Characterizing the development of biofilm in polyethylene pipes in the non-chlorinated Danish drinking-water distribution system.

In newly commissioned drinking-water polyethylene (PE) pipes, biofilm develops on the inner pipe surface. The microbial community composition from colonization to the establishment of mature biofilms is less known, including the effect on the distributed water quality. Biofilm development was followed through 1.5 years in PE-pipe side streams at two locations of a full-scale, non-chlorinated drinking-water distribution system (leaving a waterworks versus 5-6 km from a waterworks) along with inlet and outlet water quality. Mature biofilms were established after ∼8-9 months, dominated by Proteobacteria, Actinobacteria and Saccharibacteria (61-93% relative abundance), with a higher diversity (OTUs/Shannon Index/16S rRNA gene amplicon sequencing) in pipes in the far end of the distribution system. Comamonadaceae, and specifically Aquabacterium (>30% of reads), dominated young (∼1.5-month-old) biofilms. Young biofilms were linked to increased microbiological counts in drinking water (HPC/ATP/qPCR), while the establishment of mature biofilms led to a drop in HPC and benefited the water quality, highlighting the importance of optimizing commissioning procedures for rapidly achieving mature and stable biofilms.

Management and control of microbiologically influenced corrosion (MIC) in the oil and gas industry-Overview and a North Sea case study.

Microbiologically influenced corrosion (MIC) is the terminology applied where the actions of microorganisms influence the corrosion process. In literature, terms such as microbial corrosion, biocorrosion, microbially influenced/induced corrosion, and biodegradation are often applied. MIC research in the oil and gas industry has seen a revolution over the past decade, with the introduction of molecular microbiological methods: (MMM) as well as new industry standards and procedures of sampling biofilm and corrosion products from the process system. This review aims to capture the most important trends the oil and gas industry has seen regarding MIC research over the past decade. The paper starts out with an overview of where in the process stream MIC occurs - from the oil reservoir to the consumer. Both biotic and abiotic corrosion mechanisms are explained in the context of managing MIC using a structured corrosion management (CM) approach. The corrosion management approach employs the elements of a management system to ensure that essential corrosion control activities are carried out in an effective, sustainable, well-planned and properly executed manner. The 3-phase corrosion management approach covering of both biotic and abiotic internal corrosion mechanisms consists of 1) corrosion assessment, 2) corrosion mitigation and 3) corrosion monitoring. Each of the three phases are described in detail with links to recent field cases, methods, industry standards and sampling protocols. In order to manage the corrosion threat, operators commonly use models to support decision making. The models use qualitative, semi-quantitative or quantitative measures to help assess the rate of degradation caused by MIC. The paper reviews four existing models for MIC Threat Assessment and describe a new model that links the threat of MIC in the oil processing system located on an offshore platform with a Risk Based Inspection (RBI) approach. A recent field case highlights and explains the conflicting historic results obtained through serial dilution of culture media using the most probable number (MPN) method as compared to data obtained from corrosion monitoring and the quantitative polymerase chain reaction (qPCR) method. Results from qPCR application in the field case have changed the way MIC is monitored on the oil production facility in the North Sea. A number of high quality resources have been published as technical conference papers, books, educational videos and peer-reviewed scientific papers, and thus we end the review with an updated list of state-of-the-art resources for anyone desiring to become more familiar with the topic of MIC in the upstream oil and gas sector.

Reservoir Souring - Latest developments for application and mitigation.

Sulphate-reducing prokaryotes (SRP) have been identified in oil field fluids since the 1920s. SRP reduce sulphate to sulphide, a toxic and corrosive species that impacts on operational safety, metallurgy and both capital and operational cost. Differences in water cut, temperature, pressure and fluid chemistry can impact on the observed H2S concentration, meaning that an increase in H2S concentration does not always correlate with activity of SRP. However it wasn't until the 1990s that SRP activity was accepted as the leading cause of reservoir souring (i.e. an increase in H2S concentrations) in water flooded oil fields. The process of sulphate-reduction has been well documented at the genetic, enzymatic and physiological level in pure cultures under laboratory conditions. DNA sequencing has also identified new groups of microorganisms, such as archaea which are capable of contributing to reservoir souring. This has led to some recent advances in microbial control and detection, however, despite this, many of the methods used routinely for microbial control and detection are over a century old. We therefore look towards emerging and novel mitigation technologies that may be used in mitigating against reservoir souring, along with tried and tested methods. Modelling and prediction is another important but often under-used tool in managing microbial reservoir souring. To be truly predictive, models need to take into account not only microbial H2S generation but also partitioning and mineral scavenging. The increase in 'big data' available through increased integration of sensors in the digital oil field and the increase in the DNA sequencing capabilities through next-generation sequencing (NGS) therefore offer a unique opportunity to develop and refine microbial reservoir souring models. We therefore review a number of different reservoir souring models and identify how these can be used in the future. With this comprehensive overview of the current and emerging technologies we will highlight areas where significant development effort could generate rewards that can improve detection, prediction and control of microbial reservoir souring.

Ability of Pseudoalteromonas tunicata to colonize natural biofilms and its effect on microbial community structure.

We investigated the effectiveness of surface colonization by the epiphytic marine bacterium Pseudoalteromonas tunicata firstly on a complex biofilm community on glass slides, and secondly, on the epiphytic community of Ulva australis. The effectiveness of P. tunicata was compared with the performance of Phaeobacter sp. 2.10, also a marine epiphytic isolate in the U. australis colonization experiments. Pseudoalteromonas tunicata cells were able to colonize the glass slide community at densities found naturally in the water column (9.7 x 10(4) cells mL(-1)). However, P. tunicata was a poor invader of the epiphytic community on U. australis at densities of 10(6) cells mL(-1). At densities of 10(8) cells mL(-1), P. tunicata again exerted little impact on the epiphytic community. Phaeobacter sp. 2.10 was also a poor invader at lower densities, but was able to invade and become dominant at densities of 10(8) cells mL(-1). Differences in the ability of P. tunicata and Phaeobacter sp. 2.10 to invade natural communities may be due to differences in the antibacterial compounds produced by the two species. These experiments suggest that epiphytic communities may have protective effects compared with inanimate surfaces.

Prokaryotic community structure and sulfate reducer activity in water from high-temperature oil reservoirs with and without nitrate treatment.

Sulfate-reducing prokaryotes (SRP) cause severe problems like microbial corrosion and reservoir souring in seawater-injected oil production systems. One strategy to control SRP activity is the addition of nitrate to the injection water. Production waters from two adjacent, hot (80 degrees C) oil reservoirs, one with and one without nitrate treatment, were compared for prokaryotic community structure and activity of SRP. Bacterial and archaeal 16S rRNA gene analyses revealed higher prokaryotic abundance but lower diversity for the nitrate-treated field. The 16S rRNA gene clone libraries from both fields were dominated by sequences affiliated with Firmicutes (Bacteria) and Thermococcales (Archaea). Potential heterotrophic nitrate reducers (Deferribacterales) were exclusively found at the nitrate-treated field, possibly stimulated by nitrate addition. Quantitative PCR of dsrAB genes revealed that archaeal SRP (Archaeoglobus) dominated the SRP communities, but with lower relative abundance at the nitrate-treated site. Bacterial SRP were found in only low abundance at both sites and were nearly exclusively affiliated with thermophilic genera (Desulfacinum and Desulfotomaculum). Despite the high abundance of archaeal SRP, no archaeal SRP activity was detected in [(35)S]sulfate incubations at 80 degrees C. Sulfate reduction was found at 60 degrees C in samples from the untreated field and accompanied by the growth of thermophilic bacterial SRP in batch cultures. Samples from the nitrate-treated field generally lacked SRP activity. These results indicate that (i) Archaeoglobus can be a major player in hot oil reservoirs, and (ii) nitrate may act in souring control-not only by inhibiting SRP, but also by changing the overall community structure, including the stimulation of competitive nitrate reducers.

Molecular investigation of the distribution, abundance and diversity of the genus Pseudoalteromonas in marine samples.

The genus Pseudoalteromonas has attracted interest because it has frequently been found in association with eukaryotic hosts, and because many Pseudoalteromonas species produce biologically active compounds. One distinct group of Pseudoalteromonas species is the antifouling subgroup containing Pseudoalteromonas tunicata and Ps. ulvae, which both produce extracellular compounds that inhibit growth and colonization by different marine organisms. PCR primers targeting the 16S rRNA gene of the genus Pseudoalteromonas and the antifouling subgroup were developed and applied in this study. Real-time quantitative PCR (qPCR) was applied to determine the relative bacterial abundance of the genus and the antifouling subgroup, and denaturing gradient gel electrophoresis (DGGE) was applied to study the diversity of the genus in 11 different types of marine samples from Danish coastal waters. The detection of Ps. tunicata that contain the antifouling subgroup was achieved through specific PCR amplification of the antibacterial protein gene (alpP). The Pseudoalteromonas species accounted for 1.6% of the total bacterial abundance across all samples. The Pseudoalteromonas diversity on the three unfouled marine organisms Ciona intestinalis, Ulva lactuca and Ulvaria fusca was found to be low, and Ps. tunicata was only detected on these three hosts, which all contain accessible cellulose polymers in their cell walls.

Real-time quantitative PCR for assessment of abundance of Pseudoalteromonas species in marine samples.

A real-time quantitative PCR (RTQ-PCR) method for measuring the abundance of Pseudoalteromonas species in marine samples is presented. PCR primers targeting a Pseudoalteromonas-specific region of the 16S rRNA gene were tested at three different levels using database searches (in silico), a selection of pure cultures (in vitro), and a combined denaturing gradient gel electrophoresis and cloning approach on environmental DNA (in situ). The RTQ-PCR method allowed for the detection of SYBR Green fluorescence from double-stranded DNA over a linear range spanning six orders of magnitude. The detection limit was determined as 1.4 fg of target DNA (1,000 gene copies) measured in the presence of 20 ng of nontarget DNA from salmon testes. In this study, we discuss the importance of robust post-PCR analyses to overcome pitfalls in RTQ-PCR when samples from different complex marine habitats are analyzed and compared on a nonroutine basis. Representatives of the genus Pseudoalteromonas were detected in samples from all investigated habitats, suggesting a widespread distribution of this genus across many marine habitats (e.g., seawater, rocks, macroalgae, and marine animals). Three sample types were analyzed by RTQ-PCR to determine the relative abundance of Pseudoalteromonas ribosomal DNA (rDNA) compared to the total abundance of eubacterial rDNA. The rDNA fractions of Pseudoalteromonas compared to all Eubacteria were 1.55% on the green alga Ulva lactuca, 0.10% on the tunicate Ciona intestinalis, and 0.06% on the green alga Ulvaria fusca.