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Fluid Thioglycollate Medium
For sterility testing of biologicals and for cultivation of aerobes, anaerobes and microaerophiles in accordance with US Pharmacopoeia.
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Intended Use
Fluid Thioglycollate Medium is used for sterility testing of biologicals and for cultivation of aerobes, anaerobes and microaerophiles in accordance with United States Pharmacopoeia
Composition
| Ingredients | Gms/Litre |
|---|---|
| Pancreatic digest of casein | 15.000 |
| Yeast extract | 5.000 |
| Dextrose monohydrate | 5.500 |
| Sodium chloride | 2.500 |
| L-Cystine | 0.500 |
| Sodium thioglycollate | 0.500 |
| Resazurin sodium | 0.001 |
| Agar | 0.750 |
| pH after sterilization | 7.1±0.2 |
Formula adjusted, standardized to suit performance parameters
Directions
Suspend 29.25 grams of dehydrated medium in 1000 ml purified/distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 25°C and store in a cool dark place preferably below 25°C.
Note: If more than the upper one-third of the medium has acquired a pink colour, the medium may be restored once by heating in a water bath or in free flowing steam until the pink colour
Principle And Interpretation
Brewer (1) formulated Fluid Thioglycollate Medium for rapid cultivation of aerobes as well as anaerobes by adding a reducing agent and small amount of agar. The USP (2), BP (3), EP (4)and AOAC (5) have recommended the media for sterility testing of antibiotics, biologicals and food products and for determining the phenol coefficient and sporicidal effect of disinfectants. However, it is intended for the examination of clear liquid or water-soluble materials.
Dextrose monohydrate, pancreatic digest of casein, yeast extract, L-cystine provide the growth factors necessary for bacterial multiplication. Sodium thioglycollate and L-cystine act as a reducing agent lowering the oxidation-reduction potential by removal of oxygen This condition helps to prevent the accumulation of peroxides which is toxic in nature. The SH group also neutralizes the antibacterial effect of mercurial preservatives and other heavy metal compounds which exert a bacteriostatic effect in the materials under examination. Any increase in the oxygen content is indicated by a colour change of redox indicator-resazurin; to red (6,7,8). The small amount of agar helps in maintaining low redox potential and stabilizes the medium (9).
In sterility checking, it is recommended to dilute the sample containing preservatives, with this broth to reduce the toxicity and enhance the growth of contaminants, if any.
Quality Control
Appearance
Cream to yellow homogeneous free flowing powder
Colour and Clarity of prepared medium
Light straw coloured clear to slightly opalescent solution with upper 10% or less medium pink on standing.
Reaction
After sterilization, reaction of 2.92% w/v aqueous solution. pH: 7.1±0.2
pH
6.90-7.30
Growth Promotion Test
As per United States Pharmacopoeia
Stability test
Light yellow coloured clear solution without any precipitation sedimentation at room temperature for 7 days
Cultural response
Cultural characteristics observed after an incubation at 30-35°C for not more than 3 days.
Cultural Response
| Organism | Inoculum (CFU) | Growth |
|---|---|---|
| Clostridium sporogenes ATCC 19404 | 50-100 | luxuriant |
| Clostridium sporogenes ATCC 11437 | 50-100 | luxuriant |
| Clostridium sporogenes NBRC 14293 | 50-100 | luxuriant |
| Clostridium perfringens ATCC 13124 | 50-100 | luxuriant |
| Bacteroides fragilis ATCC 23745 | 50-100 | luxuriant |
| Bacteroides vulgatus ATCC 8482 | 50-100 | luxuriant |
| Staphylococcus aureus ATCC25923 | 50-100 | luxuriant |
| Staphylococcus aureus ATCC 6538 | 50-100 | luxuriant |
| Pseudomonas aeruginosa ATCC 27853 | 50-100 | luxuriant |
| Pseudomonas aeruginosa ATCC 9027 | 50-100 | luxuriant |
| Micrococcus luteus ATCC 9341 | 50-100 | luxuriant |
| Streptococcus pneumoniae ATCC 6305 | 50-100 | luxuriant |
| Escherichia coli ATCC 25922 | 50-100 | luxuriant |
| Escherichia coli ATCC 8739 | 50-100 | luxuriant |
| Escherichia coli NCTC 9002 | 50-100 | luxuriant |
| Salmonella Typhimurium ATCC 14028 | 50-100 | luxuriant |
| Salmonella Abony NCTC 6017 | 50-100 | luxuriant |
| Bacillus subtilis ATCC 6633 | 50-100 | luxuriant |
Storage and Shelf Life
Store below 30°C in tightly closed container and the prepared medium at room temperature and away from light. Use before expiry date on the label.
| Product Name | Fluid Thioglycollate Medium |
|---|---|
| SKU | MU009 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1.Brewer, 1940, J. Am. Med. Assoc., 115:598. 2.The United States Pharmacopoeia, 2018, The United States Pharmacopoeial Convention, Rockville, MD. 3.British Pharmacopoeia, 2016, The Stationery office British Pharmacopoeia4.European Pharmacopoeia, 2017, European Dept. for the quality of Medicines. 5.Williams H., (Ed.), 2005, Official Methods of Analysis of the Association of Official Analytical Chemists, 19th Ed., AOAC,Washington, D.C 6.Marshall, Gunnison and Luxen, 1940, Proc. Soc. Exp. Biol. Med., 43:672. 7.Nungester, Hood and Warren, 1943, Proc. Soc. Exp. Biol. Med., 52:287. 8.Portwood, 1944, J. Bact., 48:255. 9.MacFaddin J.F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. 1, Williamsand Wilkins, Baltimore. 10.Federal Register, 1992, Fed. Regist., 21:640.0.. 11. Quastel and Stephenson, 1926, J.Biochem., 20:1.. 12.Isenberg, H.D. Clinical Microbiology Procedures Handb0ook. 2nd Edition.13Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015)Manual of Clinical Microbiology, 11th Edition. Vol. 1. |
| Customized Product Available | No |
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