Metabolomic and metagenomic analysis of two crude oil production pipelines experiencing differential rates of corrosion

Corrosion processes in two North Sea oil production pipelines were studied by analyzing pig envelope samples via metagenomic and metabolomic techniques. Both production systems have similar physico-chemical properties and injection waters are treated with nitrate, but one pipeline experiences severe corrosion and the other does not. Early and late pigging material was collected to gain insight into the potential causes for differential corrosion rates. Metabolites were extracted and analyzed via ultra-high performance liquid chromatography/high-resolution mass spectrometry with electrospray ionization (ESI) in both positive and negative ion modes. Metabolites were analyzed by comparison with standards indicative of aerobic and anaerobic hydrocarbon metabolism and by comparison to predicted masses for KEGG metabolites. Microbial community structure was analyzed via 16S rRNA gene qPCR, sequencing of 16S PCR products, and MySeq Illumina shotgun sequencing of community DNA. Metagenomic data were used to reconstruct the full length 16S rRNA genes and genomes of dominant microorganisms. Sequence data were also interrogated via KEGG annotation and for the presence of genes related to terminal electron accepting (TEA) processes as well as aerobic and anaerobic hydrocarbon degradation. Significant and distinct differences were observed when comparing the ‘high corrosion’ (HC) and the ‘low corrosion’ (LC) pipeline systems, especially with respect to the TEA utilization potential. The HC samples were dominated by sulfate-reducing bacteria (SRB) and archaea known for their ability to utilize simple carbon substrates, whereas LC samples were dominated by pseudomonads with the genetic potential for denitrification and aerobic hydrocarbon degradation. The frequency of aerobic hydrocarbon degradation genes was low in the HC system, and anaerobic hydrocarbon degradation genes were not detected in either pipeline. This is in contrast with metabolite analysis, which demonstrated the presence of several succinic acids in HC samples that are diagnostic of anaerobic hydrocarbon metabolism. Identifiable aerobic metabolites were confined to the LC samples, consistent with the metagenomic data. Overall, these data suggest that corrosion management might benefit from a more refined understanding of microbial community resilience in the face of disturbances such as nitrate treatment or pigging, which frequently prove insufficient to alter community structure toward a stable, less-corrosive assemblage.This work was supported in part by grants from the University of Oklahoma Biocorrosion Center, the National Science Foundation (OCE 1634630 and MCB 1329890) and BP (The Gulf of Mexico Research Initiative, Project No. 130206). The instrumentation for the metabolomic analysis was funded by ONR through a DURIP grant (Award no. N000140910797), and the method development by ONR through a MURI grant (Award no. N000141010946).Ye

Assessing the Microbial Community and Functional Genes in a Vertical Soil Profile with Long-Term Arsenic Contamination

Conceived and designed the experiments: GW. Performed the experiments: JX GL. Analyzed the data: JX JZ GW. Contributed reagents/materials/analysis tools: ST JZ GW. Wrote the paper: JX ZH JDVN JZ GW.Arsenic (As) contamination in soil and groundwater has become a serious problem to public health. To examine how microbial communities and functional genes respond to long-term arsenic contamination in vertical soil profile, soil samples were collected from the surface to the depth of 4 m (with an interval of 1 m) after 16-year arsenic downward infiltration. Integrating BioLog and functional gene microarray (GeoChip 3.0) technologies, we showed that microbial metabolic potential and diversity substantially decreased, and community structure was markedly distinct along the depth. Variations in microbial community functional genes, including genes responsible for As resistance, carbon and nitrogen cycling, phosphorus utilization and cytochrome c oxidases were detected. In particular, changes in community structures and activities were correlated with the biogeochemical features along the vertical soil profile when using the rbcL and nifH genes as biomarkers, evident for a gradual transition from aerobic to anaerobic lifestyles. The C/N showed marginally significant correlations with arsenic resistance (p = 0.069) and carbon cycling genes (p = 0.073), and significant correlation with nitrogen fixation genes (p = 0.024). The combination of C/N, NO3− and P showed the highest correlation (r = 0.779, p = 0.062) with the microbial community structure. Contradict to our hypotheses, a long-term arsenic downward infiltration was not the primary factor, while the spatial isolation and nutrient availability were the key forces in shaping the community structure. This study provides new insights about the heterogeneity of microbial community metabolic potential and future biodiversity preservation for arsenic bioremediation management.Yeshttp://www.plosone.org/static/editorial#pee

Summer activity patterns of antarctic and high alpine lichen-dominated biological soil crusts-similar but different?

Biological soil crusts (BSCs) are small-scale communities of lichens, mosses, algae, and cyanobacteria that cover much of the surface area in regions where vascular plant growth is restricted due to harsh environmental conditions, such as perpetually ice-free areas in terrestrial Antarctic environments and alpine areas above the tree line. To our knowledge, none of the available studies provides a direct Antarctic-alpine comparison of BSC activity periods and the water use, both key traits to understand their physiological behavior and therefore related growth and fitness. Here, activity patterns and water relations were studied at two sites, one in continental Antarctica (Garwood Valley 78°S) and one in the High Alps of Austria (Hochtor, Großglockner 2350m). BSCs in continental Antarctica were only rarely active, and if so, then during melt after snowfalls and by fog. In the Austrian Alps, BSCs were continuously active and additionally activated by rainfall, fog, and dew. Consequently, high alpine BSCs can be expected to have much higher photosynthetic productivity supporting higher growth rates than the same functional vegetation unit has in continental Antarctica.</p

Environmental factors influencing the nitrogen fixation activity of free-living terrestrial cyanobacteria from a high arctic area, Spitsbergen

The influence of environmental factors on the nitrogen fixation activity of free-living, terrestrial cyanobacteria from a high arctic area were investigated using experimental manipulations with two different types of field samples, including macroscopic sheets of Nostoc commune and soil samples with a cyanobacterial crust from a Puccinellia salt marsh. In addition, a cultured Anabaena sp. previously isolated from the salt marsh was examined. Nitrogen fixation activity was measured using the acetylene reduction method. The nitrogen fixation mainly took place in the light, but even after 12 h incubation in darkness, low activities were maintained. Phosphorus fertilization stimulated the nitrogen fixation activity, and the highest activities were obtained with about 300 μM phosphate, both in the field samples and the cultured Anabaena sp. Ammonium (28 mM) immediately inhibited the nitrogen fixation activity of the cultured Anabaena sp, whereas 14 mM urea and 540 μM glutamate led to a weaker and slower inhibition of the nitrogen fixation activity, showing that the cultured Anabaena sp. was able to assimilate these combined nitrogen sources. Nitrate did not have any inhibitory effect on nitrogen fixation activity, either in the field samples or in the cultured Anabaena sp. Both the field samples and the cultured Anabaena sp. showed tolerance against sodium chloride concentrations corresponding to the concentration in seawater. The temperature optimum of the nitrogen fixation activity of the cultured Anabaena sp. was about 20°C. Key words: nitrogen fixation, cyanobacteria, Nostoc commune, Anabaena sp., high arctic.</jats:p

Conversion factor between acetylene reduction and nitrogen fixation in free-living cyanobacteria from high arctic habitats

The conversion factor between acetylene reduction and15N incorporation in free-living cyanobacteria was determined in different high arctic habitats in the area of Ny-Ålesund (78.5°N, 11.6°E), Spitsbergen, in the summer of 1994. The experiments were carried out under constant conditions, 19°C and 200 µE·m-2·s-1. The nitrogen-fixation activities, measured as15N-incorporation, were in the range 4.01-6.54 mg N2fixed·gdw-1·day-1(dw, dry weight) in sheets of Nostoc commune and 778-1206 mg N2fixed·m-2·day-1in the cyanobacterial crusts. The acetylene reduction activities were in the range 0.72-1.91 mg ethylene produced·gdw-1·day-1of N. commune and 12.8-63.7 mg ethylene produced·m-2·day-1in the cyanobacterial crusts. The conversion factor of N. commune ranged from 0.11 to 0.48 for ethylene produced to nitrogen fixed, whereas the cyanobacterial crusts covering the soil surface gave conversion factors in the range 0.022-0.073 for ethylene produced to nitrogen fixed. An Anabaena sp., isolated from one of the habitats investigated, gave conversion factors near the theoretical factor of 4, when determined at 14.0 and 17.3°C. It was concluded that the acetylene reduction activity of free-living cyanobacteria in high arctic habitats results in underestimates of the real nitrogen-fixation activity in these environments.Key words: nitrogen fixation, acetylene reduction, conversion factor, cyanobacteria, Nostoc commune, high arctic.</jats:p

Seasonal and site-specific variations in nitrogen fixation in a high arctic area, Ny-Ålesund, Spitsbergen

Nitrogen fixation was measured in different habitats in the area of Ny-Ålesund, Spitsbergen, using the acetylene reduction method on intact soil cores and Nostoc commune growing in macroscopic sheet communities. The samples were incubated both under constant conditions (19 °C and 200 μE∙m−2∙s−1) and under in situ conditions. Cyanobacteria were considered to be the major nitrogen-fixing organisms. The nitrogen fixation rates showed a seasonal variation during the growing season of 1994, with low activities just after the snow melt, increasing until the middle of August and showing a rapid decline after the snow fell on August 29. The soil temperature at the time of sampling showed a positive, linear correlation with the nitrogen fixation activities measured on intact soil cores, whereas the nitrogen fixation activities measured in situ of N. commune showed a positive, linear dependence on the moisture content in the sheets and the incubation temperatures inside the incubation vessels during the experiments. The optimal temperature of the nitrogen fixation activity was about 20 °C, both for N. commune and a Puccinellia salt marsh. The highest nitrogen fixation rate measured in situ was at a patterned ground, which had the highest pH, the highest concentrations of extractable calcium and magnesium, and the highest C:N ratio measured.Key words: nitrogen fixation, cyanobacteria, Nostoc commune, high arctic. </jats:p

Metabolomic and Metagenomic Analysis of Two Crude Oil Production Pipelines Experiencing Differential Rates of Corrosion

Corrosion processes in two North Sea oil production pipelines were studied by analyzing pig envelope samples via metagenomic and metabolomic techniques. Both production systems have similar physico-chemical properties and injection waters are treated with nitrate, but one pipeline experiences severe corrosion and the other does not. Early and late pigging material was collected to gain insight into the potential causes for differential corrosion rates. Metabolites were extracted and analyzed via ultra-high performance liquid chromatography/high-resolution mass spectrometry with electrospray ionization (ESI) in both positive and negative ion modes. Metabolites were analyzed by comparison with standards indicative of aerobic and anaerobic hydrocarbon metabolism and by comparison to predicted masses for KEGG metabolites. Microbial community structure was analyzed via 16S rRNA gene qPCR, sequencing of 16S PCR products, and MySeq Illumina shotgun sequencing of community DNA. Metagenomic data were used to reconstruct the full length 16S rRNA genes and genomes of dominant microorganisms. Sequence data were also interrogated via KEGG annotation and for the presence of genes related to terminal electron accepting (TEA) processes as well as aerobic and anaerobic hydrocarbon degradation. Significant and distinct differences were observed when comparing the 'high corrosion' (HC) and the 'low corrosion' (LC) pipeline systems, especially with respect to the TEA utilization potential. The HC samples were dominated by sulfate-reducing bacteria (SRB) and archaea known for their ability to utilize simple carbon substrates, whereas LC samples were dominated by pseudomonads with the genetic potential for denitrification and aerobic hydrocarbon degradation. The frequency of aerobic hydrocarbon degradation genes was low in the HC system, and anaerobic hydrocarbon degradation genes were not detected in either pipeline. This is in contrast with metabolite analysis, which demonstrated the presence of several succinic acids in HC samples that are diagnostic of anaerobic hydrocarbon metabolism. Identifiable aerobic metabolites were confined to the LC samples, consistent with the metagenomic data. Overall, these data suggest that corrosion management might benefit from a more refined understanding of microbial community resilience in the face of disturbances such as nitrate treatment or pigging, which frequently prove insufficient to alter community structure toward a stable, less-corrosive assemblage