BACTERIA AND FUNGI DETECTION BY CULTURE

LABORATORY PROCEDURES FOR ANIMAL & HUMAN CELL LINES

REFERENCE NO: AHC/1998/3/3.3/2.1

TITLE: BACTERIA AND FUNGI DETECTION BY CULTURE

INTRODUCTION

This protocol describes the detection of bacteria and fungi in cell cultures and reagents by isolation by culture.

PROCEDURE

Notes:

All cultures should be submitted for testing in antibiotic and cryoprotectant free medium. In addition cultures should not be passaged for three days prior to testing.

Inoculate 2 Fluid Thiogylcollate Medium and 2 Soyabean Casein Digest (Tryptone Soya Broth) with 1.5ml of test article.
Inoculate 2 Fluid Thiogylcollate Medium and 2 Soyabean Casein Digest (Tryptone Soya Broth) with 0.1ml of Candida albicans

Inoculate 2 Fluid Thiogylcollate Medium and 2 Soyabean Casein Digest (Tryptone Soya Broth) with Bacillus subtilis

Inoculate 1 Fluid Thiogylcollate Medium (Tryptone Soya Broth) with 0.1ml of Clostridium sporogenes

Leave 2 of each media type un-inoculated as negative controls.
For Tryptone Soya Broths, incubate one broth of each pair at 32+2⁰C for 14 days and one broth of each pair at 22+2⁰C for 14 days
For Fluid Thioglycollate Medium broths, incubate the broth inoculated with Clostridium sporogenes and one broth of each pair at 32+2⁰C for 14 days and one broth of each pair at 22+2⁰C for 14 days
Examine broths for turbidity after 14 days incubation

Record the results on the appropriate worksheet

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