Revival and Growth of Lactic Acid Bacteria

In This Article

Abstract

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Source: Ayivi, R. et al., The Cultivation, Growth, and Viability of Lactic Acid Bacteria: A Quality Control Perspective. J. Vis. Exp. (2022)

This video demonstrates the method for recovering cold-stressed lactic acid bacteria from a frozen glycerol stock, restoring bacterial viability through anaerobic incubation, and isolating active colonies for culture expansion.

Protocol

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1. Protocol for the activation and quality control of Lactic acid bacteria (LAB) cultures

Take the prepared glycerol stock of LAB strains (in 2 mL centrifuge tubes) from the -80 °C ultra-low freezer, and do not allow them to thaw before use.
Clean and disinfect the opening of the centrifuge tubes with 70% alcohol, and gently vortex before use.
Pipette about 250 µL (0.25 mL) of the stock LAB culture from the centrifuge tubes into fresh 2 mL MRS (deMan, Rogosa, and Sharpe media) test tubes.
Gently vortex, parafilm the test tubes, and anaerobically incubate them overnight at 42 °C for 12-16 h.
Take about 500 µL (....

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Materials

List of materials used in this article

Name

Company

Catalog Number

Comments

Aniline Blue

Thermo Scientific

R21526

25 g

Beef extract

Research Products International

50-197-7509

500 g

Yeast extract

Fisher Scientific

BP1422-500

500 g

Calcium Chloride dihydrate

Fisher Scientific

C79-500

500 g

Dextrose Anhydrous

Fisher Scientific

BP350500

500 g

D-Fructose

ACROS Organics

AC161355000

500 g

Difco agar powder

Difco

DF0812-07-1

2 kg

TPY agar

Difco

211921

500 g

Eppendorf microcentrifuge tube (Snap-Cap Microcentrifuge Safe-Lock)

Fisher Scientific

05-402-12

2 mL

Glycerol

Thermo Scientific

PI17904

500 mL

Infrared CO₂ Incubator

Forma Scientific