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Cyt <br /> Culture <br /> ENVIRONMENTAL <br /> BIOTECH NOLOG Y <br /> CytoCulture International,Inc <br /> 249 Tewksbury Avenue <br /> Pt Richmond,CA 94801 USA <br /> Clearwater Group Reporting Date September 26,2003 <br /> Project Name Barnes Trucking CytoCulture Lab Login 03-162 <br /> Project Manager Brian Pierskalla P O number <br /> Address 229 Tewksbury Ave Project number ZB178C <br /> Point Richmond, CA 94801 Tel 510-307-9943 Fax 510-232-2823 <br /> Samples: Four water samples on ice were received on 09/17/03 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 gasoline No 2 and diesel <br /> were dissolved into agar plates as the sole carbon and energy source for the growth of aerobic <br /> hydrocarbon-degrading bacteria Sterile agar plates (100x 15 mm) were prepared with minimal <br /> salts medium at pH 6 8 with agar and hydrocarbons, without any other carbon sources or <br /> nutrients added Triplicate plates were inoculated with 1 0 ml of each sample (log dilution 10a) <br /> or log dilutions of each sample at 10 1, 10-2, and 10 3 Hydrocarbon plates were counted after 7 <br /> days incubation at 30°C The plate count data is reported as colony forming units (cfu) per <br /> 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 10 ml of each sample at log <br /> dilutions 101, 10 z, and 10 3 The heterotrophic plates were counted after 3 days of incubation at <br /> 30°C The plate count data is reported as colony forming units (cfu)per milliliter (ml) of sample <br /> . Each enumeration value represents a statistical average of two of the four inoculating log <br /> dilutions assayed <br /> i <br />