Bifidobacterium bifidum|BNCC337499 |BNCC

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Bifidobacterium bifidum

Culture medium	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 (excluding liquid), distilled water 1.0L,pH value 7.0 0.1. 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 ℃;3-5d; Anaerobic
Growth characteristics	G +, rod-shaped cells, 0.55 μm × 1.82~2.12 μm, solitary, paired, V-shaped or pile-shaped arrangement, no spores. Strictly anaerobic, fermentation of glucose to produce lactic acid.
Storage conditions	2-8 ℃
morphology	Bifidobacterium bifidum, Gram-positive bacilli, is pleomorphic, BBL liquid culture medium culture bacteria fluid turbid, plate strain growth is weak, it is recommended to use liquid culture.
Separation substrate	Baby feces
application	Taxonomic studies, probiotics.
Sharing mode	Public welfare sharing

Bifidobacterium bifidum

Storage conditions: 2~8 ℃

No. 337499

Product format: freeze dried,200ul

Validity period: 6 years

Biosafety level: 1, handle in ultra-clear 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, BBL liquid medium, 3-5 days. BBL liquid 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 5.0g, liver extract 2.0g, Tween 80 1.0g, distilled water 1L,pH6.6-7.0. 115℃

Recovery steps:

(1)Prepare a φ18mm test tube of 10ml liquid medium; and two agar plates（place in anaerobic environment for 24hours before usage）；

(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 days liquid medium become turbid
colony morphology: (above)	Bifidobacterium bifidum, Gram-positive bacilli, is pleomorphic, BBL liquid culture medium is turbid, plate strain is weak, it is recommended to use liquid culture.
Conclusion	good viability， no abnormal colony morphology, qualified

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