Mucophilic Akkermansia|341917 |BNCC

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Akkermansia muciniphila

Subculture procedure	① Dissolve the frozen tube in a 37 ℃ water bath and shake quickly to dissolve; ② Wipe the outer wall of the Frozen vial with an alcohol swab for disinfection, and then transfer it to a safe cabinet for operation; ③ Unscrew the tube cap, aspirate all the dissolved solution, and transfer it into 1-2 agar plates (each Agar plate contains approximately 200 μ L, and for anaerobic bacteria cultivation, the culture medium needs to be placed in an anaerobic environment for 24 hours to remove oxygen); ④ After uniform coating, transfer to the above cultivation conditions for cultivation.
Growth conditions	37 ℃; 4-5 days; anaerobic; (Liquid culture medium FT)
Storage conditions	-80 ℃
Safety level	1
morphology	Colony diameter: 0.5–1 mm, circular, with neat margins; opaque, grayish-white on the upper surface, convex in the center, smooth surface, moist texture; G– (red); cocci; Purity: Pure
Sharing mode	Public welfare sharing

DSM 22959 Akkermansia muciniphila

Storage conditions : 2~8 ℃

No. : 341917

Product format: freeze dried, 200ul

Validity period: 6 years

Biosafety level : 1 , handle in ultra-clean 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, thioglycolate liquid medium, 5 days. Thioglycolate liquid medium: caseptone (trypsin hydrolysis) 15.0g, yeast extract 5.0g, glucose 5.0g, sodium thioglycolate 0.5g,L-cystine 0.5g, sodium chloride 2.5g, azure 0.001g, agar 0.75g, distilled water 1.0L,pH 7.1 ± 0.2. Sterilization at 121 ℃ for 15min.

Recovery steps:
(1)Prepare a φ18mm test tube of 10ml liquid medium(put in the anaerobic environment for deoxygenation for 24h in advance);

(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 5 days, liquid medium become turbid
colony morphology: (above)	Akkermansia muciniphila are gram-negative bacteria, oval bacteria that do not produce spores, and the bacterial liquid culture medium is turbid. Liquid culture is recommended to use liquid culture.
conclusion	good viability, no abnormal colony morphology, qualified

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