Bifidobacterium bifidum|336858 |BNCC

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Bifidobacterium bifidum

Subculture procedure	① Prepare two of the abovementioned agar plates (pre-placed in an anaerobic environment for 24 hours for oxygen removal).② Surface-sterilize the ampoule vial, then open it inside a biological safety cabinet. Flame the top of the vial with an alcohol burner, and immediately apply a drop of sterile water to crack it. Then, use forceps to carefully break it open.③ Aspirate 0.5 mL of sterile water (pre-placed in an anaerobic environment for 24 hours for oxygen removal) and inject it into the lyophilized vial. Thoroughly resuspend the bacterial powder. Then, transfer the suspension onto the two prepared plates, 200 μL per Agar plate, and spread evenly.④ Incubate the plates under the specified culture conditions mentioned above. The bacterial strain is ready for use once growth is observed.
Growth conditions	37 ℃; 3-4 days; anaerobic
Storage conditions	2-8 ℃
Safety level	1
morphology	The colony diameter is 1-2mm, circular, with neat edges, milky white on the front, raised in the middle, smooth surface, bright surface, moist texture, no pigment, G+(blue purple), Bacillus, purity: pure
Sharing mode	Public welfare sharing

Bifidobacterium bifidum

Storage conditions : 2~8 ℃

No. : 336858

Product format: freeze dried, 200ul

Validity : 6 years

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

Notes: 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. 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 ℃, anaerobic, bifid enrichment medium, 3-4 days. Bifid enrichment medium: 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.

Recovery steps:
(1) Prepare a φ18mm test tube of 10ml liquid medium(put in an anaerobic environment for deoxygenation in advance for 24 hours);
(2) Sterilizing the ampoule, open it in 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 the freeze dried ampoule, fully rehydrate and transfer the solution to the liquid test tube, mix evenly;
(4) Put the test tube under the above culture conditions for cultivation. The culture solution is obviously turbid or mass growth at the solution bottom occur, this indicates the bacterial grows well.
Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability:	good viability,in 3-4 days liquid medium become turbid
colony morphology: (above)	Bifidobacterium bifidus, gram-positive bacilli, straight rod, rod, y-shaped, Bifidobacterium enrichment medium culture bacterial fluid is turbid, liquid culture is recommended
Conclusion:	good viability, no abnormal colony morphology, qualified

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