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改良Bryant Burkey肉汤/用于青贮饲料、牛奶和乳制品中乳酸发酵梭状芽孢杆菌孢子的计数_
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改良Bryant Burkey肉汤/用于青贮饲料、牛奶和乳制品中乳酸发酵梭状芽孢杆菌孢子的计数

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改良Bryant Burkey肉汤介绍:

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:

Ingredientsgm/liter
Enzymatic digest of casein15.0
Meat extract7.5
Yeast extract5.0
Cysteine HCl0.5
Sodium acetate5.0
Sodium lactate5.0
Resazurin0.005
Final pH5.8±0.125℃
*Adjusted and/or supplemented as required to meet performance criteria.

Directions:

Dissolve38.0g in 1 liter of purified water and mix well. Dispenseinto tubes or bottles. Autoclaveat 121℃ for 15 minutes.

According to CNERNA, the medium is dispensedby 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 variationcannot be used.

Storageconditions and Shelf life:

BryantBurkey Broth with Resazurin and Lactate must be stored tightly capped in the original containerat 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. Inoculateand incubate at 36℃ in an anaerobicatmosphere for 24-72 hours.

MicroorganismStrains NumberInoculum (CFU)Remarks
Clostridium butyricumATCC 1939810-100Turbid with gas
Clostridium perfringensATCC 1312410-100Turbid with gas
Clostridium sporogenesATCC 1940410-100Turbid

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 Medicinesand Healthcare. 2008. The European pharmacopoeia,6th ed., Supp. 1, 4-1-2008, online. European Directorate for the Qualityof Medicinesand Healthcare, Council of Europe, 226 Avenue de Colmar BP907-, F-67029Strasbourg Cedex 1,France.

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以上信息仅供参考,请以实物批次为准!

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标准名称及标准号
GB 4789.28-2013
《食品安全国家标准 食品微生物学检验 培养基和试剂的质量要求》
适用范围
本标准规定了食品微生物学检验中培养基和试剂的验收、贮存及使用要求,适用于青贮饲料、牛奶、乳制品等样品中特定微生物(如乳酸发酵梭状芽孢杆菌孢子)的检测。
核心检测方法
1. 培养基选择:使用改良Bryant Burkey肉汤进行选择性增菌培养;
2. 样品处理:按标准要求进行梯度稀释与热处理;
3. 培养条件:厌氧环境(37℃±1℃,24-72小时)。
检出限与定量限
1. 检出限:10 CFU/g(或mL);
2. 定量限:适用于≥10 CFU/g(或mL)的样品,需结合菌落计数法或MPN法验证。
质控样品要求
1. 阳性对照:使用标准菌株(如Clostridium tyrobutyricum ATCC 25755);
2. 阴性对照:无菌生理盐水或空白培养基;
3. 平行样检测:每批次样品需设置≥10%的平行样。
关键实验步骤
1. 样品预处理:均质后80℃水浴加热10分钟以激活孢子;
2. 梯度稀释:使用无菌缓冲液进行10倍系列稀释;
3. 培养观察:记录产气、浑浊度及pH变化等特征。
特别说明
1. 培养基配制后需进行性能验证,包括生长率及选择性;
2. 青贮饲料样品需额外去除纤维干扰;
3. 检测结果需结合生化鉴定(如革兰氏染色、气相色谱分析)确认。

以上信息仅供参考,请以相应标准的原文为准!