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b. Procedure <br /> (i) Use aseptic techniques to transfer growth from each colony into individual tubes. For 10 colonies, <br /> there should be 10 tubes total, for each verification test media. <br /> (ii) See Table 2 for the method requirements for fecal coliform and Enterococci. Follow the method <br /> requirements for all others, <br /> Table 2 : Verification Tests <br /> Method Verify with the wing. <br /> BHI Agar Slant BHI Gram stain BHI BEA BHI w/6 .5% <br /> NaCl <br /> 35 f 0. 5 °C 35 t 0. 5 °C (gram 45 t 0 . 5 °C 35 t 0 . 5 °C <br /> Enterococci 48 f 3 h 24 f 2 h positive 48 f 3 h 48 t 3 h 35 f, 0 . 5 °C <br /> EPA 1600 (use growth for (turbidity) cocci) (turbidity) (growth w/ 48 t 3 h <br /> gram staining) black brown (turbidity) <br /> precipitate) <br /> LTB EC <br /> Fecal 35 f 0 . 5 °C 44 . 5 t 0 .2°C <br /> Coliforms 48 t 3 h 24 f 2 h <br /> SM9222D (turbidity and (turbidity and <br /> gas) gas) <br /> The response bolded in parentheses indicate the positive result for each test. <br /> c . Reporting - <br /> (i) Adjust colony counts for the original positive sample based on percent of colonies verified positive . <br /> ( 1 ) For example : A sample dilution has a colony count of 30 fecal coliforms. Ten of these positive <br /> blue colonies were used in the verification tests, but only 8 verified positive for both EC and LTB . <br /> This means only 80% were verified positive and therefore the final count of 30 is adjusted by 80% <br /> to 24 colonies. <br /> (ii) For verified samples, report as "Verified" in the comment field, <br /> 10. Data Qualifier Codes - The use of Data Qualifier Codes is not discussed in the methods, but the Department ' s <br /> QA-Rule 62460 F .A.C . requires that data qualifiers be used when the data is being submitted to the State. The <br /> purpose of the Data Qualifier Codes is to communicate the reliability of the reported data to the consumer. <br /> Table 3 identifies those Data Qualifier codes that are generally associated with microbiological data reporting. <br /> Others may apply. The laboratory must apply any applicable data qualifiers as listed in Table 1 of the Quality <br /> Assurance Rule (62- 160, F.A.C .) <br /> a. Any data point which is derived from any analysis other than the direct calculation of the number <br /> of <br /> colonies on a membrane filter that were within the recommended range of the method must be qualified <br /> with one or more of the Data Qualifier Codes listed below. <br /> b. Any result associated with a failed QC test must be reported with applicable data qualifiers. <br /> c . Any result that is associated with a failure to meet test requirements (e .g. , holding time, incubation time, <br /> etc .) shall also be qualified with applicable data qualifiers. <br /> d. Failure to report data with appropriate data . qualifier codes will be returned to the laboratory <br /> without <br /> payment for services until corrections are made . <br /> REMAINDER OF PAGE INTENTIONALLY LEFT BLANK <br /> Revision Date: 02/09 <br /> DEP Agreement No. G0353 , Attachment J, Page 12 of 14 <br />