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r <br /> cyto <br /> Culture <br /> E N V I R O N M E N T A L <br /> B I O T E C H N O L O G Y <br /> CytoCulture International, Inc 1986 <br /> April 7, 1998 <br /> Client Upgradient Environmental Consultants <br /> P O Box 8002 <br /> Stockton, CA 95208 Project Description Valley Motors <br /> Project No 97023 <br /> Contact Victor B Cherven, Ph D Tel: 209-772-3620 Fax 209-772-3620 <br /> SAMPLES: Three soil samples delivered on Ice on 3/27/98 and processed on 3/30/98 Samples stored at 4 <br /> Deg C for possible follow-up work <br /> Hydrocarbon-Degrading Bacteria Enumeration Assays <br /> ANALYSIS REQUEST: Bacterial enumeration for total petroleum hydrocarbon-degraders (broad range <br /> petroleum hydrocarbons diesel/gasoline, and waste oil) <br /> CARBON SOURCE Petroleum hydrocarbons were added as the sole carbon and energy sources for the <br /> 40, growth of hydrocarbon-degrading aerobic bacteria on agar plates Gasoline <br /> (Chevron Reg ) was added to the lids to provide petroleum hydrocarbon vapors <br /> Diesel (Chevron#2) and motor oil were blended in a 50 50 ratio and dissolved <br /> into the agar to provide additional aliphatic hydrocarbons in the growth matrix <br /> PROTOCOL: <br /> Hydrocarbon Degraders Stenle agar plates (100 x 15 mm) were prepared with <br /> minimal salts medium at pH 6 8 with 15% noble agar, without any other carbon <br /> sources or nutrients added A 200 pl aliquot of pasteurized gasoline was added to <br /> absorbent paper in the plate lids to provide the vapor source of light fraction <br /> hydrocarbons Plates were Inoculated with 10 ml of sample or a log dilution of <br /> each water sample Triplicate plates were inoculated with sample log dilutions of <br /> 10°, 10-', 10-2 and 10-3 The hydrocarbon plates were poured on 3/30198 and <br /> counted after 8 days on 4/7/98 The plate count data are reported as colony <br /> forming units (cfu)per gram(g)of sample Each bacteria population value <br /> represents a statistical average of the plate count data obtained with inoculations <br /> for two of the three log dilutions tested <br /> Vox 510 233 0102 Fax 510 233 3777 Email Cyto®CytoCulture com <br /> 249 Tewksbury Avenue Point Richmond California 94801 3829 USA <br />