Clostridium perfringens|185933 |BNCC

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Clostridium perfringens

Subculture procedure	① Prepare 2 of the above plates (placed in an anaerobic environment for 24 hours of equilibration); ② After disinfecting the surface of the ampoule tube, open it in a 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 sterile water (placed in an anaerobic environment for 24 hours) and transfer it into a freeze-drying tube. After fully dissolving the bacterial powder, transfer it into 2 plates at a rate of 200 μ L per Agar plate and apply evenly; ④ Place the petri dish under the above cultivation conditions for cultivation, and once the bacterial strain grows, it can be used.
Growth conditions	37 ℃; 24-48 hours; anaerobic;
Storage conditions	2-8 ℃
Safety level	2
morphology	The colonies are 2-4 mm in diameter, circular with irregular edges, opaque, grayish-white on the obverse side, convex with a rough and dull surface, easily picked. They are Gram-positive (blue-violet), rod-shaped, and of pure culture.
Sharing mode	Public welfare sharing

Clostridium perfringens

Storage conditions: 2~8 ℃

No.: 185933

Product format: freeze dried, 200ul

Validity period: 6 years

Biosafety level: 2 , handle in 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, 24-48h, liquid thioglycolate 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, blade 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; and two agar plates(put in an anaerobic environment for deoxygenation 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 result
viability	good viability, in 24-48h, the liquid medium is turbid, and the strain grow slowly on the plate.
colony morphology: (above)	Clostridium perfringens is a gram-positive bacterium. FT liquid culture is turbid and plate strains are weak. Liquid culture is recommended.
conclusion	good viability, no abnormal colony morphology, qualified

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