Application of Denaturing High-Performance Liquid Chromatography for Monitoring Sulfate-Reducing Bacteria in Oil Fields

Abstract

Sulfate-reducing bacteria (SRB) participate in microbially induced corrosion (MIC) of equipment and H 2 S-driven reservoir souring in oil field sites. Successful management of industrial processes requires methods that allow robust monitoring of microbial communities. This study investigated the applicability of denaturing high-performance liquid chromatography (DHPLC) targeting the dissimilatory sulfite reductase ß-subunit (dsrB) gene for monitoring SRB communities in oil field samples from the North Sea, the United States, and Brazil. Fifteen of the 28 screened samples gave a positive result in real-time PCR assays, containing 9 ؋ 10 1 to 6 ؋ 10 5 dsrB gene copies ml ؊1 . DHPLC and denaturing gradient gel electrophoresis (DGGE) community profiles of the PCR-positive samples shared an overall similarity; both methods revealed the same samples to have the lowest and highest diversity. The SRB communities were diverse, and different dsrB compositions were detected at different geographical locations. The identified dsrB gene sequences belonged to several phylogenetic groups, such as Desulfovibrio, Desulfococcus, Desulfomicrobium, Desulfobulbus, Desulfotignum, Desulfonatronovibrio, and Desulfonauticus. DHPLC showed an advantage over DGGE in that the community profiles were very reproducible from run to run, and the resolved gene fragments could be collected using an automated fraction collector and sequenced without a further purification step. DGGE, on the other hand, included casting of gradient gels, and several rounds of rerunning, excising, and reamplification of bands were needed for successful sequencing. In summary, DHPLC proved to be a suitable tool for routine monitoring of the diversity of SRB communities in oil field samples.

A Post-Genomic View of the Ecophysiology, Catabolism and Biotechnological Relevance of Sulphate-Reducing Prokaryotes

“…The mechanism of oil degradation under sulphate-reducing conditions, which is responsible for oil field souring, remains largely unknown, because this putative oil-degrading consortium of syntrophs and hydrogenotrophic and acetotrophic SRB has not yet been studied in any detail. A recently introduced method to monitor SRB in oil fields complementary to long established DGGE (Muyzer, 1999) is the so-called denaturing HPLC targeting the dsrB gene coding for the β-subunit of dissimilatory sulphite reductase (Priha et al, 2013).…”

Glycoside hydrolase DisH from Desulfovibrio vulgaris degrades the N‐acetylgalactosamine component of diverse biofilms

“…Annual global corrosion costs are around 3.4% of the global gross domestic product [$2.5 trillion (Koch et al, 2016)], and biocorrosion is responsible for up to 30% of all corrosion losses, especially those in oil production, drinking water systems, pipelines, steel piling in quays, harbours and jetties (Beech and Sunner, 2007;Koch et al, 2016). For example, SRB can corrode buried gas transmission pipelines as well as cause reservoir souring by their H 2 S production (Priha et al, 2013;Enning and Garrelfs, 2014). As MIC is commonly caused by SRB biofilms (Lee et al, 1995), inhibiting and dispersing their biofilms is imperative.…”

Inhibition of biogenic hydrogen sulfide produce by Sulfate Reducing Bacteria isolated from oil fields in Basra by nitrate based treatment

“…Tables (1and 2) revealed reduced in number of SRB at second day of treatment at all of nitrate and nitrite treatments in comparison with control the viable count of SRB reached a maximum number of >1100 cell/ml, then the numbers were rise at all concentrations except of 1000 mg/l nitrate where the growth was completely inhibited and the highest number of cells was 34.8 cell/ml. [27]. In recent years, some papers have examined the microbial community and souring activity of oil fields [2,28].…”