Lactobacillus delbrueckii subsp. bulgaricus|BNCC137361 |BNCC

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Lactobacillus delbrueckii subsp. bulgaricus

Culture medium	MRS medium (MRS): peptone 10.0g, beef paste 10.0g, yeast powder 4.0g, glucose 20.0g, magnesium sulfate 0.2g, sodium acetate 5.0g, triammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, manganese sulfate 0.04g, Tween 80 1.0g, agar 20.0g (not included in liquid medium), distilled water 1.0L. pH 5.7±0.2. Sterilization at 121 ℃ for 15min.
Subculture procedure	(1) Prepare a test tube containing 5~10mL of liquid medium and 2 plates; (2) Open it in the safety 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 ℃;24-48h; Anaerobic;
Storage conditions	2-8 ℃
morphology	Lactobacillus delmarii subsp. Bulgaria, Gram-positive bacilli, MRS liquid culture bacterial fluid is turbid, and there is a large amount of precipitation at the bottom of the test tube, Plate strains are weak in growth and liquid culture is recommended.
Separation substrate	bulgarian yoghourt
application	Type strain
Sharing mode	Public welfare sharing

Lactobacillus delbrueckii subsp.

Storage conditions: 2~8 ℃

No.: 137361

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. 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, MRS medium, 48-72h. MRS medium: peptone 10.0g, beef paste 10.0g, yeast powder 4.0g, glucose 20.0g, magnesium sulfate 0.2g, sodium acetate 5.0g, triammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, manganese sulfate 0.04g, Tween 80 1.0g, agar 20.0g (not included in liquid medium), distilled water 1.0L. pH 5.7 & plusmn;0.2. Sterilization at 121 ℃ for 15min.

Recovery steps:

(1)Prepare a φ18mm test tube of 10ml liquid medium;

(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 48-72h a large number of bacteria precipitate at the bottom of the liquid medium, and the plate strain does not grow.
Colony morphology: (pictured above)	Lactobacillus delkii subsp. Bulgaria, a gram-positive bacterium, the bottom of MRS liquid culture is turbid, the plate strain does not grow, build discuss the use of liquid culture.
Conclusion:	good viability, no abnormal colony morphology, qualified