Bryant Burkey Broth with Resazurin and Lactate is also known as BB broth and BBMB broth. It’s used for the enumeration of the spores of lactic acid-fermenting Clostridia spp. in silage, milk and dairy products.
Approximate Formula:
Ingredients
|
gm/liter
|
Enzymatic digest of casein
|
15.0
|
Meat extract
|
7.5
|
Yeast extract
|
5.0
|
Cysteine HCl
|
0.5
|
Sodium acetate
|
5.0
|
Sodium lactate
|
5.0
|
Resazurin
|
0.005
|
Final pH5.8±0.1
|
25℃
|
*Adjusted and/or supplemented as required to meet performance criteria.
Directions:
Dissolve
38.0g in 1 liter of purified water and mix well. Dispense
into tubes or bottles. Autoclave
at 121℃ for 15 minutes.
According to CNERNA, the medium is dispensed
by 10ml into 16x160mm tubes before sterilization.
Principle and Interpretation:
Enzymatic digest of casein, meat extract, yeast extract and cysteine HCl create the conditions for a rapid growth of lactate fermenting Clostridium spp. Resazurin is a redox indicator and monitors the oxygen level. Sodium lactate acts as the carbon source to allow selective growth of lactate-fermenting spore formers and is the substrate for the gas production of Clostridium spp. Sodium acetate facilitates the lactate fermentation by C. tyrobutyricum. Sodium acetate also promotes the spore germination, which is activated by the heat treatment of the sample.
Appearance:
Dehydrated medium is a free-flowing yellowish powder. The prepared medium is a kind of yellowish-red turbid liquid.
Precautions:
This medium is for laboratory use only. Dried medium which is past shelf life, caking or color variation
cannot be used.
Storage
conditions and Shelf life:
Bryant
Burkey Broth with Resazurin and Lactate must be stored tightly capped in the original container
at 5-30°C. The dehydrated medium has a shelf life of 3 years from date of manufacturing. Prepared medium may be stored, out of direct light at 2-8°C.
Quality control:
Prepare the medium per label directions. Inoculate
and incubate at 36℃ in an anaerobic
atmosphere for 24-72 hours.
Microorganism
|
Strains Number
|
Inoculum (CFU)
|
Remarks
|
Clostridium butyricum
|
ATCC 19398
|
10-100
|
Turbid with gas
|
Clostridium perfringens
|
ATCC 13124
|
10-100
|
Turbid with gas
|
Clostridium sporogenes
|
ATCC 19404
|
10-100
|
Turbid
|
Reference:
1. United States Pharmacopeial Convention, Inc. 2008. The United States pharmacopeia 31/The nationalformulary 26, Supp. 1, 8-1-08, online. United States Pharmacopeial Convention, Inc., Rockville, Md.
2. European Directorate for the Quality of Medicines
and Healthcare. 2008. The European pharmacopoeia,6th ed., Supp. 1, 4-1-2008, online. European Directorate for the Quality
of Medicinesand Healthcare, Council of Europe, 226 Avenue de Colmar BP907-, F-67029
Strasbourg Cedex 1,France.