CN105733974B

CN105733974B - Selective enterococcus enriched medium

Info

Publication number: CN105733974B
Application number: CN201410755242.1A
Authority: CN
Inventors: 明儒成; 李汝期
Original assignee: Individual
Current assignee: Individual
Priority date: 2014-12-11
Filing date: 2014-12-11
Publication date: 2019-02-22
Anticipated expiration: 2034-12-11
Also published as: CN105733974A

Abstract

The present invention provides selective enterococcus enriched medium, its technical solution are as follows:: Shi peptone 5.0g composed of the following components, 2.5 g of yeast extract powder, 3.0 g of beef extract, 20.0 g of glucose, 4.0 g of lactose, 5.0 g of sodium chloride, 2.5 g of Sodium azide, 10.0 g of bovine bile, 0.1% bromocresol green 20% ethanol water 6.0ml, distilled water 1000ml.Cheng Qu Shi peptone 5.0 g, 2.5 g of yeast extract powder, 3.0 g of beef extract, 20.0 g of glucose, 4.0 g of lactose, 5.0 g of sodium chloride, 2.5 g of Sodium azide, 10.0 g of bovine bile, in 1000ml conical flask, 1000ml distilled water is measured in above-mentioned conical flask, heating is dissolved, and adjusts pH value to 6.0, add 0.1% bromocresol green, 20% ethanol water 6.0ml, after packing 50.0ml is wrapped up in 150ml conical flask, at 121 DEG C of temperature, sterilizing 15 minutes spare.The invention has the advantages that: compared with AC broth bouillon, color change is become apparent from, and is easier to observation result.

Description

Selective enterococcus enriched medium

Technical field

The present invention relates to microbiological culture media technical field, especially selective enterococcus enriched medium.

Background technique

It has recently been demonstrated that most enterococcus can produce bacteriocin, it is considered to be conditioned pathogen can cause: urinary tract Infection, abdominal cavity, pelvic infection, burnt degree wound infection etc. is the major hospital infection pathogen for being only second to staphylococcus.I State reported that food poisoning caused by enterococcus was a lot of in recent years, was mostly by enterococcus faecalis and mutation, enterococcus faecium, quail chicken intestines ball Bacterium causes；Foreign countries have been reported that food poisoning caused by the sugared enterococcus of solution and class enterococcus avium.If the intestines ball from clinical samples Bacterium amount is few or the enterococcus of food sample is damaged in the production process, is not easy to be separately cultured, and needs first to be increased bacterium, It is separately cultured again.Current existing AC broth bouillon and the enterococcus meat soup (culture medium of all hydrolysis aesculin positives All blackening), effect is not highly satisfactory (Fig. 1, Fig. 2).Therefore, there is a need in the art for a kind of preferable enterococcus to increase bacterium Culture medium.

Summary of the invention

The present invention provides selective enterococcus enriched medium, this culture medium is named as MRCH2 culture medium.

Its technical solution are as follows:: Shi peptone 5.0g composed of the following components, 2.5 g of yeast extract powder, 3.0 g of beef extract, 20.0 g of glucose, 4.0 g of lactose, 5.0 g of sodium chloride, 2.5 g of Sodium azide, 10.0 g of bovine bile, 0.1% bromocresol green 20% Ethanol water 6.0ml, distilled water 1000ml.

Cheng Qu Shi peptone 5.0 g, 2.5 g of yeast extract powder, 3.0 g of beef extract, 20.0 g of glucose, 4.0 g of lactose, 5.0 g of sodium chloride, 2.5 g of Sodium azide, 10.0 g of bovine bile measure 1000ml distilled water in upper in 1000ml conical flask It states in conical flask, heating is dissolved, and is adjusted pH value to 6.0, is added 0.1% bromocresol green, 20% ethanol water 6.0ml, is dispensed After 50.0ml is wrapped up in 150ml conical flask, at 121 DEG C of temperature, sterilizing 15 minutes spare.

MRCH2 media components Zhong Shi peptone 5.0g, yeast extract powder 2.5g derive from plate count agar culture Base.

Beef extract 3.0g, sodium chloride 5.0g in MRCH2 media components derive from nutrient broth medium.

Sodium azide 2.5g in MRCH2 media components derives from Pficer enterococcosel agar culture medium.

Bovine bile 10.0g in MRCH2 media components derives from enterococcus broth bouillon.

Glucose 20.0g, lactose 4.0g in MRCH2 media components are to optimize the knot obtained using the method compared Fruit.

The ethanol water 6.0ml of 0.1% bromocresol green 20% in MRCH2 media components is answered from acid-base indicator Use guidance method.

Beneficial effects of the present invention: MRCH2 culture medium of the present invention, compared with AC broth bouillon, color change is become apparent from, Observation is easier to as a result, popularity is suitble to use.

Detailed description of the invention

Fig. 1 is enterococcus effect picture on enterococcus broth bouillon；

Fig. 2 is enterococcus effect picture on AC broth bouillon；

Fig. 3 is enterococcus 37 DEG C of cultures, 96 hours growing state effects on MRCH1, MRCH2, MRCH3, MRCH4 culture medium Fruit figure；

Fig. 4 is enterococcus effect picture on MRCH2 culture medium.

Specific embodiment

Shi peptone 5.0g, yeast extract 2.5g, beef extract 3.0g, sodium chloride 5.0g, bovine bile 10.0g, Sodium azide In 2.5g, 0.1% bromocresol green 20% ethanol water 6ml, distilled water 1000ml, the amount that glucose and lactose is added is shown in

Table one

Glucose and lactose magnitude table are added in (table one) culture medium

4 kinds of culture mediums so are made, are referred to as MRCH1, MRCH2, MRCH3, MRCH4 culture medium.PH to 6.0 is adjusted, point 50ml to 150ml conical flask is filled, 121 DEG C of wrapping sterilizing, 15 minutes spare.

Take bacterium to be made bacterium solution from plate count agar plate, draw bacterium solution 1ml be inoculated into respectively MRCH1, MRCH2, MRCH3, MRCH4 set 37 DEG C and cultivate 96 hours, and observation result (result of culture 96 hours is shown in Fig. 3) simultaneously measures its pH value.

The pH value application PHSJ -4A type laboratory PH meter for measuring culture measures (the 37 DEG C of cultures 96 of each culture respectively Hour) pH value.

The pH value for measuring each culture is shown in Table two

The pH value of (table two) each culture

	MRCH1	MRCH2	MRCH3	MRCH4
					PH value	4.82	4.49	4.43	4.25

Because not between 4.2~4.6, illustrating that sugar is added from table two it is not difficult to find out that the pH value of MRCH1 culture is 4.82 Amount it is insufficient, produced subacidity is to drop to 4.2~4.6.In line with the viewpoint of saving, glucose 20.0g, lactose 4g are determined.

It is the research of the influence according to the initial pH of Li Lijun to thalli growth that culture medium MRCH2, which finally adjusts PH to 6.0, As a result: enterococcus can well grow in the range of PH5.0~7.5, and PH grows best and determine when being 6.0.

Enterococcus effect picture (Fig. 4) on MRCH2 culture medium

Embodiment

Strain: (1) ATCC29212 is purchased from Qingdao Lv Gu commerce and trade Co., Ltd.

Reagent and culture medium: (1) PROTEOSE PEPTONE (Shi peptone) it is purchased from MADE IN ENGLAND BY OXOID LIMITED LONDON. S.E.1；(2) bispin microbiological culture media products factory in yeast extract Beijing produces；(3) beef The production of medicinal extract Shanghai City milk company integrated plant；(4) glucose Beijing China of state chemical reagent factory produces；(5) lactose Shanghai Learn chemical reagent work's production；(6) sodium chloride Zibo chemical reagent factory produces；(7) Sodium azide does not mark place of production label substance: Art.822335 Natriumazid Zur Synthese NaN3 MERCK-schuchardt schuchardt,8011 Hohenbrunn bei Mǖnchen Sehr giftig Verschlucken. Entwickelt bei Berǖhrung saure sehr giftige Gase；(8) bovine bile Shijiazhuang City Lang Te biological products Co., Ltd produces.(9) nutrient meat Soup culture medium is that laboratory is homemade；(10) AC broth bouillon is the production of Qingdao GaoKeYuan Hai Bo Bioisystech Co., Ltd； (11) plate count agar is trained

Feeding base is the neat space preventive medicine technology development co. production of Shandong Center for Disease Control & Prevention；(12) enterococcus meat Soup culture medium is

The production of Qingdao GaoKeYuan Hai Bo Bioisystech Co., Ltd；

Key instrument: (1) DHP -9270B Intelligent electric heating constant incubator, Shanghai Lang ?experimental facilities Co., Ltd it is raw It produces.

(2) YXQG02 type electrothermal steam sterilizer, Xinhua Medical Apparatus Co., Ltd. Shandong's production.(3) PHSJ -4A type laboratory PH meter, Shanghai Precision Scientific Apparatus Co., Ltd's production.

Actication of culture: take bacterium in nutrient broth (ingredient: beef extract 3.0g, peptone 10g, yeast extract 2.5g, chlorination Sodium 5g, distilled water 1000ml) in 37 DEG C cultivate 48 hours.Therefrom take bacterium streak inoculation PCA plate count agar (ingredient: pancreas egg White peptone 5.0g, yeast extract 2.5g, glucose 1.0g, agar 15.0g, distilled water 1000ml) plate, it is small to be placed in 37 DEG C of cultures 48 When.

Claims

1. selective enterococcus enriched medium, it is characterised in that:: Shi peptone 5.0g composed of the following components, yeast leach 2.5 g of powder, 3.0 g of beef extract, 20.0 g of glucose, 4.0 g of lactose, 5.0 g of sodium chloride, 2.5 g of Sodium azide, bovine bile 10.0 g, 0.1% bromocresol green 20% ethanol water 6.0ml, distilled water 1000ml.

2. selectivity enterococcus enriched medium according to claim 1, it is characterised in that: 5.0 g of Cheng Qu Shi peptone, 2.5 g of yeast extract powder, 3.0 g of beef extract, 20.0 g of glucose, 4.0 g of lactose, 5.0 g of sodium chloride, 2.5 g of Sodium azide, 10.0 g of bovine bile measures 1000ml distilled water in above-mentioned conical flask in 1000ml conical flask, and heating is dissolved, and is adjusted PH value adds 0.1% bromocresol green, 20% ethanol water 6.0ml to 6.0, and packing 50.0ml is wrapped in 150ml conical flask After bundle, at 121 DEG C of temperature, sterilizing 15 minutes spare.