Lactobacillus brevis|BNCC189801 |BNCC

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Lactobacillus brevis

Culture medium	MRS medium (MRS): peptone 10.0g, beef paste 10.0g, yeast powder 4.0g, glucose 20.0g, magnesium sulfate 0.2g, sodium acetate 5.0g, triammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, manganese sulfate 0.04g, Tween 80 1.0g, agar 20.0g (not included in liquid medium), distilled water 1.0L. pH 5.7±0.2. Sterilization at 121 ℃ for 15min.
Subculture procedure	(1) Prepare a test tube containing 5~10mL of liquid medium and 2 plates; (2) Open it in the safety cabinet, heat the tip of ampoule in a flame, quickly drop sterile water to creak it, then break it with forceps; (3) draw 0.5mL of liquid culture medium into a freeze dried ampoule, fully rehydrate and transfer the solution to the liquid test tube, mix evenly; (4) inoculate a plate with 0.2mL of suspension liquid, repeat the step to obtain two plates; ⑤ Put all the liquid test tubes and plates under the above culture conditions for cultivation, and the strains can be used when they grow.
Growth conditions	37 ℃;48-72h; Micro aerobic (3%-5% O2,10% CO2)
Storage conditions	2-8 ℃
Separation substrate	Silage
application	Making silage.
Sharing mode	Public welfare sharing

1. Description

1. Name: Lactobacillus brevis

2. BNCC No.: 189801

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. MRS medium: cheese peptone 10.0g, beef powder 8.0g, yeast powder 4.0g, glucose 20.0g, magnesium sulfate 0.2g, sodium acetate 5.0g, triammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, manganese sulfate 0.05g, Tween 80 1.0g, distilled water 1000mL. pH6.2±0.2. 121 ℃,15min.

2. Atmosphere: micro aerobic

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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