Phylogenetic Analysis of Sulphidogenic Bacteria Isolated From Nigerian Crude Oil and Its Produce Water

Phylogenetic analysis of sulphidogenic bacterial isolates from Nigerian Bonny light crude oil and its produce water samples was carried out following preliminary detection of the bacterial species. Samples were enriched in mixed carbon postgate’s (MCP) medium and incubated at 55 ºC for 28 days. DNA was extracted from isolated culture and amplified by Polymerase Chain Reaction (PCR) using 16S rRNA eubacterial primers with a GC–clamp. Successful amplification was confirmed by ethidium bromide fluorescence in 1 % agarose gelafter agarose gel electrophoresis. PCR-amplified DNA was separated by Denaturing Gradient Gel Electrophoresis (DGGE) analysis, to show the relationship between samples based on their GC content. Following successful DGGE analysis, DGGE bands were excised, purified and sequenced to display sequence homology and affiliation to related genera of sulphide producers. The DGGE products sequencing was successful, displaying 98 % and 99 % similarity and homology to Petrotogamexicana (AY125964.1) and Petrotogaolearia (AJ311703.1) respectively. Phylogenetic analysis showed a relationship between the sulphidogenic bacteria with other members of the Thermotogales.