Bifidobacterium brevis|185972 |BNCC

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Bifidobacterium breve

Subculture procedure	① Prepare 2 of the above plates; ② After disinfecting the surface of the ampoule tube, open it in a safety cabinet, burn the top with an alcohol lamp, quickly drip sterile water to break it, and then use tweezers to crush it; ③ Take 0.5mL of sterile water (placed in an anaerobic environment for 24 hours) and transfer it into a freeze-drying tube. After fully dissolving the bacterial powder, transfer it into 2 plates at a rate of 200 μ L per Agar plate and apply evenly; ④ Place the petri dish under the above cultivation conditions for cultivation, and once the bacterial strain grows, it can be used.
Growth conditions	37 ℃; 3-5 days; anaerobic;
Storage conditions	2-8 ℃
Safety level	1
morphology	The colonies measure 0.5-1 mm in diameter, are circular with regular margins, and are opaque. They appear white on the surface, with a raised center, and are smooth and easy to lift. The organisms are Gram-positive (G⁺, appearing blue-violet) and rod-shaped (bacilli).
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1. Description

1. Name: Bifidobacterium breve

2. BNCC No.: 185972

3. Biosafety level: 4

2. Storage conditions:

Storage of freezed dried ampoule and agar slant at 2°C to 8°C.

3. Growth Conditions:

1. Bifid enrichment medium (BBL): peptone 15.0g, glucose 20.0g, yeast extract 2.0g, soluble starch 0.5g, sodium chloride 5.0g,L-cysteine 0.5g, tomato powder 5.0g, liver powder 2.0g, Tween 80 1mL, agar 20.0g (not included in liquid medium), distilled water 1.0L,pH 7.0±0.1. Sterilization at 121 ℃ for 15min.

2. Atmosphere: anaerobic

3. Temperature: 37 ℃

4. Notes:

1.Normal culturing time, 24-48hours for bacterial, 72 hours for yeast, 5-7days for mould, 7-10days for fungal.

2.Agar slant shall be inoculated asap, and do not keep the storage for more than 3 months.

3.Please recover the strains in strict accordance with this instruction, otherwise the replacement of the strain are not be available in case of viability loss caused by different media or growth conditions.

4.Waste generated from the handling process should be discarded after high-pressure sterilization.

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