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Production of insensitive munitions (IM) creates wastewater containing 3-nitro-1,2,4-triazol-5-one (NTO). As NTO is an anion in neutral pH wastewaters it can be removed via ion exchange (IX), released during IX brine recharge and may be subsequently degraded by halophiles. IX brine regeneration through halophilic NTO biotransformation was observed over 210 cycles in a sequencing batch reactor (SBR) and averaged 64.8 ± 20.2% NTO removal. NTO formed 3-amino-1,2,4-triazol-5-one (ATO) under sulfate- and nitrate-reducing conditions. Variable NTO reductions were observed over four phases including anaerobic enrichment (80.5 ± 7.7%, cycle 0 to 41); anaerobic followed by aerobic (42.2 ± 23.5%, cycle 42 to 81); anaerobic supplemented with nitrate (64.7 ± 21.5%, cycle 82 to 177); and anaerobic stress test with nitrate and perchlorate (91.0 ± 9.7%, cycle 178 to 210). Sequencing results show most of the genera to be halophiles and sulfate-reducing bacteria and archaea including Desulfovibrio and Halanaerobium. The results of these studies demonstrate that NTO biotransformation by halophiles in brine solutions is feasible.

Chloroacetamide herbicides, namely acetochlor and metolachlor, are common herbicides used on corn and soybean fields. Dichloroacetamide safeners, namely benoxacor and furilazole, are commonly used in formulations containing chloroacetamide herbicides. Extensive reports on adsorption of chloroacetamide herbicides are available, yet little information exists regarding adsorption potential of co-applied safeners. Herein, the adsorption and desorption characteristics of selected herbicide safeners to granular activated carbon (GAC) and in agricultural soils are reported. Further, soil column studies were performed to understand the leaching behaviour of the herbicide Dual II Magnum. Equilibrium sorption experiments of safeners to three agricultural soils and one GAC showed that adsorption was best fitted by the Freundlich isotherm. The Freundlich adsorption constant, Kf, for benoxacor and furilazole sorption onto three agricultural soils ranged from 0.1 to 0.27 and 0.1 to 0.13 (mg/g) × (mg/L)ˆ(1/n), respectively. The Kf for benoxacor and furilazole to GAC was 6.4 and 3.4 (mg/g) × (mg/L)ˆ(1/n), respectively, suggesting more favorable sorption of benoxacor to GAC than furilazole to GAC. The sorption to soils was reversible as almost 40%–90% of both safeners was desorbed from three soils. These results were validated in four replicated soil column studies, where S-metolachlor was shown to leach similarly to the safener benoxacor, originating from the herbicide formulation. The leaching of S-metolachlor and benoxacor was influenced by soil texture. Cumulatively, these results show that safeners will move through the environment to surface waters similarly to the active ingredients in herbicides, but may be removed during drinking water treatment via GAC.

Diffusivities of several explosive compounds, as well as other complex organic compounds were experimentally derived using a peak parking methodology. High performance liquid chromatography (HPLC) with stop flow ‘parking’ was used to experimentally determine diffusivity based on band broadening associated with chemical diffusion within the HPLC column. This research builds on prior methods by determining an obstruction factor through comparing benzene diffusion in methanol in a C18 column to the previously published capillary column. The method is useful for structures not easily described by computational chemistry methods or for solvents that do not have association coefficients in published diffusivity models. Using the peak parking method, diffusivities (cm2/s) ranged from 6.29 × 10-7 to 1.06 × 10-6 for hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), 4.82 × 10-5 to 1.26 × 10-4 for 3-nitro-1,2,4-triazol-5-one (NTO), 1.63 × 10-5 to 7.20 × 10-6 for nitroguanidine (NQ), 7.07 × 10-6 to 2.09 × 10-5 for pyrimidine-2-carboxylic acid, and 1.02 × 10-6 to 2.27 × 10-6 for streptomycin. Compared to diffusivities estimated computationally, the empirical diffusivities reported herein estimated by two different methods have percent differences averaging 142%, 174%, 27%, 85%, and 33% for RDX, NTO, NQ, pyrimidine-2-carboxylic acid, and streptomycin, respectively. NTO had experimental diffusivities greater than the computational values, while RDX and streptomycin had experimental diffusivities lower than computational values.

The explosive 3-nitro-1,2,4-triazol-5-one (NTO) presents a physiochemical challenge for treatment of munitions wastewater. Leveraging NTO's ionic character in neutral pH wastewater allows for expanded treatment options. Four commercial drinking water anion exchange resins specific for NO

An innovative process consisting of Fe(II)-dosed anaerobic bioreactors and an oxidizing basin was used to continuously treat a synthetic wastewater (COD/sulfate mass ratio 2:1 and Fe/S molar ratio 1:1). Sludge recycling effects were evaluated on ten occasions, in which anaerobic sludge was collected, biochemically oxidized with mechanical aeration in the oxidizing basin before being mixed with the wastewater influent. The sludge recycling resulted in better effluent quality compared to the baseline operation without recycling. More Fe and S were retained as sludge in the bioreactors with sludge recycling (Fe 94%, S 91%) than those when the bioreactors were operated without sludge recycling (Fe 76%, S 86%). Scanning electron microscopic analysis showed that bacterial cells and iron sulfide intermixed in the anaerobic sludge and the presence of microbial exopolymeric substances. X-ray spectroscopic analyses showed amorphous FeS formed from the dosed ferrous iron and biogenic bisulfide, and suggested long-term conversion of the amorphous FeS to more stable crystalline FeS and FeS2 in the anaerobic bioreactors. In the oxidizing basin, oxidation of iron sulfides was of both chemical and biological nature, and their oxidized forms including amorphous FeO/Fe2O3 mixture and partially/fully oxidized sulfurs. Experimental results also indicated amorphous FeS was more readily oxidized than FeS2. Phylogenetic analysis revealed microorganisms related to Desulfomonile tiedjei (sulfur reducing) and Alkaliphilus metalliredigens (iron reducing) in the anaerobic bioreactors, and Thiobacter subterraneus (sulfur oxidizing) and Rubrivivax gelatinosus (iron oxidizing) related microorganisms in the oxidizing basin.

Rooftop harvested rainwater has become an alternative, potable, and non-potable water source used around the world. In the United States, rooftop harvested rainwater is most commonly used for irrigation. Rooftop harvested rainwater may contain contaminants from bird or animal feces that may present a risk to water users. Different roofing materials may influence the survival of fecal bacteria on the rooftop prior to runoff during rainfall. In this study, three pathogen groups (E. coli, enterococci and Salmonella enterica) in rooftop runoff from three, replicated roof types (asphalt shingle, synthetic slate, and wood shake) were quantified in multiple rain events. Matched roofs were selected from locations with differing amounts of tree cover. Enterococci were the most frequently detected bacteria from all roof types. Wood shake and asphalt shingle roofing materials had the poorest microbial water quality. Rainwater runoff from two of the six buildings failed to meet United States Food and Drug Administration microbial standards for irrigation water. A quantitative microbial risk assessment indicated that the annual probability of illness from consuming lettuce irrigated with rooftop harvested rainwater varied by roofing material, irrigation water withholding period, and exposure frequency. Consuming lettuce immediately after irrigation with rooftop rainwater presented the highest human health risk based on the probability of illness from E. coli and enterococci exposure. Withholding irrigation by 1 day prior to harvest decreased the annual probability of illness from E. coli by 2 log, but had a minimal effect on the risk from enterococci.

As a cost-effective and objective population-wide surveillance tool, wastewater-based epidemiology (WBE) has been widely implemented worldwide to monitor the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA concentration in wastewater. However, viral concentrations or loads in wastewater often correlate poorly with clinical case numbers. To date, there is no reliable method to back-estimate the coronavirus disease 2019 (COVID-19) case numbers from SARS-CoV-2 concentrations in wastewater. This greatly limits WBE in achieving its full potential in monitoring the unfolding pandemic. The exponentially growing SARS-CoV-2 WBE dataset, on the other hand, offers an opportunity to develop data-driven models for the estimation of COVID-19 case numbers (both incidence and prevalence) and transmission dynamics (effective reproduction rate). This study developed artificial neural network (ANN) models by innovatively expanding a conventional WBE dataset to include catchment, weather, clinical testing coverage and vaccination rate. The ANN models were trained and evaluated with a comprehensive state-wide wastewater monitoring dataset from Utah, USA during May 2020 to December 2021. In diverse sewer catchments, ANN models were found to accurately estimate the COVID-19 prevalence and incidence rates, with excellent precision for prevalence rates. Also, an ANN model was developed to estimate the effective reproduction number from both wastewater data and other pertinent factors affecting viral transmission and pandemic dynamics. The established ANN model was successfully validated for its transferability to other states or countries using the WBE dataset from Wisconsin, USA.

SUMMARY Animal well-being, evolving feed manufacturing regulations, human health, and consumer perception drive the continued need to provide safe feed to food animals. Two experiments were conducted to determine the effects of antimicrobial inclusion and conditioning temperature variations on feed manufacturing metrics (EXP1) and the inactivation of Enterococcus faecium, a Salmonella surrogate (EXP2). Both short-term (ST) and long-term (LT) steam conditioning techniques were utilized in this split-plot design where ST (10 s) or LT (60 s) steam conditioning served as whole plot units. Whole plots contained either 2 × 3 (EXP1) or 2 × 4 (EXP2) factorial arrangements varying in mixer-added antimicrobial inclusion (with or without) and degree of thermal processing (unprocessed mash, 71°C, 82°C, and 88°C). Manufacturing of dietary treatments was replicated 3 times. A basal diet was used to obtain desired conditioning temperature before Salmonella surrogate-inoculated feed was exposed to either ST or LT conditioning. In EXP1, conditioning time and antimicrobial inclusion interacted (P = 0.03) by decreasing pellet durability when antimicrobial was included in diets subjected to ST conditioning. Pellet durability was not affected when LT conditioning was utilized. In EXP2, conditioning time, antimicrobial inclusion, and degree of thermal processing interacted (P

Detection of low abundance human health pathogens in environmental samples is a challenge for water monitoring. This limitation can be overcome by the introduction of multiple displacement amplification (MDA) where a minute amount of genetic material can be amplified using a phi-29 DNA polymerase. However, the genetic makeup and the concentration of the polynucleotides might influence the amplification process due to inherent assay bias. Herein, a series of experiments were designed to demonstrate the effect of genome length, guanidine and cytosine content, and template concentration on the efficiency of MDA. Quantitative polymerase chain reaction (qPCR) was performed to quantify pre- and post-MDA concentrations of selected genes. Linear regression between pre- and post-MDA log gene copies L−1 of both environmental and lab-grown samples showed a positive correlation (F = 77.59, P

Fast-settling, anoxic sludge (FAS) was cultivated and utilized in this study to simultaneously reduce elevated levels of perchlorate and nitrate in an anaerobic sequencing batch reactor (AnSBR). Average perchlorate and nitrate removal efficiencies of 96.5 ± 8.44 % and 99.8 ± 0.32 %, respectively, were achieved from an average perchlorate and nitrate loading rate of 159 ± 101 g ClO4−/m3·d and 10.8 ± 7.25 g NO3−-N/m3·d, respectively, throughout long-term operation (>500-d). Batch activity tests revealed a preferential utilization of nitrate over perchlorate, where significant perchlorate reduction inhibition occurred when nitrate was present as a competing electron acceptor under carbon-limiting conditions. Specific perchlorate and nitrate reduction rates were shown to increase as the hydraulic retention time (HRT) of the AnSBR was step-wise decreased and subsequently the perchlorate and nitrate loading rates were step-wise increased. Functional, mRNA-based expression of the nitrite reductase (nirS and nirK), nitrous oxide reductase (nosZ), perchlorate reductase subunit A (pcrA), and the chlorite dismutase (cld) genes illustrated the simultaneous activity of heterotrophic denitrification and perchlorate reduction occurring throughout a complete standard reactor operational cycle, and allowed for expression trends to be documented as the HRT of the AnSBR was reduced from 5-d to 1.25-d. Nitrous oxide (N2O) production was detected as a result of incomplete denitrification, where the largest N2O production occurred at the highest nitrate loading rates investigated in this study. Thauera species were heavily enriched at a longer HRT of 5-d, but were out-competed by Dechloromonas species as the HRT of the AnSBR was step-wise reduced to 1.25-d.

Poor urban water quality has been linked to diminished source water quality, poorly functioning water treatment systems and infiltration into distribution lines after treatment resulting in microbiological contamination. With limited funding to rehabilitate distribution lines, developing nations need tools to identify the areas of greatest concern to human health so as to target cost effective remediation approaches. Herein, a case study of Hyderabad, Pakistan was used to demonstrate the efficacy of combining quantitative microbial risk assessment (QMRA) for multiple pathogens with spatial distribution system modeling to identify areas for pipe rehabilitation. Abundance of Escherichia coli, Enterococcus (enterococci), Salmonella spp., Shigella spp., Giardia intestinalis, Vibrio cholera, norovirus GI and adenovirus 40/41, were determined in 85 locations including the source water, treatment plant effluent and the city distribution lines. Bayesian statistics and Monte Carlo simulations were used in the QMRA to account for left-censored microbial abundance distributions. Bacterial and viral abundances in the distribution system samples decreased as follows: 9400 ± 19,800 norovirus gene copies/100 mL (average ± standard deviation, 100% of samples positive); 340 ± 2200 enterococci CFU/100 mL (94%), 71 ± 97 Shigella sp. CFU/100 mL (97%), 60 ± 360 E. coli CFU/100 mL (89%), 35 ± 79 adenovirus gene copies/100 mL (100%), and 21 ± 46 Salmonella sp. CFU/100 mL (76%). The QMRA revealed unacceptable probabilities of illness (>1 in 10,000 illness level) from the four exposure routes considered (drinking water, or only showering, tooth brushing, and rinsing vegetables consumed raw). Disease severity indices based on the QMRA combined with mapping the distribution system revealed areas for targeted rehabilitation. The combined intensive sampling, risk assessment and mapping can be used in low- and middle-income countries to target distribution system rehabilitation efforts and improve health outcomes.

Keenjhar Lake in Pakistan provides several ecosystems services including drinking water to millions of people. The seasonal variation (pre-, during, and post-monsoon) in Keenjhar Lake water quality was investigated during nine sampling rounds at ten locations. Water samples (n = 270) were analyzed for As, Cd, Cr(III), Cu, Fe, Hg, Mn, Ni, and Pb using atomic absorption spectrophotometry, Cr(VI) with a UV spectrophotometer, and water quality parameters (solids, oxygen, conductivity, and pH). Several elements exceeded WHO or Pakistan safe limits for unrestricted use including As (0.038 mg·L−1, 95th upper confidence limit), Cr(III) (0.15 mg·L−1), Cd (0.11 mg·L−1), Fe (0.42 mg·L−1), Ni (0.037 mg·L−1), and Pb (0.04 mg·L−1). Monsoon levels of chromium, Cu, Fe, Mn, and Pb were higher than other seasons, while As was highest during post-monsoon, and Ni was highest during pre and post-monsoon. There were significant spatial differences in metals concentrations in the lake. As, Cu, and Fe significantly declined in concentration with distance from the lake inlet. Using the lake as a source of drinking water presented unacceptable carcinogenic risk and non-carcinogenic hazards. This study suggests measures should be taken to limit discharge of pollution to the lake and future monitoring should consider seasonal monsoon effects on lake quality.

The Keenjhar Lake in the southern part of Pakistan is exposed to heavy metal contamination, mainly due to the lack of proper upstream industrial effluent treatment. In this study, accumulation of heavy metals in five frequently consumed fish species Cirrhinus mrigala (Morakhi), Labeo rohita (Kur’ro), Solea solea (Mundi), Sperata seenghala (Singharo), and Anguilla bengalensis (Baam) from Keenjhar Lake were investigated. A total of 30 fish samples were collected from fishermen at the lake. Fish tissues were digested using standard methods and analyzed for metals using an atomic absorption spectrophotometer (AAS). Heavy metal (Cu, Ni, Pb, total Cr, Cr(VI), and Cd) concentrations were detected both in fish muscles and liver. Among the five targeted species, Cirrhinus mrigala appeared to be the most bioaccumulative fish, with Cr (219.78) > Pb (34.20) > Cu (22.49) > Ni ((16.24) > Cr(VI) (15.25) > Cd (0.01). The target hazard quotient (THQ) and hazard index (HI) were calculated to assess human health risk. The concentration of metals showed a significant difference among fish muscles and liver in the three studied locations. The total HI for heavy metals were estimated to be less than 1, but the cancer risk (CR) for Pb and Cr(VI) exceeded acceptable limits for fish consumption. From the human health perspective, this study informs local populations, i.e., fishing communities and population consuming approximately 150 g/day of Keenjhar Lake fish regularly are chronically exposed to Pb and Cr(VI) contamination with (10ˉ4 to 10ˉ6) carcinogenic risks.

Millions of tons of fecal-contaminated poultry litter are applied to U.S. agricultural fields annually. Precipitation and irrigation facilitate transport of fecal-derived pathogens and fecal indicator bacteria (FIB) to groundwater. The goal of this study was to compare transport of pathogens, FIB, and a microbial source tracking marker gene for poultry litter (LA35) in a simulated soil-to-groundwater system. Nine laboratory soil columns containing four different soil types were used to evaluate microbial transport to groundwater via infiltration. Quantitative polymerase chain reaction was used to monitor Salmonella enterica Typhimurium, Escherichia coli, Enterococcus spp., Brevibacterium sp. LA35 and Bacteroidales leached from soil columns inoculated with poultry litter. S. enterica was correlated with LA35 poultry litter marker gene and FIB concentrations in column soils containing organic matter, but not in acid washed sands. In contrast, S. enterica was found to correlate with LA35 and FIB in the leachate from columns containing sand, but not with leachate from organic soil columns. The majority of recovered DNA was found in leachate of predominately sandy soil columns, and in the soil of loamy columns. At least 90% of the DNA retained in soils for each microbial target was found in the top 3cm of the column. These studies suggest that poultry litter associated pathogens and FIB are rapidly released from litter, but are influenced by complex attenuation mechanisms during infiltration, including soil type. This study advances our understanding of the potential for subsurface transport of poultry litter associated pathogens and FIB, and support the use of the LA35 marker gene for evaluating poultry litter impacts on groundwater.

This study aimed to evaluate the microbiological quality/safety of fresh produce from farmers’ markets (FM) and assess the post-harvest washing practice with antimicrobials to inactivate Salmonella and Listeria monocytogenes on fresh produce. In study I, 212 produce samples were tested for the presence of Salmonella and Listeria spp. using modified FDA-BAM methods. Aerobic plate counts (APCs), total coliforms (TCCs), and yeast/molds were analyzed on petri-films. Among the 212 samples, the APCs, TCCs, and yeast/molds were 3.72–5.63, 3.67–5.47, and 3.07–4.13 log CFU/g, respectively, with spinach containing the highest ( P Salmonella enterica spp. enterica was detected on 18.6% of spinach, 10.9% of tomatoes, 18.5% of peppers, and 56.3% of cantaloupes, which is much higher than previous reported. Only 3.78% of the samples were confirmed for Listeria spp., and 50% of them were identified as L. monocytogenes , based on multiplex PCR results. Due to the high percentage of pathogens detected on the farmers marker produce an evaluation of post-harvest produce washing with various antimicrobials was conducted in study II. Specifically, spinach, tomatoes, green peppers and cucumbers were inoculated with S . Typhimurium and Tennessee or L. monocytogenes and washed in tap water, vinegar water (10%), lactic acid (5%), a lactic and citric acid blend (2.5%), and sodium hypochlorite (200 ppm) for 30 s or unwashed. Vinegar water (10%) showed better ( P S. Typhimurium and Tennessee on tomatoes and cucumbers, and L. monocytogenes on tomatoes and peppers than tap water. The three antimicrobials reached an additional reduction level of 0.9–2.7 ( S. Typhimurium and Tennessee) and 0.2 to 1.4 log CFU/g ( L . monocytogenes ) compared to tap water. Lactic acid indicated the best ( P S. Typhimurium and Tennessee on spinach and green peppers, and sodium hypochlorite showed the best ( P L. monocytogenes on cucumbers. The results supplied important information for FM vendors to develop post-harvest protocols to control foodborne pathogens.

There are an estimated 30,000 chemicals in commerce used in quantities >1016kg per year in the US. Unfortunately there is limited information on the chemicals partitioning and reactivity properties. These orphaned or understudied chemicals are viewed as non-hazardous but can still pose serious economic, health, environmental and societal impacts as evidenced by the January 2014 spill of 37,900L of crude-MCHM (primarily 4-methylcyclohexanemethanol) and stripped-PPH (primarily dipropylene glycol phenyl ether and propylene glycol phenyl ether) into the Elk River near Charleston, WV. Using the Elk River spill as a case study of orphaned or understudied chemicals, experiments were undertaken to evaluate the adequacy of standard approaches to emergency drinking water treatment (oxidation and sorption to activated carbons). Further available sorption models for estimating the potential of these compounds to sorb to Elk River sediments and to activated carbons in drinking water systems were investigated. The results showed that powdered activated carbon (PAC) was the most effective sorbent. The trans-MCHM isomer was found to preferentially sorb compared to cis-MCHM. For MCHM concentrations ranging from 2 to 5mgL-1 in the treatment plant, PAC concentrations of 0.1-1.4mgL-1 would be required to lower both MCHM isomers to the CDC screening level. In most cases, published linear solvation energy relationships and quantitative structure activity relationships were inadequate to estimate the a priori likelihood of sorption of MCHM and PPH to sediments and GAC, but did fit the PAC results well. Permanganate and chlorine oxidation of the compounds showed limited to insignificant removal. The studies presented herein suggest that there are still inherent vulnerabilities to drinking water treatment systems that need to be addressed.

The kinetics and microbial ecology in sulfidogenic bioreactors used in a novel two-stage process for co-treatment of acid mine drainage (AMD) and municipal wastewater (MWW) were investigated. Michaelis–Menten modeling of COD oxidation by sulfate reducing bacteria (SRB) (Vmax = 0.33 mg L−1 min−1, Km = 4.3 mg L−1) suggested that the Vmax can be reasonably achieved given the typical COD values in MWW and anticipated mixing with AMD. Non-competitive inhibition modeling (Ki = 6.55 mg L−1) indicated that excessive iron level should be avoided to limit its effects on SRB. The COD oxidation rate was positively correlated to COD/sulfate ratio and SRB population, as evidenced by dsrA gene copies. Phylogenetic analysis revealed diverse microbial communities dominated by sulfate reducing delta-proteobacteria. Microbial community and relative quantities of SRB showed significant differences under different COD/sulfate ratios (0.2, 1 and 2), and the highest dsrA gene concentration and most complex microbial diversity were observed under COD/sulfate ratio 2. Major species were associated with Desulfovirga, Desulfobulbus, Desulfovibrio, and Syntrophus sp. The reported COD kinetics, SRB abundances and the phylogenetic profile provide insights into the co-treatment process and help identify the parameters of concerns for such technology development.

Pathogen identification and microbial source tracking (MST) to identify sources of fecal pollution improve evaluation of water quality. They contribute to improved assessment of human health risks and remediation of pollution sources. An MST microarray was used to simultaneously detect genes for multiple pathogens and indicators of fecal pollution in freshwater, marine water, sewage-contaminated freshwater and marine water, and treated wastewater. Dead-end ultrafiltration (DEUF) was used to concentrate organisms from water samples, yielding a recovery efficiency of >95% for Escherichia coli and human polyomavirus. Whole-genome amplification (WGA) increased gene copies from ultrafiltered samples and increased the sensitivity of the microarray. Viruses (adenovirus, bocavirus, hepatitis A virus, and human polyomaviruses) were detected in sewage-contaminated samples. Pathogens such as Legionella pneumophila , Shigella flexneri , and Campylobacter fetus were detected along with genes conferring resistance to aminoglycosides, beta-lactams, and tetracycline. Nonmetric dimensional analysis of MST marker genes grouped sewage-spiked freshwater and marine samples with sewage and apart from other fecal sources. The sensitivity (percent true positives) of the microarray probes for gene targets anticipated in sewage was 51 to 57% and was lower than the specificity (percent true negatives; 79 to 81%). A linear relationship between gene copies determined by quantitative PCR and microarray fluorescence was found, indicating the semiquantitative nature of the MST microarray. These results indicate that ultrafiltration coupled with WGA provides sufficient nucleic acids for detection of viruses, bacteria, protozoa, and antibiotic resistance genes by the microarray in applications ranging from beach monitoring to risk assessment.

Approximately 242 million kg of herbicides are applied in the United States (US) annually. While herbicides prevent the growth of weeds, they can damage crops resulting in so called “herbicide injury.” To increase herbicidal tolerance of crops, herbicide safeners are commonly added to formulations. Herbicidal safeners are labeled as inert ingredients in herbicide formulations and typically minimal fate and transport information is available. The objective of this study is to experimentally derive the aqueous solubility, octanol-water partition coefficient, and photo-degradation and permanganate oxidation of two dichloroacetamide safeners, benoxacor and furilazole. These results can be used to understand the environmental chemodynamics of these compounds and their degradation via common drinking water treatment systems containing permanganate. The aqueous solubility of benoxacor and furilazole was 22 and 235 mg L−1 respectively while the estimated solubility from EPI Suite was 102.7 and 255 mg L−1 respectively. The log octanol-water partition coefficient of benoxacor and furilazole was 2.23 and 1.96 respectively. The selected safeners were not photodegradable to any great extent. Both benoxacor and furilazole oxidation were found to be second order with respect to initial KMnO4 concentrations. The results of this study can be used to predict distribution of safeners after application in agricultural fields and aid in the design of treatment systems.

Fecal indicator bacteria (FIB) are the basis for water quality regulations and are considered proxies for waterborne pathogens when conducting human health risk assessments. The direct detection of pathogens in water and simultaneous identification of the source of fecal contamination are possible with microarrays, circumventing the drawbacks to FIB approaches. A multigene target microarray was used to assess the prevalence of waterborne pathogens in a fecally impaired mixed-use watershed. The results indicate that fecal coliforms have improved substantially in the watershed since its listing as a 303(d) impaired stream in 2002 and are now near United States recreational water criterion standards. However, waterborne pathogens are still prevalent in the watershed, as viruses (bocavirus, hepatitis E and A viruses, norovirus, and enterovirus G), bacteria ( Campylobacter spp., Clostridium spp., enterohemorrhagic and enterotoxigenic Escherichia coli , uropathogenic E. coli , Enterococcus faecalis , Helicobacter spp., Salmonella spp., and Vibrio spp.), and eukaryotes ( Acanthamoeba spp., Entamoeba histolytica , and Naegleria fowleri ) were detected. A comparison of the stream microbial ecology with that of sewage, cattle, and swine fecal samples revealed that human sources of fecal contamination dominate in the watershed. The methodology presented is applicable to a wide range of impaired streams for the identification of human health risk due to waterborne pathogens and for the identification of areas for remediation efforts. IMPORTANCE The direct detection of waterborne pathogens in water overcomes many of the limitations of the fecal indicator paradigm. Furthermore, the identification of the source of fecal impairment aids in identifying areas for remediation efforts. Multitarget gene microarrays are shown to simultaneously identify waterborne pathogens and aid in determining the sources of impairment, enabling further focused investigations. This study shows the use of this methodology in a historically impaired watershed in which total maximum daily load reductions have been successfully implemented to reduce risk. The results suggest that while the fecal indicators have been reduced more than 96% and are nearing recreational water criterion levels, pathogens are still detectable in the watershed. Microbial source tracking results show that additional remediation efforts are needed to reduce the impact of human sewage in the watershed.

Pathogen detection and the identification of fecal contamination sources are challenging in environmental waters. Factors including pathogen diversity and ubiquity of fecal indicator bacteria hamper risk assessment and remediation of contamination sources. A custom microarray targeting pathogens (viruses, bacteria, protozoa), microbial source tracking (MST) markers, and antibiotic resistance genes was tested against DNA obtained from whole genome amplification (WGA) of RNA and DNA from sewage and animal (avian, cattle, poultry, and swine) feces. Perfect and mismatch probes established the specificity of the microarray in sewage, and fluorescence decrease of positive probes over a 1:10 dilution series demonstrated semiquantitative measurement. Pathogens, including norovirus, Campylobacter fetus, Helicobacter pylori, Salmonella enterica, and Giardia lamblia were detected in sewage, as well as MST markers and resistance genes to aminoglycosides, beta-lactams, and tetracycline. Sensitivity (percentage true positives) of MST results in sewage and animal waste samples (21-33%) was lower than specificity (83-90%, percentage of true negatives). Next generation DNA sequencing revealed two dominant bacterial families that were common to all sample types: Ruminococcaceae and Lachnospiraceae. Five dominant phyla and 15 dominant families comprised 97% and 74%, respectively, of sequences from all fecal sources. Phyla and families not represented on the microarray are possible candidates for inclusion in subsequent array designs.

Water quality monitoring techniques that target microorganisms in the order Bacteroidales are potential alternatives to conventional methods for detection of fecal indicator bacteria. Bacteroidales and members of the genus Bacteroides have been the focus of microbial source tracking (MST) investigations for discriminating sources of fecal pollution (e.g., human or cattle feces) in environmental waters. For accurate source apportionment to occur, one needs to understand both the abundance of Bacteroides in host feces and the survival of these host-associated microbial markers after deposition in the environment. Studies were undertaken to evaluate the abundance, persistence, and potential for growth of Bacteroidales originating from poultry litter under oxic and anoxic environmental conditions. Bacteroidales abundance, as determined by quantitative PCR (qPCR) with GenBac primers and probe, increased 2 to 5 log gene copies ml −1 and 2 log gene copies g litter −1 under most conditions during incubation of poultry litter in a variety of laboratory microcosm and field mesocosm studies. DNA sequencing of the Bacteroidales organisms in the litter identified taxa with sequences corresponding exactly to the GenBac primer and probe sequences and that were closely related to Bacteroides uniformis , B. ovatus , and B. vulgatus . These results suggest that MST studies using qPCR methods targeting Bacteroidales in watersheds that are affected by poultry litter should be interpreted cautiously. Growth of Bacteroidales originating from poultry litter in environmental waters may occur while Bacteroidales growth from other fecal sources declines, thus confounding the interpretation of MST results.

The insensitive munition, IMX-101 approved for use in the USA, contains 2,4-dinitroanisole (DNAN), 3-nitro-1,2,4-triazol-5-one (NTO), and nitroguanidine (NQ) and is designed to be less sensitive to shock and sympathetic detonation. Given the estimated future use of IMX-101, an understanding of IMX-101 constituent attenuation mechanisms on testing and training ranges is needed. Studies were conducted to determine (1) the rates of IMX-101 fragment dissolution during simulated rainfall, (2) DNAN and NTO soil sorption coefficients, (3) ability of grasses to germinate in and phytoremediate IMX-101 contaminated soil, and (4) effect of the addition of IMX-101 degrading enrichment cultures on phytoremediation. The IMX-101 particles were found to dissolve slowly under simulated rainfall conditions with NQ and NTO dissolving first, leaving DNAN crystals. DNAN and NTO sorption to soils fit Freundlich isotherms and limited desorption was observed. DNAN and NQ were shown to be taken up into the roots and shoots of a mixture of big bluestem grass (Andropogon gerardii), Nash Indiangrass (Sorghastrum nutans), and switchgrass (Panicum virgatum) during phytoremediation of soils contaminated with up to 50 mg kg−1 IMX-101. Complete degradation of IMX-101 to below detection limits occurred over 225 days. The addition of an IMX-101 degrading enrichment culture to the treatments significantly increased the root and shoot mass.

Defense agencies are increasingly using insensitive munitions (IM) in place of explosives such as 2,4,6-trinitrotoluene. In this study simultaneous aerobic degradation of the IMX-101 formulation constituents 2,4-dinitroanisole (DNAN), 3-nitro-1,2,4-triazol-5-one (NTO), and nitroguanidine (NQ) was observed and degradation products were examined. Degradation products over four days of incubation included: nitrourea, 1,2-dihydro-3H-1,2,4-triazol-3-one, and 2,4-dinitrophenol. The enrichment culture maximum specific growth rate of 0.12 h −1 and half saturation constant of 288 mg L −1 during degradation of IMX-101 as a sole nitrogen source suggest that enrichment culture growth kinetics may closely relate to those of other explosive and nitroaromatic compounds.

A mixture of coal processing agents, namely crude-MCHM (primarily 4-methylcyclohexanemethanol) and stripped-PPH (primarily dipropylene glycol phenyl ether and propylene glycol phenyl ether) spilled into the Elk River near Charleston, West Virginia in January 2014. The spill resulted in a do-not-use ban on drinking water for up to 10 days. Studies were undertaken to determine the potential for aerobic biodegradation of crude-MCHM and PPH in the riverine system and in the sewage treatment plant postflushing of affected water lines. Complete biotransformation of crude-MCHM occurred only when MCHM was initially

Chemical spills and accidents contaminate the environment and disrupt societies and economies around the globe. In the United States there were approximately 172,000 chemical spills that affected US waterbodies from 2004 to 2014. More than 8000 of these spills involved non-petroleum-related chemicals. Traditional emergency responses or incident command structures (ICSs) that respond to chemical spills require coordinated efforts by predominantly government personnel from multiple disciplines, including disaster management, public health, and environmental protection. However, the requirements of emergency response teams for science support might not be met within the traditional ICS. We describe the US ICS as an example of emergency-response approaches to chemical spills and provide examples in which external scientific support from research personnel benefitted the ICS emergency response, focusing primarily on nonpetroleum chemical spills. We then propose immediate, near-term, and long-term activities to support the response to chemical spills, focusing on nonpetroleum chemical spills. Further, we call for science support for spill prevention and near-term spill-incident response and identify longer-term research needs. The development of a formal mechanism for external science support of ICS from governmental and nongovernmental scientists would benefit rapid responders, advance incident- and crisis-response science, and aid society in coping with and recovering from chemical spills.

Aim To validate the distribution of a poultry litter-specific marker gene in faecally contaminated environmental waters of an intensive poultry litter rearing region. Methods and Results A TaqMan®-based qPCR assay for Brevibacterium sp. LA35 16S rRNA (LA35 gene), which was previously shown to be associated with poultry litter and faeces, was tested on 126 nontarget faecal samples and 28 poultry litter and faecal samples. The TaqMan assay was sensitive (76%) and specific (100%) to the LA35 gene and exhibited a detection limit for poultry litter in water samples that is sufficiently low (2·5 × 10−2 mg litter l−1) to be applicable for environmental monitoring. The LA35 gene was detected in 43% of water samples (n = 30) collected in an intensive poultry rearing region of West Virginia which drains to the Chesapeake Bay. Conclusions The poultry-specific TaqMan qPCR method for the LA35 gene is more specific than previously published methods and can be used to identify regions impacted by poultry rearing activities. Significance and Impact of the Study The LA35 gene appears to have a broad geographical distribution as it has been found in poultry litter and faeces from Delaware and West Virginia, in this study and from Arkansas, Georgia, Florida, Minnesota, Oklahoma and Utah previously.

N-Nitrosodimethylamine (NDMA) is an emerging contaminant of concern. N-nitrodimethylamine (DMNA) is a structural analog to NDMA. NDMA and DMNA have been found in drinking water, groundwater, and other media and are of concern due their toxicity. The authors evaluated biotransformation of NDMA and DMNA by cultures enriched from contaminated groundwater growing on benzene, butane, methane, propane, or toluene. Maximum specific growth rates of enriched cultures on butane (μmax = 1.1 h−1) and propane (μmax = 0.65 h−1) were 1 to 2 orders of magnitude higher than those presented in the literature. Growth rates of mixed cultures grown on benzene (μmax = 1.3 h−1), methane (μmax = 0.09 h−1), and toluene (μmax = 0.99 h−1) in these studies were similar to those presented in the literature. NDMA biotransformation rates for methane oxidizers (υmax = 1.4 ng min−1 mg−1) and toluene oxidizers (υmax = 2.3 ng min−1 mg−1) were comparable to those presented in the literature, whereas the biotransformation rate for p...

Afieldpilottestinwhichhydraulicfracturingwasusedtoemplacegranularremediationamendment (a mixture of zero-valent iron [ZVI] and organic carbon) into fine-grained sandstone to remediate dissolved trichloroethene (TCE)-contaminated groundwater was performed at a former intercontinental ballistic missile site in Colorado. Hydraulic fracturing was used to enhance the permeability of the aquifer with concurrent emplacement of amendment that facilitates TCE degradation. Geophysical monitoring and inverse modeling show that the network of amendment-filled fractures extends throughout the aquifer volume targeted in the pilot test zone. Two years of subsequent groundwater monitoring demonstrate that amendment addition resulted in development of geochemical conditions favorable to both abiotic and biological TCE degradation, that TCE concentrations were substantially reduced (i.e., greater than 90 percent reduction in TCE mass), and that the primary degradation processes are likely abiotic. The pilot-test data aided in re-evaluating the conceptual site model and in designing the full-scale remedy to address a larger portion of the TCE-contaminated groundwater plume. O c 2012 Wiley Periodicals, Inc.

Identification of bacteria that can utilize a wide range of nitroaromatic compounds will allow development of more effective biological treatment methods in industrial wastewater treatment processes and environmental remediation efforts. A new strain of Rhodococcus opacus capable of mineralizing or transforming nitroaromatic and nitramine compounds of importance was isolated. Specifically, the bacterium were found to utilize 2,4,6-trinitrophenol (TNP) as a sole carbon and nitrogen source and have a yield coefficient of 0.16 g cells-N/g TNP-N. The Edwards model was found to provide the best fit to the data and the estimated inhibited growth parameters μmax , KS , and KI were 0.58 h−1 , 25, and 112 mg/L , respectively. It was found that the TNP-degraders could degrade 2,4-dinitrophenol as the sole carbon and nitrogen source and utilize 1,3,5-trinitrobenzene as the sole nitrogen source. Additionally, the results show that the isolates are able to cometabolize 2,4,6-trinitrotoluene (TNT), hexahydro-1,3,5-tri...

A bench scale sequencing batch reactor (SBR) was designed and tested for degradation of 2,4,6-trinitrophenol (TNP) or picric acid-contaminated wastewater or groundwater. Under varying temperatures (25, 15 and 10°C) and influent concentrations (40–200 mg/L TNP) a stable biomass was developed that was consistently capable of degrading the explosive compound to below regulatory drinking water limits (0.057 mg/L). The reactor was initially seeded with a nitroaromatic and nitramine degrading isolate Rhodococcus opacus strain JW01. Kinetic growth modeling was conducted revealing µmax values at 25, 15 and 10°C of 0.14, 0.08 and 0.04 d−1, while the modeled Ks values were 0.68, 1.11 and 1.24 mg/L, respectively. Overall TNP removal efficiency in the SBR was on average > 99.9% over the 2000 hours of operation. Removal of TNP to below drinking water standards, with low residual dissolved carbon and significant release of nitrogen from the parent compound was accomplished. Biotechnol. Bioneg. 2007;97: 1408–1414. © 2007 Wiley Periodicals, Inc.

The determination of fecal pollution sources in waters is essential in the management of catchments. Although traditional microbial water analyses using indicator microorganisms have been used for water-health management for more than a century, it is known that they cannot provide information about the origin of fecal pollution. The distinction between anthropogenic and non-anthropogenic (animal) fecal pollution in water would greatly support assessment of health risks based on knowledge of the host-specificity of many pathogens. For example, human sewage could constitute a higher health risk to humans than wastewater of animal origin. However, there are some exceptions because some pathogens can infect and cause clinical disease in both humans and animals. Therefore, the fecal pollution source assessment could support different water management strategies, treatment measures and policies to prevent or decrease fecal inputs in water and remediation at the source. In this chapter, proposed chemical and biological MST indicators for the determination of animal fecal sources are discussed. The biological indicators are grouped based on the phylogenetic description of the proposed target (eukarya, bacteria, and virus). A comprehensive description for each proposed target is provided and the developed methodologies employed are presented. Emphasis is placed on the validation and applicability of each proposed method and animal-MST indicator. New molecular approaches for animal-NST targets based on metagenomics are also presented. Finally, MST assay implementation, their contribution to the assessment of maximum fecal load of water bodies and their relationship to traditional microbial indicators and water-borne pathogens is examined.

Disposal of fecally contaminated poultry litter by land application can deliver pathogens and fecal indicator bacteria (FIB) into receiving waters via runoff. While water quality is regulated by FIB enumeration, FIB testing provides inadequate information about contamination source and health risk. This microbial source tracking (MST) study compared the persistence of the Brevibacterium sp. strain LA35 16S rRNA gene (marker) for poultry litter with that of pathogens and FIB under outdoor, environmentally relevant conditions in freshwater, marine water, and sediments over 7 days. Salmonella enterica , Campylobacter jejuni , Campylobacter coli , Bacteroidales , and LA35 were enumerated by quantitative PCR (qPCR), and Enterococcus spp. and E. coli were quantified by culture and qPCR. Unlike the other bacteria, C. jejuni was not detectable after 48 h. Bacterial levels in the water column consistently declined over time and were highly correlated among species. Survival in sediments ranged from a slow decrease over time to growth, particularly in marine microcosms and for Bacteroidales. S. enterica also grew in marine sediments. Linear decay rates in water ( k ) ranged from −0.17 day −1 for LA35 to −3.12 day −1 for C. coli . LA35 levels correlated well with those of other bacteria in the water column but not in sediments. These observations suggest that, particularly in the water column, the fate of LA35 in aquatic environments is similar to that of FIB, C. coli , and Salmonella , supporting the hypothesis that the LA35 marker gene can be a useful tool for evaluating the impact of poultry litter on water quality and human health risk.

Aims To determine whether poultry litter marker gene LA35 is correlated with pathogens and fecal indicator bacteria (FIB) in run-off from poultry litter-amended plots. Methods and Results A rainfall simulator with various vegetative filter strip lengths was employed to evaluate the correlation of a microbial source tracking (MST) marker for poultry feces/litter (the 16S rRNA gene of Brevibacterium sp. LA35 [LA35] measured by quantitative PCR) with pathogens and FIB in run-off. LA35 was correlated with Staphylococcus aureus, Escherichia coli, Enterococcus spp. and Bacteroidales levels. Salmonella was present at low concentration in litter, but became undetectable by qPCR in run-off. Escherichia coli, LA35 and Staph. aureus exhibited mass-based first flush behaviour in the run-off. Conclusions Correlation of LA35 with FIB and pathogens in run-off from poultry litter-amended fields suggest comparable transport mechanisms and that LA35 is a useful tracer for harmful bacteria in the environment released from poultry litter. Significance and Impact of the Study To protect human health, an effective marker for poultry fecal contamination should exhibit similar fate and transport characteristics compared to pathogens. This study is among the first to demonstrate such a relationship in run-off for a MST marker.

Aerobic biotransformation of N-nitrosodimethylamine (NDMA), an emerging contaminant of concern, and its structural analog N-nitrodimethylamine (DMN), was evaluated in benzene and methane amended groundwater passed through laboratory scale soil columns. Competitive inhibition models were used to model the kinetics for NDMA and DMN cometabolism accounting for the concurrent degradation of the growth and cometabolic substrates. Transformation capacities for NDMA and DMN with benzene (13 and 23 μg (mg cells) − 1 ) and methane (0.14 and 8.4 μg (mg cells) − 1 ) grown cultures, respectively are comparable to those presented in the literature, as were first order endogenous decay rates estimated to be 2.1 × 10 − 2 ± 1.7 × 10 − 3 d − 1 and 6.5 × 10 − 1 ± 7.1 × 10 − 1 d − 1 for the methane and benzene amended cultures, respectively. These studies highlight possible attenuation mechanisms and rates for NDMA and DMN biotransformation in aerobic aquifers undergoing active remediation, natural attenuation or managed aquifer recharge with treated wastewater (i.e., reclaimed water).

The impact of fecal contamination from human and agricultural animal waste on water quality is a major public health concern. Identification of the dominant source(s) of fecal pollution in a watershed is necessary for assessing the safety of recreational water and protecting water resources. A field study was conducted using quantitative PCR (qPCR) for the 16S rRNA gene of Brevibacterium sp. LA35 to track feces-contaminated poultry litter in environmental samples. Based on sensitivity and specificity characteristics of the qPCR method, the Bayesian conditional probability that detection of the LA35 marker gene in a water sample represented a true-positive result was 93%. The marker's covariance with fecal indicator bacteria (FIB) and metals associated with poultry litter was also assessed in litter, runoff, surface water, and groundwater samples. LA35 was detected in water and soil samples collected throughout the watershed, and its concentration covaried with concentrations of Escherichia coli , enterococci, As, Cu, P, and Zn. Significantly greater concentrations of FIB, As, Cu, P, and Zn were observed in edge-of-field runoff samples in which LA35 was detected, compared to samples in which it was not detected. Furthermore, As, Cu, P, and Zn concentrations covaried in environmental samples in which LA35 was detected and typically did not in samples in which the marker gene was not detected. The covariance of the poultry-specific LA35 marker gene with these known contaminants from poultry feces provides further evidence that it is a useful tool for assessing the impact of poultry-derived fecal pollution in environmental waters.

Aim: To identify a DNA sequence specific to a bacterium found in poultry litter that was indicative of faecal contamination by poultry sources. Methods and Results: Faecally contaminated poultry litter and soils were used as source material for the development of a quantitative polymerase chain reaction (qPCR) method targeting the 16S rRNA gene of a Brevibacterium sp. The identified sequence had 98% nucleotide identity to the 16S rRNA gene of Brevibacterium avium. The qPCR method was tested on 17 soiled litter samples; 40 chicken faecal samples; and 116 nontarget faecal samples from cattle, swine, ducks, geese, and human sewage collected across the United States. The 571-bp product was detected in 76% of poultry-associated samples, but not in 93% of faecal samples from other sources. Marker concentrations were 107–109 gene copies per gram in soiled litter, up to 105 gene copies per gram in spread-site soils, and 107 gene copies per litre in field run-off water. Results were corroborated by a blinded study conducted by a second laboratory. Conclusion: The poultry-specific PCR product is a useful marker gene for assessing the impact of faecal contamination as a result of land-applied poultry litter. Significance and Impact of the Study: This study describes the first quantitative, sensitive and specific microbial source tracking method for the detection of poultry litter contamination.