Feature Engineering and Supervised Machine Learning to Forecast Biogas Production during Municipal Anaerobic Co-Digestion.

Municipalities with excess anaerobic digestion capacity accept offsite wastes for co-digestion to meet sustainability goals and create more biogas. Despite the benefits inherent to co-digestion, the temporal and compositional heterogeneity of external waste streams creates operational challenges that lead to upsets or conservative co-digestion. Given the complex microbial bioprocesses occurring during anaerobic digestion, prediction and modeling of the outcomes can be challenging, and machine learning has the potential to improve understanding and control of co-digestion processes. Biogas flows are a surrogate for process health, and here, we predicted biogas production from historical data collected by a water resource recovery facility (WRRF) during normal operation. We tested a daily lab and operational data set (n = 1089 after cleaning) and a minute-by-minute supervisory control and data acquisition (SCADA) operational data set (n = 491,761 after cleaning) to determine if forecasting biogas flow for a 24 h time horizon is feasible without collecting additional data. We found that a multilayer perceptron (MLP) neural network model outperformed tree-based and multiple linear regression models. Using a high-resolution SCADA data set for the first time, we showed that MLP neural networks could predict biogas production with an adjusted coefficient of determination (R2) of 0.78 and a mean absolute percentage error of 13.4% on a holdout test set. Adding daily laboratory analyses to the model did not appreciably improve the prediction of biogas flows. Feature engineering was essential to an accurate prediction, and 11 of the 15 most important features in the SCADA model were calculated from raw SCADA outputs. In summary, this paper demonstrates that minute-scale SCADA information collected at a municipal co-digestion facility can forecast biogas production, as a first step toward a digital twin model, without additional data collection.

Performance of Community Water Board-Managed Passive In-Line Chlorinators Supported by a Circuit Rider Program in Rural Honduras.

This study evaluated the ability of passive chlorinators and the associated kinds of external support necessary to provide adequate free chlorine residual (FCR) for community distribution systems in rural Honduras. We found that 77% of samples, from distribution systems with passive chlorinators installed by EOS International at storage tanks within these distribution systems, had FCR concentrations that met or exceeded the World Health Organization minimum threshold of 0.2 mg/L for point-of-use or piped systems. In EOS-supported communities, passive chlorinators delivered FCR ≥ 0.2 mg/L in 90% of tank samples, 83% of middle-house samples, and 79% of last-house samples. Technical issues accounted for only 26% of all lapses in chlorination (i.e., FCR = 0 mg/L). Occasional and habitual errors of the local water board accounted for 24 and 15% of all lapses. Visit frequency by EOS circuit riders was strongly correlated with positive chlorination outcomes, and technical assistance visits were the most valuable of all visit types. It was also shown that monitoring visits were negatively correlated with other visit types, indicating that monitoring may take place at the expense of more valuable visit types, which highlights the potential need for remote FCR monitoring approaches.

Congener-Specific Emissions from Floors and Walls Characterize Indoor Airborne Polychlorinated Biphenyls.

To reconcile the federal regulation of material polychlorinated biphenyl (PCB) concentrations with recently implemented state regulations of airborne PCBs, there is a need to characterize the relationship between PCB emissions from surfaces and air concentrations. We hypothesized that the magnitude and congener distribution of emissions from floors and walls fully account for the airborne PCBs measured in rooms constructed during the height of PCB production and sales. We measured emissions of PCB congeners from various wall and floor materials using polyurethane foam passive emission samplers before and after hexane wiping. Our results revealed that PCB emissions from flooring adequately predicted the magnitude and congener distribution of PCBs observed in the room air. Emissions varied by material within a single building (5 × 103 ng m-2 day-1 from wood panel walls to 3 × 104 ng m-2 day-1 from vinyl tile) and within the same room. Yet congener distributions between material emission PCB profiles and room air PCB profiles were statistically similar. Hexane wiping significantly reduced PCB emissions (>60%), indicating the importance of surface films as an ongoing source of airborne PCBs. The magnitude and congener distribution of material bulk concentrations did not explain that of material emissions or air concentrations. Passive measurements of polychlorinated biphenyl emissions from floors in a university building predict the concentrations of PCBs in room air.

Reconnaissance of Oxygenic Denitrifiers in Agriculturally Impacted Soils.

Row crop production in the agricultural Midwest pollutes waterways with nitrate, and exacerbates climate change through increased emissions of nitrous oxide and methane. Oxygenic denitrification processes in agricultural soils mitigate nitrate and nitrous oxide pollution by short-circuiting the canonical pathway to avoid nitrous oxide formation. Furthermore, many oxygenic denitrifiers employ a nitric oxide dismutase (nod) to create molecular oxygen that is used by methane monooxygenase to oxidize methane in otherwise anoxic soils. The direct investigation of nod genes that could facilitate oxygenic denitrification processes in agricultural sites is limited, with no prior studies investigating nod genes at tile drainage sites. Thus, we performed a reconnaissance of nod genes at variably saturated surface sites, and within a variably to fully saturated soil core in Iowa to expand the known distribution of oxygenic denitrifiers. We identified new nod gene sequences from agricultural soil and freshwater sediments in addition to identifying nitric oxide reductase (qNor) related sequences. Surface and variably saturated core samples displayed a nod to 16S rRNA gene relative abundance of 0.004% to 0.1% and fully saturated core samples had relative nod gene abundance of 1.2%. The relative abundance of the phylum Methylomirabilota increased from 0.6% and 1% in the variably saturated core samples to 3.8% and 5.3% in the fully saturated core samples. The more than 10-fold increase in relative nod abundance and almost 9-fold increase in relative Methylomirabilota abundance in fully saturated soils suggests that potential oxygenic denitrifiers play a greater nitrogen cycling role under these conditions. IMPORTANCE The direct investigation of nod genes in agricultural sites is limited, with no prior studies investigating nod genes at tile drains. An improved understanding of nod gene diversity and distribution is significant to the field of bioremediation and ecosystem services. The expansion of the nod gene database will advance oxygenic denitrification as a potential strategy for sustainable nitrate and nitrous oxide mitigation, specifically for agricultural sites.

Photolysis of 3-Nitro-1,2,4-triazol-5-one: Mechanisms and Products.

Insensitive munitions formulations that include 3-nitro-1,2,4-triazol-5-one (NTO) are replacing traditional explosive compounds. While these new formulations have superior safety characteristics, the compounds have greater environmental mobility, raising concern over potential contamination and cleanup of training and manufacturing facilities. Here, we examine the mechanisms and products of NTO photolysis in simulated sunlight to further inform NTO degradation in sunlit surface waters. We demonstrate that NTO produces singlet oxygen and that dissolved oxygen increases the NTO photolysis rate in deionized water. The rate of NTO photolysis is independent of concentration and decreases slightly in the presence of Suwannee River Natural Organic Matter. The apparent quantum yield of NTO generally decreases as pH increases, ranging from 2.0 × 10-5 at pH 12 to 1.3 × 10-3 at pH 2. Bimolecular reaction rate constants for NTO with singlet oxygen and hydroxyl radical were measured to be (1.95 ± 0.15) × 106 and (3.28 ± 0.23) × 1010 M-1 s-1, respectively. Major photolysis reaction products were ammonium, nitrite, and nitrate, with nitrite produced in nearly stoichiometric yield upon the reaction of NTO with singlet oxygen. Environmental half-lives are predicted to span from 1.1 to 5.7 days. Taken together, these data enhance our understanding of NTO photolysis under environmentally relevant conditions.

Passive In-Line Chlorination for Drinking Water Disinfection: A Critical Review.

The world is not on track to meet Sustainable Development Goal 6.1 to provide universal access to safely managed drinking water by 2030. Removal of priority microbial contaminants by disinfection is one aspect of ensuring water is safely managed. Passive chlorination (also called in-line chlorination) represents one approach to disinfecting drinking water before or at the point of collection (POC), without requiring daily user input or electricity. In contrast to manual household chlorination methods typically implemented at the point of use (POU), passive chlorinators can reduce the user burden for chlorine dosing and enable treatment at scales ranging from communities to small municipalities. In this review, we synthesized evidence from 27 evaluations of passive chlorinators (in 19 articles, 3 NGO reports, and 5 theses) conducted across 16 countries in communities, schools, health care facilities, and refugee camps. Of the 27 passive chlorinators we identified, the majority (22/27) were solid tablet or granular chlorine dosers, and the remaining devices were liquid chlorine dosers. We identified the following research priorities to address existing barriers to scaled deployment of passive chlorinators: (i) strengthening local chlorine supply chains through decentralized liquid chlorine production, (ii) validating context-specific business models and financial sustainability, (iii) leveraging remote monitoring and sensing tools to monitor real-time chlorine levels and potential system failures, and (iv) designing handpump-compatible passive chlorinators to serve the many communities reliant on handpumps as a primary drinking water source. We also propose a set of reporting indicators for future studies to facilitate standardized evaluations of the technical performance and financial sustainability of passive chlorinators. In addition, we discuss the limitations of chlorine-based disinfection and recognize the importance of addressing chemical contamination in drinking water supplies. Passive chlorinators deployed and managed at-scale have the potential to elevate the quality of existing accessible and available water services to meet "safely managed" requirements.

Room-to-Room Variability of Airborne Polychlorinated Biphenyls in Schools and the Application of Air Sampling for Targeted Source Evaluation.

Airborne polychlorinated biphenyl (PCB) concentrations are higher indoors than outdoors due to their historical use in building materials and their presence in modern paints and surface treatments. For some populations, including school children, PCB levels indoors result in inhalation exposures that may be greater than or equivalent to exposure through diet. In a school, PCB exposure may come from multiple sources. We hypothesized that there are both Aroclor and non-Aroclor sources within a single school and that PCB concentration and congener profiles differ among rooms within a single building. To evaluate this hypothesis and to identify potential localized sources, we measured airborne PCBs in nine rooms in a school. We found that schoolroom concentrations exceed outdoor air concentrations. Schoolroom concentrations and congener profiles also varied from one room to another. The concentrations were highest in the math room (35.75 ng m-3 ± 8.08) and lowest in the practice gym (1.54 ng m-3 ± 0.35). Rooms in the oldest wing of the building, originally constructed between 1920 and 1970, had the highest concentrations. The congener distribution patterns indicate historic use of Aroclor 1254 as well as modern sources of non-Aroclor congeners associated with paint pigments and surface coatings. Our findings suggest this noninvasive source identification method presents an opportunity for targeted source testing for more cost-effective prioritization of materials remediation in schools.

Evaluating Collaborative Readiness for Interdisciplinary Flood Research.

Increasing trends in global flood risk are driven by a complex web of interactions among natural, built environment, and social systems. As a result, flood resilience research is an ideal topic for an interdisciplinary approach. Core characteristics of interdisciplinary research are team collaboration and the systematic integration of disciplinary knowledge, in both problem formulation and analytical methods. Indicators of interdisciplinarity tend to focus on scholarly outcomes, but collaborative processes may be even more important for knowledge integration. In this Perspective piece, we outline and advocate a two-pronged approach to enhance potential for integrating knowledge: using collaborative proximity to assess team readiness to conduct interdisciplinary research and employing program evaluation to assess change in proximity components over time. To do so, we draw on scholarship in economic geography, team science, and program evaluation. We then connect the findings to a case study of collaboration within our interdisciplinary team of flood researchers, program evaluators, and local stakeholders, as we navigate a multi-institutional project on flood resilience.

Impacts of ammonia loading and biofilm age on the prevalence of nitrogen-cycling microorganisms in a full-scale submerged attached-growth reactor.

This study reports the impacts of seasonal ammonia load changes and biofilm age on the quantity of biomass and on the prevalence of ammonia- and nitrite-metabolizing organisms within a submerged attached-growth reactor (SAGR™) following lagoon treatment. Ammonia (NH3 ) loadings (0.12-3.17 kg/d) in the primary SAGR were measured over 223 days from May to December in 2017. Adjustment of the wastewater flow path on September 1 successfully increased NH3 loading to the primary SAGR, which subsequently caused reactor biomass to increase. The NH3 removal rate in October (0.5 kg/d) was greater than rates in June and July (0.3 and 0.2 kg/d) despite a water temperature decrease from >24 to 15.6°C. This elevated removal rate in October, and the sustained removal rate in December (0.4 kg/d, 5.3°C) were associated with a measured increase in microbial biomass. The relative abundance of the anammox organism C. Brocadia was 5 times greater in the mature biofilm after 686 days of growth, and the genus Pseudomonas increased sevenfold. The presence of Pseudomonas, which contains denitrifying species, and anammox suggests a high potential for removal of total nitrogen in SAGRs. PRACTITIONER POINTS: Pseudomonas prevalence and the presence of anammox suggest a high potential for total nitrogen removal in mature SAGR biofilms. The abundance of the anammox microorganism C. Brocadia was greater after 686 days of biofilm growth compared with 33 days. Simple operational changes can increase biomass in the SAGR to maintain, or even increase, NH3 transformation rates during cold weather.

Metagenomic analysis of nitrogen-cycling genes in upper Mississippi river sediment with mussel assemblages.

We investigated the impact of native freshwater mussel assemblages (order Unionoida) on the abundance and composition of nitrogen-cycling genes in sediment of an upper Mississippi river habitat. We hypothesized that the genomic potential for ammonia and nitrite oxidation would be greater in the sediment with mussel assemblages, presumably due to mussel biodeposition products, namely ammonia and organic carbon. Regardless of the presence of mussels, upper Mississippi river sediment microbial communities had the largest genomic potential for nitrogen fixation followed by urea catabolism, nitrate metabolism, and nitrate assimilation, as evidenced by analysis of nitrogen cycling pathway abundances. However, genes encoding nitrate and nitrite redox reactions, narGHI and nxrAB, were the most abundant functional genes of the nitrogen cycling gene families. Using linear discriminant analysis (LDA), we found nitrification genes were the most important biomarkers for nitrogen cycling genomic potential when mussels were present, and this presented an opposing effect on the abundance of genes encoding nitric oxide reduction. The genes involved in nitrification that increased the most were amoA associated with comammox Nitrospira and nxr homologs associated with Nitrospira. On the other hand, the most distinctive biomarkers of microbial communities without mussels were norB and nrfA, as part of denitrification and dissimilatory nitrate reduction to ammonium pathways, respectively. Ultimately, this research demonstrates the impact of native mollusks on microbial nitrogen cycling in an aquatic agroecosystem.

Submerged attached-growth reactors as lagoon retrofits for cold-weather ammonia removal: performance and sizing.

Small towns that operate wastewater treatment lagoons struggle to meet ammonia limits in cold weather. Here we report the performance of a lagoon, retrofitted with submerged attached growth reactors (SAGRsTM), to provide insight on ammonia effluent compliance and optimal SAGR sizing as functions of water temperature. The lagoon-SAGR water resource recovery facility (WRRF) removed 95% of incoming ammonia with 94% attributed to the SAGRs. The high treatment capacity of the two primary SAGRs, evidenced by nearly continuous dissolved oxygen saturation and exceedingly high ammonia removals, suggested the two secondary SAGRs were essentially unnecessary and that all four SAGRs should be reduced in size. Furthermore, without the secondary SAGRs, the primary SAGR effluent would have exceeded the permitted ammonia discharge limit only four times in the 2.5-year study. At its current size, the lagoon-SAGR WRRF never exceeded permitted ammonia limits, but size reductions should be used for future retrofits.

The Genomic Potentials of NOB and Comammox Nitrospira in River Sediment Are Impacted by Native Freshwater Mussels.

Freshwater mussel assemblages of the Upper Mississippi River (UMR) sequester tons of ammonia- and urea-based biodeposits each day and aerate sediment through burrowing activities, thus creating a unique niche for nitrogen (N) cycling microorganisms. This study explored how mussels impact the abundance of N-cycling species with an emphasis on Candidatus Nitrospira inopinata, the first microorganism known to completely oxidize ammonia (comammox) to nitrate. This study used metagenomic shotgun sequencing of genomic DNA to compare nitrogen cycling species in sediment under a well-established mussel assemblage and in nearby sediment without mussels. Metagenomic reads were aligned to the prokaryotic RefSeq non-redundant protein database using BLASTx, taxonomic binning was performed using the weighted lowest common ancestor algorithm, and protein-coding genes were categorized by metabolic function using the SEED subsystem. Linear discriminant analysis (LDA) effect sizes were used to determine which metagenomes and metabolic features explained the most differences between the mussel habitat sediment and sediment without mussels. Of the N-cycling species deemed differentially abundant, Nitrospira moscoviensis and "Candidatus Nitrospira inopinata" were responsible for creating a distinctive N-cycling microbiome in the mussel habitat sediment. Further investigation revealed that comammox Nitrospira had a large metabolic potential to degrade mussel biodeposits, as evidenced the top ten percent of protein-coding genes including the cytochrome c-type biogenesis protein required for hydroxylamine oxidation, ammonia monooxygenase, and urea decomposition SEED subsystems. Genetic marker analysis of these two Nitrospira taxons suggested that N. moscoviensis was most impacted by diverse carbon metabolic processes while "Candidatus Nitrospira inopinata" was most distinguished by multidrug efflux proteins (AcrB), NiFe hydrogenase (HypF) used in hydrogen oxidation and sulfur reduction coupled reactions, and a heme chaperone (CcmE). Furthermore, our research suggests that comammox and NOB Nitrospira likely coexisted by utilizing mixotrophic metabolisms. For example, "Candidatus Nitrospira inopinata" had the largest potentials for ammonia oxidation, nitrite reduction with NirK, and hydrogen oxidation, while NOB Nitrospira had the greatest potential for nitrite oxidation, and nitrate reduction possibly coupled with formate oxidation. Overall, our results suggest that this mussel habitat sediment harbors a niche for NOB and comammox Nitrospira, and ultimately impacts N-cycling in backwaters of the UMR.

Metabolism and Photolysis of 2,4-Dinitroanisole in Arabidopsis.

New insensitive munitions explosives, including 2,4-dinitroanisole (DNAN), are replacing traditional explosive compounds to protect soldiers and simplify transport logistics. Despite the occupational safety benefits of these new explosives, feasible strategies for cleaning up DNAN from soil and water have not been developed. Here, we evaluate the metabolism of DNAN by the model plant Arabidopsis to determine whether phytoremediation can be used to clean up contaminated sites. Furthermore, we evaluate the role of photodegradation of DNAN and its plant metabolites within Arabidopsis leaves to determine the potential impact of photolysis on the phytoremediation of contaminants. When exposed to DNAN for three days, Arabidopsis took up and metabolized 67% of the DNAN in hydroponic solution. We used high resolution and tandem mass spectrometry in combination with stable-isotope labeled DNAN to confirm ten phase II DNAN metabolites in Arabidopsis. The plants separately reduced both the para- and ortho-nitro groups and produced glycosylated products that accumulated within plant tissues. Both DNAN and a glycosylated metabolite were subsequently photolyzed within leaf tissue under simulated sunlight, and [15N2]DNAN yielded 15NO2- in leaves. Therefore, photolysis inside leaves may be an important, yet under-explored, phytoremediation mechanism.

Effect of freshwater mussels on the vertical distribution of anaerobic ammonia oxidizers and other nitrogen-transforming microorganisms in upper Mississippi river sediment.

Targeted qPCR and non-targeted amplicon sequencing of 16S rRNA genes within sediment layers identified the anaerobic ammonium oxidation (anammox) niche and characterized microbial community changes attributable to freshwater mussels. Anammox bacteria were normally distributed (Shapiro-Wilk normality test, W-statistic =0.954, p = 0.773) between 1 and 15 cm depth and were increased by a factor of 2.2 (p < 0.001) at 3 cm below the water-sediment interface when mussels were present. Amplicon sequencing of sediment at depths relevant to mussel burrowing (3 and 5 cm) showed that mussel presence reduced observed species richness (p = 0.005), Chao1 diversity (p = 0.005), and Shannon diversity (p < 0.001), with more pronounced decreases at 5 cm depth. A non-metric, multidimensional scaling model showed that intersample microbial species diversity varied as a function of mussel presence, indicating that sediment below mussels harbored distinct microbial communities. Mussel presence corresponded with a 4-fold decrease in a majority of operational taxonomic units (OTUs) classified in the phyla Gemmatimonadetes, Actinobacteria, Acidobacteria, Plantomycetes, Chloroflexi, Firmicutes, Crenarcheota, and Verrucomicrobia. 38 OTUs in the phylum Nitrospirae were differentially abundant (p < 0.001) with mussels, resulting in an overall increase from 25% to 35%. Nitrogen (N)-cycle OTUs significantly impacted by mussels belonged to anammmox genus Candidatus Brocadia, ammonium oxidizing bacteria family Nitrosomonadaceae, ammonium oxidizing archaea genus Candidatus Nitrososphaera, nitrite oxidizing bacteria in genus Nitrospira, and nitrate- and nitrite-dependent anaerobic methane oxidizing organisms in the archaeal family "ANME-2d" and bacterial phylum "NC10", respectively. Nitrosomonadaceae (0.9-fold (p < 0.001)) increased with mussels, while NC10 (2.1-fold (p < 0.001)), ANME-2d (1.8-fold (p < 0.001)), and Candidatus Nitrososphaera (1.5-fold (p < 0.001)) decreased with mussels. Co-occurrence of 2-fold increases in Candidatus Brocadia and Nitrospira in shallow sediments suggests that mussels may enhance microbial niches at the interface of oxic-anoxic conditions, presumably through biodeposition and burrowing. Furthermore, it is likely that the niches of Candidatus Nitrososphaera and nitrite- and nitrate-dependent anaerobic methane oxidizers were suppressed by mussel biodeposition and sediment aeration, as these phylotypes require low ammonium concentrations and anoxic conditions, respectively. As far as we know, this is the first study to characterize freshwater mussel impacts on microbial diversity and the vertical distribution of N-cycle microorganisms in upper Mississippi river sediment. These findings advance our understanding of ecosystem services provided by mussels and their impact on aquatic biogeochemical N-cycling.

Simulated mussel mortality thresholds as a function of mussel biomass and nutrient loading.

A freshwater "mussel mortality threshold" was explored as a function of porewater ammonium (NH4+) concentration, mussel biomass, and total nitrogen (N) utilizing a numerical model calibrated with data from mesocosms with and without mussels. A mortality threshold of 2 mg-N L-1 porewater NH4+ was selected based on a study that estimated 100% mortality of juvenile Lampsilis mussels exposed to 1.9 mg-N L-1 NH4+ in equilibrium with 0.18 mg-N L-1 NH3. At the highest simulated mussel biomass (560 g m-2) and the lowest simulated influent water "food" concentration (0.1 mg-N L-1), the porewater NH4+ concentration after a 2,160 h timespan without mussels was 0.5 mg-N L-1 compared to 2.25 mg-N L-1 with mussels. Continuing these simulations while varying mussel biomass and N content yielded a mortality threshold contour that was essentially linear which contradicted the non-linear and non-monotonic relationship suggested by Strayer (2014). Our model suggests that mussels spatially focus nutrients from the overlying water to the sediments as evidenced by elevated porewater NH4+ in mesocosms with mussels. However, our previous work and the model utilized here show elevated concentrations of nitrite and nitrate in overlying waters as an indirect consequence of mussel activity. Even when the simulated overlying water food availability was quite low, the mortality threshold was reached at a mussel biomass of about 480 g m-2. At a food concentration of 10 mg-N L-1, the mortality threshold was reached at a biomass of about 250 g m-2. Our model suggests the mortality threshold for juvenile Lampsilis species could be exceeded at low mussel biomass if exposed for even a short time to the highly elevated total N loadings endemic to the agricultural Midwest.

Biotransformation of 2,4-dinitroanisole by a fungal Penicillium sp.

Insensitive munitions explosives are new formulations that are less prone to unintended detonation compared to traditional explosives. While these formulations have safety benefits, the individual constituents, such as 2,4-dinitroanisole (DNAN), have an unknown ecosystem fate with potentially toxic impacts to flora and fauna exposed to DNAN and/or its metabolites. Fungi may be useful in remediation and have been shown to degrade traditional nitroaromatic explosives, such as 2,4,6-trinitrotoluene and 2,4-dinitrotoluene, that are structurally similar to DNAN. In this study, a fungal Penicillium sp., isolated from willow trees and designated strain KH1, was shown to degrade DNAN in solution within 14 days. Stable-isotope labeled DNAN and an untargeted metabolomics approach were used to discover 13 novel transformation products. Penicillium sp. KH1 produced DNAN metabolites resulting from ortho- and para-nitroreduction, demethylation, acetylation, hydroxylation, malonylation, and sulfation. Incubations with intermediate metabolites such as 2-amino-4-nitroanisole and 4-amino-2-nitroanisole as the primary substrates confirmed putative metabolite isomerism and pathways. No ring-cleavage products were observed, consistent with other reports that mineralization of DNAN is an uncommon metabolic outcome. The production of metabolites with unknown persistence and toxicity suggests further study will be needed to implement remediation with Penicillium sp. KH1. To our knowledge, this is the first report on the biotransformation of DNAN by a fungus.

Modeling sustainable reuse of nitrogen-laden wastewater by poplar.

Numerical modeling was used to simulate the leaching of nitrogen (N) to groundwater as a consequence of irrigating food processing wastewater onto grass and poplar under various management scenarios. Under current management practices for a large food processor, a simulated annual N loading of 540 kg ha(-1) yielded 93 kg ha(-1) of N leaching for grass and no N leaching for poplar during the growing season. Increasing the annual growing season N loading to approximately 1,550 kg ha(-1) for poplar only, using "weekly", "daily" and "calculated" irrigation scenarios, yielded N leaching of 17 kg ha(-1), 6 kg ha(-1), and 4 kg ha(-1), respectively. Constraining the simulated irrigation schedule by the current onsite wastewater storage capacity of approximately 757 megaliters (Ml) yielded N leaching of 146 kg ha(-1) yr(-1) while storage capacity scenarios of 3,024 and 4,536 Ml yielded N leaching of 65 and 13 kg ha(-1) yr(-1), respectively, for a loading of 1,550 kg ha(-1) yr(-1). Further constraining the model by the current wastewater storage volume and the available land area (approximately 1,000 hectares) required a "diverse" irrigation schedule that was predicted to leach a weighted average of 13 kg-N ha(-1) yr(-1) when dosed with 1,063 kg-N ha(-1) yr(-1).

Enhanced vadose zone nitrogen removal by poplar during dormancy.

A pilot-scale, engineered poplar tree vadose zone system was utilized to determine effluent nitrate (NO3(-)) and ammonium concentrations resulting from intermittent dosing of a synthetic wastewater onto sandy soils at 4.5°C. The synthetic wastewater replicated that of an industrial food processor that irrigates onto sandy soils even during dormancy which can leave groundwater vulnerable to NO3(-) contamination. Data from a 21-day experiment was used to assess various Hydrus model parameterizations that simulated the impact of dormant roots. Bromide tracer data indicated that roots impacted the hydraulic properties of the packed sand by increasing effective dispersion, water content and residence time. The simulated effluent NO3(-) concentration on day 21 was 1.2 mg-N L(-1) in the rooted treatments compared to a measured value of 1.0 ± 0.72 mg-N L(-1). For the non-rooted treatment, the simulated NO3(-) concentration was 4.7 mg-N L(-1) compared to 5.1 ± 3.5 mg-N L(-1) measured on day 21. The model predicted a substantial "root benefit" toward protecting groundwater through increased denitrification in rooted treatments during a 21-day simulation with 8% of dosed nitrogen converted to N2 compared to 3.3% converted in the non-rooted test cells. Simulations at the 90-day timescale provided similar results, indicating increased denitrification in rooted treatments.

Partial nitritation ANAMMOX in submerged attached growth bioreactors with smart aeration at 20 °C.

Submerged attached growth bioreactors (SAGBs) were operated at 20 °C for 30 weeks in smart-aerated, partial nitritation ANAMMOX mode and in a timer-controlled, cyclic aeration mode. The smart-aerated SAGBs removed 48-53% of total nitrogen (TN) compared to 45% for SAGBs with timed aeration. Low dissolved oxygen concentrations and cyclic pH patterns in the smart-aerated SAGBs suggested conditions favorable to partial nitritation ANAMMOX and stoichiometrically-derived and numerically modeled estimations attributed 63-68% and 14-44% of TN removal to partial nitritation ANAMMOX in these bioreactors, respectively. Ammonia removals of 36-67% in the smart-aerated SAGBs, with measured oxygen and organic carbon limitations, further suggest partial nitritation ANAMMOX. The smart-aerated SAGBs required substantially less aeration to achieve TN removals similar to SAGBs with timer-controlled aeration. Genomic DNA testing confirmed that the dominant ANAMMOX seed bacteria, received from a treatment plant utilizing the DEMON® sidestream deammonification process, was a Candidatus Brocadia sp. (of the Planctomycetales order). The DNA from these bacteria was also present in the SAGBs at the conclusion of the study providing evidence for attached growth and limited biomass washout.