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Cyto <br /> Culture <br /> ENVIRON MEN TAL <br /> B I O T E C H N O L O G Y <br /> C;yrnC:ulturt tntcrnati,mat tnc 1986 <br /> October 31, 1996 <br /> Client: Upgradrent Environmental <br /> Contact: Vic Cherven Fax: (209) 772 - 3620 <br /> Client Code: Upgrad Project Description: Gillies Trucknig <br /> SAMPLES: Two soil samples in 2" brass sort sleezes were received on 10/17196 The samples were stored at <br /> 4°C <br /> Hydrocarbon-Degrading Bacteria Plate Enumeration Assay Results <br /> ANALYSIS REQUEST: Bacterial enumeration for total petroleum hydrocarbon-degraders (target <br /> hydrocarbons Gasoline/Diesel and Motor Oil) <br /> CARBON SOURCE Diesel(Chevron #2) and motor oil were blended in a 50 50 ratio as the sole carbon <br /> and energy sources for the growth of hydrocarbon-degrading aerobic bacteria on <br /> agar plates Gasoline (Chevron Reg ) was added to the lids to provide petroleum <br /> hydrocarbon vapors <br /> PROTOCOL: Sterile agar plates(100 x 15 mm)were prepared with minimal salts medium at pH <br /> 6 8 with 1 5%noble agar, without any other carbon sources or nutrients added A <br /> 200 gl aliquot of pasteurized gasoline was added to absorbent paper in the plate <br /> lids to provide a vapor source of light fraction hydrocarbons Plates for the soil <br /> samples received on 10117/96 were inoculated with 100 gl of a soil extract, or a <br /> log dilution of the extract Triplicate plates were inoculated at sample dilutions of <br /> 10°, 10"i, and 10-2 The two soil sample plates were spread on I0/17/96 and <br /> counted after 14 days on 10/31/96 The plate count data are reported as colony <br /> forming units (efu) per gram (gm) for the soil samples Each enumeration value <br /> represents a statistical average of the plate count data obtained with inoculations <br /> at the 3 dilutions <br /> Vox 51C 233 0102 Fax 510 2333777 Ecniil C'vr,)1nc@ jot c,mi <br /> 24'�Tewk.bury Avenue Poinr Rldi mond Cflibirmt 94801-3829 U,A <br />