Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Fluid Selenite Cystine Medium (Twin Pack)
For isolation of Salmonellae from food, dairy products, materials of sanitary importance and clinical specimens, as per IP.
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Intended Use
Recommended as an enrichment medium for isolation of Salmonellae from food, dairy products and materials of sanitary importance in accordance with Indian Pharmacopoeia.
Composition
| Ingredients | Gms / Litre |
|---|---|
| Part A | |
| Tryptone # | 5.000 |
| Lactose | 4.000 |
| Sodium phosphate | 10.000 |
| L-Cystine | 0.010 |
| Part B | |
| Sodium hydrogen selenite | 4.000 |
| Final pH (at 25°C) | 7.0±0.2 |
**Formula adjusted, standardized to suit performance parameters
# Pancreatic digest of casein
Directions
Suspend 4.0 grams of Part B in 1000 ml purified/distilled water. Add 19.01 grams of Part A. Mix well. Warm to dissolve the medium completely. Distribute in sterile test tubes. Sterilize in a boiling water bath or free flowing steam for 15 minutes. DO NOT AUTOCLAVE. Excessive heating is detrimental. Discard the prepared medium if large amount of selenite is reduced (indicated by red precipitate at the bottom of tube/bottle).
Caution: Sodium hydrogen selenite (Sodium bi-selenite) is very toxic, corrosive agent and causes teratogenicity. Handle with great care. Upon contact with skin, wash immediately with a lot of water.
Principle And Interpretation
Selective inhibitory effects of selenite were first demonstrated by Klett (1). Guth (2) used it to isolate Salmonella Typhi. Leifson studied selenite and formulated a medium. Fluid Selenite Cystine Medium is a modification of Leifsons (3) formula with added cystine by North and Bartram (4). It is employed for the detection of Salmonellae in foodstuff, particularly egg products. It is included by APНА (5,6), IP (7). Selenite Cystine Broth is useful for detecting Salmonella in the non acute stages of illness when organisms occur in low numbers in test samples and for epidemiological studies to enhance the detection of low numbers of organisms from asymptomatic or convalescent patients (8). Tryptone provide nitrogenous substances. Lactose maintains the pH in medium as selenite is reduced by bacterial growth and alkali is produced. An increase in pH lessens the toxicity of the selenite and results in overgrowth of other bacteria. The acid produced by bacteria due to lactose fermentation serves to maintain a neutral pH. Phosphate maintains a stable pH and also lessens the toxicity of selenite. L-cystine improves recovery of Salmonellae. Enriched broth is subcultured on solid medium.
Type of specimen
Food & dairy samples; Pharmaceutical samples.
Specimen Collection and Handling
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (5,6). After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions
Read the label before opening the container. Wear protective gloves/protective clothing/eye protection/face protection. Follow good microbiological lab practices while handling specimens and culture. Standard precautions as per established guidelines should be followed while handling clinical specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations
- Some organisms may show poor growth due to variable nutritional requirement.
- Do not incubate the broth longer than 24 hours as inhibitory effect of selenite reduces after 6 - 12 hours of incubation (9).
Performance and Evaluation
Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at recommended temperature.
Quality Control
Appearance: Part A: White to cream homogeneous free flowing powder Part B: White to cream Crystalline powder
Colour and Clarity of prepared medium: Light yellow coloured, clear to slightly opalescent solution of complete medium
pH: 6.80-7.20
Growth Promotion Test
In accordance with the harmonized method of IP. Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours when sub cultured on MacConkey Agar (M081).
| Organism | Inoculum (CFU) | Recovery | Colour of Colony |
|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | 50-100 | little-none(no increase in numbers) | pink with bile precipitate |
| Salmonella Choleraesuis ATCC 12011 | 50-100 | luxuriant | colourless |
| Salmonella Typhimurium ATCC 14028 (00031*) | 50-100 | luxuriant | colourless |
| Salmonella Typhi ATCC 6539 | 50-100 | luxuriant | colourless |
| Escherichia coli NCTC 9002 | 50-100 | little-none (no increase in numbers) | pink with bile precipitate |
| Escherichia coli ATCC 8739 (00012*) | 50-100 | little-none (no increase in numbers) | pink with bile precipitate |
Key: *Corresponding WDCM numbers.
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 15-25°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle in order to prevent lump formation due to the hygroscopic nature of the product. Improper storage of the product may lead to lump formation. Store in dry ventilated area protected from extremes of temperature and sources of ignition. Seal the container tightly after use. Product performance is best if used within stated expiry period.
Disposal
User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow established laboratory procedures in disposing of infectious materials and material that comes into contact with clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (10,11)
| Product Name | Fluid Selenite Cystine Medium (Twin Pack) |
|---|---|
| SKU | MM025 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Klett A., 1900, Zeitsch Fer Hyg. Und. Infekt, 33: 137. 2.Guth F., 1916, Zbl. Bakt. I. Orig., 77:487. 3.Leifson E., 1936, Am. J. Hyg., 24(2) : 423. 4.North W.R. and Bartram M.T., 1953, Appl. Microbiol., 1:130. 5.Downes F P and Ito K(Eds.), 2001, Compendium of Methods For The Microbiological Examination of Foods, 4th ed.,APHA, Washington, D.C. 6.Wehr H M and Frank J H., 2004, Standard Methods for the Examination of Dairy Products, 17th ed., APHA Inc., Washington,D.C. 7.Indian Pharmacopoeia, 200 8.Murray PR, Baren EJ, Jorgensen JH, Pfaller MA, Yolken RH (editors) 2003, Manual of clinical Microbiology, 8th ed.,ASM, Washington, D.C. 9.Chattopadhyay W. and Pilford J. N., 1976, Med. Lab. Sci., 33:191. |
| Customized Product Available | No |
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