SU1382850A1 - Method of determining mobility of non-fermenting gram-negative microorganisms - Google Patents

Description

(21) 4103384 / 28-13

(22) 08/07/86

(46) 03/23/88. Bul. No. 11

(71) Moscow Research Institute of Hygiene. F.F. Erisman

(72) G.M.Trukhina, N.B.Komzolov and G.P. Kalina

(53) 576.8.093 (088.8)

(56) Labinsk A.S. Microbiologists with the technique of microbiological research .- M .: Medicine, 1978,

p.350, 60.

(54) METHOD FOR DETERMINING THE MOBILITY OF NONFERNENTIUM TYPE OF GRADEMARKET MICROORGANISMS

(57) The invention relates to medical microbiology and can be applied in the diagnosis of diseases caused by bacteria of the non-fermentative group of gram-negative

microorganisms (UFHOM) in the pathological material, with hygienic and epidemiological studies of the environment. The purpose of the invention is the accuracy of the method. The method is characterized by the use of a two-layer medium in bacteriological cups, in which the lower layer consists of 1.2-1.8 wt.% Agar in distilled water, and the upper layer — 1.2-1.8 wt.% Agar in meat infusion. with the addition of 0.8-1.2 wt.% peptone, 0.75-1.25 wt.% sodium chloride, 0.00018-0.00022 wt.% crystal violet. When sowing a prick 12-18 test strains in one pack and incubating at room temperature for 1-2 days, the immovable species grow as a small colony at the site of a prick with a diameter of 1-3 mm, and mobile species grow as wide plaques with a diameter of 12-20 mmo 2 table .

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The invention relates to medical microbiology and may not be used in the diagnosis of diseases caused by non-fermentative negative microorganisms. The aim of the invention is to improve the accuracy of the method.

Example 1. In a bacteriological cup with a double lid (paper and glass), pour 20-25 ml of the medium (bottom layer), rods, mass%: agar 1.5; water is distilled the rest, sterilized at 1 atm for 20 min. After 15 | freezing of the medium under the paper surface, the upper layer of the medium is poured. The top layer is prepared as follows. Peptone (1.0 wt.%) 5, agar (0.35 wt.%), NaCe (1.0 wt.%) Is added to the meat infusion, the volume of the medium is adjusted to 1 l, sterilized at 1 ATM 20 min. Separately, a 0.01% solution of crystal violet in distilled water is prepared and injected into the upper layer at the rate of 0.0002 per 1 l of medium.

The resulting nutrient medium in the amount of 20-25 ml layered on the frozen bottom layer. After solidification of the medium, the cultures or colonies inoculated were sowed from dense media to the bottom of the cup. It is possible to sow up to 12,218 strains at the same time. After sowing 5, the cup is closed on top of the glass of paper. Incubate crops at 20-22 C for 12-18 h (pre-counting) and an additional 20-22 h (final metering). Then, 30-40 of the Seed is determined.

, Positive result: a growth zone around the seeding with a puncture in the form of a rim with a diameter of 12 mm or more.

Negative result: small size of the colon or point growth at the injection site (diameter 3 mm or less),

The mobility of microorganisms is shown in table 1,

Example 2. The method is carried out in accordance with Example 1, but the components of the medium are taken in the following

thirty

45

50

ratios, wt.%: bottom layer - agar 1,2, distilled water - up to 1 l; the top layer is agar 0,3; peptone 0.8; NaC 0.75; crystal violet 0.00018, m infusion up to 1 liter.

Example 3. The method is carried out in accordance with Example 1, but the components of the medium are taken in the following ratios, wt%: the lower layer — agar 1.8; distilled water - up to 1 l, the top layer - agar 0.4; peptone. 1.2; NaCP 1, -5; crystal violet 0.00022; mew infusion up to 1 l.

Comparative data to determine the mobility of non-fermentative gram-negative bacteria are given in table 2.

Using the proposed method allows to increase the accuracy of determining the mobility of non-fermentative gram-negative bacteria.

Claims (1)

Invention Formula The method for determining the mobility of non-fermentative gram-negative microorganisms by seeding the culture under study by injecting into a nutrient medium containing meat infusion, peptone, MaSb, agar, followed by incubating the crops and determining the mobility according to the growth pattern of the microorganisms, o. t l and chu and with the fact that, in order to improve the accuracy of the method, use a nutrient medium consisting of two layers, while the additional bottom layer contains, wt.%: Agar1,2-1,8 Distilled water the rest and the top layer additionally contains crystal violet in the following proportion components, wt.%: Agar 0.3-0.4 Peptone 0.8-1.2 NaCfi0.75-1.25 Crystalline Violet 0.00018-0.00022 M a clear infusion The rest of the incubation of the cultures is carried out at. 20-22 ° C, and the mobility of bacteria is determined by the presence of 1 non-thin plaques with a diameter of 12 to 20 mm at the injection site. Kind of bacteria Citrobacter spp. Enterobacter spp E.coli Ps. aeruginosa Klebsiella pneumonia Klebsiela ozenae Str. faecalis Acihetobacter caleoaceticus table 2 Correct answer. Type of bacteria Number-,. % for method property predla strains known iss rs 1 day 2 day 1 day 2 day Pseudomonas aemginosa80OO96 100 Acinetobacter spp. 320OO100 100 I Compiled by A.Zavadska Editor N.Gunko Tekhred L.Oliynyk Proofreader I.Erdeyi Order 1263/22 Circulation 520 Subscription VNIIPI USSR State Committee for inventions and discoveries 113035, Moscow, Zh-35, Raushsk nab., 4/5 Production and printing company, Uzhgorod, Projecto st., 4 Table 1 Immobility + + + + +