 28-13

(22) 04/07/86

(71) All-Union Research Anti-Plague Institute Microb and Rostov Research Anti-Plague Institute

(72) T.A.Kudr Kova, V.P. Moroz, N.M.Ostroumova, O.S.min, I.E. Ushakov, and N.P. Konnov

(53) 576, .8.093 (088.8)

(56) USSR author's certificate

No 701148, cl. C 12 N 7/00, 1980.

(54) VIBRIO CHOLERAE BAKTERIUM STRAIN, 042 SEROVARA, USED TO RECEIVE VIRULENT PHAGA 781 (781c) C- 170 042/1 CELVARI Behavi. phage 781 (781v) C-170 042/1 serovar. The purpose of the invention is to find out the strain of the new 042 serovar, spontaneously producing dego phage 042/1 serovar. The V. cholerae strain is derived from environmental objects and deposited under the number CM-14 (781). Phage .042 / 1 of the serovar produced by the strain 14 (781) can be used. van for identification of enteropathogenic vibrios 042 serovar. 1 tab.

i

(L

This invention relates to medical microbiology and can be used to identify hobby vibrios.

The purpose of the invention is to find out the strain of the new 042 serovar, spontaneously producing phage 042/1 of the serovar.

The strain of Vibrio cholerae deposited at the Microbe Institute under the number KM-14 (781). The strain is derived from environmental objects,

Cultural and morphological. Motile, slightly curved rods 1.5–3 µm in length and 0.2–0.6 µm in width, with one polarly arranged flagellum, do not form spores and capsules, gram-negative.

On dense nutrient media (Martin agar, Hrttinger agar, alkaline m and septone agar pH 7.6 + 0.2) forms round, with a diameter of 3.5i ± 0.5 mm, translucent colonies with a bluish tinge, smooth edge, slightly compacted center after 18 h of cultivation at 37 + 1 C,

On liquid nutrient media (Boules he Martin, 1% - peptone water, Hottinger broth pH 7.8 + 0.2) forms a uniform turbidity and delicate film on the surface of a part of 6 hours of growth at 37 + 1 ° C,

Physiological and biochemical properties. The oxidase test is positive.

Decomposes to acid without gas man

nose, sucrose, glucose, mannitol, n.

decomposes arabinose, ioic, lactose, oxides, and ferments gluchoe into Hugh-Leifson's medium,

Forms lysine decarboxylase, does not form arginine dihydrolase.

Decomposes. Starch, ferments gelatin. Does not grow in the presence of 7-10% sodium chloride.

Serological properties. It is not agglutinated by species O and typical (Inaba, Ogawa) cholera serums, agglutinated by monovar vibratory serum 042 of the serovar, is not agglutinated by monovar serums of others (02-041, 043-056) of serovars,.

It is capable of spontaneously producing a moderate phage of a previously unknown serotype, designated 042/1, whose virulence mutants are used for laboratory diagnostics of enteropathogenic vibrios of V. cholerae 042 serovar.

Q

five

0 5

l

five

0

50

five

The moderate phage 042/1 serotype has the following electron microscopic structure: the head of a hexagonal shape is 300 + 22 A in diameter, has no appendix, i.e., belongs to the small phages II of the morphological group on Systematics A, SaTihokenko (1968), His single The development cycle is characterized by a minimum latency period of 31+ minutes and an average yield of 20 + 3 particles per infected cell.

At the pen of the indicator strain forms negative colonies of round shape 5 + 1 mm. In diameter with uneven edges, dense center and narrow zone of incomplete lysis at the periphery. Virulent mutants differ in the structure of the colonies and the spectrum of lytic action: colonies of the same size, but with a transparent center.

A moderate phage lyses only the indicator strain (K, M 13), virulent mutants of 86-90% of the strains of vibrios of the homologous serovar.

Strain 781 is propagated in broth and marten pH 7.6 + 0.2 on agar, stored in a lyophilized state.

The use of the strain is illustrated with the following example.

- PRI me R. To detect the spontaneous production of moderate phage 042/1 serotype and its virulent mutax, strain-14 (781) is seeded in Martin's broth pH 7.6 + 0.2 and grown at 37 + 1 ° C for 20 + 2 h, and then filtered through F-5 porcelain bacterial filters. The resulting filtrate in an amount of 1.0 ml is added to 5.0 ml of 0.7% Märten agar pH 7.6 + 0.2, melted and cooled to a test tube. 0.2 ml broth culture of a three-hour growth of a special indicator gum (K.M - 13) is added thereto, mixed thoroughly and drunk on an agar plate (Marten's agar) in a Petri dish. After incubation at 37 + 1 ° C for 20 + 2 h on the plate detect negative colonies of moderate phage 041/1 and virulent mutants of the latter.

Using a platinum needle, inject a phage into the center of a transparent negative colony in 10 test tubes from 1.0 ml of Martin's broth, pH 7.6 + 0.2, warmed to 3-7 seconds.

0.2 ml broth culture of strain strainer (K.M. -13) and 5.0 nl 0.7% agar Martin. The contents of each tube are mixed and drunk on a separate plate (Marten's agar) in a Petri dish and allowed to solidify with the second agar layer. After refluxing for 20 + 2 hours, 10 ml of Martin's broth is poured into each dish and mixed with a spatula with the upper agar layer, and a part is transferred into a vial and filtered as indicated.

The resulting filtrate of the virulent mutant is propagated in a liquid medium (Marten broth pH 7.6 + 0.2), 3.0 ml of the filtrate and 1.0 ml broth of a three-hour culture of the strain mentor are added to 100.0 ml of broth (K, M- 13), incubated for 2, + ± 0.5 h before enlightenment, disinfected by filtration through bacteriologists -0

five

about

a sterile filter (as indicated), put the control of sterility and use it for its intended purpose. The table characterizes the specificity and activity of the virulent mutant 781 (781b) C-170 phage 042/1, from which it is clear that the virulent mutants of this phage have a wide spectrum The lytic effect on cultures homologous O serovar is enteropathogenic vibrios and highly specific.

The use of the invention for identifying enteropathogenic vibrioes O42., A serovar increases the efficiency of laboratory diagnostics of NAG infections,

Claims (1)

Invention Formula Bacterial strain Vibrio cholerae 042 serovar, KM-14 (781), used to produce virulent phage 781 (781c) C-170 042/1 serovar. The spectrum for the specificity of the unreth phage serotype 042/1, it is its vi  iutvkta 781 (781в) С-170 50 SO 782 34 36 20 39 fft