Bifidobacterium adolescentis|186535 |BNCC

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Bifidobacterium adolenscentis

Subculture procedure	① Prepare two plates as described above; ② Disinfect ampoule surface, open in safety cabinet, cauterize top with alcohol lamp, then rapidly add sterile water to rupture. Crush with forceps; ③ Pipette 0.5mL sterile water (pre-equilibrated in anaerobic conditions for 24h) into freeze-dried vial. Thoroughly dissolve bacterial powder, then inoculate 2 plates (200μL/Agar plate) with uniform spread; ④ Incubate plates under the specified conditions. The culture is ready for use once bacterial growth appears.
Growth conditions	37°C; 3-5 days; anaerobic;
Storage conditions	2-8°C
Safety level	1
morphology	Colony diameter: 1-2 mm; circular with irregular margins; opaque; pale yellow upper surface; central elevation; rough, dull surface; dry texture; Gram-positive (blue-violet); bacilli; purity: pure
Sharing mode	Public welfare sharing

Bifidobacterium adolensentis

Storage conditions: 2~8 ℃

No. : 186535

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. 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, MRS medium, 48-72h. MRS medium: cheese peptone 10.0g, beef powder 8.0g, yeast powder 4.0g, glucose 20.0g, magnesium sulfate 0.2g, sodium acetate 5.0g, triammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, manganese sulfate 0.05g, Tween 80 1.0g, distilled water 1000mL. pH6.2± 0.2. 121 ℃,15min.

Recovery steps:

(1)Prepare a φ18mm test tube of 25ml liquid medium. Silica gel plug is used for liquid test tube culture medium . Please put the liquid medium and plate in an anaerobic environment for 24h 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 result
viability	good viability, in 48-72h the liquid medium become turbid, and the plate strain does not grow.
colony morphology: (above)	Bifidobacterium adolescence, a gram-positive bacilli, MRS liquid culture bacterial liquid is turbid, plate strains do not grow, it is recommended to use liquid culture.
Conclusion:	good viability, no abnormal strain morphology, qualified

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