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• Each enumeration value represents a statistical average of two of the four inoculating log <br /> dilutions assayed <br /> AEROBIC <br /> Hydrocarbon-Degrading and Total Heterotrophic <br /> Bacteria Enumeration Assay Results <br /> Client Sample Hydrocarbon Target Total <br /> Number Sample Date Degraders Hydrocarbons Heterotrophs <br /> cfu/ml) Tested cfu/ml <br /> MW-1 12/16/03 6 x 102 Gasoline/Diesel 7x 103 <br /> MW-4 12/16/03 < 1 x 101 Gasoline/Diesel 3 x 103 <br /> MW-5B 12/16/03 3 x 103 Gasoline/Diesel 9X 103 <br /> MW-8 12/16/03 1 x 102 Gasoline/Diesel 6x 102 <br /> Sterile Water 12/17/03 0 Gasoline/Diesel 0 <br /> Air Control 12/17/03 0 Gasoline/Diesel < 1 x 101 <br /> Positive Control 12/17/03 6 x 10" Gasoline/Diesel 8X 109 <br /> Reporting Limit for enumeration data is 1 0 x 101 cfu/ml <br /> ANAEROBIC <br /> Hydrocarbon-Degrading and Total Heterotrophic <br /> Bacteria Enumeration Assays <br /> Analysis Request Enumeration of anaerobic total heterotrophic bacteria and for anaerobic <br /> petroleum hydrocarbon-degrading bacteria (broad range petroleum hydrocarbons derived from <br /> diesel and gasoline) by method 9215A (HPC) / Standard Methods 9215B modified for anaerobic <br /> conditions at 30° C <br /> Protocol for Anaerobic Total Heterotrophic Bacteria- Growth medium was prepared with <br /> Brewer's Anaerobic Agar (Difco) containing a wide range of carbon sources derived from yeast <br /> extract, tryptone, pancreatic digest of casein and glucose The medium includes alternative <br /> terminal electron acceptors such as sulfate, nitrate and iron Sterile petri dish plates (100 x 15 <br /> mm) were prepared with Difco Brewer Anaerobic agar and minimal salts medium at pH 6 8 <br /> without any other carbon sources or nutrients added Plates were set up and poured in a Coy <br /> anaerobic glove box under strict anaerobic conditions (atmosphere of 80% nitrogen, 15% carbon <br /> dioxide and 5% hydrogen) Triplicate sets of plates were inoculated with 1 0 ml of the sample <br /> (log dilution 100) or with log dilutions of the sample at 10-1, 10-2, and 10-3 The heterotrophic <br /> plates were counted after 13 days of incubation at ambient temperature inside the anaerobic <br /> 2 <br />