High diversity, abundance, and expression of hydrogenases in groundwater.

Hydrogen may be the most important electron donor available in the subsurface. Here we analyse the diversity, abundance and expression of hydrogenases in 5 proteomes, 25 metagenomes, and 265 amplicon datasets of groundwaters with diverse geochemistry. A total of 1545 new [NiFe]-hydrogenase gene sequences were recovered, which considerably increased the number of sequences (1999) in a widely used database. [NiFe]-hydrogenases were highly abundant, as abundant as the DNA-directed RNA polymerase. The abundance of hydrogenase genes increased with depth from 0 to 129 m. Hydrogenases were present in 481 out of 1245 metagenome-assembled genomes. The relative abundance of microbes with hydrogenases accounted for ~50% of the entire community. Hydrogenases were actively expressed, making up as much as 5.9% of methanogen proteomes. Most of the newly discovered diversity of hydrogenases was in "Group 3b", which has been associated with sulfur metabolism. "Group 3d", facilitating the interconversion of electrons between hydrogen and NAD, was the most abundant and mainly observed in methanotrophs and chemoautotrophs. "Group 3a", associated with methanogenesis, was the most abundant in proteomes. Two newly discovered groups of [NiFe]-hydrogenases, observed in Methanobacteriaceae and Anaerolineaceae, further expanded diversity. Our results highlight the vast diversity, abundance and expression of hydrogenases in groundwaters, suggesting a high potential for hydrogen oxidation in subsurface habitats.

Global diversity and inferred ecophysiology of microorganisms with the potential for dissimilatory sulfate/sulfite reduction.

Sulfate/sulfite-reducing microorganisms (SRM) are ubiquitous in nature, driving the global sulfur cycle. A hallmark of SRM is the dissimilatory sulfite reductase encoded by the genes dsrAB. Based on analysis of 950 mainly metagenome-derived dsrAB-carrying genomes, we redefine the global diversity of microorganisms with the potential for dissimilatory sulfate/sulfite reduction and uncover genetic repertoires that challenge earlier generalizations regarding their mode of energy metabolism. We show: (i) 19 out of 23 bacterial and 2 out of 4 archaeal phyla harbor uncharacterized SRM, (ii) four phyla including the Desulfobacterota harbor microorganisms with the genetic potential to switch between sulfate/sulfite reduction and sulfur oxidation, and (iii) the combination as well as presence/absence of different dsrAB-types, dsrL-types and dsrD provides guidance on the inferred direction of dissimilatory sulfur metabolism. We further provide an updated dsrAB database including > 60% taxonomically resolved, uncultured family-level lineages and recommendations on existing dsrAB-targeted primers for environmental surveys. Our work summarizes insights into the inferred ecophysiology of newly discovered SRM, puts SRM diversity into context of the major recent changes in bacterial and archaeal taxonomy, and provides an up-to-date framework to study SRM in a global context.

Frequency of change determines effectiveness of microbial response strategies.

Nature challenges microbes with change at different frequencies and demands an effective response for survival. Here, we used controlled laboratory experiments to investigate the effectiveness of different response strategies, such as post-translational modification, transcriptional regulation, and specialized versus adaptable metabolisms. For this, we inoculated replicated chemostats with an enrichment culture obtained from sulfidic stream microbiomes 16 weeks prior. The chemostats were submitted to alternatingly oxic and anoxic conditions at three frequencies, with periods of 1, 4 and 16 days. The microbial response was recorded with 16S rRNA gene amplicon sequencing, shotgun metagenomics, transcriptomics and proteomics. Metagenomics resolved provisional genomes of all abundant bacterial populations, mainly affiliated with Proteobacteria and Bacteroidetes. Almost all these populations maintained a steady growth rate under both redox conditions at all three frequencies of change. Our results supported three conclusions: (1) Oscillating oxic/anoxic conditions selected for generalistic species, rather than species specializing in only a single condition. (2) A high frequency of change selected for strong codon usage bias. (3) Alignment of transcriptomes and proteomes required multiple generations and was dependent on a low frequency of change.

Performance, microbial growth and community interactions of iron-dependent denitrification in freshwaters.

Iron-dependent denitrification is a safe and promising technology for nitrogen removal in freshwaters. However, the understanding of microbial physiology and interactions during the process was limited. Denitrifying systems inoculated with freshwater samples were operated with and without iron(II) at a low C/N ratio for 54 days. Iron addition improved nitrogen removal. Batch experiments confirmed that microbially mediated reaction rather than abiotic reaction dominated during the process. Metagenomics recovered genomes of the five most abundant microorganisms, which accounted for over 99% of the community in every triplicate of the iron-based system. Based on codon usage bias, all of them were fast-growing organisms. The total abundance of fast-growing organisms was 38% higher in the system with iron than in the system without iron. Notably, the most abundant organism Diaphorobacter did not have enzymes for asparagine and aspartate biosynthesis, whereas Rhodanobacter could not produce serine and cobalamin. Algoriphagus and Areminomonas lost synthesis enzymes for more amino acids and vitamins. However, they could always obtain these growth-required substances from another microorganism in the community. The two-partner relationship minimized the limitation on microbial reproduction and increased community stability. Our results indicated that iron addition improved nitrogen removal by supplying electron donors, promoting microbial growth, and building up syntrophic interactions among microorganisms with timely communications. The findings provided new insights into the process, with implications for freshwater remediation.

Hydrogen and dark oxygen drive microbial productivity in diverse groundwater ecosystems.

Around 50% of humankind relies on groundwater as a source of drinking water. Here we investigate the age, geochemistry, and microbiology of 138 groundwater samples from 95 monitoring wells (<250 m depth) located in 14 aquifers in Canada. The geochemistry and microbiology show consistent trends suggesting large-scale aerobic and anaerobic hydrogen, methane, nitrogen, and sulfur cycling carried out by diverse microbial communities. Older groundwaters, especially in aquifers with organic carbon-rich strata, contain on average more cells (up to 1.4 × 107 mL-1) than younger groundwaters, challenging current estimates of subsurface cell abundances. We observe substantial concentrations of dissolved oxygen (0.52 ± 0.12 mg L-1 [mean ± SE]; n = 57) in older groundwaters that seem to support aerobic metabolisms in subsurface ecosystems at an unprecedented scale. Metagenomics, oxygen isotope analyses and mixing models indicate that dark oxygen is produced in situ via microbial dismutation. We show that ancient groundwaters sustain productive communities and highlight an overlooked oxygen source in present and past subsurface ecosystems of Earth.

Succession of bacteria and archaea involved in the nitrogen cycle of a seasonally stratified lake.

Human-driven changes affect nutrient inputs, oxygen solubility, and the hydrodynamics of lakes, which affect biogeochemical cycles mediated by microbial communities. However, information on the succession of microbes involved in nitrogen cycling in seasonally stratified lakes is still incomplete. Here, we investigated the succession of nitrogen-transforming microorganisms in Lake Vechten over a period of 19 months, combining 16S rRNA gene amplicon sequencing and quantification of functional genes. Ammonia-oxidizing archaea (AOA) and bacteria (AOB) and anammox bacteria were abundant in the sediment during winter, accompanied by nitrate in the water column. Nitrogen-fixing bacteria and denitrifying bacteria emerged in the water column in spring when nitrate was gradually depleted. Denitrifying bacteria containing nirS genes were exclusively present in the anoxic hypolimnion. During summer stratification, abundances of AOA, AOB, and anammox bacteria decreased sharply in the sediment, and ammonium accumulated in hypolimnion. After lake mixing during fall turnover, abundances of AOA, AOB, and anammox bacteria increased and ammonium was oxidized to nitrate. Hence, nitrogen-transforming microorganisms in Lake Vechten displayed a pronounced seasonal succession, which was strongly determined by the seasonal stratification pattern. These results imply that changes in stratification and vertical mixing induced by global warming are likely to alter the nitrogen cycle of seasonally stratified lakes.

Magnetic reed biochar materials as adsorbents for aqueous copper and phenol removal.

Organics and heavy metals are common pollutants in many wastewaters and water bodies. Adsorption processes by magnetic materials can rapidly remove these pollutants from water and effectively recycle adsorbent. In this study, magnetic analyzer, X-ray diffraction, Flourier transform infrared spectroscopy, and granulometry were used to characterize the synthesized magnetic reed biochar materials (ZnFe2O4/biochar). Influences of adsorption time, pH, temperature, initial solution concentration, and adsorption equilibrium concentration on adsorption performances were investigated for Cu2+ and phenol adsorption by ZnFe2O4/biochar. Adsorption kinetic and isotherm models were used to describe the adsorption processes. Adsorption of phenol and Cu2+ by ZnFe2O4/biochar reached saturation within 45 min and increased slightly with the increase of temperature from 15 to 45 °C. Adsorption of Cu2+ increased with the increase of pH, while the adsorption of phenol peaked at pH = 6. The adsorption processes fit the pseudo-second order kinetics model, and both conformed to the Langmuir model. The fitting results show that the maximum single-component adsorption capacity of phenol and Cu2+ by ZnFe2O4/biochar is 63.29 and 12.20 mg/g, and the maximum bi-component adsorption capacity reaches 40.16 and 9.48 mg/g, respectively. All the findings demonstrate that ZnFe2O4/biochar has good adsorption performance for phenol and Cu2+.

Distinct oxygen isotope fractionations driven by different electron donors during microbial nitrate reduction in lake sediments.

Microbial nitrate reduction can be driven by organic carbon oxidation, as well as by inorganic electron donors, such as reduced forms of sulfur and iron. An apparent inverse oxygen isotope fractionation effect was observed during nitrate reduction in sediment incubations from five sampling sites of a freshwater lake, Hongze Lake, China. Incubations with organic and inorganic electron donor additions were performed. Especially, the inverse oxygen isotope effect was intensified after glucose addition, whereas the incubations with sulfide and Fe2+ showed normal fractionation factors. Nitrate reductase encoding genes, napA and narG, were analysed with metagenomics. Higher napA/narG ratios were associated with higher oxygen fractionation factors. The most abundant clade (59%) of NapA in the incubation with glucose was affiliated with Rhodocyclales. In contrast, it only accounted for 8%-9% of NapA in the incubations with sulfide and Fe2+ . Differences in nitrate reductases might explain different oxygen isotope effects. Our findings also suggested that large variance of O-nitrate isotope fractionations might have to be considered in the interpretation of natural isotope records.

Responses of phytoremediation in urban wastewater with water hyacinths to extreme precipitation.

Climate change not only intensifies eutrophication and enhances the rainfall, but also elevates the contents of greenhouse gases, which can further increase the intensity and frequency of extreme precipitation events. The effectivity of phytoremediation of urban wastewaters by water hyacinths under an extreme rainfall event (up to 380 mm d-1) was investigated using self-designed fabrications with six flow rates (2-15 m3 d-1) in situ on pilot scale for 30 days. The results suggest that water hyacinths had high N and P removal capacities even under adverse conditions such as low dissolved oxygen concentrations (DO, <1 mg L-1) and high ammonium concentrations (NH4+-N, >7 mg L-1). Specifically, the highest removal yields of N and P were 13.14 ± 0.47 g N·m-2·d-1 and 2.12 ± 0.04 g P·m-2·d-1, respectively. The results indicate that water hyacinths can be used for water treatment to reduce the amounts of NH4+-N, dissolved organic nitrogen (DON) and phosphate (PO43-) even during extreme precipitation events. Moreover, DO increased due to wet deposition, runoff and surface flows during the extreme rainfall event, resulting in shifts between nitrification and denitrification processes which significantly altered nitrogen forms in urban wastewater. Results of this study suggest that water hyacinths could be recommended as a cost-effective and eco-friendly technology for urban wastewater phytoremediation in areas suffering from frequent extreme precipitation events.

Enhancement of synchronous bio-reductions of vanadium (V) and chromium (VI) by mixed anaerobic culture.

The co-occurrence of toxic vanadium (V) and chromium (VI) in groundwater receives incremental attention while knowledge on their interactions in biogeochemical processes is limited, with lack of efficient removal means. This study is the first to realize synchronous bio-reductions of V(V) and Cr(VI) with high efficiency by mixed anaerobic culture. After 72-h operation, 97.0 ±â€¯1.0% of V(V) and 99.1 ±â€¯0.7% of Cr(VI) were removed, respectively, with initial concentration of 1 mM for both V(V) and Cr(VI). Cr(VI) bio-reduction took priority while V(V) detoxification was inhibited. V(IV) and Cr(III) were the identified reduction products, both of which could precipitate naturally. Initial Cr(VI) and acetate concentrations as well as pH affected this process significantly. High-throughput 16S rRNA gene sequencing analysis indicated the accumulation of Anaerolineaceae, Spirochaeta and Spirochaetaceae, which could contribute to V(V) and Cr(VI) bio-reductions. The new knowledge obtained in this study will facilitate understanding the biogeochemical fate of co-existing V(V) and Cr(VI) in groundwater and development of bioremediation strategy for their induced combined pollution.

Spatio-temporal dynamics of sulfur bacteria during oxic--anoxic regime shifts in a seasonally stratified lake.

Sulfate-reducing bacteria (SRB) and sulfur-oxidizing bacteria drive major transformations in the sulfur cycle, and play vital roles in oxic--anoxic transitions in lakes and coastal waters. However, information on the succession of these sulfur bacteria in seasonally stratified lakes using molecular biological techniques is scarce. Here, we used 16S rRNA gene amplicon sequencing to study the spatio-temporal dynamics of sulfur bacteria during oxic--anoxic regime shifts in Lake Vechten. Oxygen and sulfate were mixed throughout the water column in winter and early spring. Meanwhile, SRB, green sulfur bacteria (GSB), purple sulfur bacteria (PSB), and colorless sulfur bacteria (CSB) exclusively inhabited the sediment. After the water column stratified, oxygen and nitrate concentrations decreased in the hypolimnion and various SRB species expanded into the anoxic hypolimnion. Consequently, sulfate was reduced to sulfide, stimulating the growth of PSB and GSB in the metalimnion and hypolimnion during summer stratification. When hypoxia spread throughout the water column during fall turnover, SRB and GSB vanished from the water column, whereas CSB (mainly Arcobacter) and PSB (Lamprocystis) became dominant and oxidized the accumulated sulfide under micro-aerobic conditions. Our results support the view that, once ecosystems have become anoxic and sulfidic, a large oxygen influx is needed to overcome the anaerobic sulfur cycle and bring the ecosystems back into their oxic state.

Oxic-anoxic regime shifts mediated by feedbacks between biogeochemical processes and microbial community dynamics.

Although regime shifts are known from various ecosystems, the involvement of microbial communities is poorly understood. Here we show that gradual environmental changes induced by, for example, eutrophication or global warming can induce major oxic-anoxic regime shifts. We first investigate a mathematical model describing interactions between microbial communities and biogeochemical oxidation-reduction reactions. In response to gradual changes in oxygen influx, this model abruptly transitions between an oxic state dominated by cyanobacteria and an anoxic state with sulfate-reducing bacteria and phototrophic sulfur bacteria. The model predictions are consistent with observations from a seasonally stratified lake, which shows hysteresis in the transition between oxic and anoxic states with similar changes in microbial community composition. Our results suggest that hysteresis loops and tipping points are a common feature of oxic-anoxic transitions, causing rapid drops in oxygen levels that are not easily reversed, at scales ranging from small ponds to global oceanic anoxic events.The role of microbial communities in regime shifts is poorly understood. Here, the authors use a mathematical model and field data from a seasonally stratified lake to show that gradual environmental changes can induce oxic-anoxic regime shifts mediated by microbial community dynamics and redox processes.

Spatial distribution of vanadium and microbial community responses in surface soil of Panzhihua mining and smelting area, China.

Spatial distribution of vanadium in surface soils from different processing stages of vanadium-bearing titanomagnetite in Panzhihua mining and smelting area (China) as well as responses of microbial communities including bacteria and fungi to vanadium were investigated by fieldwork and laboratory incubation experiment. The vanadium contents in this region ranged from 149.3 to 4793.6 mg kg-1, exceeding the soil background value of vanadium in China (82 mg kg-1) largely. High-throughput DNA sequencing results showed bacterial communities from different manufacturing locations were quite diverse, but Bacteroidetes and Proteobacteria were abundant in all samples. The contents of organic matter, available P, available S and vanadium had great influences on the structures of bacterial communities in soils. Bacterial communities converged to similar structure after long-term (240 d) cultivation with vanadium containing medium, dominating by bacteria which can tolerate or reduce toxicities of heavy metals. Fungal diversities decreased after cultivation, but Ascomycota and Ciliophora were still the most abundant phyla as in the original soil samples. Results in this study emphasize the urgency of investigating vanadium contaminations in soils and provide valuable information on how vanadium contamination influences bacterial and fungal communities.

Effects of phenol on physicochemical properties and treatment performances of partial nitrifying granules in sequencing batch reactors.

This study attempts to investigate the effect of phenol on physicochemical properties and treatment performances of partial nitrifying granules (PNGs). Two sequencing batch reactors (SBRs) fed with synthetic ammonium wastewaters were operated in absence (R1) or presence (R2) of phenol. The PNGs in R1 maintained excellent partial nitrification performance and relatively stable physicochemical properties, and exhibited compact and regular shaped structure with a cocci-dominant surface. However, as phenol concentration was stepwise increased from 0 to 300 mg/L in R2, filamentous bacteria appeared and gradually dominated within granules, which in turn resulted in settleability deterioration. Most notably, granules in R2 got easier to agglomerate in the reactor walls and then been washed out with effluent, leading to significant biomass loss, frequent outflow pipe blockage, and eventual system failure. The extracellular polymeric substances (EPS) contents including proteins and polysaccharides in R2 reached 1.8 and 1.7 times of that in R1, respectively, indicating that the presence of phenol played an important role on EPS production. Removal efficiency of ammonium and phenol remained high, but dropped sharply when phenol concentration reached 300 mg/L. Moreover, the failed maintenance of partial nitrification was observed due to the revival of nitrite oxidizing bacteria (NOB) within granules after phenol exposure, which was confirmed by quantitative fluorescence in situ hybridization (FISH) analysis. Overall this study demonstrates that phenol had negative effects on PNGs, and pretreatment to eliminate phenolic substances is recommended when using PNGs for wastewater treatment.

Succession of Bacterial Communities in a Seasonally Stratified Lake with an Anoxic and Sulfidic Hypolimnion.

Although bacteria play key roles in aquatic food webs and biogeochemical cycles, information on the seasonal succession of bacterial communities in lakes is still far from complete. Here, we report results of an integrative study on the successional trajectories of bacterial communities in a seasonally stratified lake with an anoxic hypolimnion. The bacterial community composition of epilimnion, metalimnion, and hypolimnion diverged during summer stratification and converged when the lake was mixed. In contrast, bacterial communities in the sediment remained relatively stable over the year. Phototrophic Cyanobacteria and heterotrophic Actinobacteria, Alphaproteobacteria and Planktomycetes were abundant in the aerobic epilimnion, Gammaproteobacteria (mainly Chromatiaceae) dominated in the metalimnion, and Chlorobi, Betaproteobacteria, Deltaproteobacteria, and Firmicutes were abundant in the anoxic sulfidic hypolimnion. Anoxic but nonsulfidic conditions expanded to the surface layer during fall turnover, when the epilimnion, metalimnion and upper hypolimnion mixed. During this period, phototrophic sulfur bacteria (Chromatiaceae and Chlorobi) disappeared, Polynucleobacter (Betaproteobacteria) and Methylobacter (Gammaproteobacteria) spread out from the former meta- and hypolimnion to the surface layer, and Epsilonproteobacteria dominated in the bottom water layer. Cyanobacteria and Planktomycetes regained dominance in early spring, after the oxygen concentration was restored by winter mixing. In total, these results show large spatio-temporal changes in bacterial community composition, especially during transitions from oxic to anoxic and from sulfidic to nonsulfidic conditions.

Enhanced aerobic nitrifying granulation by static magnetic field.

One of the main challenging issues for aerobic nitrifying granules in treating high strength ammonia wastewater is the long granulation time required for activated sludge to transform into aerobic granules. The present study provides a novel strategy for enhancing aerobic nitrifying granulation by applying an intensity of 48.0mT static magnetic field. The element analysis showed that the applied magnetic field could promote the accumulation of iron compounds in the sludge. And then the aggregation of iron decreased the full granulation time from 41 to 25days by enhancing the setting properties of granules and stimulating the secretion of extracellular polymeric substances (EPS). Long-term, cycle experiments and fluorescence in-situ hybridization (FISH) analysis proved that an intensity of 48.0mT magnetic field could enhance the activities and growth of nitrite-oxidizing bacteria (NOB). These findings suggest that magnetic field is helpful and reliable for accelerating the aerobic nitrifying granulation.

Sorption and biodegradation of tetracycline by nitrifying granules and the toxicity of tetracycline on granules.

This paper examines the simultaneous sorption and biodegradation performance of tetracycline (TC) by the nitrifying granular sludge as well as the short-term exposure toxicity of TC. The removal of TC was characterized by a quick sorption and a slow process of biodegradation. The adsorption process fits pseudo-second-order kinetic model, with a complex mechanism of surface adsorption and intra-particle diffusion. Both temperature and mixed liquor suspended solid (MLSS) influenced TC sorption to the granules. TC biodegradation was enhanced with the increase of COD and NH(4)(+)-N concentrations, with except of the NH(4)(+)-N concentrations higher than 150 mg/L. With the ATU addition, TC degradation was weakened remarkably, indicating a synergistic effect of multiple microbes. Results of the short-term exposure (12h) effects showed that the respirometric activities of the microbes decreased greatly. The addition of TC also decreased the rate of NH(4)(+)-N utilization considerably, with the half saturation constant (K(s)) increasing from 297.7 to 347.2 mg/L.