Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Tryptose Sulphite Neomycin Agar is used for selective isolation of Clostridium perfringens in foods or other specimens.
Composition
| Ingredients | Gms / Litre |
|---|---|
| Tryptose | 15.000 |
| Yeast extract | 10.000 |
| Sodium sulphite | 1.000 |
| Ferric citrate | 0.500 |
| Neomycin sulphate | 0.050 |
| Polymyxin B sulphate | 0.020 |
| Agar | 13.500 |
| Final pH (at 25°C) | 7.2±0.2 |
Formula adjusted, standardized to suit performance parameters
Directions
Suspend 40.07 grams in 1000 ml distilled water. Heat to boiling to dissolve the medium completely. Dispense in screw capped containers. Sterilize by autoclaving with caps loose at 118°C for 12 minutes. Close the caps while the medium is still hot. 5 ml of sterile buffered thioglycollate solution may be added to every 200 ml of medium if desired. The buffered aqueous thioglycollate solution contains 35 ml buffer mixture (5.7% dipotassium phosphate and 28% sodium carbonate) and 15 ml sodium thioglycollate solution (13.3%).
Principle And Interpretation
Clostridium perfringens food poisoning is one of the most common types of human foodborne illness (1). The foods usually involved are cooked meat or poultry containing large numbers of viable cells. A heat labile enterotoxin produced only by sporulation cells (2) induces the major symptoms of diarrhea in perfringens poisoning (6). Tryptose Sulphite Neomycin Agar is a modification of Mossel Medium (3) developed by Marshall et al (4) for the selective isolation and enumeration of C.perfringens from food. Thioglycollate addition is recommended if the cultured medium is to be incubated anaerobically (4, 5).
Tryptose and yeast extract provide nitrogenous compounds, vitamin B complex and other growth nutrients. The antibiotics neomycin and polymyxin B sulphate inhibit gram-negative enteric bacilli. Neomycin is also lethal for C.bifermentans The colonies of C.perfringens are black due to the ferric sulphide formed after the sulphite reduction. The high incubation temperature of 46°C renders the medium specific for C.perfringens. The presumptive black colonies of C.perfringens should be confirmed biochemically. The selectivity of the medium results in the inhibition of some strains of C.perfringens (6).
Quality Control
Appearance
Cream to yellow homogeneous free flowing powder
Gelling
Firm, comparable with 1.35% Agar gel.
Colour and Clarity of prepared medium
Medium amber coloured clear to slightly opalescent gel forms in Petri plates.
Reaction
Reaction of 4.0% w/v aqueous solution at 25°C. pH : 7.2±0.2
pH
7.00-7.40
Cultural Characteristics
M634: Cultural characteristics observed under anaerobic conditions, after an incubation at 46°C for 18-48 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of colony |
|---|---|---|---|---|
| Clostridium perfringens ATCC 12924 | 50-100 | good-luxuriant | >=50% | black |
| Escherichia coli ATCC 25922 | >=103 | inhibited | 0% | |
| Staphylococcus aureus ATCC 25923 | >=103 | inhibited | 0% |
Storage and Shelf Life
Store dehydrated medium and prepared medium at 2-8°C. Use before expiry period on the label.
| Product Name | Tryptose Sulphite Neomycin Agar |
|---|---|
| SKU | M634 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Doyle M. P., (Ed.), 1989, Foodborne Practical Pathogens, Marell Dekker, New York , N. Y. 2.Dunean C. L., 1973, A. J. Bacteriol., 113: 932 3.Mossel, 1959, J. Sci. Food Agric., 10:662. 4.Marshall, Steenbergen and McClung, 1965, Appl. Microbiol., 13:559. 5.MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. I, Williamsand Wilkins, Baltimore. 6.Vanderzant C. and Splittstoesser D. F., (Eds.), 1992, Compendium of Methods for the Microbiological Examination ofFoods, 3rd Ed., APHA, Washington, D.C. |
| Customized Product Available | No |
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