Boronella bronchisepticus|BNCC360694 |BNCC

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Boronella bronchisepticus

Culture medium	Nutritional gravy medium (English name: NA/NB): beef paste 3.0g, peptone 10.0g,NaCl 5.0g, agar 20.0g (not included in liquid medium), distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min. [Note] Adding 5 mg MnSO4 · H2O to culture Bacillus is beneficial to produce spores.
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 ℃;18-24h; Aerobic
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-(red), bacilli, purity: pure
Sharing mode	Public welfare sharing

Bordetella bronchiseptica

Strain number: BNCC 360694

Batch number: 220608

Biosafety level: 2, handle in safety cabinet

Provide form: Freeze-dried powder beveled bacterial liquid plate

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.

Medium: Nutrient gravy medium (English name: NA/NB): beef extract 3.0g, peptone 10.0g, NaCl 5.0g, agar 20.0g (not included in liquid medium), distilled water 1.0L, pH 7.0. Sterilize at 121°C for 15min . [Note] When culturing Bacillus, adding 5mg MnSO4·H2O is conducive to the production of spores.

Growth conditions: 37 ℃;18-24h; Aerobic

Recovery steps:

(1)Prepare a test tube containing 5-10mL of liquid culture medium and 2 plates;

(2)Open the ampoule 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.

Viability: good viability, in 24 hours, liquid medium become turbid, obvious strain layer occurs on the plate; colony is typical on marked plate characteristics of colony: colony diameter is 1-2mm, round, edge neat, opaque, front off-white, back off-white, middle convex, smooth surface, bright surface, moist texture, easy to stir up, G-(red), bacilli, purity: pure

Quality inspection chart:

Conclusion: good viability, no abnormal colony morphology, qualified

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