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Fluid Selenite Cystine HiVeg™ Medium (Selenite Cystine HiVeg™ Broth) (Twin Pack)
An enrichment medium for isolation of Salmonellae in food, dairy products and materials of sanitary importance and clinical specimens, animal free.
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Intended Use
Recommended as an enrichment medium for isolation of Salmonellae from foods, dairy products, materials of sanitary specimens.
Composition**
| Ingredients | g / L |
|---|---|
| Part A | |
| HiVeg hydrolysate | 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
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 10 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).
Note: Instead of Part B, DD056 (Sterile Sodium Biselenite Discs) or DB001 (Sterile Sodium Biselenite Bud) can also be used. It has to be added to the dissolved Part A medium after boiling as above in appropriate quantities. Recommended to use; one disc of DD056 per 10 ml of Part A medium or one bud of DB001 per 100 ml of Part A medium.
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 using selenite. Fluid Selenite Cystine Medium is a modification of Leifsons (3) formula with added cystine (4). The formulation corresponds to that recommended by AOAC (5) for the detection of Salmonella in foodstuff, particularly egg products. It is also recommended by APHA (6,7) and USP (8).
Selenite Cystine Broth is useful for detecting Salmonella in the non-acute stages of illness when organisms occur in the faeces in low numbers and for epidemiological studies to enhance the detection of low numbers of organisms from asymptomatic or convalescent patients (9). Salmonella are also injured during various food processing procedures, including exposure to low temperatures, sub-marginal heat, drying, radiation, preservatives or sanitizers. Recovery of Salmonella involves pre- enrichment, selective enrichment and selective plating since Salmonella may be present in low numbers in food sample in an injured conditions. Fluid Selenite Cystine HiVeg® Medium is same as Fluid Selenite Cystine Medium except that the animal based peptones are completely replaced with vegetable peptones to avoid BSE/TSE risks associated with animal peptones. Fluid Selenite Cystine HiVeg® Medium is used as selective enrichment medium for the cultivation of Salmonella species. This medium is formulated to allow the proliferation of Salmonella while inhibiting the growth of competing non-Salmonella organisms.
HiVeg® hydrolysate provides nitrogenous substances. Lactose is the fermentable carbohydrate and 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 is the reducing agent, improving the recovery of Salmonella. Enriched broth is subcultured on solid medium. Do not incubate the broth longer than 24 hours as inhibitory effect of selenite reduces after 6 - 12 hours of incubation (10). Inoculate the food sample into recommended pre-enrichment broth, and then transfer 1 ml of mixture to 10 ml of Fluid Selenite Cystine HiVeg® Medium and also to 10 ml Fluid Tetrathionate HiVeg® Medium w/o Iodine and BG (MV032). Incubate and subsequently subculture on to Bismuth Sulphite HiVeg® Agar (MV027), XLD HiVeg® Agar (MV031), Hektoen Enteric HiVeg®Agar (MV467) or MacConkey HiVeg® Agar (MV081).
Type of specimen
Food and dairy samples.
Specimen Collection and Handling:
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (6,7,11). 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 specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations :
- Some organisms may show poor growth due to variable nutritional requirement.
Quality Control
Appearance
Part A:Cream to yellow homogeneous free flowing powder. Part B: White to cream homogeneous free flowing powder
Colour and Clarity of prepared medium
Light yellow coloured, clear to slightly opalescent solution of complete medium
Reaction
Reaction of medium [(1.9% w/v) Part A and (0.4% w/v) Part B] at 25°C. pH: 7.0±0.2
pH
6.80-7.20
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours when sub cultured on XLD HiVeg™ Agar (MV031).
| Organism | Inoculum (CFU) | Recovery | Colour of Colony |
|---|---|---|---|
| Salmonella Choleraesuis ATCC 12011 | 50-100 | luxuriant | red w/black centre |
| Salmonella Typhimurium ATCC 14028 (00031*) | 50-100 | luxuriant | red w/black centre |
| Salmonella Typhi ATCC 6539 | 50-100 | luxuriant | red w/black centre |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | luxuriant | red w/black centre |
| Pseudomonas aeruginosa ATCC 27853 (00025*) | 50-100 | luxuriant | red |
| Escherichia coli ATCC 8739 (00012*) | 50-100 | little-none (no increase in numbers) | yellow |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | little-none(no increase in numbers) | yellow |
| Enterococcus faecalis ATCC 29212 (00087*) | >=104 | Inhibited |
Key: (*) Corresponding WDCM numbers
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 2-8°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle inorder 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 sample must be decontaminated and disposed of in accordance with current laboratory techniques (12,13).
| Product Name | Fluid Selenite Cystine HiVeg™ Medium (Selenite Cystine HiVeg™ Broth) (Twin Pack) |
|---|---|
| SKU | MV025 |
| Product Type | HiVeg™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg), Lactose |
| 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.FDA Bacteriological Analytical Manual, 2005, 18th Ed., AOAC, Washington, DC. 6.Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2015, Compendium of Methods for the Microbiological Examination ofFoods, 5th Ed.,APHA, Washington, D.C. 7.Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th Ed.,APHA Inc., Washington, D.C. 8.The United States Pharmacopeia, 2017, USP29/NF24, The United States Pharmacopeial Convention, Rockville, M. D. 9.Murray P. R., Baron E. J., Jorgensen J. H., Pfaller M. A., Yolken R. H., (Eds.), 8th Ed., 2003, Manual of ClinicalMicrobiology, ASM, Washington, D.C. 10.Chattopadhyay W. and Pilford J. N., 1976, Med. Lab. Sci., 33:191. 11. Hartman P. A. and S. A., Munich, 1981, J. FoodPract., 44: 385-386 12.Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition.1 3.Jorgensen,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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