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•i <br /> AEROBIC <br /> . Hydrocarbon-Degrading and Total Heterotrophic Bacteria Enumeration Assay <br /> Results <br /> Client Sample Sample Hydrocarbon Target Total <br /> Number Date Degraders Hydrocarbons Heterotrophs <br /> (cfu/ml) Tested (cfu/ml) <br /> MW-1 06/05/03 9 x 102 Gasoline/Diesel 4x 103 <br /> MW-4 06/05/03 4X 103 Gasoline/Diesel 7x 103 <br /> MW-5B 06/05/03 5X 103 Gasoline/Diesel 1 x 104 <br /> MW-8 06/05/03 6x 103 Gasoline/Diesel 9x 103 <br /> Sterile Water 06/06/03 Zero Gasoline/Diesel Zero <br /> Air Control 06/06/03 Zero Gasoline/Diesel Zero <br /> Positive Control 06/05/03 1 x 108 Gasoline/Diesel 3 x 108 <br /> Reporting Limit for enumeration data is 1 0 x 101 cfu/ml <br /> A positive control sample of hydrocarbon-degrading bacteria was run concurrently with <br /> these samples using a mixed flask culture of bacteria isolated from contaminated <br /> groundwater sites in Northern California <br /> ANAEROBIC <br /> Hydrocarbon-Degrading Bacteria Enumeration Assay <br /> Analysis Request: Enumeration of anaerobic petroleum hydrocarbon-degrading bacteria <br /> (broad range petroleum hydrocarbons derived from diesel and gasoline) by method <br /> 9215A (HPC) / Standard Methods 9215B modified for anaerobic conditions <br /> Protocol for Anaerobic Hydrocarbon-Degrading Bacteria- Pasteurized Chevron No 2 <br /> diesel and gasoline were dissolved into agar plates as the sole carbon and energy sources <br /> for the growth of hydrocarbon-degrading anaerobic bacteria The medium includes <br /> alternative terminal electron acceptors such as sulfate, nitrate, and iron Sterile agar <br /> plates (100 x 15 mm)were prepared with minimal salts medium at pH 6 8 with agar and <br /> hydrocarbons, without any other carbon sources or nutrients added Plates were setup <br /> and poured in a Coy anaerobic glove box under strict anaerobic conditions (atmosphere <br /> of nitrogen, carbon dioxide and hydrogen) Triplicate plates were inoculated with 10 ml <br /> of each sample (log dilution 10°) or log dilutions of the sample at 1Ot, 10-2,and 10 3 <br /> Hydrocarbon plates were counted after 12 days incubation in the glove box at ambient <br /> temperature The plate count data is reported as colony farming units (cfu) per milliliter <br /> (ml) Each enumeration value represents a statistical average of two of the four <br /> inoculating log dilutions assayed <br />