Indicators of petroleum hydrocarbon biodegradation in anaerobic granitic groundwater

• The aim of this study was to find indicators of petroleum biodegradation in granitic groundwater. Both pristine and contaminated groundwaters from boreholes around petroleum storage vaults located approximately 40 m below the surface in granite and with storage capacities of up to 120,000 m(3) were sampled. Total numbers of microorganisms, "most probable numbers" (MPN) of anaerobic bacteria, and chemical indications of microbial activity were determined in the groundwater. Hydrocarbon contaminants and metabolites were detected using gas chromatographymass spectrometry (GC-MS). In contaminated groundwater, the total number of microorganisms was 2-4 x 10(6) ml(-1), which was significantly higher than the 6 x 10(4) ml(-1) found in pristine groundwater. This microbial abundance was also reflected in the MPN analysis. Up to 7 x 10(4) nitrate-, 2 x 10(3) iron-, and 3 x 10(4) sulfate-reducing bacteria were detected in contaminated groundwaters. In such groundwaters, depletion of anaerobic electron acceptors and detection of reduced species could be established. We also proposed using a high alkalinity/hardness of water quota (A/H quota) as an indicator of microbial activity. In contaminated groundwaters the A/H quota averaged 2.8, while in pristine groundwater the same was only 1.3. Moreover, the presence of 20 oxidized petroleum hydrocarbons, i.e., putative metabolites of which 9 were strictly intracellular, was detected in the contaminated groundwaters. Phylogenetic neighbor-joining analysis of 16S rRNA genes provided information about the bacterial communities. The bacteria in contaminated groundwater were found to be strikingly similar to bacteria in other hydrocarbon-contaminated environments.

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