Animal Bifidobacterium lactis subspecies|134318 |BNCC

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Bifidobacterium animalis subsp. lactis

Subculture procedure	①Prepare 1-2 fresh agar plates (placed in an anaerobic environment for 24 hours of deoxygenation) with typical bacterial colonies; ② Open the 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 liquid culture medium or sterile water (placed in an anaerobic environment for 24 hours of deoxygenation in advance) and transfer it into a freeze-drying tube. Dissolve it thoroughly and mix well; ④ Transfer 0.2mL of bacterial suspension into a Agar plate, apply evenly, and repeat twice to obtain two plates; ⑤ Cultivate under the above conditions, and the strain can be used once it grows.
Growth conditions	37 ℃; 24-48 hours; anaerobic
Storage conditions	2-8 ℃
Safety level	1
morphology	Colonies were 1-2 mm in diameter, circular with an entire margin, and opaque. The colonies appeared creamy-white, convex in elevation, with a smooth, glistening, and moist surface that was easily liftable. No soluble pigment was produced. Microscopic examination revealed Gram-positive (blue-violet), rod-shaped cells. The culture was pure.
Sharing mode	Public welfare sharing

Bifidobacterium animalis subsp. lactis

Storage conditions: 2~8 ℃

No. : 134318

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 ℃, anaerobic, bifid enrichment medium, 24-48h, 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 801 mL, 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 test tube containing 5-10mL of liquid culture medium and 2 plates(put them in an anaerobic environment for 24 hours in advance);

(2)Open it 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 result
viability:	good viability, in 24-48h liquid medium become turbid, obvious strain layer occurs on the plate,colony is typical on marked plate.
colony morphology: (above)	Size: 1-2mm Shape: Round Edge: Irregular Transparency: Opaque Color: milky white uplift: middle raised surface: rough texture: dry
Conclusion:	good viability, no abnormal colony morphology, qualified

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