Ready-to-use media
TCBS Agar ISO
Solid medium for the selective isolation of Vibrio spp. and Vibrio parahaemolyticus according to the ISO standard.
- Cat. No.
- LC-957
- Category
- Ready-to-use media
- Format
- 20 plates, 90 mm
- Storage
- 2 to 14 °C
- Minimum guaranteed shelf life
- 1 month
- Standards
- ISO 21872
- Also available
- Dehydrated medium, 1 format (LC-1074)
TCBS Agar is universally accepted as the medium of choice for differential isolation of enteropathogenic vibrios, whilst inhibiting all the accompanying organisms. This formulation provides high growth of Vibrio cholerae, V. parahaemolyticus, V. alginolyticus and NAG-vibrios. Enterobacteria are strongly inhibited by high concentrations of citrate, thiosulfate, bile and sodium chloride.
Key features
Applications
Selective isolation of Vibrio.
Industry
Clinical, food.
Standards
ISO 21872.
Presentation
20 prepared plates, 90 mm, with 21 ± 2 mL.
Description and technique
Description
Although some enteric bacteria may also grow in this medium, their colony morphology is quite different to that of Vibrio spp.
The organisms that can be confused with vibrios are some biotypes of Proteus and Pseudomonas. There are some resistant enterococci which may form exceptionally small and yellow colonies on this medium. Usually, colonies are selected or chosen and then identified with primary tests (oxidase reaction, Kligler Iron Agar, MRVP Broth and antibiotic sensitivity test) before performing serological identification and phage typing.
Due to its high selectivity, the medium can be seeded with a large inoculum of pathological material. Once solidified and cooled, the medium is turbid, but the observations are not affected.
This medium is very thermolabile and so it must not be autoclaved, overheated or re-melted.
Colonial appearance on TCBS Agar after 24 hours at 37 °C:
- Vibrio alginolyticus and Vibrio cholerae: large, yellow colonies.
- Vibrio parahaemolyticus: small, yellow, without halo and with a green core.
- Streptococcus faecalis: very small and convex, yellow with yellow halo.
- Enterobacteria generally: small and transparent.
- Pseudomonas, Aeromonas, Proteus: medium size and blue.
- Some strains of Vibrio cholerae and Vibrio parahaemolyticus carry out delayed sucrose fermentation so they produce medium sized colonies, and are colourless or dirty yellow with a dark nucleus.
Presentation
| Presentation | 20 prepared plates, 90 mm |
|---|---|
| Filling | 21 ± 2 mL |
| Packaging | 1 box with 2 packs of 10 plates/pack. Single cellophane. |
Composition
| Ingredient | g/L |
|---|---|
| Peptone from casein | 5.00 |
| Meat peptone | 5.00 |
| Yeast extract | 5.00 |
| Sodium citrate | 10.0 |
| Sodium thiosulfate | 10.0 |
| Ox bile | 8.00 |
| Sucrose | 20.0 |
| Sodium chloride | 10.0 |
| Ferric citrate | 1.00 |
| Thymol blue | 0.04 |
| Bromthymol blue | 0.04 |
| Agar | 14.0 |
Quality control
Physical/chemical control
| Color | Green |
|---|---|
| pH | 8.6 ± 0.2 at 25 °C |
Microbiological control
Previous enrichment: 6 ± 1 h (ASPW). Streak isolation.
Aerobiosis. Incubation at 37 ± 1 °C, reading after 24-48 ± 2 h.
| Microorganism | Growth |
|---|---|
| Vibrio alginolyticusATCC 17749 | Good - opaque yellowish colonies |
| Vibrio parahaemolyticusATCC 17802, WDCM 00037 | Good / green to dark green |
| Vibrio furnissiiNCTC 11218, WDCM 00186 | Good - opaque yellowish colonies |
| Escherichia coliATCC 8739, WDCM 00012 | Inhibited |
Sterility control
Incubation 48 hours at 30-35 °C and 48 hours at 20-25 °C: NO GROWTH.
Check at 7 days after incubation in same conditions.
Bibliography
- ATLAS, R.M. & L.C. PARKS (1993) Handbook of Microbiological Media. CRC Press. Boca Raton. Fla. USA.
- BHATTACHARYA, M.K., S.K. BATTACHARYA, S. GARG, P.K. SAHA, D. DUTTA, G.B. NAIR, B.C. DEB & K.P. DAS (1993) Outbreak of Vibrio cholerae non-01 in India and Bangladesh. Lancet, 341:1345-1347.
- DOWNES, F.P. & K. ITO (2001). Compendium of Methods for the Microbiological Examination of Foods. 4th ed. APHA. Washington. DC. USA.
- FORBES, B.A., D.F. SAHM & A.S. WEISSFELD (Eds) (1998) Bailey & Scott's Diagnostic Microbiology 10th ed. Mosby. St. Louis, MO. USA.
- HORWITZ, W. (Ed) (2000) Official Methods of Analysis of AOAC International. 17th ed. Gaithersburg. MD. USA.
- ISO 21872-1 Technical Specification (2017) Microbiology of Food chain - Horizontal method for the detection of potentially enteropathogenic Vibrio spp. - Part 1: Detection of Vibrio parahaemolyticus and Vibrio cholerae and Vibrio vulnificus.
- ISO 11133:2014/Amd 1:2018. Microbiology of food, animal feed and water. Preparation, production, storage and performance testing of culture media.
- KOBAYASHI, T., ENOMOTO, S. SAKAZARI, R. and KUWAHARA, S. (1963) A new selective medium for pathogenic vibrios: TCBS (modified Nakanishi Agar). Jap. J. Bact. 18:387.
- MacFADDIN, J.F. (1985) Media for isolation-cultivation-identification-maintenance of medical bacteria. Williams & Wilkins. Baltimore. MD. USA.
- MURRAY, P.R., E.J. BARON, J.H. JORGENSEN, M.A. PFALLER & R.H. YOLKEN (Eds) (2003) Manual of Clinical Microbiology 8th ed. ASM Press. Washington. DC. USA.
- PASCUAL ANDERSON, Mª R. (1992) Microbiología Alimentaria. Díaz de Santos, S.A. Madrid.
- US FDA (Food and Drug Administration) (1998) Bacteriological Analytical Manual 8th ed. Rev. A. AOAC International Inc. Gaithersburg. MD. USA.
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