Mueller Kauffman Tetrathionate Broth Base, Granulated

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SKU:
GM876
For improved enrichment and isolation of Salmonellae.


Composition

Ingredients Gms/Litre
Casein enzymic hydrolysate 7.000
Papaic digest of soyabean meal 2.300
Sodium chloride 2.300
Calcium carbonate 25.000
Sodium thiosulphate 40.700
Ox bile 4.750

Formula adjusted, standardized to suit performance parameters

Directions

Suspend 82.05 grams in 1000 ml distilled water. Heat just to boiling. DO NOT AUTOCLAVE. Cool and just before use aseptically add 19 ml of iodine solution (20 g iodine and 25 g potassium iodide in 100 ml sterile distilled water) and 9.5 ml of 0.1% brilliant green solution. Mix well before dispensing in sterile tubes to disperse calcium carbonate uniformly .

Note: Due to presence of calcium carbonate, the prepared media forms opalescent solution with white precipitate

Principle And Interpretation

The examination of various types of food products for Salmonella requires methods different from those used in clinical laboratories. The need for such method is due to the generally low numbers of Salmonellae in foods and the frequently poor physiological state of these pathogens following exposure to stressful conditions during food processing or storage. Injured Salmonella are resuscitated in non-selective broth medium, which facilitates detection of sublethally injured Salmonella . The ideal pre-enrichment broth should provide for the repair of cell damage, dilute toxic or inhibitory substances and nutritive enough to favour growth of Salmonella. In the analysis of food for Salmonella, pre-enrichment cultures are usually incubated at 35-37°C for 18-24 hours and then a portion is subcultured to one or more selective enrichment broths. Normally 1 ml of pre-enrichment culture is inoculated to 9 ml of selective enrichment broth. Selective enrichment media contains selective ingredients that allow the proliferation of Salmonella and inhibit the growth of competing non-salmonella microorganisms. Lactose Broth (M1003) is recommended by BAM for pre-enrichment of Salmonella from food. Selective enrichment is done in Tetrathionate Broth and Rappaport Vassiliadis Medium. For the detection of foodborne Salmonella, various modifications of Tetrathionate Broth have generally found wider applications (7).

Mueller (1) recommended Tetrathionate Broth as a selective medium for the isolation of Salmonella. Kauffman (2) modified the formula to include ox bile and brilliant green as selective agents to suppress bacteria such as Proteus species. The British Standard Specification specifies Brilliant Green Tetrathionate Broth for isolating Salmonella from meat and meat products and from poultry and poultry products (3). It is also a recommended selective broth for isolating Salmonella from animal feces and sewage-polluted water (4). Selectivity is conferred by tetrathionate (from the reaction of thiosulphate and iodine). Using more than one selective broth increases the isolation of Salmonella from samples with multiple serotypes (5). Mueller Kauffman Tetrathionate Broth Base conforms to ISO specifications (9).

Mueller Kauffman Tetrathionate Broth Base contains casein enzymic hydrolysate and papaic digest of soyabean meal as sources of carbon, nitrogen, vitamins and minerals. Ox bile and added brilliant green are selective agents, which inhibit gram-positive and other gram-negative organisms. Calcium carbonate is the buffer. Sodium chloride maintains osmotic equilibrium. Sodium thiosulphate is a source of sulfur. The tetrathionate (S4O6) anions constitute the principle selective agent in these enrichment media. If desired, 4 mg of novobiocin per litre of broth can be added to suppress Proteus species (6).

Add approximately 10 grams of sample to 100 ml of broth. Shake well and place the flask in a 45°C water bath for 15 minutes. Remove the flasks and place in an incubator or water bath at 43°C. Several studies have shown increased recovery of Salmonella following incubation of selective enrichment at 43°C (8). After an incubation for 18-24 hours and 48 hours, subculture on Brilliant Green Agar, Modified (M016). This medium is not suitable for the growth of Salmonella Typhi, Salmonella Sendai, and Salmonella Pullorum etc.

The complete medium is unstable and should be used immediately. It may be stored at 2-8°C in the dark for no more than 7 days.

Organisms other than Salmonellae, such as Morganella morganii and some Enterobacteriaceae may grow in the medium. Therefore, confirmatory tests should be carried out on all presumptive Salmonella colonies that are recovered.

Quality Control

Appearance: Cream to yellow coloured granular medium

Colour and Clarity of prepared medium: With added brilliant green and iodine solution - Light green coloured opalescent solution forms with heavy white precipitate

Cultural Response: Cultural characteristics observed, when subcultured on Soyabean Casein digest Agar, after an incubation at 43°C for 18-24 hours with added iodine and brilliant green solution.

Organism Inoculum
(CFU)
Recovery
Salmonella Typhimurium ATCC 14028 50-100 excellent
Salmonella Enteritidis ATCC 13076 50-100 excellent
Salmonella Paratyphi A ATCC 9150 50-100 excellent
Salmonella Paratyphi B ATCC 8759 50-100 excellent
Salmonella Typhi ATCC 6539 >=10³ inhibited
Escherichia coli ATCC 25922 50-100 none-poor
Proteus vulgaris ATCC 13315 50-100 none-poor
Shigella flexneri ATCC12022 >=10³ inhibited

Storage and Shelf Life

Store below 30°C in tightly closed container and use the freshly prepared medium. Use before expiry date on the label.

More Information
Product Name Mueller Kauffman Tetrathionate Broth Base, Granulated
SKU GM876
Customized Product Available No
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