TITAN MEDIA NIH THIOGLYCOLLATE MEDIUM (ALTERNATIVE THIOGLYCOLLATE MEDIUM) (as per IP) For sterility testing of biological

products Composition Ingredients Casein enzymatic hydrolysate Dextrose (Anh.) Yeast extract Sodium chloride L- Cystine Sodium thioglycollate Gms/Ltr. 15.00 5.50 5.00 2.50 0.50 0.50 TM 302

* 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 29gms in 1000ml distilled water. Gently heat to boiling with gentle swirling and dissolve the medium completely. Dispense into culture tubes in 15 to 18 ml volumes. Sterilize by autoclaving at 15 psi (121C) for 15 minutes. Note: Prepared medium should be store at 15-300C. If more than 30% of the medium is pink before use, reheat once (1000C) in a water bath or in free flowing steam until the pink colour disappears and to drive off absorbed oxygen. Appearance: Light yellow colour, slightly opalescent solution pH (at 25C): 7.1 0.2 Principle NIH THIOGLYCOLLATE MEDIUM is used for sterility testing of biological products. The media is also used for sterility testing of antibiotics, biologicals and foods and also for determining the phenol coefficient and sporicidal effect of disinfectants. However, it is intended for the examination of clear liquid or water-soluble materials. Fluid Thioglycollate Medium is also routinely used to check the sterility of stored blood in blood banks. The medium contains Casein enzymatic hydrolysate and Yeast extract those provides the nitrogen and essential growth enhancers necessary for bacterial multiplication. Sodium thioglycollate and L- Cystine acts as a reducing agent which maintains a low oxygen tension by removing molecular oxygen from the environment. Dextrose is a fermentable energy source. On incubation period of 30 - 350 C for 48- 72 hours shown luxuriant growth in all the culture strains tested. Sodium chloride helps maintaining the osmotic balance. The NIH Thioglycollate Medium without Indicator is suitable for diagnostic work of viscous products because the lack of indicator avoids possible toxicity to organisms. Interpretation Cultural characteristics observed after inoculating (103CFU/ml) in culture tubes, tightly caped on incubation at 30-350C for 18-48 hours (or upto 72 hours, if necessary) under anaerobic conditions.

Microorganisms

ATCC

Inoculum (CFU/ml) 10 10 10 10

Growth

Bacillus subtilis Clostridium sporogenes Staphylococcus aureus Bacteroides fragilis

6633 19404 25923 23745

Luxuriant Luxuriant Luxuriant Luxuriant

Clostridium sporogenes incubated at 35C for 24 - 48 hours under aerobic conditions. References 1. Brewer, J. Am. Med. Assoc., 115:598. (1940). 2. Marshall, Gunnison and Luxen, Proc. Soc. Exp. Biol. Med., 43:672. (1940). 3. Williams H., (Ed.), Official Methods of Analysis of the Association of Official Analytical Chemists, 4. 19th Ed., AOAC, Washington, D.C. (2005). 5. N.I.H. Memorandum. Culture Media for Sterility Tests, 4th Revision. (1955). For more details visit:- http://www.titanmedia.in/images/upload/td/TM %20302_NIHTHIOGLYCOLLATEMEDIUMTM302.pdf or http://www.titanmedia.in