SU1298245A1 - Nutrient medium for differentiating enterobacteria - Google Patents

Abstract

This invention relates to medical microbiology and can be applied in the bacteriological diagnosis of dysbacteriosis. The purpose of the invention is to enhance the differentiating properties of the medium. For this, the nutrient medium for differentiation of the bacterium contains, g / l: as a nutrient base, dry tryptic protein hydrolyzate 6.0-9.0, dry extract of fodder yeast 0.4-0.6, agar 15.6-23 , 4, sodium alkylbenzenesulfonate 0.6-0.75 as a surfactant, neutral red-O as an indicator, 025-0.036, sodium tartrate acid 0.9-1.1, lactose 10.0 - 12.0, sodium carbonate 0.5 - 1.0, distilled water - the rest. The medium is boiled at 2-3. min, filtered, poured into sterile Petri dishes. Then the cups with medium are dried in a thermostat. Medium ready for use. 1 tab. Q S CL N3 (G) 00 y Ni ;; cl

Description

The invention relates to medical microbiology and may be used in the bacteriological diagnosis of dysbiosis,

The purpose of the invention is to enhance the differential properties of the medium,

Example 1.9 g of dry tryptic hydrolyzate of belcosine, 0.6 g of dry extract of fodder yeast, 0.75 g of sodium alkylbenzenesulfonate, 0.036 g of neutral red, 23.4 g of agar-agar, 1 g of sodium wine-sour acid and 12.0 g of lactose is dissolved in 1 liter of distilled water in a flask. The addition of 1.0 sodium carbonate establish pH 7,5 tO, l

The medium is boiled for 2-3 minutes, filtered and poured into 25 ml of sterile petri dishes and left on the table for cpejpj to solidify with the lids closed. Then the cups with the medium and the lids are separately dried in a thermostat until the moisture condenses on the lids of the cups, and the latter are closed with the lids and (: the water is ready for use. The color of the finished medium is pink.

Example 2. Prepare the medium according to Example 1, but 7.5 g of dry tryptic Belkozin hydrolyzate, 0.5 g of dry yeast extract, 0.65 g of sodium alkyl benzene, 0.03 g are added to 1 liter of distilled water. neutral red, 19.5 g of agar-agar, 1.0 sodium tartaric acid, 11.0 g of lactose, and 0.8 g of sodium carbonate.

PRI me R 3. Prepare the medium as in Example 1, but in 1 liter of distilled water 6.0 g of dry tryptic hydrolyzate of belcosine, 0.4 g of dry extract of fodder yeast, 0.6 g of sodium alkyl benzene sulfonate, 0.025 g neutral red, 15.6 g agar-agar, 0.9 g sodium tartaric acid, 10 g lactose and 0.5 g sodium carbonate.

Example 4. The medium can be prepared in a dry form.

A ball mill with a capacity of 1 kg is loaded with 75 g of dry tryptic hydrolyzate of belcosine, 5 g of dry extract of fodder yeast, 6.5 g of alkylbenzenesulfonate sodium, 0.3 g of neutral red, 195 g of agar-agar, 10 g of sodium tartaric acid , 110 g lactose, 3 g sodium carbonate

five

0

0 5

l

go The mixture is stirred for 45 minutes.

A portion of the substance 40 g (it contains the optimal amount of the indicated ingredients) is added to 1000 ml of water, boiled for 2-3 minutes, filtered, poured into 25 ml in Petri dishes.

Similarly receive drugs with a content of minimum and maximum concentrations of ingredients. The weight is 3.4 and 4.8 g, respectively, per liter of medium.

The medium is used as follows.

EXAMPLE 5 For the preparation of microbial suspensions, daily cultures of test strains, grown on stubble nutrient agar at 37 ° C, are washed off with 0.9% isotonic sodium chloride solution, and the suspension is adjusted to 10 units. optical turbidity standard, which corresponds to 10 microbial cells in 1 ml.

Then, serial tenfold dilutions are brought to a content of 10 thousand (lO), 1 thousand (10) and (10), 100 microbial cells in 1 ml of suspension.

A mixture of cultures is also prepared by mixing lactose-negative strains (Shigella, Salmonella, Prote) with lactose positive Escherichia coli from a dilution of 10 in a 1: 1 ratio, which corresponds to 5 thousand microbial cells of each culture in I ml.

From dilutions 10 and 10 of each suspension monocultures, with the exception of staphylococcus, which is sown from a dilution of 10 ° (100 million microbial cells in 1 ml), and mixtures of cultures are sown 0.1 ml per cup with the proposed and known media.

Comparison media are; Wednesday HS and Levin Wednesday.

Recording of results is carried out after 18-20 hours of incubation of crops at 37 C. Sensitivity, stability of the initial biological properties, and growth rate of the tested test strains on the experimental and known media of the same order: growth of test strains is noted (S. flexneri No. 8516, S. sonnei, S-form, S. dysent. 1, S. typ-hi H901, E. coli 3912/41) when seeding 10 microbial cells (10).

Differentiating properties: when sowing a mixture of crops on a test environment, there is a clear differentiation

pathogenic enterobacteriaceae (Salmonella, Shigella), non-lactose-fermenting colorless colonies from Escherichia coli, fermenting lactose and stained in raspberry.

Diffuse staining of the medium, including in the accumulation of colonies of Escherichia coli, is not noted, which is a positive sign of the quality of the medium that ensures the isolation and further study of the cultures.

On the comparison medium of VShD diffuse yellowing of the medium is noted, on the formula

the invention

Nutrient medium for the differentiation of eterobacteria,;, containing 5 nutrient base, carbohydrate, indicator, surfactant agar, distilled water, in particular, in order to enhance the differentiating properties, it also contains sodium wine sour acid and sodium carbonate, dry extract of fodder yeast as a nutrient base; it contains dry tryptic hydroline

some of which are pathogenic microorganisms of belkoin as a carbohydrate — we differ from non-pathogenic only lactose, as an indicator in the degree of transparency and diameter neutral red, in the quality of colonies, but not in the main adverbially active substance — al- ku, pledged on Wednesday - according to sodium colored benzene sulphonate during guardianship of colonies, which is much more difficult for the ratio of components, g / l:

There is no allocation of the desired culture.

The inhibition index determined when sowing staphylococcus (10 million microbial cells) and vulgar protea (1000 a crab cells) on the experimental environment and the HSV, paBHqpHa4eH: the staphylococcus does not grow on both environments and the protey does not grow.

On Levin's medium, taken as control, there is a confluent growth of staphylococcus and the phenomenon of protein

The results of approbation of the differential-diagic medium (neutralized lactose agar, 1,2,3 series average data) at the 11th stage (in the study of feces) are shown in the table.

Wednesday Levin 105

15 14.2 30 28.5 Differentiation

missing in U.2a

Editor N.Rogulich

Compiled by G.Smirnova

Tehred A. Kravchuk Proofreader and. Erdegsh

Order 862/26 Circulation 500 Subscription

VNIIPI USSR State Committee

for inventions and discoveries 113035, Moscow,, Raushsk nab., 4/5

Production and printing company, Uzhgorod, Projecto st., 4

rmula

the invention

Nutrient medium for the differentiation of eterobacteria,;, containing a nutrient base, carbohydrate, indicator, surfactant, agar, distilled water, about one hundred percent, that, in order to enhance the differentiating properties, it additionally contains sodium sour wine acid and sodium carbonate, dry extract of fodder yeast, it contains dry tryptic hydrol as a nutrient base

for belkozina as a carbohydrate - lactose, as an indicator - neutral red, as a surface-active substance - sodium alkyl benzene sulfonate with a lower ratio of components, g / l:

with her

6.0-9.0

0.4-0.6 15.6-23.4

0.6-0.75 0.025-0.036

0.9-1.1 10.0-12.0

0.5-1.0 Else

No growth, X

8.6 3.8

Claims (1)

The formula of the invention is that the selection and further study of cultures. In the comparison medium, the HFS marks a diffuse yellowing of the medium, for which pathogenic microorganisms differ from non-pathogenic only in the degree of transparency and diameter, not by colony, but not by the main feature laid in the medium - by colony coloration, which makes it difficult to isolate the desired culture. A nutrient medium for the differentiation of enterobacteria ^ containing a nutrient base, carbohydrate, indicator, surfactant, agar, distilled water, characterized in that, in order to enhance the differentiating properties, it additionally contains sodium tartaric acid and sodium carbonate, dry extract fodder yeast, as a nutrient base it contains dry tryptic hydrolyzate of proteinosin as a lactose carbohydrate, neutral red as an indicator, as a surfactant - al _with sodium kipbenzenesulfonate in the following ratio of components, g / l: The inhibition rate, determined by seeding staphylococcus (10 million microbial cells) and vulgar protea (1000 microbial cells) in the experimental medium and the HCV medium, is equivalent: staphylococcus does not grow on both media and does not swarm. On Levin's medium, taken as a control, there is a confluent growth of staphylococcus and the phenomenon of swarming of the protea. The results of testing the differential diagnostic environment (neutralrot lactose agar, series 1,2,3 -35 average data) at stage 11 (during the study of bowel movements) are shown in the table. Dry triptych hydrolyzate cue belcosine 6.0-9.0 Dry extract fodder yeast 0.4-0.6 Agar 15.6-23.4 Apkylbenzene- sulfonate sodium 0.6-0.75 Neutral red 0,025-0,036 Sodium Vin sour sour 0.9-1.1 Lactose 10.0-12.0 Sodium carbon sour 0.5-1.0 Distilliro Rest bathroom water --- Wednesday Total analyzes performed ' ι · ι * ~ ·· »» »............ Swarm of Proteus Growth of coccal flora Differentiation properties Lack of growth,% Neutropyr Lactoic Agar ¹⁰⁵ _ in. Clear 9 8.6 Wednesday Levin 105 15 14.2 30 28.5 Differentiation 6 3.8 absent in 14.2X Corrector I. Erdeyi