Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended used for selective isolation and cultivation of fastidious anaerobic bacteria.
Composition**
| Ingredients | g/L |
|---|---|
| Tryptone | 15.000 |
| Soya peptone | 5.000 |
| Yeast extract | 5.000 |
| Sodium chloride | 5.000 |
| Phenylethyl alcohol | 2.500 |
| L-Cystine | 0.400 |
| Vitamin K1 | 0.010 |
| Hemin | 0.005 |
| Agar | 20.000 |
| Blood | 50ml |
| Final pH (at 25°C) | 7.5±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically on the plate.
Principle And Interpretation
Phenylethyl alcohol is a chemical agent that exhibits inhibitory action against gram-negative and certain gram-positive bacteria Phenylethyl Blood Agar Base (Anaerobic) is used for the isolation of obligate anaerobic gram-positive and gram-negative bacteria (1). Supplementation of medium with L-cystine permits growth of certain thiol-dependent or sulphur containing amino acids- requiring bacteria (1,2) and fastidious Streptococci. This medium inhibits facultative anaerobic gram-negative bacteria such as E. coli and Proteus species.
Tryptone and Soya peptone provide nitrogen, carbon, sulfur and trace elements to the growing organisms. Addition of sheep blood provides many growth factors. Sodium chloride maintains osmotic equilibrium. Addition of phenylethanol to a nutritive medium permits the growth of gram-positive organisms but inhibits the gram-negative organisms found in the same specimen (3). Phenylethyl alcohol exerts inhibitory bacteriostatic action on gram-negative bacteria by inhibiting their DNA synthesis (4). Addition of hemin, vitamin K1 and L-cystine makes the medium more nutritious and suitable for the growth of fastidious anaerobic bacteria.
Type of specimen
Clinical samples - faeces, pus, etc.
Specimen Collection and Handling:
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (5,6).
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 pack. 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 :
- 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 further identification.
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 Phenyl Blood Agar Plate w/ 5% Sheep Blood in 90 mm disposable plates with smooth surface and absence of
black particles/cracks/bubbles
Colour of medium
Cherry red coloured medium
Quantity of medium
25 ml of medium in 90 mm disposable plates.
pH
7.30-7.70
Sterility Check
Passes release criteria
Cultural Response
Cultural characteristics observed in an anaerobic condition after an incubation at 35-37°C for 48-72 hours (longer if
necessary).
| Organism | Inoculum (CFU) | Growth | Recovery |
|---|---|---|---|
| Bacteroides fragilis ATCC 25285 | 50-100 | good-luxuriant | >=50% |
| Clostridium perfringens ATCC 13124 (00007*) | 50-100 | good-luxuriant | >=50% |
| Clostridium butyricum ATCC 9690 | 50-100 | good-luxuriant | >=50% |
| Clostridium sporogenes ATCC 11437 | 50-100 | good-luxuriant | >=50% |
| Proteus mirabilis ATCC 25933 | 50-100 | fair-good | 30-40% |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | 50-100 | none-poor | <=10% |
Key : (*) Corresponding WDCM numbers.
Storage and Shelf Life
On receipt store between 2-8°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 sample must be decontaminated and disposed of in accordance with current laboratory techniques (5,6).
Reference
- MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. 1, Williams and Wilkins, Baltimore
- Allen S. D., Lombard G. L, Armfield A. Y., Thompson F. S. and Stargel M. D., Abstracts of the Annual Meeting of the American Society for Microbiology, Abstract C142, 1977, p. 59
- Lilley B. D. and Brewer J. H., 1953, J. Am. Pharm. Assoc., 42:6.
- Dowell, Hill and Altemeier, 1964, J. Bacteriol., 88:1811.
- Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
- 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.
| Product Name | Phenyl Blood Agar Plate w/ 5% Sheep Blood |
|---|---|
| SKU | MP540 |
| Customized Product Available | No |
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