Clostridium sporogenes|266047 |BNCC

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Clostridium sporogenes (Metchnikoff) Bergey et al.

Subculture procedure	① Prepare 1-2 fresh agar plates as described above (pre-conditioned in an anaerobic environment for 24 h to remove oxygen).② In a biosafety cabinet, open the vial and briefly flame the top with an alcohol burner, then immediately add sterile water to break the seal. Use sterile forceps to crush the vial.③ Aspirate 0.5 mL of liquid medium or sterile water (pre-conditioned in an anaerobic environment for 24 h to remove oxygen) into the freeze‑dried vial, thoroughly dissolve the contents, and mix well.④ Transfer 0.2 mL of the resulting bacterial suspension onto an agar plate, spread evenly, and repeat to inoculate a second Agar plate.⑤ Incubate the plates under the specified culture conditions. The strain is ready for use once growth appears.
Growth conditions	37 ℃; 18-24h; anaerobic
Storage conditions	2-8 ℃
Safety level	1
morphology	The colonies measure 1-2 mm in diameter, are irregular in shape with filamentous margins, and are opaque. They exhibit a grayish-white pigmentation on the surface, have a flat morphology, and are rough and dull in appearance. The organisms are Gram-positive (G⁺, appearing blue-violet) and rod-shaped (bacilli). Purity: pure.
Sharing mode	Public welfare sharing

Clostridium sporogenes

Storage conditions: 2~8 ℃

No. 266047

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, 18-24h, ①Columbia blood plate (ready-to-use) ②Liquid thioglycolic acid medium: caseptone (trypsin hydrolyzed) 15.0g, yeast extract 5.0g, glucose 5.0g, thioglycolic acid Sodium 0.5g, L-cystine 0.5g, sodium chloride 2.5g, resazurin 0.001g, agar 0.75g, distilled water 1.0L, pH 7.1 ± 0.2. Sterilize at 121°C for 15min.

Recovery steps:

(1)Prepare sterile test tube of 5-10ml liquid medium and 2 plates; Silica gel plug is used for liquid test tube culture medium to ensure ventilation and facilitate deoxygenation. Please put the liquid medium and plate in an anaerobic environment for 24h before usage.

(2) Open it in the biosafety cabinet, sterilize of the ampoule, heat the tip 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) Put the plates under the above culture conditions for cultivation for 24-48 hours.

Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability	good viability,in 18-24h strain layer become obvious, colony is typical on the marked plate
colony morphology: (above)	Size: 2-4mm Shape: Round Edge: Neat Transparency: Opaque color: milky white uplift: middle convex surface: bright and smooth texture: moist and easy to stir
Conclusion	good viability, no abnormal colony morphology, qualified

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