CN102492765A

CN102492765A - Agar culture medium and preparation method thereof

Info

Publication number: CN102492765A
Application number: CN2011104140801A
Authority: CN
Inventors: 王立超
Original assignee: JIANGMEN KAILIN TRADE CO Ltd
Current assignee: JIANGMEN KAILIN TRADE CO Ltd
Priority date: 2011-12-13
Filing date: 2011-12-13
Publication date: 2012-06-13

Abstract

The invention discloses an agar culture medium. Each 1,000 milliliters of nutrient solution comprises the following components by mass: 17 to 23 grams of peptone, 7 to 13 grams of lactose, 4 to 6 grams of sodium chloride, 4 to 6 grams of cholate, 0.04 to 0.06 gram of neutral red and 13 to 17 grams of agar. The preparation method comprises the following steps of: 1) dissolving 20 grams of peptone, 10 grams of lactose, 5 grams of sodium chloride, 5 grams of cholate and 15 grams of agar in 1,000 milliliters of nutrient solution; 2) heating; 3) regulating the pH value to be between 7.2+/-0.2, and adding 0.05 gram of neutral red; 4) heating to the temperature of 115 DEG C, keeping the temperature for 20 minutes, cooling to the temperature of about 50 DEG C, and pouring into a disposable sterile plastic plate; and 5) refrigerating.

Description

Nutrient agar and preparation method thereof

Technical field

The present invention relates to a kind of agar culture technique field, nutrient agar of especially a kind of separation and discriminating intestinal bacteria and preparation method thereof.

Background technology

The effect of like product in the market mainly is to differentiate intestinal bacteria; This is one of this one of them index of product; And being non-fermenting bacterial, the another one important indicator of this product can on this substratum, grow; That this is that like product is not accomplished on the market or accomplish that DeGrain and colour developing are not easy to differentiate, this index also is the sole criterion of this product of check.

Summary of the invention

The object of the invention just provides a kind of nutrient agar and preparation method thereof, has improved the phenomenon to non-fermenting bacterial omission.

In order to reach above-mentioned purpose of design, the technical scheme that the present invention adopts is following:

A kind of nutrient agar, its moity and total mass number are: in every 1000ml nutritive medium: peptone 17-23g, lactose 7-13g, sodium-chlor 4-6g, cholate 4-6g, toluylene red 0.04-0.06g, agar 13-17g.

Preferably, its moity and total mass number are: in every 1000ml nutritive medium: peptone 20g, lactose 10g, sodium-chlor 5g, cholate 5g, toluylene red 0.05g, agar 15g.

Preferably, the staple of said nutritive medium is amino acid, VITAMINs, growth factor.

A kind of making method of nutrient agar may further comprise the steps:

1) peptone 20g, lactose 10g, sodium-chlor 5g, cholate 5g, agar 15g are dissolved in the 1000ml nutritive medium;

2) behind the mixing, heated and boiled is fully dissolved;

3) regulate pH to 7.2 ± 0.2, add toluylene red 0.05g;

4) be heated to 115 ℃, and keep 20min, be cooled to about 50 ℃, be poured in the disposable sterilized plastics plate;

5) behind the steriling test in 2 ℃ of-8 ℃ of environment refrigeration subsequent use.

Preferably, the method for said adjustment PH to 7.2 ± 0.2 is to regulate pH to 7.2 ± 0.2 with 1N-10N sodium hydroxide or 1N-10N hydrochloric acid soln.

Its principle is: peptone provides nitrogenous source and other nutritive substance in substratum; Lactose is as fermentable carbon source, and after lactose was fermented and utilizes, partial pH value reduces made bacterium colony take on a red color; Form the cholate deposition simultaneously, cholate suppresses the Gram-positive bacteria growing.

The beneficial effect of nutrient agar of the present invention and preparation method thereof is: the required special nutrition material that on original basis, adds non-fermenting bacterial (Acinetobacter bauamnnii); Through the supply of nutritive substance to bacterium; The result shows that not merely right and wrong fermenting bacterial growing state on this product is fabulous; And that other intestinal bacteria grow on this product is also fabulous; Time of bacterium colony size and sentence read result even surpassed similar products at home and abroad, the improvement of this product has improved the phenomenon to non-fermenting bacterial omission greatly.

The bacterium colony size is better than similar products at home and abroad; The time ratio like product of sample sentence read result is shorter; The growth of non-fermenting bacterial has improved on the market similar cultivation and has not gone out or cultivate bad phenomenon; The discriminating of non-fermenting bacterial is more prone to judge through colour developing.

Embodiment

Embodiment 1,

A kind of nutrient agar, its moity and total mass number are: in every 1000ml nutritive medium: peptone 17g, lactose 7g, sodium-chlor 4g, cholate 6g, toluylene red 0.06g, agar 17g.

Embodiment 2,

A kind of nutrient agar, its moity and total mass number are: in every 1000ml nutritive medium: peptone 23g, lactose 13g, sodium-chlor 6g, cholate 4g, toluylene red 0.04g, agar 13g.

Embodiment 3,

A kind of nutrient agar, its moity and total mass number are: in every 1000ml nutritive medium: peptone 20g, lactose 10g, sodium-chlor 5g, cholate 5g, toluylene red 0.05g, agar 15g.

In the foregoing description, the staple of said nutritive medium is amino acid, VITAMINs, growth factor.

Embodiment 4,

A kind of making method of nutrient agar may further comprise the steps:

1) peptone 17g, lactose 7g, sodium-chlor 4g, cholate 6g, agar 17g are dissolved in the 1000ml nutritive medium;

2) behind the mixing, heated and boiled is fully dissolved;

3) regulate pH to 7.2 ± 0.2, add toluylene red 0.06g;

Embodiment 5,

A kind of making method of nutrient agar may further comprise the steps:

1) peptone 23g, lactose 13g, sodium-chlor 6g, cholate 4g, agar 13g are dissolved in the 1000ml nutritive medium;

2) behind the mixing, heated and boiled is fully dissolved;

3) regulate pH to 7.2 ± 0.2, add toluylene red 0.04g;

Embodiment 6,

A kind of making method of nutrient agar may further comprise the steps:

2) behind the mixing, heated and boiled is fully dissolved;

3) regulate pH to 7.2 ± 0.2, add toluylene red 0.05g;

In the foregoing description, regulate pH to 7.2 ± 0.2 with 1N-10N sodium hydroxide or 1N-10N hydrochloric acid soln.

This embodiment the preferred embodiments of the present invention can not limit the present invention, and concrete each item rights protection scope is defined by the claims.

Claims

1. nutrient agar, it is characterized in that: its moity and total mass number are: in every 1000ml nutritive medium: peptone 17-23g, lactose 7-13g, sodium-chlor 4-6g, cholate 4-6g, toluylene red 0.04-0.06g, agar 13-17g.

2. nutrient agar according to claim 1 is characterized in that: its moity and total mass number are: in every 1000ml nutritive medium: peptone 20g, lactose 10g, sodium-chlor 5g, cholate 5g, toluylene red 0.05g, agar 15g.

3. nutrient agar according to claim 1 and 2 is characterized in that: the staple of said nutritive medium is amino acid, VITAMINs, growth factor.

4. the making method of a nutrient agar is characterized in that: may further comprise the steps:

2) behind the mixing, heated and boiled is fully dissolved;

3) regulate pH to 7.2 ± 0.2, add toluylene red 0.05g;

4) be heated to 115 ℃, and keep 20min, be cooled to about 50, be poured in the disposable sterilized plastics plate;

5. the making method of nutrient agar according to claim 4 is characterized in that: the method for said adjustment PH to 7.2 ± 0.2 is, with 1N-10N sodium hydroxide or 1N-10N hydrochloric acid soln adjusting pH to 7.2 ± 0.2.