Bifidobacterium bifidum|186522 |BNCC

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Bifidobacterium bifidum

Subculture procedure	① Prepare 1 tube containing 5-10mL liquid medium and 2 plates; ② Open in a safety cabinet, cauterize the top with a酒精灯, then quickly add sterile water to rupture it; break it into pieces with forceps; ③ Pipette 0.5mL liquid medium into the freeze-dried vial, dissolve completely, then return to the liquid test tube and mix thoroughly; ④ Pipette 0.2mL bacterial suspension onto plates, spread evenly; repeat twice to obtain two plates; ⑤ Incubate all liquid tubes and plates under the specified conditions. The culture is ready for use once the strain grows.
Growth conditions	37°C; 3-4 days; anaerobic;
Storage conditions	2-8°C
Safety level	1
morphology	Colony diameter: 0.5–1 mm; circular with irregular margins; upper surface grayish-white, centrally convex; surface dull gray, dry texture; Gram-positive (blue-violet), bacilli; purity: pure
Sharing mode	Public welfare sharing

Bifidobacterium bifidum

Storage conditions: 2~8 ℃

No. 186522

Product format： freeze dried, 200ul

Validity period: 6 years

Biosafety level: 1, handle in ultra-clear table or safety cabinet

Receiving notice: if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. If it is overdue, it is deemed that the bacteria are well. The freeze dried culture shall be used up once and shall not be retained. The bacterial viability will resume after 1-2 generation of recovery and can be used normally. Handling instructions of agar slant and suspension liquid is enclosed. Please operate in strict accordance with this instruction, otherwise the replacement of bacteria are not be available in case of aberrant growth and loss of viability.

Growth conditions: 37°C, anaerobic, BBL agar medium, 3-4 days. BBL agar medium: peptone 15.0g, glucose 20.0g, yeast extract 2.0g, soluble starch 0.5g, sodium chloride 5.0g, L-cysteine 0.5g, tomato extract powder 5.0g, liver extract powder 2.0g, Tween 80 1mL, agar 20.0g (not included in liquid medium), distilled water 1.0L, pH 7.0±0.1. Sterilize at 121°C for 15min.

Recovery steps:

(1)Prepare a test tube containing 5-10mL of liquid culture medium and 2 plates（ place in anaerobic environment for 24hours before usage）;

(2) Open the ampoule in the biosafety 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.

Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability	good viability，in 4 days, the liquid culture medium is turbid, and the plate grows bacteria.
colony morphology:	Bifidobacterium bifidum, a gram-positive bacilli with a bifurcation at one end, BBL liquid culture bacterial liquid is turbid, and there is a large amount of precipitation at the bottom of the test tube
Conclusion:	good viability, no abnormal colony morphology, qualified

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