Leuconostoc enterica|193296 |BNCC

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Leuconostoc mesenteroides

Subculture procedure	① Prepare 1-2 of the above plates; ② Open the 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 and transfer it into a freeze-drying tube. After fully dissolving the bacterial powder, transfer it into two plates, each Agar plate containing about 200 μ L, and apply evenly; ④ Place the petri dish under the above cultivation conditions for cultivation. Once the strain grows, it can be used.
Growth conditions	37 ℃; 24-48 hours; microaerophilic (3% -5% O2, 10% CO2)
Storage conditions	vacuum freeze-drying method
Safety level	1
morphology	Size: 1-2 mm Shape: Circular Margin: Entire Transparency: Opaque Color: Cream-white Elevation: Convex (raised center) Surface: Smooth and shiny Texture: Moist and easily picked up Gram Stain: Gram-positive (blue-purple), cocci
Sharing mode	Public welfare sharing

Leuconostoc mesenteroides

No. 193296

Product format： freeze dried, 200ul

Storage conditions: 2~8 ℃

Validity period: 6 years

Biosafety level: 1, handle in ultra-clean 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 ℃, slightly aerobic, 48-72h,MRS medium. 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

Recovery steps:

(1)Prepare a φ18mm test tube of 10ml 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.

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 liquid medium become turbid, obvious strain layer occurs on the plate,colony is typical on marked plate.
colony morphology: (above)	size: 1-2mm shape: round edge: neat transparency: opaque color: milky white uplift: middle raised surface: bright and smooth texture: moist and easy to stir
conclusion:	good viability, no abnormal colony morphology, qualified

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