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cyto <br /> Culture <br /> ENVIRON MlNT ,tL <br /> 13101 LCH NOL06Y <br /> CytoCulture International, Inc <br /> 249 Tewksbury Avenue <br /> Pt RLchmond,CA 94801 USA <br /> Clearwater Group Reporting Date December 30, 2003 <br /> Project Name Barnes Trucking CytoCulture Lab Login 03-210 <br /> Project Manager Floyd Barnes Project number ZB178C <br /> Address 229 Tewksbury Ave <br /> Point Richmond, CA 94801 <br /> Tel 510-307-9943 Fax 510-232-2823 <br /> Samples Four water samples on ice were received on 12/17/03 They were stored at 4°C <br /> Bacterial and chemical assays were performed the same day Please see attached chain of <br /> custody <br /> AEROBIC <br /> Hydrocarbon-Degrading and Total Heterotrophic <br /> Bacteria Enumeration Assays <br /> Analysis Request: Enumeration of aerobic petroleum hydrocarbon-degrading bacteria (broad <br /> range petroleum derived from gasoline and diesel) and aerobic total heterotrophic bacteria by <br /> method 9215A (HPC)/ Standard Methods 9215B modified <br /> Protocol for Hydrocarbon-Degrading Bacteria. Pasteurized Chevron regular gasoline and <br /> diesel No 2 were dissolved into agar plates as the sole carbon and energy source for the growth <br /> of aerobic hydrocarbon-degrading bacteria Sterile agar plates (100x 15 mm) were prepared with <br /> minimal salts medium at pH 6 8 with agar and hydrocarbons, without any other carbon sources <br /> or nutrients added Triplicate plates were inoculated with 1 0 ml of each sample (log dilution <br /> °) or log dilutions of each sample at 10 1, 102, and 10.3 <br /> 10 Hydrocarbon plates were counted <br /> after 7 days incubation at 30°C The plate count data is reported as colony forming units (efu) <br /> per milliliter (ml) Each enumeration value represents a statistical average of the plate count data <br /> obtained from two of the four inoculating log dilutions assayed <br /> Protocol for Total Heterotrophic Bacteria. Sterile agar plates (100 x 15 mm) were prepared <br /> with minimal salts and 2 35% heterotrophic plate count agar at pH 6 8 without any other carbon <br /> source or nutrients added Triplicate plates were inoculated with 1 0 ml of each sample at log <br /> dilutions 10-1, 10 2, and 10.3 The heterotrophic plates were counted after 5 days of incubation at <br /> 30°C The plate count data is reported as colony forming units (cfu) per milliliter (ml) of sample <br /> 1 <br />