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Endospore Production in Response to Microbiocides in the Microbiologically Influenced Corrosion Implicated Genus Clostridium

Endospore-producing bacteria are often detected in oilfield produced waters via 16S rRNA identification. Although prevalent in oilfield brines little is known about their significance and potential implications on microbial control efforts. The presence of noxious chemicals and subsequent chemical damage can force a capable cell into an endospore a dormant structure that preserves the cellular genetic material. As extracellular conditions improve such as the dilution or removal of antimicrobials the endospore again becomes a vegetative cell. These endospores are largely impervious to and are not readily eradicated by antimicrobial agents. Here commonly used microbiocide chemistries are applied in differing concentrations and contact times in order to evaluate the cellular response scored as cell survival endospore formation or cell death. Outcomes were determined utilizing the schaeffer-fulton method for endospore staining and a fluorescence assay of bacterial viability based upon membrane integrity. Treatments are applied to cultured Bacillus subtilis the laboratory model organism commonly utilized to study endospore formation as well as unadulterated produced water containing members of the Phylum Firmicutes including the endospore capable Geobacillus stearothermophilus. Results indicate that the presence of a large population of endospore-forming bacteria may explain the failure of some microbiocide applications.

Product Number: 51319-13126-SG
Author: Jodi Wrangham
Publication Date: 2019
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Characterization of Microbiologically Influenced Corrosion Potential in Nitrate Injected Produced Waters

Product Number: 51319-13198-SG
Author: Mohita Sharma
Publication Date: 2019
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Microorganisms are notorious for being involved in serious metal infrastructure damage popularly known as microbiologically influenced corrosion (MIC). Long term corrosion incubations (~2 years) with carbon steel (CS) beads were established using produced water collected from a Canadian oilfield where nitrate was routinely used for souring mitigation.Experiments were set up under methanogenic sulfate-reducing and nitrate-reducing conditions to stimulate electrical MIC (EMIC) with iron present as the sole electron donor.Microbial community analysis chemical measurements metal weight loss and surface analysis were conducted to assess EMIC under these different conditions. After 2 years incubations in the nitrate-reducing environment did not show surface damage to the carbon steel beads nor substantial weight loss (2-3%). However incubations in the sulfate-reducing environment showed 10-16% weight loss with severe pitting on the CS beads and community sequencing revealed the predominance of known acid producers (<em>Mesotoga</em>and<em>Acetobacterium</em>) and methanogens (<em>Methanosaeta</em>). Incubations in the methanogenic environment showed comparatively less weight loss (2-6%) though surface analysis revealed an abundance of pinhole-like pits; microbial communities were dominated by putative syntrophs (<em>Petrimonas</em>and<em>Pseudomonas</em>) and a known methanogen(<em>Methanosaeta</em>). In addition to the establishment of new EMIC enrichment cultures this study demonstrated that the localized effect of MIC cannot be accurately assessed solely using weight loss corrosion assays but additionally requires microscopic and surface studies along with an understanding of the microbial community composition.