TITAN MEDIA ANAEROBIC THIOGLYCOLLATE MEDIUM BASE For cultivation of anaerobes Composition Ingredients Casein enzymatic hydrolysate Meat

extract D- Glucose Papaic digest of Soyabean meal Liver hydrolysate Sodium chloride Sodium sulphite Sodium thioglycollate L-Cystine Agar Methylene blue Gms/Ltr. 17.00 7.50 6.00 3.00 3.00 2.50 0.100 0.50 0.25 0.70 0.002 TM 1499

* Dehydrated powder, hygroscopic in nature, store in a dry place, in tightly-sealed containers below 25C and protect from direct Sunlight. Instructions for Use Dissolve 40.50gms in 900 ml of distilled water. Gently heat to boiling with gentle swirling and dissolve the medium completely. Sterilize by autoclaving at 15 psi (121C) for 15 minutes. Mix well. Cool to 45500C and pour into sterile test tubes. Appearance: Light amber to dark amber colour, clear to slightly opalescent gel pH (at 25C): 7.3 0.2 Principle ANAEROBIC THIOGLYCOLLATE MEDIUM BASE is used for cultivation of anaerobes. Medium contains Casein enzymatic hydrolysate, Papaic digest of Soyabean meal, Liver hydrolysate and Meat extract are the source of nitrogen, carbon, vitamins & minerals for the growth of anaerobic bacteria. D- Glucose is a carbon energy source. D-glucose in plant extracts usually occurs together with maltose, malto saccharides, starch, sucrose and/or -linked gluco oligosaccharides. L- cysteine act as reducing agents and maintain a low oxygen tension in the medium. This enables the obligate anaerobes to multiply. The small amount of Agar helps in anaerobiosis. Methylene blue is a dye which provides a visual indication and presence of oxygen. Methylene blue is blue when oxidized and colorless when reduced, a suitable environment for anaerobes to grow. Both facultative anaerobes and aerotolerant anaerobes will grow throughout the tube; however, facultative anaerobes will grow most densely where Oxygen is present. Sodium thiglycollate and Sodium sulpthite provides a low oxidation-reduction potential, and securing anaerobic conditions. This sulphydryl groups also inactivates arsenic, mercury and other metal compounds. Therefore, the medium containing thioglycollate are suitable for examine heavy metals in samples. Sodium chloride helps to maintain the osmotic balance of the medium.

Interpretation Cultural characteristics observed under anaerobic conditions after inoculating (103CFU/ml), on incubation at 35 2C for 48 - 72 hours.

Microorganisms

ATCC

Inoculum (CFU/ml) 10 10 10

Growth

Clostridium perfringens Clostridium sporogenes Bacteroides vulgatus

12924 19404 8482

Good Good Good

References 1.Brewer J. H., Science, 95:587. (1942). 2. Baron E. J., Peterson and Finegold S. M., Bailey & Scotts Diagnostic Microbiology, 9th Ed., MosbyYear Book, Inc., St. Louis, Mo. (1994). 3. Bulletin, National Institute of Health, (1941). 4. United States Pharmacopeia XXVI, Chapter "Microbial Limit Tests", (2003). For more details visit:- http://www.titanmedia.in/images/upload/td/TM %201499_ANAEROBICTHIOGLYCOLLATEMEDIUMBASETM1499.pdf or http://www.titanmedia.in