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' Cyl% ture <br /> ENVIRONMENTAL <br /> BIOTECHNOLOGY <br /> CytoCulture International,Inc. <br /> 249 Tewksbury Avenue <br /> Pt.Richmond,CA 94501 USA <br /> ' PSC Industrial Outsourcing, LP Reporting Date: July 7,2009 <br /> 210 West Sand Bank Road CytoCulture lab login: 09-151 <br /> Columbia, IL 62236 Project number: 62403335-024522 <br /> Tel: (618) 281-1543 Fax: <br /> Project Manager: Paul Anderson <br /> ' E-mail: panderson@pscnow.com <br /> ' Samples: Two soil samples on ice were received 06/26/09.They were stored at 4°C and assayed on <br /> the same day. Please see attached chain of custody. <br /> ' AEROBIC <br /> ' Hydrocarbon-Degrading and Heterotrophic Bacteria Enumeration Assays <br /> Analysis Request: Enumeration of aerobic petroleum hydrocarbon-degrading bacteria (broad range <br /> ' petroleum derived from gasoline and diesel) and aerobic total heterotrophic bacteria by method <br /> 9215A (HPC)/ Standard Methods 9215B modified. <br /> ' Carbon Source for Aerobic Hydrocarbon-Degrading Bacteria: Pasteurized Chevron No. 2 <br /> gasoline and diesel were dissolved into agar plates as the sole carbon and energy source for the <br /> growth of aerobic hydrocarbon-degrading bacteria. <br /> Carbon Source for Aerobic Total Heterotrophic Bacteria: Growth medium was prepared with <br /> standard methods total plate count agar (Difco) containing a wide range of carbon sources derived <br /> from yeast extract,tryptone, pancreatic digest of casein and glucose. <br /> Protocol for Hydrocarbon-Degrading Bacteria: Sterile agar plates (100 x 15 mm) were prepared <br /> with 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 a 0.2% sodium pyrophosphate <br /> extract of the sample (log dilution 10°) or log dilutions of the extract at 10 3, 10 4, 10 s, 10-6 and <br /> 10�. Hydrocarbon plates were counted after 7 days incubation at 30°C. The plate count data are <br /> reported as colony forming units (cfu)per gram (g)of soil. Each enumeration value represents a <br /> statistical average of the plate count data obtained from two of the four inoculating log dilutions <br /> ' assayed. <br /> Protocol for Total Heterotrophic Bacteria: Sterile agar plates (100 x 15 mm)were prepared with <br /> minimal salts medium and 2.35%Difco heterotrophic plate count agar at pH 6.8 without any other <br /> 1 <br />