Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended for selective isolation of Salmonellae from faeces, urine, sewage and other materials in accordance with USP.
Composition**
| Ingredients | g/L |
|---|---|
| Peptone | 5.000 |
| HM peptone B # | 5.000 |
| Tryptone | 5.000 |
| Dextrose (Glucose) | 5.000 |
| Sodium phosphate | 4.000 |
| Ferrous sulphate | 0.300 |
| Bismuth sulphite indicator | 8.000 |
| Brilliant green | 0.025 |
| Agar | 20.000 |
| Final pH (at 25°C) | 7.6±0.2 |
**Formula adjusted, standardized to suit performance parameters
#Equivalent to Beef extract
Directions
Suspend 52.32 grams in 1000 ml purified/distilled water. Heat to boiling to dissolve the medium completely. DO NOT OVERHEAT OR STERILIZE IN AUTOCLAVE or by fractional sterilization since overheating may destroy the selectivity of the medium. Transfer to a water bath maintained at about 50°C.The sensitivity of the medium depends largely upon uniform dispersion of precipitated bismuth sulphite in the final gel, which should be dispersed before pouring into the sterile Petri plates.
Principle And Interpretation
Bismuth Sulphite Agar Medium is prepared in accordance with USP (1) and is employed for the isolation and preliminary identification of Salmonella Typhi and other Salmonellae from pathological materials, sewage, water, food and other products. Bismuth Sulphite Agar is recommended by various Associations (2,3,4,5,6) for the isolation and preliminary identification of Salmonella Typhi and other Salmonellae from pathological materials, sewage, water, food, pharmaceutical and other products. It is a modification of Wilson and Blair medium.
Brilliant green and bismuth sulphite incorporated into the medium inhibit the intestinal gram-negative and gram-positive bacteria, peptone, tryptone and HM peptone B are rich source for supplying essential nutrients for growth of the organism. The fermentable source of carbohydrate in this medium is dextrose, which provides energy for enhanced microbial growth. Phosphates incorporated in the medium act as a good buffering agent. The bismuth ions are reduced to metallic bismuth, which impart the metallic sheen around the colonies. Sulphite is reduced to black ferric sulphide giving the black colour with release of H2S.
Salmonella Enteritidis and Salmonella Typhimurium typically grow as black colonies (rabbit eye colonies) with a surrounding metallic sheen. Salmonella Paratyphi A grow as light green colonies. This medium also favors use of larger inoculum and heavily contaminated samples as compared to other selective media, as it has unique inhibitory action towards gram-positive and coliform organisms. The medium may be inhibitory to some strains of Salmonella species and therefore should not be used as the sole selective medium for these organisms. Shigella species are mostly inhibited on this medium and also some Salmonellae like S.Sendai, S.Berta, S. Gallinarum, S. Abortus-equi are equally inhibited. Proteus species are inhibited but few strains give dull green or brown colonies with metallic sheen.
Type of specimen
Pharmaceutical samples
Specimen Collection and Handling
For pharmaceutical products, follow appropriate techniques for sample processing in case of viscous materials as mentioned under sterility (1). 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
- DO NOT AUTOCLAVE OR OVERHEAT THE MEDIUM, as it destroys the selectivity of the medium.
- S. Typhi and S.Arizonae exhibit typical brown colonies, with or without metallic sheen.
- This medium is highly selective and must be used in parallel with less selective media for isolation.
- With certain Salmonella species, typical black colonies with metallic sheen is observed near heavy inoculation and isolated colonies may show green colonies.
- Shigella species are mostly inhibited on this medium; exceptions being S. flexneri and S. sonnei (7)
- Some Salmonella like S. Sendai, S. Berta, S. Gallinarum, S. Abortus-equi are also inhibited (7).
Quality Control
Appearance
Light yellow to greenish yellow homogeneous free flowing powder
Gelling
Firm, comparable with 2.0% agar gel.
Colour and clarity of prepared medium
Yellow to greenish yellow opalescent with flocculant precipitate
Reaction
Reaction of 5.23% w/v aqueous solution. pH : 7.6±0.2
pH
7.40-7.80
Growth Promotion Test
Growth Promotion is carried out in accordance with the harmonized method of USP. Cultural response was observed after an incubation at 30-35°C for 24-48 hours. Recovery rate is considered as 100% for bacteria growth on Soyabean Casein Digest Agar.
Cultural Response
Cultural characteristics observed after incubation at 30-35 °C for 24-48 hours. Recovery rate is considered as 100%for bacteria growth on Soyabean Casein Digest Agar.
| Organism | Inoculum (CFU) | Growth | Lot value (CFU) | Recovery | Colour of Colony |
|---|---|---|---|---|---|
| Salmonella Typhimurium ATCC 14028 (00031*) | 50-100 | luxuriant | 25-100 | >=50% | black or greenish-grey may have sheen |
| Salmonella Abony NCTC 6017 (00029*) | 50-100 | good-luxuriant | 25-100 | >=50% | black with metallic sheen |
| Additional Microbiological testing | |||||
| #Klebsiella aerogenes ATCC 13048 (00175*) | 50-100 | none-poor | 0-10 | 0-10% | brown-green (depends on the inoculum density) |
| Enterococcus faecalis ATCC 29212 (00087*) | >=103 | inhibited | 0 | 0% | |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | luxuriant | 25-100 | >=50% | black with metallic sheen |
| Salmonella Typhi ATCC 6539 | 50-100 | luxuriant | 50-100 | >=50% | black with metallic sheen |
| Shigella flexneri ATCC 12022 (00126*) | 50-100 | none-poor | 0-10 | <=10% | brown |
| Escherichia coli ATCC 8739 (00012*) | 50-100 | none-poor | 0-10 | <=10% | Brown to green, depends on inoculum density |
Key: *Corresponding WDCM numbers.
#- Formerly known as Enterobacter aerogenes
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 20-30°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 sample must be decontaminated and disposed of in accordance with current laboratory techniques (8,9).
| Product Name | Bismuth Sulphite Agar Medium |
|---|---|
| SKU | MU027 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Tindall B. J., Crimont P. A. D., Gorrity G. M., EUZESY B. P., 2005, Int. J. Sys. Evol. Microbiol., 55:521 2.Koneman E. W., Allen S. D., Janda W. M., Schreckenberger P. C., Winn W. C. Jr., 1992, Colour Atlas and Textbook ofDiagnostic Microbiology, 4 th Ed., J. B. Lippinccott Company 3.Mandell G. L., Douglas R. G. Jr., Bennet J. E., (Eds.) , 1985, Principles and Practice of Infectious Diseases, 2nd Ed., 660-669,John Wiley & Sons New York. 4.Gunter and Tuft, 1939, J. Lab. Clin. Med., 24:461. 5.Wilson and Blair, 1926, J. Pathol. Bateriol., 29:310. 6.Wilson and Blair, 1927, J. Hyg., 26:374 9.Baird R.B., Eaton A.D., and Rice E.W., (Eds.), 2015, Standard Methods for the Examination of Water andWastewater, 23rd ed., APHA, Washington, D.C. 10.FDA Bacteriological Analytical Manual, 2005, 18th Ed., AOAC, Washington, D.C. 11. Murray P. R., Baron J. H., Pfaller M. A., Tenover F. C. and Yolken R. H., (Eds.). 1999, Manual of Clinical Microbiology,7th Ed., American Society for Microbiology, Washington, D.C. 12.Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2015, Compendium of Methods for the Microbiological Examination ofFoods, 5th Ed., American Public Health Association, Washington, D.C.1 3.Indian Pharmacopoeia, 1996, Ministry of Health and Family Welfare, Govt. of India, Volume 2.1 4.MacFaddin J. F., 2000, (Ed.), Biochemical Tests for Identification of Medical Bacteria, 3rd Edition, Lippincott, Williamss& Wilkins, New York.1 5.American Public Health Association, Standard Methods for the Examination of Dairy Products, 1978, 14th Ed.,Washington D.C.1 6.Isenberg, H.D. Clinical Microbiology Procedures Handb0ook. 2nd Edition.1 |
| Customized Product Available | No |
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