Page 67 - MNU PM502 Ph. Micreo practical

Pharm D- Clinical Pharmacy Program Third Level Pharmaceutical Microbiology& Antimicrobials (PM 502)

Practical mission:

You are provided with sterile water for injection and required to test its sterility

Equipment:
1. Two test tubes, one containing Fluid thioglycolate medium and the other one

Fluid sabouraud’s medium.

2. 1ml pipette.

Steps:
1- Under aseptic conditions transfer 1ml of the sample to the thioglycolate media
and transfer 1 ml of the sample to the sabouraud’s media.

2- Incubate the thioglycolate media for 7 days at 30-35 ºC

Incubate the sabouraud’s media for 10 days at 22-25 ºC

Results:
– If thioglycolate media tubes are

▪ Clear and transparent: sterile sample.
▪ Turbid: bacteria present.

.– If sabouraud's media tubes are

▪ Clear and transparent: sterile sample.

▪ Turbid: fungi present

Comment:

• Preparation of positive and negative controls as:

Negative controls contain media only to ensure sterility of the media.

Positive controls contain media and M.O to ensure suitability of media to M.O.

• Example of fungi used to inoculate Sabouraud’s media: Candida albicans.

• Example of aerobic bacteria used to inoculate thioglycolate media: E. coli or
Staphylococcus aureus.

• Example of anaerobic bacteria used to inoculate thioglycolate media: Clostridium

species.

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