EPA-OW: Bacantr-SG: Bacillus anthracis - Sentinel Procedure
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Official Method Name
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Epidemiologic and Laboratory Investigations of Bovine Anthrax in Two Utah Counties in 1975. |
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Current Revision
| 1977 |
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Media
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WATER |
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Instrumentation
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Plate Count |
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Method Subcategory
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Microbiological |
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Method Source
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Citation
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Sentinel Laboratory Guidelines from Suspected Agents of Bioterrorism: American Society for Microbiology. Final 8/8/03. 23 p. |
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Brief Method Summary
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This method describes procedures for analysis of water and solid samples and may be adapted for assessment of particulate and aerosol samples. After dilution and suspension in phosphate buffer with Tween® 80, soil samples are centrifuged at low speed (400 × g) for 5 minutes to remove sediment, followed by a high-speed (6,000 × g) centrifugation of the supernatant for 30 minutes to pellet spores. The heat-shocked pellet is plated directly on Polymyxin, Lysozyme, EDTA, Thallous acetate (PLET) and sheep blood agar (SBA). Water samples are subjected only to the low speed centrifugation, followed by passing the supernatant through a 0.45 µm filter. The filter is placed in buffer, heat-shocked, and the supernatant plated on PLET and SBA. PLET and SBA plates are examined after 24 and 48 hours incubation at 37°C for colonies with typical morphology. Cultures (isolates) that cannot be ruled out as B. anthracis based on the characteristics noted above should be referred to an appropriate reference laboratory for confirmation. |
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Scope and Application
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This culture method was developed for identifying B. anthracis in soil and water samples. |
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Applicable Concentration Range
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Interferences
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Cross contamination could interfer with results. |
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Quality Control Requirements
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SAM lists these procedures for detection in solid, particulate, aerosol, liquid, and water samples. Further research is needed to develop and standardize the procedures for environmental sample types. At a minimum, the following QC checks should be performed and evaluated when using this protocol: positive control, negative control, and blank. Ongoing analysis of QC samples to ensure reliability of the analytical results should also be performed. |
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Sample Handling
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Maximum Holding Time
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Relative Cost
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Less than $50 |
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Sample Preparation Methods
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