Bilophila wadsworthia|BNCC358991 |BNCC

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Bilophila wadsworthia

Culture medium	Columbia blood plate (CA-B): casein pancreatin digest 10.0g, cardiopancreatin digest 3.0g, corn starch 1.0g, meat gastric enzyme digest 5.0g, yeast extract 5.0g, sodium chloride 5.0, agar 20.0g, distilled water 1.0L,pH 7.3±0.2. After sterilization at 121 ℃ for 15min, cool to 55 ℃, add 5% sterile defibrated sheep blood, shake well, pour the plate and cool it for later use.
Subculture procedure	(1) Prepare a test tube containing 5~10mL of liquid medium and 2 plates; (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.
Growth conditions	37 ℃;3-4 days; Anaerobic;
Storage conditions	2-8 ℃
morphology	The colony diameter is 1-2mm, round, with neat edges, opaque, gray-white on the front, gray-white on the back, convex in the middle, smooth and bright surface, moist texture, easy to stir up, G + (blue-purple), bacilli, purity: pure
Separation substrate	Human blood
Sharing mode	Public welfare sharing

Bilophila wadsworthia

Storage conditions: 2~8 ℃

No. 358991

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 ℃, strictly anaerobic, Colombian blood plate (ready-to-use type),3-4 days.

Recovery steps:

①Prepare 1-2 pieces of Blood agar plates;

②sterilizing 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;

③draw 0.5mL of liquid culture medium into the freeze dried ampoule,make the strain pellet fully dissolved and inoculate a plate with the solution, then distribute it well in 200ml/plate;

④Put the agar plates under the above culture conditions for cultivation.

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

Item	test results
viability:	good viability. in 72 hour obvious strain layer occurs on the plate; colony is typical
Colony morphology: (pictured above)	size: 1-2mm ; Shape: round ; Edge: irregular ; Transparency: opaque ; Color: off-white uplift: middle bulge ; Surface: gray ; Texture: moist and easy to stir
conclusion: good viability, no abnormal colony morphology, qualified

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