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Antibiotic Assay Medium No. 36
Recommended as a general purpose medium used with or without blood or other enrichment, for isolating a wide variety of fastidious microorganisms.
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Description
Antibiotic Assay Medium No. 36 is used as a general purpose medium for cultivation of a wide variety of fastidious microorganisms.
Composition**
| Ingredients | Gms / Litre |
|---|---|
| Casein enzymic hydrolysate | 15.000 |
| Papaic digest of soyabean meal | 5.000 |
| Sodium chloride | 5.000 |
| Agar | 15.000 |
Final pH ( at 25°C): 7.3±0.2
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 40.0 grams in 1000 ml purified / distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. If desired aseptically add 5% v/v defibrinated blood in previously cooled medium at 45 - 50°C. Mix well before pouring.
Principle And Interpretation
The composition of this medium is in accordance to CFR (1). This medium is recommended for sterility testing (2). Antibiotic Assay Medium No. 36 is widely employed as seed agar for agar diffusion assay for antibiotic bleomycin. The test organism Mycobacterium smegmatis is also maintained in this medium. This medium is employed for cultivation and isolation of fastidious or nonfastidious microorganisms. This medium is also used as maintenance medium of Psuedomonas aeruginosa for plate assay of ticarcillin. The medium like the conventional medium is used for a multitude of purposes including maintenance of stock cultures, plate counting, isolation of microorganisms from a variety of specimen types and a base for media containing blood (3,4).
Casein enzymic hydrolysate and papiac digest of soyabean gives the essential nutrients for maintenance and growth of the test organisms used. Osmotic balance is maintained by sodium chloride. Agar provides excellent medium for antibiotic diffusion and gives well defined zones of inhibition. Freshly prepared plates should be used for antibiotic assays. Test organisms are inoculated in sterile seed agar pre-cooled to 40-45°C and spread evenly over the surface of solidified base agar. All conditions in the microbiological assay must be controlled carefully. The use of standard culture media in the test is one of the important step for the good results.
Quality Control
Appearance: Cream to yellow homogeneous free flowing powder
Gelling: Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium: Basal Medium : Light yellow coloured clear to slightly opalescent gel. After addition of 5-7%w/v sterile defibrinated blood : Cherry red coloured opaque gel forms in Petri plates
Reaction: Reaction of 4.0% w/v aqueous solution at 25°C (after sterilization). pH : 7.3±0.2
pH: 7.10-7.50
Cultural Response: M1666: Cultural characteristics observed with added 5-7%w/v sterile defibrinated blood after an incubation at 35-37°C for 18-48 hours.
Cultural Response
| Organism | Inoculum (CFU) | Growth | Recovery | Growth w/ blood | Recovery w/ blood |
|---|---|---|---|---|---|
| Growth at 30-35°C for <= 3 days | |||||
| Bacillus subtilis ATCC 6633 | 50-100 | luxuriant | >=70% | luxuriant | >=70% |
| Staphylococcus aureus ATCC 25923 | 50-100 | luxuriant | >=70% | luxuriant | >=70% |
| Staphylococcus aureus ATCC 6538 | 50-100 | luxuriant | >=70% | luxuriant | >=70% |
| Escherichia coli ATCC 25922 | 50-100 | luxuriant | >=70% | luxuriant | >=70% |
| Escherichia coli ATCC 8739 | 50-100 | luxuriant | >=70% | luxuriant | >=70% |
| Escherichia coli NCTC 9002 | 50-100 | luxuriant | >=70% | luxuriant | >=70% |
| Pseudomonas aeruginosa ATCC 27853 | 50-100 | luxuriant | >=70% | luxuriant | >=70% |
| Pseudomonas aeruginosa ATCC 9027 | 50-100 | luxuriant | >=70% | luxuriant | >=70% |
| Salmonella Abony NCTC 6017 | 50-100 | luxuriant | >=70% | luxuriant | >=70% |
| Micrococcus luteus ATCC 9341 | 50-100 | luxuriant | >=70% | luxuriant | >=70% |
| Salmonella Typhimurium ATCC 14028 | 50-100 | luxuriant | >=70% | luxuriant | >=70% |
| Streptococcus pneumoniae ATCC 6305 | 50-100 | luxuriant | >=70% | luxuriant | >=70% |
| Growth at 20-25°C for <= 5 days | |||||
| *Aspergillus brasiliensis ATCC 16404 | 50-100 | luxuriant | luxuriant | ||
| Candida albicans ATCC 2091 | 50-100 | luxuriant | >=70% | luxuriant | >=70% |
| Candida albicans ATCC 10231 | 50-100 | luxuriant | >=70% | luxuriant | >=70% |
Storage and Shelf Life
Store below 30°C in tightly closed container and the prepared medium at 2 - 8°C. Use before expiry date on the label.
Reference
- Tests and Methods of Assay of Antibiotics and Antibiotic containing Drugs, FDA, CFR, 1983 Title 21, Part 436,Subpart D, Washington, D.C.: U.S. Government Printing Office, paragraphs 436, 100-436, 106, p. 242-259, (April1).
- Wright and Welch, 1959-60, Antibiotic Ann., 61.
- MacFaddin 1985, Media for isolation-cultivation-identification-maintenance medical bacteria Vol, I, Williams, & Wilkins, Baltimore, MD
- Forbes BA, Sahm DF, Weissfeld AS, 2002, Bailey and Scott's Diagnostic Microbiology, 11th ed.
| Product Name | Antibiotic Assay Medium No. 36 |
|---|---|
| SKU | M1666 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Tests and Methods of Assay of Antibiotics and Antibiotic containing Drugs, FDA, CFR, 1983 Title 21, Part 436,Subpart D,Washington, D.C.: U.S. Government Printing Office, paragraphs436, 100-436, 106, p. 242-259, (April1). |
| Customized Product Available | No |
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