Bidirectional Enhancement of Nitrogen Removal by Indigenous Synergetic Microalgal-Bacterial Consortia in Harsh Low-C/N Wastewater.

Conventional microalgal-bacterial consortia have limited capacity to treat low-C/N wastewater due to carbon limitation and single nitrogen (N) removal mode. In this work, indigenous synergetic microalgal-bacterial consortia with high N removal performance and bidirectional interaction were successful in treating rare earth tailing wastewaters with low-C/N. Ammonia removal reached 0.89 mg N L-1 h-1, 1.84-fold more efficient than a common microalgal-bacterial system. Metagenomics-based metabolic reconstruction revealed bidirectional microalgal-bacterial interactions. The presence of microalgae increased the abundance of bacterial N-related genes by 1.5- to 57-fold. Similarly, the presence of bacteria increased the abundance of microalgal N assimilation by 2.5- to 15.8-fold. Furthermore, nine bacterial species were isolated, and the bidirectional promotion of N removal by the microalgal-bacterial system was verified. The mechanism of microalgal N assimilation enhanced by indole-3-acetic acid was revealed. In addition, the bidirectional mode of the system ensured the scavenging of toxic byproducts from nitrate metabolism to maintain the stability of the system. Collectively, the bidirectional enhancement system of synergetic microalgae-bacteria was established as an effective N removal strategy to broaden the stable application of this system for the effective treatment of low C/N ratio wastewater.

Resourceful application and mechanism of oyster shell-microalgae synergistic system：Sustainable treatment of harsh low carbon nitrogen ratio actual wastewater.

Microalgal technology holds great promise for both low C/N wastewater treatment and resource recovery simultaneously. Nevertheless, the advancement of microalgal technology is hindered by its reduced nitrogen removal efficiency in low C/N ratio wastewater. In this work, microalgae and waste oyster shells were combined to achieve a total inorganic nitrogen removal efficiency of 93.85% at a rate of 2.05 mg L-1 h-1 in low C/N wastewater. Notably, over four cycles of oyster shell reuse, the reactor achieved an average 85% ammonia nitrogen removal extent, with a wastewater treatment cost of only $0.092/ton. Moreover, microbial community analysis during the reuse of oyster shells revealed the critical importance of timely replacement in inhibiting the growth of non-functional bacteria (Poterioochromonas_malhamensi). The work demonstrated that the oyster shell - microalgae system provides a time- and cost-saving, environmental approach for the resourceful treatment of harsh low C/N wastewater.

Microalgae-bacteria tandem-type ponds simultaneously removing ammonia nitrogen and phosphorus towards municipal wastewater advanced treatment.

Low C/N municipal wastewater is difficult to be treated effectively via traditional biological methods, leading to concentrations of pollutants in effluent far exceeding increasingly strict standards. In this work, we propose a novel microalgae-bacteria tandem-type process to simultaneously remove ammonia nitrogen (NH4+-N) and phosphorus (P) from municipal wastewater. A 4.5 L microalgae-bacteria tandem-type reactor was constructed and operated stably for 40 days. The removal efficiencies of NH4+-N and P reached 97.5% and 92.9%, respectively, effluent concentrations were 0.53 and 0.17 mg/L on average, which met the Environmental quality standards for surface water in China (GB 3838-2002). Remarkably, microalgae ponds accounted for 69.3% and 76.3% of the overall NH4+-N and P removal via microalgae assimilation. Furthermore, 16 S rRNA gene amplicon sequencing revealed the abundance of bacteria changed, suggesting that the presence of microalgae leads to some species extinction and low-abundance bacteria increase. This work demonstrated that the microalgae-bacteria tandem-type processes can be efficient and widely applied in the advanced treatment of municipal wastewater.

Biotransformation of 4-Hydroxybenzoic Acid under Nitrate-Reducing Conditions in a MEC Bioanode.

4-Hydroxybenzoic acid (HBA) is commonly found at high concentrations in waste streams generated by the thermochemical conversion of lignocellulosic biomass to bio-oils and biofuels. The objective of this study was to systematically assess the biotransformation of HBA in the bioanode of a microbial electrolysis cell (MEC) for the production of renewable cathodic H2. A mixed, denitrifying culture, enriched with HBA as the sole electron donor, was used as the anode inoculum. MEC electrochemical performance, H2 yield, HBA biotransformation pathways and products, and the bioanode suspended and biofilm microbial communities were examined. In the absence of nitrate, 60%-100% HBA was converted to phenol, which persisted, resulting in very limited exoelectrogenesis. Under nitrate-reducing conditions, complete HBA degradation was achieved in the MEC bioanode with very low phenol production, resulting in the production of cathodic H2. The predominant bacterial genus in the MEC bioanode (relative abundance 33.4%-41.9%) was the denitrifier Magnetospirillum, which uses the benzoyl-CoA pathway to degrade aromatic compounds. Geobacter accounted for 5.9-7.8% of the MEC bioanode community. Thus, active nitrate reduction in the MEC bioanode led to complete HBA degradation, resulting in a higher extent of exoelectrogenesis and cathodic H2 production. The results of this study provide mechanistic insights into a productive use of HBA and other phenolic compounds typically found in waste streams resulting from the thermochemical conversion of lignocellulosic biomass to biofuels.

Transformation and Mobility of Cu, Zn, and Cr in Sewage Sludge during Anaerobic Digestion with Pre- or Interstage Hydrothermal Treatment.

Anaerobic digestion (AD) combined with hydrothermal treatment (HT) is an attractive technology for sewage sludge treatment and resource recovery. The fate and distribution of heavy metals in the sludge during combined HT/AD significantly affect the sludge final disposal/utilization options, yet such information is still lacking. This study systematically characterizes the transformation of important heavy metals Cu, Zn, and Cr in sewage sludge during AD with pre- or interstage HT (i.e., HT-AD or AD-HT-AD, respectively). Complementary sequential chemical extraction and X-ray absorption spectroscopy were used to characterize the speciation and mobility of metals. For the HT-AD system, both Cu and Zn predominantly occur as sulfides in HT hydrochars. Subsequent AD favors the formation of Cu2S and partial transformation of nano-ZnS to adsorbed and organo-complexed Zn species. HT favors the formation of Cr-bearing silicates in hydrochars, whereas Fe(III)-Cr(III)-hydroxide and Cr(III)-humic complex are the predominant Cr species in AD solids. Similar reaction pathways occur in the AD-HT-AD system with some minor differences in metal species and contents, as the first-stage AD changed the sludge matrix. These findings have important implications for understanding the fate and mobility of heavy metals in sludge-derived hydrochars and AD solids.

New insights in correlating greenhouse gas emissions and microbial carbon and nitrogen transformations in wetland sediments based on genomic and functional analysis.

Greenhouse gas (GHG) emissions from constructed wetlands (CWs) lower the environmental and ecological benefits of CWs and thus have raised increasing environmental concern. To prevent GHGs emissions, it is important to assess and quantify the correlation of GHGs emission and microbial carbon and nitrogen transformations. In this study, two typical wetland substrate samples (mud sampled from Xiaomei River CW and sand sampled from Dongwen River CW) were used to build lab-scale vertical subsurface flow CW microcosms, labeled as XRCW and DRCW, respectively. The mean COD removal rate of the DRCW group (76.1%) was higher than that of XRCW group (60.6%). Both groups achieved a high extent of nitrogen nutrient removal, indicating a higher metabolic activity of nitrifying and denitrifying microorganisms in the system, especially in XRCW. The mean emission fluxes of N2O, CH4 and CO2 in the XRCW group were 52.7 µg/m2-h, 1.6 mg/m2-h and 100.4 mg/m2-h, which were higher than that in the DRCW group (30.0 µg/m2-h, 1.0 mg/m2-h and 28.0 mg/m2-h, respectively). The relation of GHG emissions to microbial carbon and nitrogen transformation was assessed by genomics and functional analysis. The release of GHGs by the XRCW group had a positive correlation with the relative abundance of Proteobacteria, while for the DRCW group a positive correlation was found with the relative abundance of Cyanobacteria. Nitrogen fixation by Cyanobacteria could be an approach to reduce GHG emissions. The release of CH4 and CO2 was positively correlated with glucose metabolism. N2O gas emission was affected by the species of denitrifiers. This study is of great importance to clarify the emissions of GHGs in vertical subsurface flow CWs, as it is relating to microbial carbon and nitrogen transformation. The connection is of great significance to control the emission of GHGs in wetlands.

Comparison of Carbon Dioxide with Anaerobic Digester Biogas as a Methanogenic Biocathode Feedstock.

BES biogas upgrading studies have typically used bicarbonate or commercial gas mixtures as a biocathode substrate instead of anaerobic digester biogas. Therefore, the objective of this study was to (i) compare the performance of a methanogenic BES between CO2-fed and biogas-fed cycles; (ii) investigate possible factors that may account for observed performance differences; and (iii) assess the performance of a biogas-fed biocathode at various applied cathode potentials. The maximum 1-d CH4 production rate in a biogas-fed biocathode (3003 mmol/m2-d) was 350% higher than in a CO2-fed biocathode (666 mmol/m2-d), and the biogas-fed biocathode was capable of maintaining high performance despite a variable biogas feed composition. Anode oxidation of reduced gases (e.g., CH4 and H2S) from biogas may theoretically contribute 4% to 35% of the total charge transfer from anode to cathode at applied cathode potentials of -0.80 to -0.55 V (vs SHE). The introduction of biogas did not significantly change the biocathode archaeal community (dominated by a Methanobrevibacter sp. phylotype), but the bacterial community shifted away from Bacteroidetes and toward Proteobacteria, which may have contributed to the improved performance of the biogas-fed system. This study shows that anaerobic digester biogas is a promising biocathode feedstock for BES biogas upgrading.

Energy and Nutrient Recovery from Sewage Sludge and Manure via Anaerobic Digestion with Hydrothermal Pretreatment.

Global expectation for sustainability has prompted the transition of practices in wastewater treatment plants toward not only waste management but also energy and nutrient recovery. It has been shown that low-temperature hydrotherm (HT) treatment can enhance downstream biogas production via anaerobic digestion (AD). Yet, because the application of combined HT and AD is still at an early stage, a systematic understanding of the dynamic speciation evolution of important elements is still lacking. This study investigates energy and nutrient recovery from sewage sludge and swine manure via combined HT-AD treatment. Bench-scale investigation was conducted to evaluate biogas production and understand the dynamic evolution of organic carbon (C) and phosphorus (P) speciation. C and P speciations were characterized using complementary chemical and spectroscopic techniques, including 13C nuclear magnetic resonance (NMR) spectroscopy, P X-ray absorption near edge structure (XANES) spectroscopy, and sequential chemical extraction. Results from this study suggest that low-temperature HT pretreatment can achieve enhanced biogas production for sludge compared to the minimal effect on the biogas production from manure. It also provides guidance for P recovery from liquid digestate and solid residue after the AD process.

Coevolution of Iron, Phosphorus, and Sulfur Speciation during Anaerobic Digestion with Hydrothermal Pretreatment of Sewage Sludge.

Anaerobic digestion (AD) with hydrothermal (HT) pretreatment is an emerging technology for enhanced resource recovery from sewage sludge. This study investigates the speciation of Fe, P, and S during sequential HT-AD treatment of sewage sludge using sequential chemical extraction, X-ray diffraction, and X-ray absorption spectroscopy. Results suggest strong correlations between Fe and P species as well as Fe and S species, affecting the solubility and bioavailability of each other. For instance, much vivianite formed in the hydrochars after HT treatment at low temperature, while more strengite precipitated at higher HT temperature. During the subsequent AD process, microbial reduction of strengite and other Fe(III) species led to the formation of more vivianite, with concurrent P release into the solution and adsorption onto other minerals. HT pretreatment of sewage sludge had a weak effect on the sulfidation of Fe during the AD process. This work has important implications for understanding the nutrient speciation and availability in sludge-derived hydrochars and AD solids. It also provides fundamental knowledge for the selection and optimization of HT pretreatment conditions for enhanced resource recovery through sequential HT-AD process.

Widely Used Benzalkonium Chloride Disinfectants Can Promote Antibiotic Resistance.

While the misuse of antibiotics has clearly contributed to the emergence and proliferation of resistant bacterial pathogens, with major health consequences, it remains less clear if the widespread use of disinfectants, such as benzalkonium chlorides (BAC), a different class of biocides than antibiotics, has contributed to this problem. Here, we provide evidence that exposure to BAC coselects for antibiotic-resistant bacteria and describe the underlying genetic mechanisms. After inoculation with river sediment, BAC-fed bioreactors selected for several bacterial taxa, including the opportunistic pathogen Pseudomonas aeruginosa, that were more resistant to several antibiotics than their counterparts in a control (no BAC) bioreactor. A metagenomic analysis of the bioreactor microbial communities, confirmed by gene cloning experiments with the derived isolates, suggested that integrative and conjugative elements encoding a BAC efflux pump together with antibiotic resistance genes were responsible for these results. Furthermore, the exposure of the P. aeruginosa isolates to increasing concentrations of BAC selected for mutations in pmrB (polymyxin resistance) and physiological adaptations that contributed to a higher tolerance to polymyxin B and other antibiotics. The physiological adaptations included the overexpression of mexCD-oprJ multidrug efflux pump genes when BAC was added in the growth medium at subinhibitory concentrations. Collectively, our results demonstrated that disinfectants promote antibiotic resistance via several mechanisms and highlight the need to remediate (degrade) disinfectants in nontarget environments to further restrain the spread of antibiotic-resistant bacteria.IMPORTANCE Benzalkonium chlorides (BAC) are biocides broadly used in disinfectant solutions. Disinfectants are widely used in food processing lines, domestic households, and pharmaceutical products and are typically designed to have a different mode of action than antibiotics to avoid interfering with the use of the latter. Whether exposure to BAC makes bacteria more resistant to antibiotics remains an unresolved issue of obvious practical consequences for public health. Using an integrated approach that combines metagenomics of natural microbial communities with gene cloning experiments with isolates and experimental evolution assays, we show that the widely used benzalkonium chloride disinfectants promote clinically relevant antibiotic resistance. Therefore, more attention should be given to the usage of these disinfectants, and their fate in nontarget environments should be monitored more tightly.

Genomic and Transcriptomic Insights into How Bacteria Withstand High Concentrations of Benzalkonium Chloride Biocides.

Benzalkonium chlorides (BAC) are commonly used biocides in broad-spectrum disinfectant solutions. How microorganisms cope with BAC exposure remains poorly understood, despite its importance for disinfection and disinfectant-induced antibiotic resistance. To provide insights into these issues, we exposed two isolates of an opportunistic pathogen, Pseudomonas aeruginosa, to increasing concentrations of BAC. One isolate was preadapted to BAC, as it originated from a bioreactor fed with subinhibitory concentrations of BAC for 3 years, while the other originated from a bioreactor that received no BAC. Replicated populations of both isolates were able to survive high concentrations of BAC, up to 1,200 and 1,600 mg/liter for the non- and preadapted strains, respectively, exceeding typical application doses. Transcriptome sequencing (RNA-seq) analysis revealed upregulation of efflux pump genes and decreased expression of porins related to BAC transport as well as reduced growth rate. Increased expression of spermidine (a polycation) synthase genes and mutations in the pmrB (polymyxin resistance) gene, which cause a reduction in membrane negative charge, suggested that a major adaptation to exposure to the cationic surfactant BAC was to actively stabilize cell surface charge. Collectively, these results revealed that P. aeruginosa adapts to BAC exposure by a combination of mechanisms and provided genetic markers to monitor BAC-resistant organisms that may have applications in the practice of disinfection.IMPORTANCE BAC are widely used as biocides in disinfectant solutions, food-processing lines, domestic households, and health care facilities. Due to their wide use and mode of action, there has been rising concern that BAC may promote antibiotic resistance. Consistent with this idea, at least 40 outbreaks have been attributed to infection by disinfectant- and antibiotic-resistant pathogens such as P. aeruginosa However, the underlying molecular mechanisms that bacteria use to deal with BAC exposure remain poorly elucidated. Elucidating these mechanisms may be important for monitoring and limiting the spread of disinfectant-resistant pathogens. Using an integrated approach that combined genomics and transcriptomics with physiological characterization of BAC-adapted isolates, this study provided a comprehensive understanding of the BAC resistance mechanisms in P. aeruginosa Our findings also revealed potential genetic markers to detect and monitor the abundance of BAC-resistant pathogens across clinical or environmental settings. This work contributes new knowledge about high concentrations of benzalkonium chlorides disinfectants-resistance mechanisms at the whole-cell genomic and transcriptomic level.

Evaluation of gas and carbon transport in a methanogenic bioelectrochemical system (BES).

Bioelectrochemical systems (BESs) may be used to upgrade anaerobic digester biogas by directly converting CO2 to CH4 . The objective of this study was to evaluate gas (N2 , CO2 , CH4 , and H2 ) and carbon transport within a methanogenic BES. Four BES configurations were evaluated: abiotic anode with abiotic cathode (AAn-ACa), bioanode with abiotic cathode (BAn-ACa), abiotic anode with biocathode (AAn-BCa), and bioanode with biocathode (BAn-BCa). Transport of N2 , a gas commonly used for flushing anoxic systems, out of the anode headspace ranged from 3.7 to 6.2 L/d-atm-m2 , normalized to the proton exchange membrane (PEM) surface area and net driving pressure (NDP). CO2 was transported from the cathode to the anode headspace at rates from 3.7 to 5.4 L/d-atm-m2 . The flux of H2 from cathode to anode headspace was 48% greater when the system had a biocathode (AAn-BCa) than when H2 was produced at an abiotic cathode (BAn-ACa), even though the abiotic cathode headspace had 75% more H2 than the AAn-BCa biocathode at the end of 1 day. A 7-day carbon balance of a batch-fed BAn-BCa BES showed transient microbial carbon storage and a net transport of carbon from anode to cathode. After a 7-day batch incubation, the CH4 production in the biocathode was 27% greater on a molar basis than the initial CO2 supplied to the biocathode headspace, indicating conversion of CO2 produced in the anode. This research expands the current understanding of methanogenic BES operation, which may be used in improving the assessment of BES performance and/or in the development of alternative BES designs and mathematical models. Biotechnol. Bioeng. 2017;114: 961-969. © 2016 Wiley Periodicals, Inc.

Zero-Valent Iron Enhances Biocathodic Carbon Dioxide Reduction to Methane.

Methanogenic bioelectrochemical systems (BESs), which convert carbon dioxide (CO2) directly to methane (CH4), promise to be an innovative technology for anaerobic digester biogas upgrading. Zero-valent iron (ZVI), which has previously been used to improve CH4 production in anaerobic digesters, has not been explored in methanogenic biocathodes. Thus, the objective of this study was to assess the effect of biocathode ZVI on BES performance at 1 and 2 g/L initial ZVI concentrations and at various cathode potentials (-0.65 to -0.80 V versus SHE). The total CH4 produced during a 7-day feeding cycle with 1 and 2 g/L initial ZVI was 2.8- and 2.9-fold higher, respectively, than the mean CH4 production in the four prior cycles without ZVI addition. Furthermore, CH4 production by the ZVI-amended biocathodes remained elevated throughout three subsequent feeding cycles, despite catholyte replacement and no new ZVI addition. The fourth cycle following a single ZVI addition of 1 g/L and 2 g/L yielded 123% and 231% more total CH4 than in the non-ZVI cycles, respectively. The higher CH4 production could not be fully explained by complete anaerobic oxidation of the ZVI and utilization of produced H2 by hydrogenotrophic methanogens. Microbial community analysis showed that the same phylotype, most closely related to Methanobrevibacter arboriphilus, dominated the archaeal community in the ZVI-free and ZVI-amended biocathodes. However, the bacterial community experienced substantial changes following ZVI exposure, with more Proteobacteria and fewer Bacteroidetes in the ZVI-amended biocathode. Furthermore, it is likely that a redox-active precipitate formed in the ZVI-amended biocathode, which sorbed to the electrode and/or biofilm, acted as a redox mediator, and enhanced electron transfer and CH4 production. Thus, ZVI may be used to increase biocathode CH4 production, assist in the start-up of an electromethanogenic biocathode, and/or maintain microbial activity during voltage interruptions.

Methanogenic Biocathode Microbial Community Development and the Role of Bacteria.

The cathode microbial community of a methanogenic bioelectrochemical system (BES) is key to the efficient conversion of carbon dioxide (CO2) to methane (CH4) with application to biogas upgrading. The objective of this study was to compare the performance and microbial community composition of a biocathode inoculated with a mixed methanogenic (MM) culture to a biocathode inoculated with an enriched hydrogenotrophic methanogenic (EHM) culture, developed from the MM culture following pre-enrichment with H2 and CO2 as the only externally supplied electron donor and carbon source, respectively. Using an adjacent Ag/AgCl reference electrode, biocathode potential was poised at -0.8 V (versus SHE) using a potentiostat, with the bioanode acting as the counter electrode. When normalized to cathode biofilm biomass, the methane production in the MM- and EHM-biocathode was 0.153 ± 0.010 and 0.586 ± 0.029 mmol CH4/mg biomass-day, respectively. This study showed that H2/CO2 pre-enriched inoculum enhanced biocathode CH4 production, although the archaeal communities in both biocathodes converged primarily (86-100%) on a phylotype closely related to Methanobrevibacter arboriphilus. The bacterial community of the MM-biocathode was similar to that of the MM inoculum but was enriched in Spirochaetes and other nonexoelectrogenic, fermentative Bacteria. In contrast, the EHM-biocathode bacterial community was enriched in Proteobacteria, exoelectrogens, and putative producers of electron shuttle mediators. Similar biomass levels were detected in the MM- and EHM-biocathodes. Thus, although the archaeal communities were similar in the two biocathodes, the difference in bacterial community composition was likely responsible for the 3.8-fold larger CH4 production rate observed in the EHM-biocathode. Roles for abundant OTUs identified in the biofilm and inoculum cultures were highlighted on the basis of previous reports.

Inhibitory Effect of Furanic and Phenolic Compounds on Exoelectrogenesis in a Microbial Electrolysis Cell Bioanode.

The objective of this study was to systematically investigate the inhibitory effect of furfural (FF), 5-hydroxymethylfurfural (HMF), syringic acid (SA), vanillic acid (VA), and 4-hydroxybenzoic acid (HBA), which are problematic lignocellulose-derived byproducts, on exoelectrogenesis in the bioanode of a microbial electrolysis cell. The five compound mixture at an initial total concentration range from 0.8 to 8.0 g/L resulted in an up to 91% current decrease as a result of exoelectrogenesis inhibition; fermentative, nonexoelectrogenic biotransformation pathways of the five compounds were not affected. Furthermore, the parent compounds at a high concentration, as opposed to their biotransformation products, were responsible for the observed inhibition. All five parent compounds contributed to the observed inhibition of the mixture. The IC50 (i.e., concentration resulting in 50% current decrease) of individually tested parent compounds was 2.7 g/L for FF, 3.0 g/L for HMF, 1.9 g/L for SA, 2.1 g/L for VA and 2.0 g/L for HBA. However, the parent compounds, when tested below their respective noninhibitory concentration, jointly resulted in significant inhibition as a mixture. Catechol and phenol, which were persistent biotransformation products, inhibited exoelectrogenesis only at high concentrations, but to a lesser extent than the parent compounds. Exoelectrogenesis recovery from inhibition by all compounds was observed at different rates, with the exception of catechol, which resulted in irreversible inhibition.

Biotransformation of Furanic and Phenolic Compounds with Hydrogen Gas Production in a Microbial Electrolysis Cell.

Furanic and phenolic compounds are problematic byproducts resulting from the breakdown of lignocellulosic biomass during biofuel production. The capacity of a microbial electrolysis cell (MEC) to produce hydrogen gas (H2) using a mixture of two furanic (furfural, FF; 5-hydroxymethyl furfural, HMF) and three phenolic (syringic acid, SA; vanillic acid, VA; and 4-hydroxybenzoic acid, HBA) compounds as the substrate in the bioanode was assessed. The rate and extent of biotransformation of the five compounds and efficiency of H2 production, as well as the structure of the anode microbial community, were investigated. The five compounds were completely transformed within 7-day batch runs and their biotransformation rate increased with increasing initial concentration. At an initial concentration of 1200 mg/L (8.7 mM) of the mixture of the five compounds, their biotransformation rate ranged from 0.85 to 2.34 mM/d. The anode Coulombic efficiency was 44-69%, which is comparable to that of wastewater-fed MECs. The H2 yield varied from 0.26 to 0.42 g H2-COD/g COD removed in the anode, and the bioanode volume-normalized H2 production rate was 0.07-0.1 L/L-d. The biotransformation of the five compounds took place via fermentation followed by exoelectrogenesis. The major identified fermentation products that did not transform further were catechol and phenol. Acetate was the direct substrate for exoelectrogenesis. Current and H2 production were inhibited at an initial substrate concentration of 1200 mg/L, resulting in acetate accumulation at a much higher level than that measured in other batch runs conducted with a lower initial concentration of the five compounds. The anode microbial community consisted of exoelectrogens, putative degraders of the five compounds, and syntrophic partners of exoelectrogens. The MEC H2 production demonstrated in this study is an alternative to the currently used process of reforming natural gas to supply H2 needed to upgrade bio-oils to stable hydrocarbon fuels.

Stacking Time and Aluminum Sulfate Effects on Polyether Ionophores in Broiler Litter.

The use of ionophores as antiparasitic drugs plays an important role in US poultry production, especially in the broiler () industry. However, administered ionophores can pass through the bird's digestive system and appear in broiler litter, which, when applied to agricultural fields, can present an environmental hazard. Stacking (storing or stockpiling) broiler litter for some time might decrease the litter ionophore concentrations before land application. Because ionophores undergo abiotic hydrolysis at low pH, decreasing litter pH with acidic aluminum sulfate (alum) might also decrease ionophore concentrations. We assessed the change in ionophore concentrations in broiler litter in response to the length of time broiler litter was stored (stacking time) and alum addition. We spiked broiler litter with monensin and salinomycin, placed alum-amended litter (∼pH 4-5) and unamended litter (∼pH 8-9) into 1.8-m bins, and repeatedly sampled each bin for 112 d. Our findings showed that stacking broiler litter alone did not have an impact on monensin concentration, but it did slowly reduce salinomycin concentration by 55%. Adding alum to broiler litter reduced monensin concentration by approximately 20% relative to unamended litter, but it did not change salinomycin concentration. These results call for continued search for alternative strategies that could potentially reduce the concentration of ionophores in broiler litter before their application to agricultural soils.

Alum and Rainfall Effects on Ionophores in Runoff from Surface-Applied Broiler Litter.

Polyether ionophores, monensin, and salinomycin are commonly used as antiparasitic drugs in broiler production and may be present in broiler litter (bird excreta plus bedding material). Long-term application of broiler litter to pastures may lead to ionophore contamination of surface waters. Because polyether ionophores break down at low pH, we hypothesized that decreasing litter pH with an acidic material such as aluminum sulfate (alum) would reduce ionophore losses to runoff (i.e., monensin and salinomycin concentrations, loads, or amounts lost). We quantified ionophore loss to runoff in response to (i) addition of alum to broiler litter and (ii) length of time between litter application and the first simulated rainfall event. The factorial experiment consisted of unamended (∼pH 9) vs. alum-amended litters (∼pH 6), each combined with simulated rainfall at 0, 2, or 4 wk after litter application. Runoff from alum-amended broiler litter had 33% lower monensin concentration ( < 0.01), 57% lower monensin load ( < 0.01), 48% lower salinomycin concentration ( < 0.01), and 66% lower salinomycin load ( < 0.01) than runoff from unamended broiler litter when averaged across all events of rainfall. Ionophore losses to runoff were also less when rainfall was delayed for 2 or 4 wk after litter application relative to applying rainfall immediately after litter application. While the weather is difficult to predict, our data suggest that ionophore losses in runoff can be reduced if broiler litter applications are made to maximize dry time after application.

Microbial community degradation of widely used quaternary ammonium disinfectants.

Benzalkonium chlorides (BACs) are disinfectants widely used in a variety of clinical and environmental settings to prevent microbial infections, and they are frequently detected in nontarget environments, such as aquatic and engineered biological systems, even at toxic levels. Therefore, microbial degradation of BACs has important ramifications for alleviating disinfectant toxicity in nontarget environments as well as compromising disinfectant efficacy in target environments. However, how natural microbial communities respond to BAC exposure and what genes underlie BAC biodegradation remain elusive. Our previous metagenomic analysis of a river sediment microbial community revealed that BAC exposure selected for a low-diversity community, dominated by several members of the Pseudomonas genus that quickly degraded BACs. To elucidate the genetic determinants of BAC degradation, we conducted time-series metatranscriptomic analysis of this microbial community during a complete feeding cycle with BACs as the sole carbon and energy source under aerobic conditions. Metatranscriptomic profiles revealed a candidate gene for BAC dealkylation, the first step in BAC biodegradation that results in a product 500 times less toxic. Subsequent biochemical assays and isolate characterization verified that the putative amine oxidase gene product was functionally capable of initiating BAC degradation. Our analysis also revealed cooperative interactions among community members to alleviate BAC toxicity, such as the further degradation of BAC dealkylation by-products by organisms not encoding amine oxidase. Collectively, our results advance the understanding of BAC aerobic biodegradation and provide genetic biomarkers to assess the critical first step of this process in nontarget environments.

Photodegradation of veterinary ionophore antibiotics under UV and solar irradiation.

The veterinary ionophore antibiotics (IPAs) are extensively used as coccidiostats and growth promoters and are released to the environment via land application of animal waste. Due to their propensity to be transported with runoff, IPAs likely end up in surface waters where they are subject to photodegradation. This study is among the first to investigate the photodegradation of three commonly used IPAs, monensin (MON), salinomycin (SAL) and narasin (NAR), under UV and solar irradiation. Results showed that MON was persistent in a deionized (DI) water matrix when exposed to UV and sunlight, whereas SAL and NAR could undergo direct photolysis with a high quantum yield. Water components including nitrate and dissolved organic matter had a great impact on the photodegradation of IPAs. A pseudosteady state kinetic model was successfully applied to predict IPAs' photodegradation rates in real water matrices. Applying LC/MS/MS, multiple photolytic transformation products of IPAs were observed and their structures were proposed. The direct photolysis of SAL and NAR occurred via cleavage on the ketone moiety and self-sensitized photolysis. With the presence of nitrate, MON was primarily degraded by hydroxyl radicals, whereas SAL showed reactivity toward both hydroxyl and nitrogen-dioxide radicals. Additionally, toxicity tests showed that photodegradation of SAL eliminated its antibiotic properties against Bacillus subtilis.