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Agar Medium for Differential Enumeration of Lactic Streptococci

[所属分类:培养基配方] [发布时间:2021-10-28] [发布人:网站管理员2] [阅读次数:] [返回]

Agar Medium for Differential

Enumeration of Lactic Streptococci

山东拓普生物工程有限公司培养基配方 http://www.topbiol.com

Composition per 1170.0mL:

Agar ..................................................................15.0g

Carboxymethylcellulose ...................................15.0g

Calcium citrate..................................................10.0g

Pancreatic digest of casein..................................5.0g

Yeast extract........................................................5.0g

L-Arginine·HCl....................................................5.0g

Casamino acids ...................................................2.5g

K2HPO4.............................................................1.25g

Calcium carbonate solution ........................ 100.0mL

Nonfat milk solution..................................... 50.0mL

Bromcresol Purple solution .......................... 20.0mL

pH 5.9 ± 0.2 at 25°C

Calcium Carbonate Solution:

Composition per 100.0mL:

CaCO3 .................................................................3.0g

Preparation of Calcium Carbonate Solution:

Add CaCO3 to distilled/deionized water and bring

volume to 100.0mL. Mix thoroughly. Autoclave for

15 min at 15 psi pressure–121°C.

Nonfat Milk Solution:

Composition per 100.0mL:

Nonfat milk.......................................................11.0g

Preparation of Nonfat Milk Solution: Add

nonfat milk to distilled/deionized water and bring

volume to 100.0mL. Mix thoroughly. Autoclave for

15 min at 15 psi pressure–121°C.

Bromcresol Purple Solution:

Composition per 20.0mL:

Bromcresol Purple ............................................0.02g

Preparation of Bromcresol Purple Solution:

Add Bromcresol Purple to distilled/deionized water

and bring volume to 20.0mL. Mix thoroughly. Filter

sterilize.

Preparation of Medium: Add agar to 500.0mL

of distilled/deionized water. Gently heat and bring to

boiling. In a separate flask, add carboxymethylcellu

lose and calcium citrate to 500.0mL of distilled/

deionized water. Gently heat while stirring until a

white, turbid suspension is formed. Combine the two

solutions. Add the pancreatic digest of casein, yeast

extract, K2HPO4, casamino acids, and arginine. Mix

thoroughly. Gently heat and bring to boiling. Adjust

pH to 5.6 with 6N HCl. Distribute into screw-capped

bottles in 100.0mL volumes. Autoclave for 15 min at

15 psi pressure–121°C. Cool to 45°–50°C. Immedi

ately prior to pouring plates, aseptically add 5.0mL

of sterile nonfat milk solution, 10.0mL of sterile cal

cium carbonate solution, and 2.0mL of sterile Brom

cresol Purple solution to each screw-capped bottle.

Mix thoroughly. The pH should be 5.9. Pour into

cold, sterile Petri dishes.

Use: For the cultivation, differentiation, and enu

meration of Lactobacillus lactis, Lactobacillus lactis

subspecies cremoris, and Lactobacillus lactis sub

species diacetylactis. Lactose-fermenting bacteria

such as Lactobacillus lactis subspecies cremoris ap

pear as yellow colonies. Arginine-utilizing bacteria

such as Lactobacillus lactis and Lactobacillus lactis

subspecies diacetylactis appear as purple colonies.

Citrate-utilizing bacteria such as Lactobacillus lactis

subspecies diacetylactis appear as colonies surround

ed by a clear zone.

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Agar Medium for Differential Enumeration of Lactic Streptococci