Assessment of the in situ biomethanation potential of a deep aquifer used for natural gas storage.

The dihydrogen (H2) sector is undergoing development and will require massive storage solutions. To minimize costs, the conversion of underground geological storage sites, such as deep aquifers, used for natural gas storage into future underground hydrogen storage sites is the favored scenario. However, these sites contain microorganisms capable of consuming H2, mainly sulfate reducers and methanogens. Methanogenesis is, therefore expected but its intensity must be evaluated. Here, in a deep aquifer used for underground geological storage, 17 sites were sampled, with low sulfate concentrations ranging from 21.9 to 197.8 µM and a slow renewal of formation water. H2-selected communities mainly were composed of the families Methanobacteriaceae and Methanothermobacteriaceae and the genera Desulfovibrio, Thermodesulfovibrio, and Desulforamulus. Experiments were done under different conditions, and sulfate reduction, as well as methanogenesis, were demonstrated in the presence of a H2 or H2/CO2 (80/20) gas phase, with or without calcite/site rock. These metabolisms led to an increase in pH up to 10.2 under certain conditions (without CO2). The results suggest competition for CO2 between lithoautotrophs and carbonate mineral precipitation, which could limit microbial H2 consumption.

Novel field-portable high-pressure adsorbent tube sampler prototype for the direct in situ preconcentration of trace compounds in gases at their working pressures: application to biomethane.

In Europe, renewable energy gases such as biomethane are aimed at substituting natural gas provided their stringent compliance to natural gas quality standards stipulating maximal levels of several chemical trace compounds (TC). Preconcentration is generally required to detect TC and inasmuch as biomethane is compressed for injection in the natural gas grid, preconcentration is commonly either done by collecting the bulk pressurized gas in a high-pressure cylinder or by first depressurizing it to collect a bulk volume in e.g. a gas sampling bag. Such whole gas samples are then transported to the lab and transferred to a preconcentration unit, entailing contamination and TC loss risks. Therefore, here a novel handy field-portable device for the direct in situ high-pressure preconcentration of TC is presented, enabling to sample gases at pressures up to 200 bara through a self-assembled Tenax®TA + Carbopack™X multibed adsorbent tube. The effect of the gas sampling pressure on the preconcentration of TC on adsorbent tubes was evaluated using a synthetic gas mixture containing 41 halogenated volatile organic compounds each at 1 ppmmol in N2. At given normalized sampled volumes and in the pressure range 5-100 bara handled in French gas transport grids, the pressure had no influence on the preconcentration when the gas circulates through the adsorbent tubes and as long as the adsorbents are not saturated. Next, for the first time, a real biomethane stream was sampled using the novel direct high-pressure preconcentration method on Tenax®TA + Carbopack™X multibed adsorbent tubes, allowing to preconcentrate, in a single sampling run, a wide range of volatile organic TC. More than 26 distinct TC were detected, belonging to seven chemical families: alkenes, aromatics, alkanes (linear, cyclic and polycyclic), sulphur-compounds and terpenes, with linear alkanes (pentane, heptane, octane) and terpenes predominating. Semi-quantification indicated pentane, dimethylcyclopropane, hexane, heptane, octane, α-pinene and camphene are present at a ≤1 ppmmol concentration threshold in the biomethane.

Biological, geological and chemical effects of oxygen injection in underground gas storage aquifers in the setting of biomethane deployment.

The last few years have seen the proliferation of anaerobic digestion plants to produce biomethane. Oxygen (O2) traces added to biogas during the desulfurization process are co-injected in the gas network and can be stored in Underground Gas Storage (UGS). However, there are no data available for the undesirable effects of O2 on these anoxic environments, especially on deep aquifers. In addition to mineral alteration, O2 can have an impact on the anaerobic autochthonous microbial life. In our study, the storage conditions of an UGS aquifer were reproduced in a high-pressure reactor and bio-geo-chemical interactions between the aqueous, gas and solid phases were studied. Sulfate was depleted from the liquid phase for three consecutive times during the first 130 days of incubation reproducing the storage conditions (36 °C, 60 bar, methane with 1% CO2). Sulfate-reducers, such as Desulfovibrionaceae, were identified from the high-pressure system. Simulations with PHREEQC were used to determine the thermodynamic equilibrium to confirm any gas consumption. CO2 quantities decreased in the gas phase, suggesting its use as carbon source by microbial life. Benzene and toluene, hydrocarbons found in traces and known to be biodegradable in storages, were monitored and a decrease of toluene was revealed and associated to the Peptococcaceae family. Afterwards, O2 was added as 1% of the gas phase, corresponding to the maximum quantity found in biomethane after desulfurization process. Re-oxidation of sulfide to sulfate was observed along with the end of sulfate reducing activity and toluene biodegradation and the disappearance of most of the community. H2 surprisingly appeared and accumulated as soon as hydrogenotrophic sulfate-reducers decreased. H2 would be produced via the necromass fermentation accomplished by microorganisms able to resist the oxic conditions of 4.42·10-4 mol.Kgw-1 of O2. The solid phase composed essentially of quartz, presented no remarkable changes.

A deep continental aquifer downhole sampler for microbiological studies.

To be effective, microbiological studies of deep aquifers must be free from surface microbial contaminants and from infrastructures allowing access to formation water (wellheads, well completions). Many microbiological studies are based on water samples obtained after rinsing a well without guaranteeing the absence of contaminants from the biofilm development in the pipes. The protocol described in this paper presents the adaptation, preparation, sterilization and deployment of a commercial downhole sampler (PDSshort, Leutert, Germany) for the microbiological studying of deep aquifers. The ATEX sampler (i.e., explosive atmospheres) can be deployed for geological gas storage (methane, hydrogen). To validate our procedure and confirm the need to use such a device, cell counting and bacterial taxonomic diversity based on high-throughput sequencing for different water samples taken at the wellhead or at depth using the downhole sampler were compared and discussed. The results show that even after extensive rinsing (7 bore volumes), the water collected at the wellhead was not free of microbial contaminants, as shown by beta-diversity analysis. The downhole sampler procedure was the only way to ensure the purity of the formation water samples from the microbiological point of view. In addition, the downhole sampler allowed the formation water and the autochthonous microbial community to be maintained at in situ pressure for laboratory analysis. The prevention of the contamination of the sample and the preservation of its representativeness are key to guaranteeing the best interpretations and understanding of the functioning of the deep biosphere.

Microbial Diversity Under the Influence of Natural Gas Storage in a Deep Aquifer.

Deep aquifers (up to 2km deep) contain massive volumes of water harboring large and diverse microbial communities at high pressure. Aquifers are home to microbial ecosystems that participate in physicochemical balances. These microorganisms can positively or negatively interfere with subsurface (i) energy storage (CH4 and H2), (ii) CO2 sequestration; and (iii) resource (water, rare metals) exploitation. The aquifer studied here (720m deep, 37°C, 88bar) is naturally oligotrophic, with a total organic carbon content of <1mg.L-1 and a phosphate content of 0.02mg.L-1. The influence of natural gas storage locally generates different pressures and formation water displacements, but it also releases organic molecules such as monoaromatic hydrocarbons at the gas/water interface. The hydrocarbon biodegradation ability of the indigenous microbial community was evaluated in this work. The in situ microbial community was dominated by sulfate-reducing (e.g., Sva0485 lineage, Thermodesulfovibriona, Desulfotomaculum, Desulfomonile, and Desulfovibrio), fermentative (e.g., Peptococcaceae SCADC1_2_3, Anaerolineae lineage and Pelotomaculum), and homoacetogenic bacteria ("Candidatus Acetothermia") with a few archaeal representatives (e.g., Methanomassiliicoccaceae, Methanobacteriaceae, and members of the Bathyarcheia class), suggesting a role of H2 in microenvironment functioning. Monoaromatic hydrocarbon biodegradation is carried out by sulfate reducers and favored by concentrated biomass and slightly acidic conditions, which suggests that biodegradation should preferably occur in biofilms present on the surfaces of aquifer rock, rather than by planktonic bacteria. A simplified bacterial community, which was able to degrade monoaromatic hydrocarbons at atmospheric pressure over several months, was selected for incubation experiments at in situ pressure (i.e., 90bar). These showed that the abundance of various bacterial genera was altered, while taxonomic diversity was mostly unchanged. The candidate phylum Acetothermia was characteristic of the community incubated at 90bar. This work suggests that even if pressures on the order of 90bar do not seem to select for obligate piezophilic organisms, modifications of the thermodynamic equilibria could favor different microbial assemblages from those observed at atmospheric pressure.

Transcriptomic Responses During Early Development Following Arsenic Exposure in Western Clawed Frogs, Silurana tropicalis.

Arsenic compounds are widespread environmental contaminants and exposure elicits serious health issues, including early developmental anomalies. Depending on the oxidation state, the intermediates of arsenic metabolism interfere with a range of subcellular events, but the fundamental molecular events that lead to speciation-dependent arsenic toxicity are not fully elucidated. This study therefore assesses the impact of arsenic exposure on early development by measuring speciation and gene expression profiles in the developing Western clawed frog (Silurana tropicalis) larvae following the environmental relevant 0.5 and 1 ppm arsenate exposure. Using HPLC-ICP-MS, arsenate, dimethylarsenic acid, arsenobetaine, arsenocholine, and tetramethylarsonium ion were detected. Microarray and pathway analyses were utilized to characterize the comprehensive transcriptomic responses to arsenic exposure. Clustering analysis of expression data showed distinct gene expression patterns in arsenate treated groups when compared with the control. Pathway enrichment revealed common biological themes enriched in both treatments, including cell signal transduction, cell survival, and developmental pathways. Moreover, the 0.5 ppm exposure led to the enrichment of pathways and biological processes involved in arsenic intake or efflux, as well as histone remodeling. These compensatory responses are hypothesized to be responsible for maintaining an in-body arsenic level comparable to control animals. With no appreciable changes observed in malformation and mortality between control and exposed larvae, this is the first study to suggest that the underlying transcriptomic regulations related to signal transduction, cell survival, developmental pathways, and histone remodeling may contribute to maintaining ongoing development while coping with the potential arsenic toxicity in S. tropicalis during early development.

Arsenic(+3) and DNA methyltransferases, and arsenic speciation in tadpole and frog life stages of western clawed frogs (Silurana tropicalis) exposed to arsenate.

Western clawed frog (Silurana tropicalis) embryos were exposed to control, low (nominally 0.5 mg L(-1)) and high (nominally 1 mg L(-1)) arsenate (As(V)) culture water concentrations to investigate the effects of arsenic (As) on different life stages, namely tadpole (Nieuwkoop and Faber stage 56, NF56) and frog stages (NF66). The effects were assessed by measuring arsenic(+3) and DNA methyltransferases (AS3MT and DNMT1), as well as As speciation in the tissues. The As content in frog tissues increased with water As concentration. The As species observed by high performance liquid chromatography - inductively coupled plasma mass spectrometry (HPLC-ICPMS) were mostly inorganic, dimethylarsinic acid (DMA) and trimethylarsine oxide (TMAO). With solid state X-ray absorption near edge structure (XANES) analysis, arsenobetaine/tetramethylarsonium ion were also seen. AS3MT levels decreased upon low As exposure in NF56, rising again to control levels at the high As exposure. In NF66 tissues, on the other hand, AS3MT decreased only with NF66 high As exposure. DNMT1 increased with exposure, and this was statistically significant only for the high As exposure at both life stages. Thus these enzymes seem to be affected by the As exposure. Methylation of As to form monomethylarsonate (MMA), DMA and TMAO in the frogs appeared to be inversely related to AS3MT levels. A possible interpretation of this finding is that when AS3MT is higher, excretion of MMA + DMA + TMAO is more efficient, leaving lower concentrations in the tissues, with the opposite effect (less excretion) when AS3MT is lower; alternatively, other enzymes or linked genes may affect the methylation of As.

Strategies of Gammarus pulex L. to cope with arsenic--Results from speciation analyses by IC-ICP-MS and XAS micro-mapping.

The invertebrate shredder Gammarus pulex L. is a key species for aquatic carbon turnover via litter decomposition and can thrive in high-arsenic (As) environments. To understand their strategies for coping with increased As concentrations while fulfilling their ecosystem functions, we analyzed the As concentration and speciation in their aquatic habitat and in leaves with heterotrophic biofilms as their natural food source. We also followed the As distribution and speciation on the cuticle and within the body of G. pulex by X-ray absorption spectroscopic imaging. Half of the total As on G. pulex was found to be associated with the cuticle but was not taken up. Removing this externally bound As yielded only arsenate in the wash solution which reflects the speciation of the surrounding aquatic phase and shows that this As does not undergo any biotransformation. The major pathway into the organism is suggested to be incorporation via food intake, but only very low amounts of As were taken up or translocated from the gut system to other tissues. In one of the main food sources, leaves, 68% arsenate and 29% monomethylarsenate were found. After ingestion into the gut system, up to 23% of the more toxic arsenite was seen, but a substantial share was methylated to dimethylarsenate (46-56%). Little arsenate and arsenite were found in the adjacent tissues. Besides 76-80% mono- and di-methylarsenate, 10-21% of the As was complexed as As(III)-S species. G. pulex plays an important role in As cycling and our results indicate that As translocation from the gut to other tissues is minimized, but a transformation to other As-species occurred.

Arsenic distribution and speciation in Daphnia pulex.

Rat Lake, Yellowknife, Northwest Territories, is situated on arsenic-rich tailings from a historical gold mine. The abundant zooplankton species Daphnia pulex in this lake was used to study the impact of arsenic at the base of the freshwater food web; the speciation and distribution of arsenic in D. pulex and its food sources; and the origin of formation of organoarsenicals in freshwater systems. The arsenic concentration in lake water was measured as 0.25 mg L(-1), while the zooplankton organisms contained up to 35 mg kg(-1) d.w. arsenic. Plankton samples were analyzed for arsenic speciation, by using X-ray Absorption Near Edge Structure (XANES) on the whole, dried samples and High Performance Liquid Chromatography coupled to Inductively Coupled Plasma Mass Spectrometry (HPLC-ICP-MS) on water extracts. XANES data suggest that D. pulex mainly contain inorganic arsenicals with 56% of arsenic with +5 oxidation state and 10% of arsenic with +3 oxidation state, but also 34% of organoarsenic compounds that were identified with HPLC-ICP-MS as monomethylarsonate (MMA), dimethylarsinate (DMA), and arsenosugars. The most abundant of the organoarsenicals was the glycerol sugar (Sugar 1). X-ray Fluorescence (XRF) mapping of D. pulex for arsenic distribution showed that arsenic was mainly distributed in the gut of the animal, where its concentration was ten times higher than in the surrounding tissues. Moreover, the analysis of residues from extractions targeting water-soluble and lipid-soluble arsenicals suggested that part of the measured arsenic signal comes from ingested sediments, phytoplankton, or other food sources. These food sources contain inorganic arsenic only, with As(V)-O in phytoplankton and As(III)-S in sediments, suggesting the possibility that the organoarsenicals compounds detected in the tissues of the organism are created by the Daphnia.

Arsenic speciation in plankton organisms from contaminated lakes: transformations at the base of the freshwater food chain.

The two complementary techniques high performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS) and X-ray absorption near edge structure (XANES) analysis were used to assess arsenic speciation in freshwater phytoplankton and zooplankton collected from arsenic-contaminated lakes in Yellowknife (Northwest Territories, Canada). Arsenic concentrations in lake water ranged from 7 µg L(-1) in a noncontaminated lake to 250 µg L(-1) in mine-contaminated lakes, which resulted in arsenic concentrations ranging from 7 to 340 mg kg(-1) d.w. in zooplankton organisms (Cyclops sp.) and from 154 to 894 mg kg(-1) d.w. in phytoplankton. The main arsenic compounds identified by HPLC-ICP-MS in all plankton were inorganic arsenic (from 38% to 98% of total arsenic). No other arsenic compounds were found in phytoplankton, but zooplankton organisms showed the presence of organoarsenic compounds, the most common being the sulfate arsenosugar, up to 47% of total arsenic, with traces of phosphate sugar, glycerol sugar, methylarsonate (MMA), and dimethylarsinate (DMA). In the uncontaminated Grace Lake, zooplankton also contained arsenobetaine (AB). XANES characterization of arsenic in the whole plankton samples showed As(V)-O as the only arsenic compound in phytoplankton, and As(III)-S and As(V)-O compounds as the two major inorganic arsenic species in zooplankton. The proportion of organoarsenicals and inorganic arsenic in zooplankton depends upon the arsenic concentration in lakes and shows the impact of arsenic contamination: zooplankton from uncontaminated lake has higher proportions of organoarsenic compounds and contains arsenobetaine, while zooplankton from contaminated area contains mostly inorganic arsenic.

Sensitivity improvement in ICP MS analysis of fuels and light petroleum matrices using a microflow nebulizer and heated spray chamber sample introduction.

Reasons for signal suppression during the analysis of light petroleum matrices by inductively coupled plasma mass spectrometry (ICP MS) were examined. A decrease of the ionization efficiency of the plasma was found to be the principal factor responsible for this loss of sensitivity. Consequently, an interface based on a total consumption micronebulizer and a heated spray chamber was constructed to alleviate this problem. A method based on flow-injection ICP MS using this interface was developed for the direct multielement analysis of undiluted fuels (gasoline, kerosene) and gas condensates offering an increase in sensitivity by at least a factor of 3-4 in comparison with the existing setups.

Monitoring the arsenic and iodine exposure of seaweed-eating North Ronaldsay sheep from the gestational and suckling periods to adulthood by using horns as a dietary archive.

Trace elements often accumulate in keratin-rich tissues. Hair, nails, and horns grow steadily but once formed are metabolically inactive and provide an archive of trace element exposure when analyzed in segments. Here we demonstrate the use of laser ablation ICP-MS for the high-resolution monitoring of trace elements in the horns of seaweed-eating sheep from North Ronaldsay, which live on grass only during lambing time. Due to this peculiar husbandry/dietary pattern and the fact that seaweed is rich in arsenic and iodine, we hoped to use iodine and arsenic as markers for seaweed ingestion. Cross sections and scans along the growing axis (representing the first 8-10 months of the sheep's life) revealed that these elements were not homogeneously distributed in the horn, with arsenic representing the amount of seaweed intake. The scans show the periods in which the lambs were fed on milk and grass and the change to seaweed ingestion with the successive replacement of milk with seaweed; this was supported by the carbon and nitrogen isotope signatures (delta13C and delta15N) of the horn and the arsenic speciation in the horn. The period of low arsenic accumulation in the horn had terrestrial isotope signatures and accumulated arsenic of mainly inorganic origin. The period of high arsenic accumulation was characterized by isotope signatures of marine origin, and the majority of accumulated arsenic in the horn was the main arsenosugar metabolite dimethylarsinic acid. Although we have investigated only four different horns of individual sheep, this study shows that arsenic is not significantly transported with milk. However, the high concentration of arsenic in the oldest part of the horn, which was formed in utero, points to a relatively high placental transport of arsenic while the ewe was eating seaweed. In contrast to arsenic, iodine is transported not only through milk ingestion but also through the placenta in large quantities.