The polyextremophile Natranaerobius thermophilus adopts a dual adaptive strategy to long-term salinity stress, simultaneously accumulating compatible solutes and K.

The bacterium Natranaerobius thermophilus is an extremely halophilic alkalithermophile that can thrive under conditions of high salinity (3.3-3.9 M Na+), alkaline pH (9.5), and elevated temperature (53°C). To understand the molecular mechanisms of salt adaptation in N. thermophilus, it is essential to investigate the protein, mRNA, and key metabolite levels on a molecular basis. Based on proteome profiling of N. thermophilus under 3.1, 3.7, and 4.3 M Na+ conditions compared to 2.5 M Na+ condition, we discovered that a hybrid strategy, combining the "compatible solute" and "salt-in" mechanisms, was utilized for osmotic adjustment dur ing the long-term salinity adaptation of N. thermophilus. The mRNA level of key proteins and the intracellular content of compatible solutes and K+ support this conclusion. Specifically, N. thermophilus employs the glycine betaine ABC transporters (Opu and ProU families), Na+/solute symporters (SSS family), and glutamate and proline synthesis pathways to adapt to high salinity. The intracellular content of compatible solutes, including glycine betaine, glutamate, and proline, increases with rising salinity levels in N. thermophilus. Additionally, the upregulation of Na+/ K+/ H+ transporters facilitates the maintenance of intracellular K+ concentration, ensuring cellular ion homeostasis under varying salinities. Furthermore, N. thermophilus exhibits cytoplasmic acidification in response to high Na+ concentrations. The median isoelectric points of the upregulated proteins decrease with increasing salinity. Amino acid metabolism, carbohydrate and energy metabolism, membrane transport, and bacterial chemotaxis activities contribute to the adaptability of N. thermophilus under high salt stress. This study provides new data that support further elucidating the complex adaptation mechanisms of N. thermophilus under multiple extremes.IMPORTANCEThis study represents the first report of simultaneous utilization of two salt adaptation mechanisms within the Clostridia class in response to long-term salinity stress.

Characterization of 1,4-dioxane degrading microbial community enriched from uncontaminated soil.

1,4-Dioxane is a contaminant of emerging concern that has been commonly detected in groundwater. In this study, a stable and robust 1,4-dioxane degrading enrichment culture was obtained from uncontaminated soil. The enrichment was capable to metabolically degrade 1,4-dioxane at both high (100 mg L-1) and environmentally relevant concentrations (300 µg L-1), with a maximum specific 1,4-dioxane degradation rate (qmax) of 0.044 ± 0.001 mg dioxane h-1 mg protein-1, and 1,4-dioxane half-velocity constant (Ks) of 25 ± 1.6 mg L-1. The microbial community structure analysis suggested Pseudonocardia species, which utilize the dioxane monooxygenase for metabolic 1,4-dioxane biodegradation, were the main functional species for 1,4-dioxane degradation. The enrichment culture can adapt to both acidic (pH 5.5) and alkaline (pH 8) conditions and can recover degradation from low temperature (10°C) and anoxic (DO < 0.5 mg L-1) conditions. 1,4-Dioxane degradation of the enrichment culture was reversibly inhibited by TCE with concentrations higher than 5 mg L-1 and was completely inhibited by the presence of 1,1-DCE as low as 1 mg L-1. Collectively, these results demonstrated indigenous stable and robust 1,4-dioxane degrading enrichment culture can be obtained from uncontaminated sources and can be a potential candidate for 1,4-dioxane bioaugmentation at environmentally relevant conditions. KEY POINTS: •1,4-Dioxane degrading enrichment was obtained from uncontaminated soil. • The enrichment culture could degrade 1,4-dioxane to below 10 µg L-1. •Low Ks and low cell yield of the enrichment benefit its application in bioremediation.

Effects of Sulfate Reduction on Trichloroethene Dechlorination by Dehalococcoides-Containing Microbial Communities.

In order to elucidate interactions between sulfate reduction and dechlorination, we systematically evaluated the effects of different concentrations of sulfate and sulfide on reductive dechlorination by isolates, constructed consortia, and enrichments containing Dehalococcoides sp. Sulfate (up to 5 mM) did not inhibit the growth or metabolism of pure cultures of the dechlorinator Dehalococcoides mccartyi 195, the sulfate reducer Desulfovibrio vulgaris Hildenborough, or the syntroph Syntrophomonas wolfei In contrast, sulfide at 5 mM exhibited inhibitory effects on growth of the sulfate reducer and the syntroph, as well as on both dechlorination and growth rates of D. mccartyi Transcriptomic analysis of D. mccartyi 195 revealed that genes encoding ATP synthase, biosynthesis, and Hym hydrogenase were downregulated during sulfide inhibition, whereas genes encoding metal-containing enzymes involved in energy metabolism were upregulated even though the activity of those enzymes (hydrogenases) was inhibited. When the electron acceptor (trichloroethene) was limiting and an electron donor (lactate) was provided in excess to cocultures and enrichments, high sulfate concentrations (5 mM) inhibited reductive dechlorination due to the toxicity of generated sulfide. The initial cell ratio of sulfate reducers to D. mccartyi (1:3, 1:1, or 3:1) did not affect the dechlorination performance in the presence of sulfate (2 and 5 mM). In contrast, under electron donor limitation, dechlorination was not affected by sulfate amendments due to low sulfide production, demonstrating that D. mccartyi can function effectively in anaerobic microbial communities containing moderate sulfate concentrations (5 mM), likely due to its ability to outcompete other hydrogen-consuming bacteria and archaea.IMPORTANCE Sulfate is common in subsurface environments and has been reported as a cocontaminant with chlorinated solvents at various concentrations. Inconsistent results for the effects of sulfate inhibition on the performance of dechlorination enrichment cultures have been reported in the literature. These inconsistent findings make it difficult to understand potential mechanisms of sulfate inhibition and complicate the interpretation of bioremediation field data. In order to elucidate interactions between sulfate reduction and reductive dechlorination, this study systematically evaluated the effects of different concentrations of sulfate and sulfide on reductive dechlorination by isolates, constructed consortia, and enrichments containing Dehalococcoides sp. This study provides a more fundamental understanding of the competition mechanisms between reductive dechlorination by Dehalococcoides mccartyi and sulfate reduction during the bioremediation process. It also provides insights on the significance of sulfate concentrations on reductive dechlorination under electron donor/acceptor-limiting conditions during in situ bioremediation applications. For example, at a trichloroethene-contaminated site with a high sulfate concentration, proper slow-releasing electron donors can be selected to generate an electron donor-limiting environment that favors reductive dechlorination and minimizes the sulfide inhibition effect.

Acetylene Fuels TCE Reductive Dechlorination by Defined Dehalococcoides/Pelobacter Consortia.

Acetylene (C2H2) can be generated in contaminated groundwater sites as a consequence of chemical degradation of trichloroethene (TCE) by in situ minerals, and C2H2 is known to inhibit bacterial dechlorination. In this study, we show that while high C2H2 (1.3 mM) concentrations reversibly inhibit reductive dechlorination of TCE by Dehalococcoides mccartyi isolates as well as enrichment cultures containing D. mccartyi sp., low C2H2 (0.4 mM) concentrations do not inhibit growth or metabolism of D. mccartyi. Cocultures of Pelobacter SFB93, a C2H2-fermenting bacterium, with D. mccartyi strain 195 or with D. mccartyi strain BAV1 were actively sustained by providing acetylene as the electron donor and carbon source while TCE or cis-DCE served as the electron acceptor. Inhibition by acetylene of reductive dechlorination and methanogenesis in the enrichment culture ANAS was observed, and the inhibition was removed by adding Pelobacter SFB93 into the consortium. Transcriptomic analysis of D. mccartyi strain 195 showed genes encoding for reductive dehalogenases (e.g., tceA) were not affected during the C2H2-inhibition, while genes encoding for ATP synthase, biosynthesis, and Hym hydrogenase were down-regulated during C2H2 inhibition, consistent with the physiological observation of lower cell yields and reduced dechlorination rates in strain 195. These results will help facilitate the optimization of TCE-bioremediation at contaminated sites containing both TCE and C2H2.

An ignored and potential source of taste and odor (T&O) issues-biofilms in drinking water distribution system (DWDS).

It is important for water utilities to provide esthetically acceptable drinking water to the public, because our consumers always initially judge the quality of the tap water by its color, taste, and odor (T&O). Microorganisms in drinking water contribute largely to T&O production and drinking water distribution systems (DWDS) are known to harbor biofilms and microorganisms in bulk water, even in the presence of a disinfectant. These microbes include T&O-causing bacteria, fungi, and algae, which may lead to unwanted effects on the organoleptic quality of distributed water. Importantly, the understanding of types of these microbes and their T&O compound-producing mechanisms is needed to prevent T&O formation during drinking water distribution. Additionally, new disinfection strategies and operation methods of DWDS are also needed for better control of T&O problems in drinking water. This review covers: (1) the microbial species which can produce T&O compounds in DWDS; (2) typical T&O compounds in DWDS and their formation mechanisms by microorganisms; (3) several common factors in DWDS which can influence the growth and T&O generation of microbes; and (4) several strategies to control biofilm and T&O compound formation in DWDS. At the end of this review, recommendations were given based on the conclusion of this review.

Efficient metabolic exchange and electron transfer within a syntrophic trichloroethene-degrading coculture of Dehalococcoides mccartyi 195 and Syntrophomonas wolfei.

Dehalococcoides mccartyi 195 (strain 195) and Syntrophomonas wolfei were grown in a sustainable syntrophic coculture using butyrate as an electron donor and carbon source and trichloroethene (TCE) as an electron acceptor. The maximum dechlorination rate (9.9 ± 0.1 µmol day(-1)) and cell yield [(1.1 ± 0.3) × 10(8) cells µmol(-1) Cl(-)] of strain 195 maintained in coculture were, respectively, 2.6 and 1.6 times higher than those measured in the pure culture. The strain 195 cell concentration was about 16 times higher than that of S. wolfei in the coculture. Aqueous H2 concentrations ranged from 24 to 180 nM during dechlorination and increased to 350 ± 20 nM when TCE was depleted, resulting in cessation of butyrate fermentation by S. wolfei with a theoretical Gibbs free energy of -13.7 ± 0.2 kJ mol(-1). Carbon monoxide in the coculture was around 0.06 µmol per bottle, which was lower than that observed for strain 195 in isolation. The minimum H2 threshold value for TCE dechlorination by strain 195 in the coculture was 0.6 ± 0.1 nM. Cell aggregates during syntrophic growth were observed by scanning electron microscopy. The interspecies distances to achieve H2 fluxes required to support the measured dechlorination rates were predicted using Fick's law and demonstrated the need for aggregation. Filamentous appendages and extracellular polymeric substance (EPS)-like structures were present in the intercellular spaces. The transcriptome of strain 195 during exponential growth in the coculture indicated increased ATP-binding cassette transporter activities compared to the pure culture, while the membrane-bound energy metabolism related genes were expressed at stable levels.

Response surface methodology for process optimization in livestock wastewater treatment: A review.

With increasing demand for meat and dairy products, the volume of wastewater generated from the livestock industry has become a significant environmental concern. The treatment of livestock wastewater (LWW) is a challenging process that involves removing nutrients, organic matter, pathogens, and other pollutants from livestock manure and urine. In response to this challenge, researchers have developed and investigated different biological, physical, and chemical treatment technologies that perform better upon optimization. Optimization of LWW handling processes can help improve the efficacy and sustainability of treatment systems as well as minimize environmental impacts and associated costs. Response surface methodology (RSM) as an optimization approach can effectively optimize operational parameters that affect process performance. This review article summarizes the main steps of RSM, recent applications of RSM in LWW treatment, highlights the advantages and limitations of this technique, and provides recommendations for future research and practice, including its cost-effectiveness, accuracy, and ability to improve treatment efficiency.

Nitrogen removal mechanisms in biochar-amended sand filters treating onsite wastewater.

The performance of biochar-amended sand filters treating septic tank effluent (STE) was investigated in bench-scale columns. Softwood biochar showed higher NH4 + -N adsorption capacity (1.3 mg N g-1 ), and its water holding capacity (0.57 g ml-1 ) was significantly higher than sand (0.26 g ml-1 ). Two biochar amendment ratios (10% and 30%) were selected for STE treatment in short-term (20 days) and long-term (8 months) studies. During the short-term experiment, the overall total nitrogen removal efficiency was greater in biochar-amended sand columns (94.7%-95.6%) than in 100% sand columns (71.2%) due to the additional NH4 + -N adsorption by biochar. Greater nitrification performance was also observed in biochar-amended columns (87.1%-96.3%) than in 100% sand columns (61.4%) during long-term operation when alkalinity was insufficient. The nitrification performance in biochar-amended columns resumed more quickly (<7 days) after sufficient alkalinity was amended. The density of total biomass and nitrifying bacteria in biochar-amended columns (30%) were significantly higher at all experimental stages, suggesting biochar served as a growth media for enhanced biomass growth. The alkalinity changes and STE composition fluctuation had little impact on the nitrification performance of the 30% biochar-amended sand columns. In addition, biochar surface functional groups and zeta potential changed little after long-term STE filtration. Collectively, the results demonstrated proper biochar amendment ratio (30%) could enhance the nitrification performance of sand filters treating STE by increasing the system hydraulic retention time, providing additional alkalinity for nitrification, and serving as a growth media for enhanced biomass growth.

Exploring the ignored role of escaped algae in a pilot-scale DWDS: Disinfectant consumption, DBP yield and risk formation.

Insufficient treatments during bloom-forming seasons allow algae to enter the subsequent drinking water distribution system (DWDS). Yet, scarce information is available regarding the role escaped algae to play in the DWDS, and how they interact with the system. Thus, three scenarios were conducted: a pilot DWDS with algae (a), pipe water (b), and pipe water with algae (c). Experimental results showed that, compared to biofilm and bulk water, escaped algae required fewer disinfectants. Competition for disinfectants varied with algal strains (Microcystis aeruginosa, MA; Pseudanabaena sp., PS) and disinfectant types (chlorine, Cl2; chloriamine, NH2Cl). Algae in the MA-Cl2 group showed the highest demand (6.25%-36.02%). However, the low-concentration disinfectants distributed to algae could trigger distinct algal status alternations. Cl2 diffused into intact MA cells and reacted with intracellular compositions. Damaged PS cells reached 100% within 2 h. Typical disinfection byproducts (DBPs), including trihalomethanes (THMs), haloacetic acids and halogenated acetonitriles were examined. Disinfectant types and algal strains affected DBP yield and distribution. Although disinfectants consumed by algae might not promote dissolved DBP formation, especially for THMs. DBP formation of the other components was affected by escaped algae via changing disinfectant assignment (reduced by 45.45% for MA-Cl2) and transformation efficiency (by 34.52%). The cytotoxicity risks were estimated. Dissolved DBP-induced risks were not added when escaped algae occurred, whereas disruption and release of intracellular substances increased risks; the maximum cytotoxicity did not occur at 12 h rather than at the end (24 h). Overall, this study provided an innovative perspective on algal-related water quality issues in water systems.

1,4-Dioxane removal in nitrifying sand filters treating domestic wastewater: Influence of water matrix and microbial inhibitors.

1,4-Dioxane is a recalcitrant pollutant in water and is ineffectively removed during conventional water and wastewater treatment processes. In this study, we demonstrate the application of nitrifying sand filters to remove 1,4-dioxane from domestic wastewater without the need for bioaugmentation or biostimulation. The sand columns were able to remove 61 ± 10% of 1,4-dioxane on average (initial concentration: 50 µg/L) from wastewater, outperforming conventional wastewater treatment approaches. Microbial analysis revealed the presence of 1,4-dioxane degrading functional genes (dxmB, phe, mmox, and prmA) to support biodegradation being the dominant degradation pathway. Adding antibiotics (sulfamethoxazole and ciprofloxacin), that temporarily inhibited the nitrification process during the dosing period, showed a minor effect in 1,4-dioxane removal (6-8% decline, p < 0.05), suggesting solid resilience of the 1,4-dioxane-degrading microbial community in the columns. Columns amended with sodium azide significantly (p < 0.05) depressed 1,4-dioxane removal in the early stage of dosing but followed by a gradual increase of the removal over time to >80%, presumably due to a shift in the microbial community toward azide-resistant 1,4-dioxane degrading microbes (e.g., fungi). This study demonstrated for the first time the resilience of the 1,4-dioxane-degrading microorganisms during antibiotic shocks, and the selective enrichment of efficient 1,4-dioxane-degrading microbes after azide poisoning. Our observation could provide insights into designing better 1,4-dioxane remediation strategies in the future.

Halomonas xianhensis sp. nov., a moderately halophilic bacterium isolated from a saline soil contaminated with crude oil.

A Gram-negative, aerobic, short rod-shaped and non-motile bacterium, strain A-1(T), was isolated from a saline soil contaminated with crude oil in Xianhe, Shangdong Province, China. Strain A-1(T) formed yellow colonies, was moderately halophilic and grew with 0.05-27.5% (w/v) total salts (optimum 5-8%), at 10-42 °C (optimum 30 °C) and at pH 5.5-9.0 (optimum pH 7.2). The dominant fatty acids (>5%) were C(16:0), summed feature 3 (comprising C(16:1)ω7c and/or iso-C(15:0) 2-OH), C(18:1)ω7c, C(19:0) cyclo ω8c and C(12:0) 3-OH and the predominant ubiquinone was Q-9. The genomic DNA G+C content was 67.1 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain A-1(T) belonged to the genus Halomonas in the class Gammaproteobacteria. The closest relatives were Halomonas lutea YIM 91125(T) (97.7% 16S rRNA gene sequence similarity), H. muralis LMG 20969(T) (95.6%), H. pantelleriensis AAP(T) (95.5%) and H. kribbensis BH843(T) (95.2%). DNA-DNA relatedness between strain A-1(T) and H. lutea CCTCC AB 206093(T) was 27±3%. On the basis of phenotypic, chemotaxonomic and phylogenetic features, strain A-1(T) should be placed in the genus Halomonas as a representative of a novel species. The name Halomonas xianhensis sp. nov. is proposed, with strain A-1(T) (=CGMCC 1.6848(T) =JCM 14849(T)) as the type strain.

Second-order chlorine decay and trihalomethanes formation in a pilot-scale water distribution systems.

It is well known that model-building of chlorine decay in real water distribution systems is difficult because chlorine decay is influenced by many factors (e.g., bulk water demand, pipe-wall demand, piping material, flow velocity, and residence time). In this paper, experiments were run to investigate the kinetic model of chlorine decay and the formation model of trihalomethanes (THMs) in pilot-scale water distribution systems. Experimental results show that the rate constants of chlorine decay, including wall decay and bulk decay, increasing with temperature. Moreover, the kinetic model of chlorine decay and the formation model of THMs describe experiment data of pilot-scale water distribution systems. The effect of different piping material on chlorine decay and THMs formation were also investigated. The rate constants of chlorine decay are ranked in order: stainless steel pipe, ductile iron pipe, and last, polyethelene pipe because wall decay is the largest in stainless steel pipe than that in other piping material. Correspondingly, the rate of THMs formation follows the order of stainless steel pipe, ductile iron pipe, and last, polyethelene pipe because of less chlorine in bulk water reacting with the trihalomethane formation potential (THMFP).

Biochar application in biofiltration systems to remove nutrients, pathogens, and pharmaceutical and personal care products from wastewater.

Although conventional on-site wastewater treatment systems (OWTSs) provide only primary treatment of domestic wastewater, removal of a limited level of nutrients (N, P), pathogens, and pharmaceuticals and personal care products (PPCPs) could be achieved by such a treatment process. Biochar has the capacity to remove various contaminants and has been widely used as an ideal soil amendment in agriculture due to its persistence, superior nutrient-retention properties, low cost, and ready availability. However, few applications on the use of biochar in onsite wastewater treatment have been explored. In this review, we systematically reviewed the applications of biochar in filtration-based OWTSs for nutrient (N, P) removal and recovery as well as pathogen and PPCP removal. Although adsorption was the main mechanism for P, pathogen, and PPCP removal, biochar can also serve as the growth media for enhanced biological degradation, improves available alkalinity, and increases water holding capacity in the OWTSs. The biochar source, surface modification methods, and preparation procedures (e.g., pyrolysis temperature change) have significant effects on contaminant removal performance in biochar-amended OWTSs. Specifically, contradictory results have been reported on the effect of pyrolysis temperature change on biochar removal performance (i.e., increased, decreased, or no change) of N, P, and PPCPs. Wastewater composition and environmental pH also play important roles in the removal of nutrients, pathogens, and PPCPs. Overall, biochar holds great potential to serve as an alternative filtration material or to be amended to the current OWTS to improve system performance in removing a variety of contaminants at low cost.

Efficient nitrogen removal from onsite wastewater by a novel continuous flow biofilter.

Soil-based passive biofiltration system is an economically feasible technology for nitrogen removal from onsite wastewater. However, the conventional design requires a large system footprint with limited treatment capacity. In this study, a novel continuous flow biofilter (CFB) with adjustable recirculation and continuous flow pattern was developed for onsite wastewater treatment with a small footprint. Efficient total nitrogen removal (80.1-97.5%) was observed at various hydraulic loadings (0.03-0.12 m3 m-2 d-1), nitrogen loadings (1.1-8.6 g N m-2 d-1) and recycle ratios (2-3) when treating septic tank effluent (STE), with low effluent TN (0.7-13.6 mg N L-1). Nitrous oxide was observed in the denitrification effluent indicating incomplete denitrification at elevated dissolved oxygen levels (3.3-5.8 mg L-1). Nitrogen removal rate (2.9-7.0 g N m-2 d-1) and ammonium removal rate (2.4-7.2 g N m-2 d-1) were positively correlated with nitrogen loadings increase (1.1-8.6 g N m-2 d-1) but were not significantly impacted by the hydraulic loading rate change (0.08-0.12 m3 m-2 d-1). The total biomass abundance and nitrifying microorganisms decreased significantly as the nitrification columns depth increased, while homogeneous microbial distribution was observed in the denitrification columns. The abundance of ammonium oxidizing archaea (AOA) increased significantly at increased hydraulic and nitrogen loading rate, while the ammonium oxidizing bacteria (AOB) abundance remained steady. The abundance of functional genes involved in denitrification process (nirS, nirK and nosZ) responded differently when hydraulic and nitrogen loading rate changes. Collectively, this study suggested the CFB could efficiently remove nitrogen from onsite wastewater with fluctuating influent compositions and various hydraulic loadings.

Removal of 1,4-dioxane during on-site wastewater treatment using nitrogen removing biofilters.

The presence and release of 1,4-dioxane to groundwater from onsite-wastewater treatment systems (OWTS), which represent 25% of the total wastewater treatment in the U.S., has not been studied to date. In this study we monitored 1,4-dioxane in six septic tank effluents (STE) and receiving OWTS installed at residences on Long Island (LI), NY, for a period of 15 months. We specifically evaluated the performance of Nitrogen Removing Biofilters (NRBs) as an innovative/alternative-OWTS, consisting of a top sand layer and a bottom woodchip/sand layer, to simultaneously remove nitrogen and 1,4-dioxane. 1,4-Dioxane levels in STE (mean: 1.49 µg L-1; range: 0.07-8.45 µg L-1; n = 37) were on average > 15 times higher than tap water from these residences, demonstrating that 1,4-dioxane primarily originated from the use of household products. NRBs were effective in removing both 1,4-dioxane and total nitrogen with an overall removal efficiency of 56 ± 20% and 88 ± 12%, respectively. The majority of 1,4-dioxane removal (~80%) occurred in the top oxic layer of the NRBs. The detection of functional genes (dxmB, prmA, and thmA), which encode for metabolic and co-metabolic 1,4-dioxane degradation, in NRBs provides the first field evidence of aerobic microbial degradation of 1,4-dioxane occurring in a wastewater system. Given that there are ~500,000 conventional OWTS on LI, the 1,4-dioxane discharge to groundwater from residential wastewater was estimated at 195 ± 205 kg yr -1, suggesting high risk of contamination to shallow aquifers. The results also demonstrate that installation of NRBs can reduce 1,4-dioxane to levels even lower than the NY State drinking water standard of 1 µg L-1.

Potential release of legacy nitrogen from soil surrounding onsite wastewater leaching pools.

This study examined whether the accumulation of nitrogen (legacy nitrogen) within and surrounding leaching pools for onsite wastewater treatment may act as a source of nitrogen contamination to groundwater upon changes to the quantity and/or composition of the influent to the pool. In this study, one concrete leaching pool with neutral pH (A, pH 6.9) and one leaching pool after acid washing (B, pH 3.7) were selected to examine the quantity and composition of legacy nitrogen in the surrounding soil, as well as evaluate the potential release of this nitrogen under two environmentally relevant leaching scenarios: (i) the concrete leaching pool serves as the final discharge unit for aerobic treatment unit (ATU) effluent; (ii) extreme weather events (flash flood/heavy rains) act to increase the quantity and dilute the composition of flow to the pool. Core sample analysis showed that organic nitrogen accounts for the majority (97.3-99.7%) of the total nitrogen (TN) at site A (4.1 ±â€¯0.6 mg N/g soil) and site B (3.0 ±â€¯0.4 mg N/g soil); while ammonium was the major form of inorganic nitrogen present at the sites. The TN accumulated under the two leaching pools was equivalent to approximately 17-39 days of nitrogen loading to the system. pH had a significant impact on the mass of TN leached from the soil, while no significant difference in leached TN was observed for the two leaching scenarios. The amount of TN leached from the soil matrix was not affected by the flow rate (18.6 mL/d in scenario i vs. 547.2 mL/d in scenario ii) or flow pattern (intermittent dosing vs. continuous flow). The quantity of TN leached from soils in both scenario (i) and (ii) was low and accounted for 2.6-8.9% of the total nitrogen in the soil.

Biodegradation of phenanthrene by a halophilic bacterial consortium under aerobic conditions.

A halophilic bacterial consortium that degraded phenanthrene was developed from oil-contaminated saline soil containing 10% salinity. The biodegradation of phenanthrene occurred at 5%, 10%, and 15% salinity, whereas no biodegradation took place at 0.1% and 20% salinity. A 16S rRNA gene analysis showed that all sequences from the denaturing gradient gel electrophoresis profile were similar to those of halophilic bacteria. This is the first report of a halophilic bacterial consortium capable of degrading phenanthrene under hypersaline conditions.

Structural dynamics and transcriptomic analysis of Dehalococcoides mccartyi within a TCE-Dechlorinating community in a completely mixed flow reactor.

A trichloroethene (TCE)-dechlorinating community (CANAS) maintained in a completely mixed flow reactor was established from a semi-batch enrichment culture (ANAS) and was monitored for 400 days at a low solids retention time (SRT) under electron acceptor limitation. Around 85% of TCE supplied to CANAS (0.13 mmol d-1) was converted to ethene at a rate of 0.1 mmol d-1, with detection of low production rates of vinyl chloride (6.8 × 10-3 mmol d-1) and cis-dichloroethene (2.3 × 10-3 mmol d-1). Two distinct Dehalococcoides mccartyi strains (ANAS1 and ANAS2) were stably maintained at 6.2 ±â€¯2.8 × 108 cells mL-1 and 5.8 ±â€¯1.2 × 108 cells mL-1, respectively. Electron balance analysis showed 107% electron recovery, in which 6.1% were involved in dechlorination. 16 S rRNA amplicon sequencing revealed a structural regime shift between ANAS and CANAS while maintaining robust TCE dechlorination due to similar relative abundances of D. mccartyi and functional redundancy among each functional guild supporting D. mccartyi activity. D. mccartyi transcriptomic analysis identified the genes encoding for ribosomal RNA and the reductive dehalogenases tceA and vcrA as the most expressed genes in CANAS, while hup and vhu were the most critical hydrogenases utilized by D. mccartyi in the community.

Membrane Complexes of Syntrophomonas wolfei Involved in Syntrophic Butyrate Degradation and Hydrogen Formation.

Syntrophic butyrate metabolism involves the thermodynamically unfavorable production of hydrogen and/or formate from the high potential electron donor, butyryl-CoA. Such redox reactions can occur only with energy input by a process called reverse electron transfer. Previous studies have demonstrated that hydrogen production from butyrate requires the presence of a proton gradient, but the biochemical machinery involved has not been clearly elucidated. In this study, the gene and enzyme systems involved in reverse electron transfer by Syntrophomonas wolfei were investigated using proteomic and gene expression approaches. S. wolfei was grown in co-culture with Methanospirillum hungatei or Dehalococcoides mccartyi under conditions requiring reverse electron transfer and compared to both axenic S. wolfei cultures and co-cultures grown in conditions that do not require reverse electron transfer. Blue native gel analysis of membranes solubilized from syntrophically grown cells revealed the presence of a membrane-bound hydrogenase, Hyd2, which exhibited hydrogenase activity during in gel assays. Bands containing a putative iron-sulfur (FeS) oxidoreductase were detected in membranes of crotonate-grown and butyrate grown S. wolfei cells. The genes for the corresponding hydrogenase subunits, hyd2ABC, were differentially expressed at higher levels during syntrophic butyrate growth when compared to growth on crotonate. The expression of the FeS oxidoreductase gene increased when S. wolfei was grown with M. hungatei. Additional membrane-associated proteins detected included FoF1 ATP synthase subunits and several membrane transporters that may aid syntrophic growth. Furthermore, syntrophic butyrate metabolism can proceed exclusively by interspecies hydrogen transfer, as demonstrated by growth with D. mccartyi, which is unable to use formate. These results argue for the importance of Hyd2 and FeS oxidoreductase in reverse electron transfer during syntrophic butyrate degradation.

Transformation of bisphenol A in water distribution systems: a pilot-scale study.

Halogenations of bisphenol A (BPA) in a pilot-scale water distribution system (WDS) of a cement-lined ductile cast iron pipe were investigated. The water in the pilot-scale WDS was chlorinated with a free chlorine concentration of 0.7 mg L(-1) using sodium hypochlorite, and with an initial BPA concentration of 100 µg L(-1) was spiked in the WDS. Halogenated compounds in the BPA experiments were identified using EI/GC/MS and GC. Several BPA congeners, including 2-chlorobisphenol A (MCBPA), dichlorobisphenol A (D2-CBPA), 2,2',6-trichlorobisphenol A (T3CBPA), 2,2',6,6'-tetrachlorobisphenol A (T4CBPA), 2-bromobisphenol A (MBBPA), and bromochlorobisphenol A (MBMCBPA) were found. Moreover, further halogenation yielded other reaction intermediates, including 2,4,6-trichlorophenol (T3CP), dichlorobisphenol A, bromodichlorophenol, and dibromochlorophenol. After halogenation for 120min, most of the abovementioned reaction intermediates disappeared and were replaced by trihalomethanes (THMs). Based on these experimental findings, the halogenation process of BPA oxidation in a WDS includes three stages: (1) halogenation on the aromatic ring; (2) chlorine or bromine substitution followed by cleavage of the α-C bond on the isopropyl moiety with a positive partial charge and a ß'-C bond on the benzene moiety with a negative partial charge; and (3) THMs and a minor HAA formation from phenolic intermediates through the benzene ring opening with a chlorine and bromine substitution of the hydrogen on the carbon atoms. The oxidation mechanisms of the entire transformation from BPA to THM/HAA in the WDS were proposed.