Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
Your enquiry has been submitted
Something problem with the server. Try again.
Intended Use:
Recommended for the selective isolation of Vibrio cholerae and other enteropathogenic Vibrio's.
Composition**
| Ingredients | g/L |
|---|---|
| Peptone, special | 10.000 |
| Yeast extract | 5.000 |
| Sodium citrate | 10.000 |
| Sodium thiosulphate | 10.000 |
| Sodium cholate | 3.000 |
| Bile# | 5.000 |
| Sucrose | 20.000 |
| Sodium chloride | 10.000 |
| Ferric citrate | 1.000 |
| Bromo thymol blue | 0.040 |
| Thymol blue | 0.040 |
| Agar | 15.000 |
| Final pH (at 25°C) | 8.8±0.2 |
**Formula adjusted, standardized to suit performance parameters
# Equivalent to Oxgall
Directions
Either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically on the plate.
Principle And Interpretation
TCBS Agar was developed by Kobayashi et al (1), who modified the selective medium of Nakanishi (2). Although this medium was originally designed for the isolation of V. cholerae and V. parahaemolyticus, most Vibrios grow to healthy large colonies with many different colonial morphologies. TCBS Agar is also recommended by APHA for the selective isolation of V.cholerae and V.parahaemolyticus (1,3). Enrichment in Alkaline Peptone Water (M618), followed by isolation on TCBS Agar is routinely used for isolation of V.cholerae (2,4,5). TCBS Agar, Selective has an additional selective ingredient i.e. sodium cholate for improved selectivity.
Peptone special and yeast extract provide nitrogenous, carbonaceous compounds, long chain amino acids, vitamin B complex and other essential growth nutrients. Bile and sodium citrate inhibit gram-positive bacteria and coliforms (6). Sodium thiosulphate serves as a good source of sulphur, which in combination with ferric citrate detects the production of hydrogen sulphide. For the metabolism of Vibrios, sucrose is added as a fermentable carbohydrate. Vibrio that is able to utilize sucrose will form yellow colonies. Bromothymol blue and thymol blue are the pH indicators. The alkaline pH of the medium improves the recovery of V.cholerae. Strains of V. cholerae produce yellow colonies on TCBS Agar because of fermentation of sucrose. V. alginolyticus also produce yellow colonies. V. parahaemolyticus is a sucrose non-fermenting organism and therefore produces blue-green colonies, as does V. vulnificus. Proteus species that are sucrose-fermenters may form yellow colonies (7). TCBS Agar is not a suitable medium for oxidase testing of Vibrio species (8). A few strains of V. cholerae may appear green or colourless on TCBS Agar due to delayed sucrose fermentation (7). TCBS Agar is highly selective for Vibrio species. However, occasional isolates of Pseudomonas and Aeromonas may also form blue green colonies on TCBS Agar (8). Any H2S negative colony of TCBS Agar can be considered presumptive positive for Vibrio.
Type of specimen
Clinical: faeces, etc; Food samples; Water samples
Specimen Collection and Handling
For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards (4). For food samples, follow appropriate techniques for sample collection and processing as per guidelines (6).
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (9,10). After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions
In Vitro diagnostic use only. For professional use only. 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
- The medium should be inoculated heavily with faecal specimens because growth of few species may be inhibited on the medium due to fermentation of sucrose and accumulation of acids.
- However, occasional isolates of Pseudomonas and Aeromonas may also form blue green colonies on TCBS Agar (8).
- Proteus species that are sucrose-fermenters may form yellow colonies (8).
- TCBS Agar is not a suitable medium for oxidase testing of Vibrio species (9).
- A few strains of V.cholerae may appear green or colourless on TCBS Agar due to delayed sucrose fermentation.
- TCBS Agar is highly selective for Vibrio species. Any H2S negative colony of TCBS Agar can be considered presumptive positive for Vibrio.
- Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
- Each lot of the medium has been tested for the organisms specified on the COA. It is recommended to users to validate the medium for any specific microorganism other than mentioned in the COA based on the user's unique requirement.
- Further biochemical and serological tests must be carried out for complete identification.
- It is recommended to store the plates at 24-30°C to avoid minimum condensation.
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
Sterile TCBS Agar in 90 mm disposable Petri plates with smooth surface and absence of black particles/cracks/bubbles
Colour of medium
Bluish green coloured medium
Quantity of medium
25ml of medium in 90mm disposable plate
pH
8.60-9.00
Sterility Check
Passes release criteria
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours.
| Organism | Inoculum (CFU) |
Growth | Recovery | Colour of colony |
|---|---|---|---|---|
| Vibrio parahaemolyticus ATCC 17802 (00037*) | 50-100 | good-luxuriant | >=50% | bluish green |
| Vibrio vulnificus ATCC 29306 | 50-100 | fair-good | >=30% | greenish yellow |
| Vibrio fluvialis ATCC 33809 (00137*) | 50-100 | good-luxuriant | >=50% | yellow |
| Vibrio cholerae ATCC 15748 | 50-100 | good-luxuriant | >=50% | yellow |
| Escherichia coli ATCC 25922 (00013*) | >=103 | inhibited | 0% | |
| Enterococcus faecalis ATCC 29212 (00087*) | >=103 | inhibited | 0% | |
| Shigella flexneri ATCC 12022 (00126*) | >=103 | inhibited | 0% |
Key: (*) Corresponding WDCM numbers
Storage and Shelf Life
On receipt store between 20-30°C. Use before expiry date on the label. 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 (11,12).
| Product Name | TCBS Agar Plate |
|---|---|
| SKU | MP870 |
| Customized Product Available | No |
Write Your Own Review