Distribution of ETBE-degrading microorganisms and functional capability in groundwater, and implications for characterising aquifer ETBE biodegradation potential

Microbes in aquifers are present suspended in groundwater or attached to the aquifer sediment. Groundwater is often sampled at gasoline ether oxygenate (GEO)-impacted sites to assess the potential biodegradation of organic constituents. However, the distribution of GEO-degrading microorganisms between the groundwater and aquifer sediment must be understood to interpret this potential. In this study, the distribution of ethyl tert-butyl ether (ETBE)-degrading organisms and ETBE biodegradation potential was investigated in laboratory microcosm studies and mixed groundwater-aquifer sediment samples obtained from pumped monitoring wells at ETBE-impacted sites. ETBE biodegradation potential (as determined by quantification of the ethB gene) was detected predominantly in the attached microbial communities and was below detection limit in the groundwater communities. The copy number of ethB genes varied with borehole purge volume at the field sites. Members of the Comamonadaceae and Gammaproteobacteria families were identified as responders for ETBE biodegradation. However, the detection of the ethB gene is a more appropriate function-based indicator of ETBE biodegradation potential than taxonomic analysis of the microbial community. The study shows that a mixed groundwater-aquifer sediment (slurry) sample collected from monitoring wells after minimal purging can be used to assess the aquifer ETBE biodegradation potential at ETBE-release sites using this function-based concept.

Acetylene-Fueled Trichloroethene Reductive Dechlorination in a Groundwater Enrichment Culture

ABSTRACT In aquifers, acetylene (C2H2) is a product of abiotic degradation of trichloroethene (TCE) catalyzed by in situ minerals. C2H2 can, in turn, inhibit multiple microbial processes including TCE dechlorination and metabolisms that commonly support dechlorination, in addition to supporting the growth of acetylenotrophic microorganisms. Previously, C2H2 was shown to support TCE reductive dechlorination in synthetic, laboratory-constructed cocultures containing the acetylenotroph Pelobacter sp. strain SFB93 and Dehalococcoides mccartyi strain 195 or strain BAV1. In this study, we demonstrate TCE and perchloroethene (PCE) reductive dechlorination by a microbial community enriched from contaminated groundwater and amended with C2H2 as the sole electron donor and organic carbon source. The metagenome of the stable, enriched community was analyzed to elucidate putative community functions. A novel anaerobic acetylenotroph in the phylum Actinobacteria was identified using metagenomic analysis. These results demonstrate that the coupling of acetylenotrophy and reductive dechlorination can occur in the environment with native bacteria and broaden our understanding of biotransformation at contaminated sites containing both TCE and C2H2. IMPORTANCE Understanding the complex metabolisms of microbial communities in contaminated groundwaters is a challenge. PCE and TCE are among the most common groundwater contaminants in the United States that, when exposed to certain minerals, exhibit a unique abiotic degradation pathway in which C2H2 is a product. C2H2 can act as both an inhibitor of TCE dechlorination and of supporting metabolisms and an energy source for acetylenotrophic bacteria. Here, we combine laboratory microcosm studies with computational approaches to enrich and characterize an environmental microbial community that couples two uncommon metabolisms, demonstrating unique metabolic interactions only yet reported in synthetic, laboratory-constructed settings. Using this comprehensive approach, we have identified the first reported anaerobic acetylenotroph in the phylum Actinobacteria, demonstrating the yet-undescribed diversity of this metabolism that is widely considered to be uncommon.

Effect of Temperature on Acetate Mineralization Kinetics and Microbial Community Composition in a Hydrocarbon-Affected Microbial Community During a Shift From Oxic to Sulfidogenic Conditions

Aquifer thermal energy storage (ATES) allows for the seasonal storage and extraction of heat in the subsurface thus reducing reliance on fossil fuels and supporting decarbonization of the heating and cooling sector. However, the impacts of higher temperatures toward biodiversity and ecosystem services in the subsurface environment remain unclear. Here, we conducted a laboratory microcosm study comprising a hydrocarbon-degrading microbial community from a sulfidic hydrocarbon-contaminated aquifer spiked with 13C-labeled acetate and incubated at temperatures between 12 and 80°C to evaluate (i) the extent and rates of acetate mineralization and (ii) the resultant temperature-induced shifts in the microbial community structure. We observed biphasic mineralization curves at 12, 25, 38, and 45°C, arising from immediate and fast aerobic mineralization due to an initial oxygen exposure, followed by slower mineralization at sulfidogenic conditions. At 60°C and several replicates at 45°C, acetate was only aerobically mineralized. At 80°C, no mineralization was observed within 178 days. Rates of acetate mineralization coupled to sulfate reduction at 25 and 38°C were six times faster than at 12°C. Distinct microbial communities developed in oxic and strictly anoxic phases of mineralization as well as at different temperatures. Members of the Alphaproteobacteria were dominant in the oxic mineralization phase at 12–38°C, succeeded by a more diverse community in the anoxic phase composed of Deltaproteobacteria, Clostridia, Spirochaetia, Gammaproteobacteria and Anaerolinea, with varying abundances dependent on the temperature. In the oxic phases at 45 and 60°C, phylotypes affiliated to spore-forming Bacilli developed. In conclusion, temperatures up to 38°C allowed aerobic and anaerobic acetate mineralization albeit at varying rates, while mineralization occurred mainly aerobically between 45 and 60°C; thermophilic sulfate reducers being active at temperatures > 45°C were not detected. Hence, temperature may affect dissolved organic carbon mineralization rates in ATES while the variability in the microbial community composition during the transition from micro-oxic to sulfidogenic conditions highlights the crucial role of electron acceptor availability when combining ATES with bioremediation.

No Clear Response in the Stormwater Phytoplankton Community to Biocide Contamination

This study investigated the responses of natural phytoplankton communities of an urban stormwater pond to biocide contamination. The biocides carbendazim, terbutryn, diuron, and irgarol 1051, and their mixture, were used in two laboratory microcosm and one outdoor mesocosm studies at concentrations of 10, 100, and 1000 ng L−1. The water samples were collected in a pond receiving significant biocide contamination. The mesocosm study was carried out in the same pond. The phytoplankton community response was evaluated after 10–15 days of exposure, with respect to its taxonomic composition, abundance and biovolume. No significant changes were observed in any of the experiments. Only at the highest applied terbutryn concentration were lower numbers of taxa identified in both microcosm and mesocosm experiments. Additionally, increases in Chlorophyta abundance and biovolume, along with an increase in irgarol concentration, were observed throughout the three experiments. Nevertheless, the statistical analysis did not confirm any significant differences among the biocide treatments. These results suggest that the biocide concentrations probably were below the harmful or toxic threshold to the stormwater pond phytoplankton. On the other hand, as the investigated pond phytoplankton taxa face biocide inputs throughout the year, they could have already adapted to the tested biocide contamination.

Inactivation of Escherichia coli enhanced by anaerobic microbial iron reduction

Microbial iron reduction (MIR) is an important and ubiquitous natural process in the biogeochemical cycling of iron and carbon in anaerobic sedimentary and subsurface environments. The objectives of this study were (1) to determine if the MIR process can enhance the inactivation of Escherichia coli cells under anaerobic conditions and (2) to identify potential inactivation mechanisms. Laboratory microcosm experiments showed that the presence of MIR activity significantly enhanced E. coli inactivation, and the inactivation rate under the MIR condition was significantly larger than those under other anaerobic redox conditions. Under anoxic condition, higher Fe2+concentrations exhibited a linear function to larger E. coli inactivation rates, indicating that the production of Fe2+by MIR was one of the important roles in E. coli inactivation. When E. coli cells were amended as the sole electron source to the MIR process, increased Fe2+ production was observed, which corresponded to decreasing TOC concentration. Together, the results suggest that MIR enhanced E. coli inactivation through the production of Fe2+ as metabolic waste, and the inactivation benefited the MIR process as the inactivated cells were used as an electron source, which represents a potential new mechanism for bacterial inter-species competition. This knowledge could further improve our understanding of the fate of fecal bacteria in natural environments where the MIR process is prevalent, and may also be explored for enhanced removal of bacterial pathogens in engineering processes.

Severe Coastal Hypoxia Interchange with Ocean Acidification: An Experimental Perturbation Study on Carbon and Nutrient Biogeochemistry

Normally atmospheric CO2 is the major driver of ocean acidification (OA); however, local discharge/degradation of organic matter (OM) and redox reactions can exacerbate OA in coastal areas. In this work we study the response of nutrient and carbon systems to pH decrease in relation to hydrographically induced intermittent characteristics and examine scenarios for future ocean acidification in a coastal system. Laboratory microcosm experiments were conducted using seawater and surface sediment collected from the deepest part of Elefsis Bay; the pH was constantly being monitored while CO2 gas addition was adjusted automatically. In Elefsis Bay surface pCO2 is already higher than global present atmospheric values, while near the bottom pCO2 reaches 1538 μatm and carbonate saturation states were calculated to be around 1.5. During the experiment, in more acidified conditions, limited alkalinity increase was observed and was correlated with the addition of bicarbonates and OM. Ammonium oxidation was decelerated and a nitrification mechanism was noticed, despite oxygen deficiency, paralleled by reduction of Mn-oxides. Phosphate was found significantly elevated for the first time in lower pH values, without reprecipitating after reoxygenation; this was linked with Fe(II) oxidation and Fe(III) reprecipitation without phosphate adsorption affecting both available dissolved phosphate and (dissolved inorganic nitrogen) DIN:DIP (dissolved inorganic phosphate)ratio.

Predator complementarity dampens variability of phytoplankton biomass in a diversity-stability trophic cascade

Trophic cascades – indirect effects of predators that propagate down through food webs – have been extensively documented, especially in aquatic ecosystems. It has also been shown that predator diversity can mediate these trophic cascades, and, separately, that herbivore biomass can impact the stability of primary producers. However, whether predator diversity can cause cascading effects on the stability of lower trophic levels has not yet been studied. We conducted a laboratory microcosm experiment and a field mesocosm experiment manipulating the presence and coexistence of two heteropteran predators and measuring their effects on zooplankton herbivores and phytoplankton basal resources. We predicted that, if the predators partitioned their herbivore prey, for example by size, then co-presence of the predators would lead to 1) increased average values and 2) decreased temporal variability of phytoplankton basal resources. We present evidence that the predators partitioned their herbivore prey and found that their simultaneous suppression of herbivore groups reduced the variability of edible (smaller) phytoplankton biomass, without affecting mean phytoplankton biomass. We also found that phytoplankton that were more resistant to herbivory were not affected by our manipulations, indicating that the zooplankton herbivores played an important role in mediating this cascading diversity-stability effect. Our results demonstrate that predator diversity may indirectly stabilize basal resource biomass via a “diversity-stability trophic cascade,” seemingly dependent on predator complementarity and the vulnerability of taxa to consumption, but independent of a classic trophic cascade in which average biomass is altered. Predator diversity, especially if correlated with diversity of prey use, may be important for regulating ecosystem stability, and this relationship suggests biological control methods for improving the reliability of microalgal yields.

Effects of Detritus on the Mosquito Culex pipiens: Phragmites and Schedonorus (Festuca) Invasion Affect Population Performance

Species interactions that influence the performance of the exotic mosquito Culex pipiens can have important effects on the transmission risk of West Nile virus (WNV). Invasive plants that alter the vegetation communities of ephemeral ground pools may facilitate or resist the spread of C. pipiens (L.) by altering allochthonous inputs of detritus in those pools. To test this hypothesis, we combined field surveys of roadside stormwater ditches with a laboratory microcosm experiment to examine relationships between C. pipiens performance and water quality in systems containing detritus from invasive Phragmites australis (Cav.) Trin. Ex Steud., introduced Schedonorus arundinaceus (Schreb.) Dumort., or native Juncus effusus L. or Typha latifolia L. In ditches, C. pipiens abundance was unrelated to detritus species but female C. pipiens were significantly larger from ditches with S. arundinaceus and smaller with J. effusus. Larger and smaller C. pipiens were also produced in microcosms provisioned with S. arundinaceus and J. effusus, respectively, yet the per capita rate of population of change did not vary. Larger females from habitats with S. arundinaceus were likely caused by faster decay rates of S. arundinaceus and resultant increases in microbial food, but lower survival as a result of fouling and higher tannin-lignin concentrations resulted in little changes to overall population performance. Larger female mosquitoes have been shown to have greater potential for transmitting arboviruses. Our findings suggest that changed community-level interactions from plant invasions in urban ephemeral ground pools can affect the fitness of C. pipiens and possibly increase WNV risk.