ABSTRACT
Microbes from oil reservoirs shape petroleum composition through processes such as biodegradation or souring. Such processes are considered economically detrimental and might pose health and safety hazards. It is therefore crucial to understand the composition of a reservoir's microbial community and its metabolic capabilities. However, such analyses are hindered by difficulties in extracting DNA from such complex fluids as crude oil. Here, we present a novel DNA extraction method from oils with a wide American Petroleum Institute (API) gravity (density) range. We investigated the ability to extract cells from oils with different solvents and surfactants, the latter both nonionic and ionic. Furthermore, we evaluated three DNA extraction methods. Overall, the best DNA yields and the highest number of 16S rRNA reads were achieved with isooctane as a solvent, followed by an ionic surfactant treatment using sodium dodecyl sulfate and DNA extraction using the PowerSoil Pro Kit (Qiagen). The final method was then applied to various oils from oil reservoirs collected in aseptic conditions. Despite the expected low cell density of 101-103 cells/ml, the new method yielded reliable results, with average 16S rRNA sequencing reads in the order of 41431 (±8860) per sample. Thermophilic, halophilic, and anaerobic taxa, which are most likely to be indigenous to the oil reservoir, were found in all samples. API gravity and DNA yield, despite the sufficient DNA obtained, did not show a correlation.
ABSTRACT
BACKGROUND: High primary productivity in the midst of high toxicity defines hydrocarbon seeps; this feature usually results in significantly higher biomass, but in lower diversity communities at seeps rather than in the surrounding non-seep benthos. Qualitative estimates indicate that this dichotomy does not necessarily hold true in high latitude regions with respect to megafauna. Instead, high latitude seeps appear to function as local hotspots of both megafaunal diversity and abundance, although quantitative studies do not exist. In this study, we tested this hypothesis quantitatively by comparing georeferenced seafloor mosaics of a seep in the southwestern Barents Sea with the adjacent non-seep seafloor. METHODS: Seafloor images of the Svanefjell seep site and the adjacent non seep-influenced background seabed in the southwestern Barents Sea were used to construct georeferenced mosaics. All megafauna were enumerated and mapped on these mosaics and comparisons of the communities at the seep site and the non-seep background site were compared. Sediment push cores were taken in order to assess the sediment geochemical environment. RESULTS: Taxonomic richness and abundance were both considerably higher at the seep site than the non-seep location. However, taxa were fewer at the seep site compared to other seeps in the Barents Sea or the Arctic, which is likely due to the Svanefjell seep site exhibiting relatively low seepage rates (and correspondingly less chemosynthesis based primary production). Crusts of seep carbonates account for the higher diversity of the seep site compared to the background site, since most animals were either colonizing crust surfaces or using them for shelter or coverage. Our results indicate that seeps in northern latitudes can enhance local benthic diversity and this effect can take place even with weak seepage. Since crusts of seep carbonates account for most of the aggregating effect of sites experiencing moderate/weak seepage such as the study site, this means that the ability of seep sites to attract benthic species extends well beyond the life cycle of the seep itself, which has important implications for the larger marine ecosystem and its management policies.
