Screening of polycyclic aromatic hydrocarbon degrading bacterial isolates from oil refinery wastewater and detection of conjugative plasmids in polycyclic aromatic hydrocarbon tolerant and multi-metal resistant bacteria

Wastewater were collected from the effluent channel in the vicinity of Mathura oil refinery, U.P. (India) and analysed for physicochemical characteristics, heavy metals as well as organic compounds including PAHs. The interaction of PAHs and heavy metals with various group of microorganisms revealed the viable count of aerobic heterotrophs, asymbiotic nitrogen fixers, actinomycetes and fungi were found to be 2.38 x 10(6), 1.89 x 10(4), 2.20 x 10(4) CFU/mL and 8.76 x 10(3) CFU/mL respectively. We have selected and screened 50 bacterial isolates for their resistance/tolerance to heavy metal and PAHs. Out of 25 multi-metal resistant isolates, 6 were able to tolerate PAHs at the concentration of 5000 mu g/mL (50 mu g/disc) to naphthalene, anthracene, phenanthrene and pyrene. The PAH degradation efficiency of the isolates was assessed using spectrophotometer with 100 mu g/mL of phenanthrene and observed different degree of degradation ranging from 34-66% after 96 h of incubation. One of the bacterial isolates KWB3 (identified as Enterobacter ludwigii by 16S rDNA sequencing) exhibited maximum degradation efficiency (66%) was further tested for phenanthrene degrading ability in the presence and absence of a co-substrate (glucose) in a mineral salt medium; and a number of metabolites were produced and detected by GC-MS which revealed the presence of benzocoumarin, phthalic acid, catechol and several low molecular weight compounds. The DNA derived from multi-metal and PAHs tolerant bacteria were PCR amplified using Inc specific primers and positive PCR products were obtained with oriT and trfA2 of the IncP group; indicates that these bacteria have gene-mobilizing capacity.