Youthful Bifidobacterium|134301 |BNCC

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Bifidobacterium adolescentis

Subculture procedure	① Prepare two of the aforementioned agar plates (pre-equilibrated in an anaerobic environment for 24 hours).② Surface-sterilize the ampoule, open it inside a biosafety cabinet, flame the top using an alcohol lamp, and promptly apply sterile water to crack it open. Then, carefully crush the ampoule using sterile forceps.③ Aspirate 0.5 mL of sterile water (pre-equilibrated in an anaerobic environment for 24 hours) and inject it into the lyophilized vial. After fully dissolving the bacterial powder, transfer the suspension onto the two prepared plates, distributing 200 µL per Agar plate, and spread evenly.④ Incubate the plates under the specified culture conditions. The bacterial strain is ready for use once growth is observed.
Growth conditions	37 ℃; 24-48 hours; anaerobic
Storage conditions	2-8 ℃
Safety level	1
morphology	The colonies are 0.5-1 mm in diameter, circular with entire edges, opaque, convex in the center, and have a dull, smooth, moist surface. They are easily picked, Gram-positive (blue-violet), and consist of rod-shaped cells.
Sharing mode	Public welfare sharing

Bifidobacterium adolescentis

Storage conditions: 2~8 ℃

No.: 134301

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 48 hours liquid medium become turbid, obvious strain layer occurs on the plate; colony is typical on marked plate
colony morphology: (see above figure)	size: 12mm, shape: round edge: neat, transparency: opaque, color: milky white, uplift: middle raised, surface: bright and smooth, texture: moist and easy to stir
Conclusion:	good viability, no abnormal colony morphology, qualified

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