Microbially influenced corrosion is a global issue influencing the premature failure of metallic infrastructure and accounting for 20% of all internal corrosion incidences (Zhu, et al., 2003).Numerous microorganisms are implicated in MIC including sulphate-reducers, methanogens, nitrate-reducers, iron-oxidisers, iron-reducers, and acid-producers.The aim of this chapter was to explore the effects of high salinity (127 g/L TDS; 204 g/L TDS) in a range of temperature incubations (15°C, 30°C,
Many factors affect the performance of microbial fuel cells (MFCs). Considerable attention has been given to the impact of cell configuration and materials on MFC performance. Much less work has been done on the impact of the anode microbiota, particularly in the context of using complex substrates as fuel. One strategy to improve MFC performance on complex substrates such as wastewater, is to pre-enrich the anode with known, efficient electrogens, such as Geobacter spp. The implication of this strategy is that the electrogens are the limiting factor in MFCs fed complex substrates and the organisms feeding the electrogens through hydrolysis and fermentation are not limiting. We conducted a systematic test of this strategy and the assumptions associated with it. Microbial fuel cells were enriched using three different substrates (acetate, synthetic wastewater and real domestic wastewater) and three different inocula (Activated Sludge, Tyne River sediment, effluent from an MFC). Reactors were either enriched on complex substrates from the start or were initially fed acetate to enrich for Geobacter spp. before switching to synthetic or real wastewater. Pre-enrichment on acetate increased the relative abundance of Geobacter spp. in MFCs that were switched to complex substrates compared to MFCs that had been fed the complex substrates from the beginning of the experiment (wastewater-fed MFCs - 21.9 ± 1.7% Geobacter spp.; acetate-enriched MFCs, fed wastewater - 34.9 ± 6.7% Geobacter spp.; Synthetic wastewater fed MFCs – 42.5 ± 3.7% Geobacter spp.; acetate-enriched synthetic wastewater-fed MFCs - 47.3 ± 3.9% Geobacter spp.). However, acetate pre-enrichment did not translate into significant improvements in cell voltage, maximum current density, maximum power density or substrate removal efficiency. Nevertheless, coulombic efficiency (CE) was higher in MFCs pre-enriched on acetate when complex substrates were fed following acetate enrichment (wastewater-fed MFCs – CE = 22.0 ± 6.2%; acetate-enriched MFCs, fed wastewater – CE =58.5 ± 3.5%; Synthetic wastewater fed MFCs – CE = 22.0 ± 3.2%; acetate-enriched synthetic wastewater-fed MFCs – 28.7 ± 4.2%.) The relative abundance of Geobacter ssp. and CE represents the average of the nine replicate reactors inoculated with three different inocula for each substrate. Efforts to improve the performance of anodic microbial communities in MFCs utilizing complex organic substrates should therefore focus on enhancing the activity of organisms driving hydrolysis and fermentation rather the terminal-oxidizing electrogens.
Unprecedented and dramatic transformations are occurring in the Arctic in response to climate change, but academic, public, and political discourse has disproportionately focussed on the most visible and direct aspects of change, including sea ice melt, permafrost thaw, the fate of charismatic megafauna, and the expansion of fisheries. Such narratives disregard the importance of less visible and indirect processes and, in particular, miss the substantive contribution of the shelf seafloor in regulating nutrients and sequestering carbon. Here, we summarise the biogeochemical functioning of the Arctic shelf seafloor before considering how climate change and regional adjustments to human activities may alter its biogeochemical and ecological dynamics, including ecosystem function, carbon burial, or nutrient recycling. We highlight the importance of the Arctic benthic system in mitigating climatic and anthropogenic change and, with a focus on the Barents Sea, offer some observations and our perspectives on future management and policy.
Over the last decade, metagenomic studies have revealed the impact of oil production on the microbial ecology of petroleum reservoirs. However, despite their fundamental roles in bioremediation of hydrocarbons, biocorrosion, biofouling and hydrogen sulfide production, oil field and oil production infrastructure microbiomes are poorly explored. Understanding of microbial activities within oil production facilities is therefore crucial for environmental risk mitigation, most notably during decommissioning. The analysis of the planktonic microbial community from the aqueous phase of a subsea oil-storage structure was conducted. This concrete structure was part of the production platform of the Brent oil field (North Sea), which is currently undergoing decommissioning. Quantification and sequencing of microbial 16S rRNA genes, metagenomic analysis and reconstruction of metagenome assembled genomes (MAGs) revealed a unique microbiome, strongly dominated by organisms related to Dethiosulfatibacter and Cloacimonadetes. Consistent with the hydrocarbon content in the aqueous phase of the structure, a strong potential for degradation of low molecular weight aromatic hydrocarbons was apparent in the microbial community. These degradation pathways were associated with taxonomically diverse microorganisms, including the predominant Dethiosulfatibacter and Cloacimonadetes lineages, expanding the list of potential hydrocarbon degraders. Genes associated with direct and indirect interspecies exchanges (multiheme type-C cytochromes, hydrogenases and formate/acetate metabolism) were widespread in the community, suggesting potential syntrophic hydrocarbon degradation processes in the system. Our results illustrate the importance of genomic data for informing decommissioning strategies in marine environments and reveal that hydrocarbon-degrading community composition and metabolisms in man-made marine structures might differ markedly from natural hydrocarbon-rich marine environments.
Reduction of CO2 to acetate in microbial electrosynthesis has been widely studied. However, the selective and quantitative production of longer chain chemicals and biofuels is still a bottleneck. Lack of sufficient energy provided by only the cathode electrode in Bio-electrochemical systems during chain elongation is one of the key challenges. It is assumed that additional electron donors than a polarised cathode is required to steer the production towards longer chain of carboxylates than acetate. In this study, formate and ethanol were supplied separately in the reactors fed by CO2 for 45 days in addition to the cathodes poised at -1.0 V vs. Ag/AgCl to investigate their effect on production. Although acetate was still the major product, supplying electron donors directed the production towards more diverse and longer chain organic chemicals than that in presence of the polarised cathode only. Significant improvement in the production of butyrate (x3.8 increase in maximum concentration) and butanol (maximum of 6.8 +/- 0.3 mmol C L-1) was observed after supplying formate, while ethanol increased the diversity of the products. Supplying formate and ethanol in reactors for another 30 days under open circuit potential clarified that only ethanol could provide sufficient energy for butyrate production from acetate in the absence of polarised cathode, which reached the highest butyrate concentration of 19.1 +/- 2.3 mmol C L-1. Formate was only consumed in presence of polarised cathode. It is proposed in our study that production of C4 products in presence of only cathodic electrode or cathodic electrode and formate could be associated to initial reduction of acetate to ethanol, consumed for production of C4 products through acetate. Trace levels of caproate and hexanol were detected in both reactors supplied with formate and ethanol only in the presence of polarised cathode.
Nitrogen-fixing bacteria (NFB) can reduce nitrogen at ambient pressure and temperature. In this study, we treated effluent from a paper mill in sequencing batch reactors (SBRs) and monitored the abundance and activity of NFB with a view to producing a sludge that could work as a biofertilizer. Four reactors were inoculated with activated sludge enriched with NFB and fed with a high C/N waste (100:0.5) from a paper mill. Though the reactors were able to reduce the organic load of the wastewater by up to 89%, they did not have any nitrogen-fixing activity and showed a decrease in the putative number of NFB (quantified with qPCR). The most abundant species in the reactors treating high C/N paper mill wastewater was identified by Illumina MiSeq 16S rRNA gene amplicon sequencing as Methyloversatilis sp. (relative abundance of 4.4%). Nitrogen fixation was observed when the C/N ratio was increased by adding sucrose. We suspect that real-world biological nitrogen fixation (BNF) will only occur where there is a C/N ratio <= 100:0.07. Consequently, operators should actively avoid adding or allowing nitrogen in the waste streams if they wish to valorize their sludge and reduce running costs. Practitioner points Efficient biological wastewater treatment of low nitrogen paper mill effluent was achieved without nutrient supplementation. The sludge was still capable of fixing nitrogen although this process was not observed in the wastewater treatment system. This high C/N wastewater treatment technology could be used with effluents from cassava flour, olive oil, wine and dairy industries.
Laminated microbial mats and microbialites have been documented from a variety of coastal marine environments. This study aims to provide the first detailed descriptions of intertidal pools, along with their hosted thrombolite and stromatolite structures, from Abu Dhabi, and to propose a model for their formation and evolution. It is proposed that the development of pools within the upper intertidal zone was initiated by localized erosion of the laminated microbial mats during high energy events. The removal of the protective mats permitted erosion of the underlying unconsolidated sediment to produce erosional scours that continued to develop to create the pools observed today. The margins of the newly-created submerged environment were colonized by a cyanobacteria dominated microbial community. The precipitation of aragonite cement, associated with the cyanobacteria, stabilized the pool walls and cemented the microbial communities to form stromatolitic and thrombolitic fabrics. Syndepositional cementation was further enhanced by the precipitation of marine cements as a result of evaporation-driven Ca(2+)and Mg(2+)supersaturation. Erosion behind and below the cemented pool wall eventually resulted in rim-collapse and the formation of the observed pool margin parallel thrombolite bands. Successive generations of lithification and erosion increased the area of the pool with the earliest thrombolites eroding and becoming increasingly isolated. In summary, the resultant microbialites developed through the complex interplay of erosion, abiotic early lithification and microbially-mediated processes, and represent a continuum between unlithified laminated microbial mats and domal microbialites. These features are most likely produced during a sea-level scenario of stillstand or transgression and, as such, may be useful as a diagnostic tool to elucidate the onset of transgression. The newly proposed model for stromatolite formation has significant implications for the recognition and interpretation of similar structures observed in the fossil record.
A previous cultivation-independent investigation of the microbial community structure of natural oil and asphalt seeps in the Kurdistan Region of Iraq (KRI) revealed the dominance of uncultured bacterial taxa belonging to the phyla Deferribacterota and Coprothermobacterota and the orders Thermodesulfobacteriales, Thermales, and Burkholderiales. Here we report on a cultivation-dependent approach to identify members of these groups involved in hydrocarbon degradation in the KRI oil and asphalt seeps. For this purpose, we set up anoxic crude oil-degrading enrichment cultures based on cultivation media known to support the growth of members of the above-mentioned taxonomic groups. During 100–200 days incubation periods, nitrate-reducing and fermentative enrichments showed up to 90% degradation of C8–C17 alkanes and up to 28% degradation of C18–C33 alkanes along with aromatic hydrocarbons. Community profiling of the enrichment cultures showed that they were dominated by diverse bacterial taxa, which were rare in situ community members in the investigated seeps. Groups initially targeted by our approach were not enriched, possibly because their members are slow-growing and involved in the degradation of recalcitrant hydrocarbons. Nevertheless, the enriched taxa were taxonomically related to phylotypes recovered from hydrocarbon-impacted environments as well as to characterized bacterial isolates not previously known to be involved in hydrocarbon degradation. Marker genes (assA and bssA), diagnostic for fumarate addition-based anaerobic hydrocarbon degradation, were not detectable in the enrichment cultures by PCR. We conclude that hydrocarbon biodegradation in our enrichments occurred via unknown pathways and synergistic interactions among the enriched taxa. We suggest, that although not representing abundant populations in situ, studies of the cultured close relatives of these taxa will reveal an unrecognized potential for anaerobic hydrocarbon degradation, possibly involving poorly characterized mechanisms.
Microbial fuel cells (MFCs) that simultaneously remove organic contaminants and recovering metals provide a potential route for industry to adopt clean technologies. In this work, two goals were set: to study the feasibility of zinc removal from industrial effluents using MFCs and to understand the removal process by using reaction rate models. The removal of Zn2+ in MFC was over 96% for synthetic and industrial samples with initial Zn2+ concentrations less than 2.0 mM after 22 h of operation. However, only 83 and 42% of the zinc recovered from synthetic and industrial samples, respectively, was attached on the cathode surface of the MFCs. The results marked the domination of electroprecipitation rather than the electrodeposition process in the industrial samples. Energy dispersive X-ray (EDX) analysis showed that the recovered compound contained not only Zn but also O, evidence that Zn(OH)2 could be formed. The removal of Zn2+ in the MFC followed a mechanism where oxygen was reduced to hydroxide before reacting with Zn2+. Nernst equations and rate law expressions were derived to understand the mechanism and used to estimate the Zn2+ concentration and removal efficiency. The zero-, first- and second-order rate equations successfully fitted the data, predicted the final Zn2+ removal efficiency, and suggested that possible mechanistic reactions occurred in the electrolysis cell (direct reduction), MFC (O2 reduction), and control (chemisorption) modes. The half-life, t1/2, of the Zn2+ removal reaction using synthetic and industrial samples was estimated to be 7.0 and 2.7 h, respectively. The t1/2 values of the controls (without the power input from the MFC bioanode) were much slower and were recorded as 21.5 and 7.3 h for synthetic and industrial samples, respectively. The study suggests that MFCs can act as a sustainable and environmentally friendly technology for heavy metal removal without electrical energy input or the addition of chemicals.
Technologies able to convert CO2 to various feedstocks for fuels and chemicals are emerging due to the urge of reducing greenhouse gas emissions and de-fossilizing chemical production. Microbial electrosynthesis (MES) has been shown a promising technique to synthesize organic products particularly acetate using microorganisms and electrons. However, the efficiency of the system is low. In this study, we demonstrated the simple yet efficient strategy in enhancing the efficiency of MES by applying continuous feeding regime. Compared to the fed-batch system, continuous operational mode provided better control of pH and constant medium refreshment, resulting in higher acetate production rate and more diverse bio-products, when the cathodic potential of -1.0 V Ag/AgCl and dissolved CO2 were provided. It was observed that hydraulic retention time (HRT) had a direct effect on the pattern of production, acetate production rate and coulombic efficiency. At HRT of 3 days, pH was around 5.2 and acetate was the dominant product with the highest production rate of 651.8 ± 214.2 ppm per day and a significant coulombic efficiency of 90%. However at the HRT of 7 days, pH was lower at around 4.5, and lower but stable acetate production rate of 280 ppm per day and a maximum coulombic efficiency of 80% was obtained. In addition, more diverse and longer chain products, such as butyrate, isovalerate and caproate, were detected with low concentrations only at the HRT of 7 days. Although microbial community analysis showed the change in the planktonic cells communities after switching the fed-batch mode to continuous feeding regime, Acetobacterium still remained as the responsible bacteria for CO2 reduction to acetate, dominating the cathodic biofilm.
Microbial Fuel Cells (MFCs) operated as biosensors could potentially enable truly low-cost, real-time monitoring of organic loading in wastewaters. The current generated by MFCs has been correlated with conventional measures of organic load such as Biochemical Oxygen Demand (BOD), but much remains to be established in terms of the reliability and applicability of such sensors. In this study, batch-mode and multi-stage, flow-mode MFCs were operated for over 800 days and regularly re-calibrated with synthetic wastewater containing glucose and glutamic acid (GGA). BOD5 calibration curves were obtained by normalising the current measured as a percentage of maximum current. There was little drift between recalibrations and non-linear Hill models of the combined dataset had R2 of 88-95%, exhibiting a stable response over time and across devices. Nonetheless, factors which do affect calibration were also assessed. Increasing external resistance (from 43.5 to 5100 0) above the internal resistance determined by polarisation curve decreased the calibration upper limit from 240 to 30 mg/l O2 BOD5. Furthermore, more fermentable carbon sources increased the detection range, as tested with samples of real wastewater and synthetic media containing GGA, glucose-only and glutamic acid-only. Biofilm acclimatisation therefore did not account for differences between aerobic oxygen demand determinations and anaerobic MFC responses; these are likely attributable to competitive processes such as fermentation. This further highlights the potential for MFCs as real-time sensors for organic load monitoring and process control in addition to BOD-compliant measurement systems.
Sulfur-oxidizing Sulfurimonas spp. are widespread in sediments, hydrothermal vent fields, aquifers and subsurface environments such as oil reservoirs where they play an important role in the sulfur cycle. We determined the genome sequence of the oil field isolate Sulfurimonas sp. strain CVO and compared its gene expression during nitrate-dependent sulfide oxidation to the coastal sediment isolate Sulfurimonas denitrificans. Formation of elemental sulfur (S-0) and high expression of sulfide quinone oxidoreductase (SQR) genes indicates that sulfide oxidation in both strains is mediated by SQR. Subsequent oxidation of S-0 was achieved by the sulfur oxidation enzyme complex (SOX). In the coastal S. denitrificans, the genes are arranged and expressed as two clusters: soxXY(1)Z(1)AB and soxCDY(2)Z(2)H, and sulfate was the sole metabolic end product. By contrast, the oil field strain CVO has only the soxCDY(2)Z(2)H cluster and not soxXY(1)Z(1)AB. Despite the absence of the soxXY(1)Z(1)AB cluster, strain CVO oxidized S-0 to thiosulfate and sulfate, demonstrating that soxCDY(2)Z(2)H genes alone are sufficient for S-0 oxidation in Sulfurimonas spp. and that thiosulfate is an additional metabolic end product. Screening of publicly available metagenomes revealed that Sulfurimonas spp. with only the soxCDY(2)Z(2)H cluster are widespread suggesting this mechanism of thiosulfate formation is environmentally significant.
Multi-stage microbial fuel cells can distinguish toxic shock events from BOD decreases, both of which result in decreased current output.
Anthropogenic nitrogen fixation is essential to sustain a global population of 7.7 billion. However, there has been a long-standing desire to find cheaper and more environmentally friendly alternatives to the Haber-Bosch process. In this study, we developed a new strategy of nitrogen fixation by enriching free-living N-2-fixing bacteria ( NFB) in reactors fed with low nitrogen wastewater, analogous to those usually found in certain industrial effluents such as paper mills. Our reactors fixed appreciable quantities of nitrogen with a rate of 11.8 mg N L-1 day(-1). This rate is comparable to recent "breakthrough" nitrogen-fixing technologies and far higher than observed in low C/N reactors (fed with organic matter and nitrogen). NFB were quantified using quantitative polymerase chain reaction (qPCR) of the nifH (marker gene used to identify biological nitrogen fixation) and 16S rRNA genes. The nifH gene was enriched by a factor of 10 in the nitrogen-fixing reactors (compared to controls) attaining 13% of the bacterial population (1:4.2 copies of nif H to 16S rRNA). The Illumina MiSeq 16S rRNA gene amplicon sequencing of reactors showed that the microbial community was dominated (19%) by Clostridium pasteurianum. We envisage that nitrogen-enriched biomass could potentially be used as a biofertilizer and that the treated wastewater could be released to the environment with very little post-treatment.
The upstream protection of the biomass present in biological treatment processes is a vital challenge as the consequences of failure could include exposure of water users to hazardous chemicals in addition to loss of treatment performance. Online detection of toxic compounds in wastewater could enable processes to be monitored in real-time and promote pro-active responses to pollution incidents. Recently, Microbial Fuel Cells (MFCs) which generate electricity from organic matter oxidation have shown potential as sensors for online detection of toxicity. In this study, the detection of a model toxicant (4-nitrophenol) was investigated using a multi-stage MFC-based toxicity sensor. MFCs were operated with synthetic wastewater to maintain realistic conditions while enabling organic carbon levels to be controlled. A positive correlation was observed between the 4-NP concentrations and the current drop area showing that the response was proportional to the toxicity level. In addition, the sensor anodic biofilm exhibited resilience to acute toxic events with recovery of 75% of the initial current following a toxic event comprising 500 mg/L 4-NP after 4 h. However, repetitive toxicity events could lead to the selection of resistant bacteria able to degrade the toxic compounds. In this study, a maximal 4-NP degradation rate of 36 mg/h was observed. This limitation could be overcome by re-calibration after a determined number of toxic events. An additional feature of the multi-stage configuration of the sensor is that a drop in output caused by the presence of a toxic compound could be distinguished from a drop in output caused by a decrease in BOD. The microbial community on the sensor anode was characterized by 16S rRNA gene sequencing and shown to comprise an anaerobic community of fermentative bacteria capable of producing volatile fatty acids and hydrogen that were consumed by electrogenic Geobacter spp (2.76 to 21.39% of the anode community) that generated the electrical signal in the sensor. The multi-stage MFC biosensor could provide an early warning system capable of alerting process operators to the presence and level of toxicity in influent wastewater.
Cathode-driven applications of bio-electrochemical systems (BESs) have the potential to transform CO2 into value-added chemicals using microorganisms. However, their commercialisation is limited as biocathodes in BESs are characterised by slow start-up and low efficiency. Understanding biosynthesis pathways, electron transfer mechanisms and the effect of operational variables on microbial electrosynthesis (MES) is of fundamental importance to advance these applications of a system that has the capacity to convert CO2 to organics and is potentially sustainable. In this work, we demonstrate that cathodic potential and inorganic carbon source are keys for the development of a dense and conductive biofilm that ensures high efficiency in the overall system. Applying the cathodic potential of −1.0 V vs. Ag/AgCl and providing only gaseous CO2 in our system, a dense biofilm dominated by Acetobacterium (ca. 50% of biofilm) was formed. The superior biofilm density was significantly correlated with a higher production yield of organic chemicals, particularly acetate. Together, a significant decrease in the H2 evolution overpotential (by 200 mV) and abundant nifH genes within the biofilm were observed. This can only be mechanistically explained if intracellular hydrogen production with direct electron uptake from the cathode via nitrogenase within bacterial cells is occurring in addition to the commonly observed extracellular H2 production. Indeed, the enzymatic activity within the biofilm accelerated the electron transfer. This was evidenced by an increase in the coulombic efficiency (ca. 69%) and a 10-fold decrease in the charge transfer resistance. This is the first report of such a significant decrease in the charge resistance via the development of a highly conductive biofilm during MES. The results highlight the fundamental importance of maintaining a highly active autotrophic Acetobacterium population through feeding CO2 in gaseous form, which its dominance in the biocathode leads to a higher efficiency of the system.
EDITORIAL article Front. Environ. Sci., 03 April 2020Sec. Microbiological Chemistry and Geomicrobiology https://doi.org/10.3389/fenvs.2020.00035
To identify hydrocarbon-degrading microorganisms contributing to the formation of heavy oil we investigated the microbial community composition in different types of crude oils from oil-production facilities and in crude oil and asphalt from different natural seeps from the Kurdistan Region of Iraq (KRI). Crude oils from five out of six production facilities did not contain microorganisms detectable by 16S rRNA gene PCR amplicon sequencing likely reflecting a low microbial abundance in these samples. Crude oil and asphalt from the natural seeps hosted diverse microbial communities. The same phylotypes of uncultivated Deferribacteres and Thermodesulfobacteraceae were predominant community members across crude oils and asphalts from separate geographical locations. Soils surrounding seeps did not contain these phylotypes suggesting that they originate from the subsurface and although they seem commonly detected in hydrocarbon-rich environments their role in hydrocarbon-degradation is unknown. GC-MS analyses showed that mainly aromatic hydrocarbons were present in the crude oil and asphalt and that they were undergoing biodegradation - likely with sulfate and nitrate as terminal oxidants. In agreement, only bssA gene, but not assA gene-carrying microorganisms were detectable in the analyzed sampled. Overall our study identified several abundant uncultivated taxa with likely roles in transformation of nitrate, sulfate and hydrocarbons.
Crude oil-amended microcosms were prepared with inocula from eleven anoxic environments (4 river sediments, 3 lake sediments, and 4 sludges from wastewater treatment reactors) to determine their ability to produce methane from the biodegradation of crude oil. Over incubation periods of up to 1150 days, oil-stimulated methanogenesis and concomitant loss of alkanes occurred in microcosms prepared with five of the inocula whereas six of the inocula did not show oil-stimulated methane production. Bacterial and archaeal communities from microcosms exhibiting high levels of oil-stimulated methanogenesis were distinct from communities where methanogenic crude oil degradation was not detected. Successional changes were consistent with the quantitative enrichment of syntrophic hydrocarbon degrading bacteria and methanogens over time. In conclusion, in oil-impacted environments methanogenic crude oil-degrading microbial consortia are present in relatively low abundance and exhibit slow growth, and while they may be ubiquitously distributed they may not be present at sufficiently high abundance to be detected.
Direct interspecies electron transfer (DIET) via electrically conductive minerals can play a role in the anaerobic oxidation of petroleum hydrocarbons in contaminated sites and can be exploited for the development of new, more effective bioremediation approaches.