Loop-mediated isothermal amplification: Development, validation and application of simple and rapid assays for quantitative detection of species of Arcobacteraceae family- and species-specific Aliarcobacter faecis and Aliarcobacter lanthieri.

AIM: The family Arcobacteraceae formerly genus Arcobacter has recently been reclassified into six genera. Among nine species of the genus Aliarcobacter, Aliarcobacter faecis and Aliarcobacter lanthieri have been identified as emerging pathogens potentially cause health risks to humans and animals. This study was designed to develop/optimize, validate and apply Arcobacteraceae family- and two species-specific (A. faecis and A. lanthieri) loop-mediated isothermal amplification (LAMP) assays to rapidly detect and quantify total number of cells in various environmental niches. METHODS AND RESULTS: Three sets of LAMP primers were designed from conserved and variable regions of 16S rRNA (family-specific) and gyrB (species-specific) genes. Optimized Arcobacteraceae family-specific LAMP assay correctly amplified and detected 24 species, whereas species-specific LAMP assays detected A. faecis and A. lanthieri reference strains as well as 91 pure and mixed culture isolates recovered from aquatic and faecal sources. The specificity of LAMP amplification of A. faecis and A. lanthieri was further confirmed by restriction fragment length polymorphism analysis. Assay sensitivities were tested using variable DNA concentrations extracted from simulated target species cells in an autoclaved agricultural water sample by achieving a minimum detection limit of 10 cells mL-1 (10 fg). Direct DNA-based quantitative detection, from agricultural surface water, identified A. faecis (17%) and A. lanthieri (1%) at a low frequency compared to family-level (93%) with the concentration ranging from 2·1 × 101 to 2·2 × 105 cells 100 mL-1 . CONCLUSIONS: Overall, these three DNA-based rapid and cost-effective novel LAMP assays are sensitive and can be completed in less than 40 min. They have potential for on-site quantitative detection of species of family Arcobacteraceae, A. faecis and A. lanthieri in food, environmental and clinical matrices. SIGNIFICANCE AND IMPACT OF THE STUDY: The newly developed LAMP assays are specific, sensitive, accurate with higher reproducibility that have potential to facilitate in a less equipped lab setting and can help in early quantitative detection and rate of prevalence in environmental niches. The assays can be adopted in the diagnostic labs and epidemiological studies.

Chlorophyll-a, dissolved organic carbon, turbidity and other variables of ecological importance in river basins in southern Ontario and British Columbia, Canada.

Optical sensing of chlorophyll-a (chl-a), turbidity, and fluorescent dissolved organic matter (fDOM) is often used to characterize the quality of water. There are many site-specific factors and environmental conditions that can affect optically sensed readings; notwithstanding the comparative implication of different procedures used to measure these properties in the laboratory. In this study, we measured these water quality properties using standard laboratory methods, and in the field using optical sensors (sonde-based) at water quality monitoring sites located in four watersheds in Canada. The overall objective of this work was to explore the relationships among sonde-based and standard laboratory measurements of the aforementioned water properties, and evaluate associations among these eco-hydrological properties and land use, environmental, and ancillary water quality variables such as dissolved organic carbon (DOC) and total suspended solids (TSS). Differences among sonde versus laboratory relationships for chl-a suggest such relationships are impacted by laboratory methods and/or site specific conditions. Data mining analysis indicated that interactive site-specific factors predominately impacting chl-a values across sites were specific conductivity and turbidity (variables with positive global associations with chl-a). The overall linear regression predicting DOC from fDOM was relatively strong (R2 = 0.77). However, slope differences in the watershed-specific models suggest laboratory DOC versus fDOM relationships could be impacted by unknown localized water quality properties affecting fDOM readings, and/or the different standard laboratory methods used to estimate DOC. Artificial neural network analyses (ANN) indicated that higher relative chl-a concentrations were associated with low to no tree cover around sample sites and higher daily rainfall in the watersheds examined. Response surfaces derived from ANN indicated that chl-a concentrations were higher where combined agricultural and urban land uses were relatively higher.

Enhanced Single-tube Multiplex PCR Assay for Detection and Identification of Six Arcobacter Species.

AIM: A single-tube multiplex PCR (mPCR) assay was developed for rapid, sensitive and simultaneous detection and identification of six Arcobacter species including two new species, A. lanthieri and A. faecis, along with A. butzleri, A. cibarius, A. cryaerophilus and A. skirrowii on the basis of differences in the lengths of their PCR products. Previously designed monoplex, mPCR and RFLP assays do not detect or differentiate A. faecis and A. lanthieri from other closely related known Arcobacter spp. METHODS AND RESULTS: Primer pairs for each target species (except A. skirrowii) and mPCR protocol were newly designed and optimized using variable regions of housekeeping including cpn60, gyrA, gyrB and rpoB genes. The accuracy and specificity of the mPCR assay was assessed using DNA templates from six targets and 11 other Arcobacter spp. as well as 50 other bacterial reference species and strains. Tests on the DNA templates of target Arcobacter spp. were appropriately identified, whereas all 61 other DNA templates from other bacterial species and strains were not amplified. Sensitivity and specificity of the mPCR assay was 10 pg µl-1 of DNA concentration per target species. The optimized assay was further evaluated, validated and compared with other mPCR assays by testing Arcobacter cultures isolated from various faecal and water sources. CONCLUSIONS: Study results confirm that the newly developed mPCR assay is rapid, accurate, reliable, simple, and valuable for the simultaneous detection and routine diagnosis of six human- and animal-associated Arcobacter spp. SIGNIFICANCE AND IMPACT OF THE STUDY: The new mPCR assay is useful not only for pure but also mixed cultures. Moreover, it has the ability to rapidly detect six species which enhances the value of this technology for aetiological and epidemiological studies.

Using AnnAGNPS to Predict the Effects of Tile Drainage Control on Nutrient and Sediment Loads for a River Basin.

Controlled tile drainage (CTD) can reduce pollutant loading. The Annualized Agricultural Nonpoint Source model (AnnAGNPS version 5.2) was used to examine changes in growing season discharge, sediment, nitrogen, and phosphorus loads due to CTD for a ∼3900-km agriculturally dominated river basin in Ontario, Canada. Two tile drain depth scenarios were examined in detail to mimic tile drainage control for flat cropland: 600 mm depth (CTD) and 200 mm (CTD) depth below surface. Summed for five growing seasons (CTD), direct runoff, total N, and dissolved N were reduced by 6.6, 3.5, and 13.7%, respectively. However, five seasons of summed total P, dissolved P, and total suspended solid loads increased as a result of CTD by 0.96, 1.6, and 0.23%. The AnnAGNPS results were compared with mass fluxes observed from paired experimental watersheds (250, 470 ha) in the river basin. The "test" experimental watershed was dominated by CTD and the "reference" watershed by free drainage. Notwithstanding environmental/land use differences between the watersheds and basin, comparisons of seasonal observed and predicted discharge reductions were comparable in 100% of respective cases. Nutrient load comparisons were more consistent for dissolved, relative to particulate water quality endpoints. For one season under corn crop production, AnnAGNPS predicted a 55% decrease (CTD) in dissolved N from the basin. AnnAGNPS v. 5.2 treats P transport from a surface pool perspective, which is appropriate for many systems. However, for assessment of tile drainage management practices for relatively flat tile-dominated systems, AnnAGNPS may benefit from consideration of P and particulate transport in the subsurface.

Long-Term Observations of Nitrogen and Phosphorus Export in Paired-Agricultural Watersheds under Controlled and Conventional Tile Drainage.

Controlled tile drainage (CTD) regulates water and nutrient export from tile drainage systems. Observations of the effects of CTD imposed en masse at watershed scales are needed to determine the effect on downstream receptors. A paired-watershed approach was used to evaluate the effect of field-to-field CTD at the watershed scale on fluxes and flow-weighted mean concentrations (FWMCs) of N and P during multiple growing seasons. One watershed (467-ha catchment area) was under CTD management (treatment [CTD] watershed); the other (250-ha catchment area) had freely draining or uncontrolled tile drainage (UCTD) (reference [UCTD] watershed). The paired agricultural watersheds are located in eastern Ontario, Canada. Analysis of covariance and paired tests were used to assess daily fluxes and FWMCs during a calibration period when CTD intervention on the treatment watershed was minimal (2005-2006, when only 4-10% of the tile-drained area was under CTD) and a treatment period when the treatment (CTD) watershed had prolific CTD intervention (2007-2011 when 82% of tile drained fields were controlled, occupying >70% of catchment area). Significant linear regression slope changes assessed using ANCOVA ( ≤ 0.1) for daily fluxes from upstream and downstream monitoring sites pooled by calibration and treatment period were -0.06 and -0.20 (stream water) (negative values represent flux declines in CTD watershed), -0.59 and -0.77 (NH-N), -0.14 and -0.15 (NO-N), -1.77 and -2.10 (dissolved reactive P), and -0.28 and 0.45 (total P). Total P results for one site comparison contrasted with other findings likely due to unknown in-stream processes affecting total P loading, not efficacy of CTD. The FWMC results were mixed and inconclusive but suggest physical abatement by CTD is the means by which nutrient fluxes are predominantly reduced at these scales. Overall, our study results indicate that CTD is an effective practice for reducing watershed scale fluxes of stream water, N, and P during the growing season.

Rainfall-induced runoff from exposed streambed sediments: an important source of water pollution.

When surface water levels decline, exposed streambed sediments can be mobilized and washed into the water course when subjected to erosive rainfall. In this study, rainfall simulations were conducted over exposed sediments along stream banks at four distinct locations in an agriculturally dominated river basin with the objective of quantifying the potential for contaminant loading from these often overlooked runoff source areas. At each location, simulations were performed at three different sites. Nitrogen, phosphorus, sediment, fecal indicator bacteria, pathogenic bacteria, and microbial source tracking (MST) markers were examined in both prerainfall sediments and rainfall-induced runoff water. Runoff generation and sediment mobilization occurred quickly (10-150 s) after rainfall initiation. Temporal trends in runoff concentrations were highly variable within and between locations. Total runoff event loads were considered large for many pollutants considered. For instance, the maximum observed total phosphorus runoff load was on the order of 1.5 kg ha. Results also demonstrate that runoff from exposed sediments can be a source of pathogenic bacteria. spp. and spp. were present in runoff from one and three locations, respectively. Ruminant MST markers were also present in runoff from two locations, one of which hosted pasturing cattle with stream access. Overall, this study demonstrated that rainfall-induced runoff from exposed streambed sediments can be an important source of surface water pollution.

Combined impacts of future climate and land use changes on discharge, nitrogen and phosphorus loads for a Canadian river basin.

Both climate and land use changes can influence water quality and quantity in different ways. Thus, for predicting future water quality and quantity trends, simulations should ideally account for both projected climate and land use changes. In this paper, land use projections and climate change scenarios were integrated with a hydrological model to estimate the relative impact of climate and land use projections on a suite of water quality and quantity endpoints for a Canadian watershed. Climatic time series representing SRES change scenario A2 were generated by downscaling the outputs of the Canadian Regional Climate Model (version 4.1.1) using a combination of quantile-quantile transformation and nearest neighbor search. The SWAT (Soil and Water Assessment Tool) model was used to simulate streamflow, nitrogen and phosphorus loading under different climate and land use scenarios. Results showed that a) climate change will drive up maximum monthly streamflow, nitrate loads, and organic phosphorus loads, while decreasing organic nitrogen and nitrite loads; and b) land use changes were found to drive the same water quality/quantity variables in the same direction as climate change, except for organic nitrogen loads, for which the effects of the two stressors had a reverse impact on loading.

Comprehensive nitrogen budgets for controlled tile drainage fields in eastern ontario, Canada.

Excessive N loading from subsurface tile drainage has been linked to water quality degradation. Controlled tile drainage (CTD) has the potential to reduce N losses via tile drainage and boost crop yields. While CTD can reduce N loss from tile drainage, it may increase losses through other pathways. A multiple-year field-scale accounting of major N inputs and outputs during the cropping season was conducted on freely drained and controlled tile drained agricultural fields under corn ( L.)-soybean [ (L.) Merr.] production systems in eastern Ontario, Canada. Greater predicted gaseous N emissions for corn and soybean and greater observed lateral seepage N losses were observed for corn and soybean fields under CTD relative to free-draining fields. However, observed N losses from tile were significantly lower for CTD fields, in relation to freely drained fields. Changes in residual soil N were essentially equivalent between drainage treatments, while mass balance residual terms were systematically negative (slightly more so for CTD). Increases in plant N uptake associated with CTD were observed, probably resulting in higher grain yields for corn and soybean. This study illustrates the benefits of CTD in decreasing subsurface tile drainage N losses and boosting crop yields, while demonstrating the potential for CTD to increase N losses via other pathways related to gaseous emissions and groundwater seepage.

Measuring and modeling the effects of drainage water management on soil greenhouse gas fluxes from corn and soybean fields.

Controlled tile drainage can boost crop yields and improve water quality, but it also has the potential to increase GHG emissions. This study compared in-situ chamber-based measures of soil CH4, N2O, and CO2 fluxes for silt loam soil under corn and soybean cropping with conventional tile drainage (UTD) and controlled tile drainage (CTD). A semi-empirical model (NEMIS-NOE) was also used to predict soil N2O fluxes from soils using observed soil data. Observed N2O and CH4 fluxes between UTD and CTD fields during the farming season were not significantly different at 0.05 level. Soils were primarily a sink for CH4 but in some cases a source (sources were associated exclusively with CTD). The average N2O fluxes measured ranged between 0.003 and 0.028 kg N ha(-1) day(-1). There were some significantly higher (p ≤ 0.05) CO2 fluxes associated with CTD relative to UTD during some years of study. Correlation analyses indicated that the shallower the water table, the greater the CO2 fluxes. Higher corn plant C for CTD tended to offset estimated higher CTD CO2 C losses via soil respiration by ∼100-300 kg C ha(-1). There were good fits between observed and predicted (NEMIS-NOE) N2O fluxes for corn (R(2) = 0.70) and soybean (R(2) = 0.53). Predicted N2O fluxes were higher for CTD for approximately 70% of the paired-field study periods suggesting that soil physical factors, such as water-filled pore space, imposed by CTD have potentially strong impacts on net N fluxes. Model predictions of daily cumulative N2O fluxes for the agronomically-active study period for corn-CTD and corn-UTD, as a percentage of total N fertilizer applied, were 3.1% and 2.6%, respectively. For predicted N2O fluxes on basis of yield units, indices were 0.0005 and 0.0004 (kg N kg(-1) crop grain yield) for CTD and UTD corn fields, respectively, and 0.0011 and 0.0005 for CTD and UTD soybean fields, respectively.

Impact of riparian zone protection from cattle on nutrient, bacteria, F-coliphage, and loading of an intermittent stream.

This 5-yr study compared, via an upstream-downstream experimental design, nutrient and microbial water quality of an intermittent stream running through a small pasture (∼2.5 animals ha) where cattle are restricted from the riparian zone (restricted cattle access [RCA]) and where cattle have unrestricted access to the stream (unrestricted cattle access [URCA]). Fencing in the RCA excluded pasturing cattle to within ∼3 to 5 m of the stream. Approximately 88% (26/32) of all comparisons of mean contaminant load reduction for lower, higher, and all stream flow conditions during the 5-yr study indicated net contaminant load reductions in the RCA; for the URCA, this percentage was 38% (12/32). For all flow conditions, mean percent load reductions in the RCA for nutrients and bacteria plus F-coliphage were 24 and 23%, respectively. These respective percentages for the URCA were -9 and -57% (positive values are reductions; negative values are increases). However, potentially as a result of protected wildlife habitat in the RCA, the mean percent load reduction for for "all flow" was -321% for the RCA and 60% for the URCA; for , these respective percentages were -209% (RCA) and 73% (URCA). For "all flow" situations, mean load reductions for the RCA were significantly greater ( < 0.1) than those from the URCA for NH-N, dissolved reactive phosphorus (DRP), total coliform, , and . For "high flow" situations, mean load reductions were significantly greater for the RCA for DRP, total coliform, and . For "low flow" conditions, significantly greater mean load reductions were in favor of the RCA for DRP, total P, total coliforms, fecal coliforms, , and . In no case were mean pollutant loads in the URCA significantly higher than RCA pollutant loads. Restricting pasturing livestock to within 3 to 5 m of intermittent streams can improve water quality; however, water quality impairment can occur if livestock have unrestricted access to a stream.

Distribution of selected virulence genes and antibiotic resistance in Enterococcus species isolated from the South Nation River drainage basin, Ontario, Canada.

AIMS: Isolate and characterize water enterococci from the South Nation River drainage basin, an area dominated by agriculture. METHODS AND RESULTS: A total of 1558 enterococci were isolated from 204 water samples from the South Nation River obtained over a 3-year period. PCR was used to identify isolates to the species level and characterize them for carriage of 12 virulence determinants. Antibiotic resistance was evaluated phenotypically. Enterococcus faecalis (36·4%), Enterococcus faecium (9·3%) and Enterococcus durans (8·5%) were the major enterococci species isolated. Enterococci carrying more than two virulence determinants were more frequently detected in the summer (59·6%) than in other seasons (≤ 37·6%). Very few isolates (≤ 2·0%) were resistant to category I antibiotics ciprofloxacin and vancomycin. CONCLUSIONS: Comparison of major water enterococci species with major faecal enterococci species obtained from various host groups (human, domesticated mammals and birds, wildlife) in this drainage basin suggest that water enterococci may have varied faecal origins. The low level of antibiotic resistance among enterococci suggests that dispersion of antibiotic resistance via waterborne enterococci in this watershed is not significant. SIGNIFICANCE AND IMPACT OF THE STUDY: The data obtained in this study suggests that water enterococci in the SNR have a faecal origin and that their potential impact on public health regarding antibiotic resistance and virulence determinants is minimal.

Mosquito Identification From Bulk Samples Using DNA Metabarcoding: a Protocol to Support Mosquito-Borne Disease Surveillance in Canada.

Approximately 80 species of mosquitoes (Diptera: Culicidae) have been documented in Canada. Exotic species such as Aedes albopictus (Skuse) (Diptera: Culicidae) are becoming established. Recently occurring endemic mosquito-borne diseases (MBD) in Canada including West-Nile virus (WNV) and Eastern Equine Encephalitis (EEE) are having significant public health impacts. Here we explore the use of DNA metabarcoding to identify mosquitoes from CDC light-trap collections from two locations in eastern Canada. Two primer pairs (BF2-BR2 and F230) were used to amplify regions of the cytochrome c oxidase subunit I (CO1) gene. High throughput sequencing was conducted using an Illumina MiSeq platform and GenBank-based species identification was applied using a QIIME 1.9 bioinformatics pipeline. From a site in southeastern Ontario, Canada, 26 CDC light trap collections of 72 to >300 individual mosquitoes were used to explore the capacity of DNA metabarcoding to identify and quantify captured mosquitoes. The DNA metabarcoding method identified 33 species overall while 24 species were identified by key. Using replicates from each trap, the dried biomass needed to identify the majority of species was determined to be 76 mg (equivalent to approximately 72 mosquitoes), and at least two replicates from the dried biomass would be needed to reliably detect the majority of species in collections of 144-215 mosquitoes and three replicates would be advised for collections with >215 mosquitoes. This study supports the use of DNA metabarcoding as a mosquito surveillance tool in Canada which can help identify the emergence of new mosquito-borne disease potential threats.

Nitrogen, phosphorus, and bacteria tile and groundwater quality following direct injection of dewatered municipal biosolids into soil.

Application of municipal biosolids (sewage) to agricultural land is a common practice to improve soil physical quality and fertility. The chosen method of land application can have a strong impact on the extent of adjacent water contamination by nutrients and bacteria. Dewatered municipal biosolids (DMB) were applied to silt-clay loam experimental field plots in Ontario, Canada using two application methods: (i) surface spreading followed by shallow incorporation (SS) and (ii) a newly developed implement that directly injects DMB into the topsoil (DI). The objective of this study was to compare N, P, and bacteria quality of tile drainage and shallow groundwater associated with each land application technique. There were no significant differences (P > 0.05) in N, P, and bacteria tile mass loads among the application treatments for time periods <100 d postapplication, when the greatest peak loads and peak tile water concentrations were observed. Both land application treatments caused groundwater Escherichia coli contamination to at least 1.2 m depth below surface after the first postapplication rainfall event, and NO(3)-N contamination to at least 2.0 m depth below surface. The DI treatment did, however, have significantly (P < 0.05) higher tile mass loads of total Kjeldahl N (TKN), total phosphorus (TP), E. coli, Enterococci, and Clostridium perfringens relative to the SS treatment for time periods >100 d postapplication. Nevertheless, relative to tile effluent data collected <100 d postapplication (no application treatment differences), peak loads, and concentrations during this time were, overall, considerably lower for both treatments. This finding, along with no significant differences in N, P, and bacteria groundwater concentrations among the application treatments, and that the direct injection technique could potentially reduce vector attraction problems and odor, suggests that the direct injection technique should be considered a dewatered municipal biosolid land application option.

Do reductions in agricultural field drainage during the growing season impact bacterial densities and loads in small tile-fed watersheds?

Predicting bacterial levels in watersheds in response to agricultural beneficial management practices (BMPs) requires understanding the germane processes at both the watershed and field scale. Controlling subsurface tile drainage (CTD) is a highly effective BMP at reducing nutrient losses from fields, and watersheds when employed en masse, but little work has been conducted on CTD effects on bacterial loads and densities in a watershed context. This study compared fecal indicator bacteria (FIB) [E. coli, Enterococcus, Fecal coliform, Total coliform, Clostridium perfringens] densities and unit area loads (UAL) from a pair of flat tile-drained watersheds (∼250-467 ha catchment areas) during the growing season over a 10-year monitoring period, using a before-after-control-impact (BACI) design (i.e., test CTD watershed vs. reference uncontrolled tile drainage (UCTD) watershed during a pre CTD intervention period and a CTD-intervention period where the test CTD watershed had CTD deployed on over 80% of the fields). With no tile drainage management, upstream tile drainage to ditches comprised ∼90% of total ditch discharge. We also examined FIB loads from a subset of tile drained fields to determine field load contributions to the watershed drainage ditches. Statistical evidence of a CTD effect on FIB UAL in the surface water systems was not strong; however, there was statistical evidence of increased FIB densities [pronounced when E. coli >200 most probable number (MPN) 100 mL-1] in the test CTD watershed during the CTD-intervention period. This was likely a result of reduced dilution/flushing in the test CTD watershed ditch due to CTD significantly decreasing the amount of tile drainage water entering the surface water system. Tile E. coli load contributions to the ditches were low; for example, during the 6-yr CTD-intervention period they amounted to on average only ∼3 and ∼9% of the ditch loads for the test CTD and reference UCTD watersheds, respectively. This suggests in-stream, or off-field FIB reservoirs and bacteria mobilization drivers, dominated ditch E. coli loads in the watersheds during the growing season. Overall, this study suggested that decision making regarding deployment of CTD en masse in tile-fed watersheds should consider drainage practice effects on bacterial densities and loads, as well as CTD's documented capacity to boost crop yields and reduce seasonal nutrient pollution.

Estimating cumulative wastewater treatment plant discharge influences on acesulfame and Escherichia coli in a highly impacted watershed with a fully-integrated modelling approach.

Wastewater treatment plant (WWTP) discharge is often considered a principal source of surface water contamination. In this study, a three-dimensional fully-integrated groundwater-surface water model was used to simulate the transport characteristics and cumulative loading of an artificial sweetener (acesulfame) and fecal indicator bacteria (Escherichia coli) from WWTPs within a 6800 km2 mixed-use, highly impacted watershed in Ontario, Canada. The model, which employed 3.5 × 106 computational nodes and 15 layers, facilitated a comprehensive assessment of groundwater-surface water interactions under high and low flow conditions; processes typically not accounted for in WWTP cumulative effects models. Simulations demonstrate that the model had significant capacity in reproducing the average and transient multi-year groundwater and surface water flow conditions in the watershed. As a proxy human-specific conservative tracer, acesulfame was useful for model validation and to help inform the representation of watershed-scale transport processes. Using a uniform WWTP acesulfame loading rate of 7.14 mg person-1 day-1, the general spatial trends and magnitudes of the acesulfame concentration profile along the main river reach within the watershed were reproduced; however, model performance was improved by tuning individual WWTP loading rates. Although instream dilution and groundwater-surface water interactions were strongly dependent on flow conditions, the main reach primarily consisted of groundwater discharge zones. For this reason, hydrodynamic dispersion in the hyporheic zone is shown as the predominant mechanism driving acesulfame into near-stream shallow groundwater, while under high flow conditions, the simulations demonstrate the potential for advective flushing of the shallow groundwater. Regarding the cumulative impact of the WWTPs on E. coli concentrations in the surface flow system, simulated transient E. coli levels downstream of WWTPs in the watershed were significantly lower than observed values, thus highlighting the potential importance of other sources of E. coli in the watershed.

Pharmaceutical and personal care products in tile drainage following land application of municipal biosolids.

Land application of municipal biosolids (sewage) is a common farming practice in many parts of the world. There is potential for transport of pharmaceuticals and personal care products (PPCPs) from agricultural fields to adjacent surface waters via tile drainage systems. In this study, liquid municipal biosolids (LMB) (total solids=11,933 mg L(-1)), supplemented with selected PPCPs and the fluorescent dye tracer rhodamine WT (RWT), were applied to tile drained fields using two land application approaches. Objectives included evaluating the relative benefits of land application practices with respect to reducing PPCP loadings to tile drains, evaluating PPCP persistence in tile water, and determining whether rhodamine WT can be used to estimate PPCP mass loads in tile. The PPCPs examined included an antibacterial agent used in personal care products (triclosan), a metabolite of nicotine (cotinine), and a variety of drugs including two sulfonamide antimicrobials (sulfapyridine, sulfamethoxazole), a beta-blocker (atenolol), an anti-epileptic (carbamazepine), an antidepressant (fluoxetine), analgesic/anti-inflammatories (acetaminophen, naproxen, ibuprofen), and a lipid-regulator (gemfibrozil). Maximum observed PPCP concentrations in the spiked LMB were about 10(3) ng g(-1) dry weight. PPCPs were shown to move rapidly via soil macropores to tile drains within minutes of the land application. Maximum observed PPCP concentrations in tile effluent associated with the LMB application-induced tile flow event were approximately 10(1) to 10(3) ng L(-1). PPCP mass loads, for the application-induced tile-hydrograph event, were significantly (p<0.1) higher for surface spreading over non-tilled soil (incorporation tillage occurring 20 h post-application), relative to aerating soil immediately prior to surface spreading using an AerWay slurry deposition system. PPCP concentrations that were detected above the limit of quantitation (LOQ) in tile water during several precipitation-induced tile flow events that occurred post-application, included: triclosan (max. approximately 1.5 x 10(2) ng L(-1)), carbamazepine (max. approximately 7 x 10(1) ng L(-1)), atenolol (max approximately 4 x 10(1) ng L(-1)), and cotinine (max approximately 2 x 10(1) ng L(-1)). In spite of their presence in biosolids, the other PPCPs were not observed above LOQ concentrations during these events. PPCP concentrations were predicted from RWT concentrations over a 40 day study period. Tile mass loads as a percent of PPCP mass applied to soil ranged from 4.2%+/-SD of 9.2% to 7.1%+/-10.9% for the AerWay system and surface spreading plus incorporation treatments, respectively.

Effect of liquid municipal biosolid application method on tile and ground water quality.

This study examined bacteria and nutrient quality in tile drainage and shallow ground water resulting from a fall land application of liquid municipal biosolids (LMB), at field application rates of 93,500 L ha(-1), to silt-clay loam agricultural field plots using two different land application approaches. The land application methods were a one-pass AerWay SSD approach (A), and surface spreading plus subsequent incorporation (SS). For both treatments, it took between 3 and 39 min for LMB to reach tile drains after land application. The A treatment significantly (p < 0.1) reduced application-induced LMB contamination of tile drains relative to the SS treatment, as shown by mass loads of total Kjeldahl N (TKN), NH(4)-N, Total P (TP), PO(4)-P, E. coli., and Clostridium perfringens. E. coli contamination resulting from application occurred to at least 2.0-m depth in ground water, but was more notable in ground water immediately beneath tile depth (1.2 m). Treatment ground water concentrations of selected nutrients and bacteria for the study period ( approximately 46 d) at 1.2-m depth were significantly higher in the treatment plots, relative to control plots. The TKN and TP ground water concentrations at 1.2-m depth were significantly (p < 0.1) higher for the SS treatment, relative to the A treatment, but there were no significant (p > 0.1) treatment differences for the bacteria. For the macroporous field conditions observed, pre-tillage by equipment such as the AerWay SSD, will reduce LMB-induced tile and shallow ground water contamination compared to surface spreading over non-tilled soil, followed by incorporation.

Tile water quality following liquid swine manure application into standing corn.

The quality of water draining fields fertilized with liquid swine (Sus scrofa) manure (LSM) sidedressed into standing corn (Zea mays L.) at rates ranging from 0 to 94 m(3) ha(-1), either topdressed (TD) onto the surface, or injected (INJ) into the soil once annually for each of three consecutive years was evaluated. Liquid swine manure application rate was a critical driver of preferential flow of LSM to tile as detected by turbidity, concentrations of NH(4)(+)-N, dissolved reactive phosphorus (DRP), and the presence of enteric bacteria (Escherichia coli). Contaminant movement to drains occurred immediately after 75 and 94 m(3) LSM ha(-1) were injected (e.g., 2.5 mg DRP L(-1), 3-yr average). With injection of 56 m(3) ha(-1) or less, drainage water was not turbid and concentrations of NH(4)(+)-N, DRP, and enteric bacteria were dramatically lower than with the higher rates, even when tiles flowed freely during manure application. Application method also affected tile water quality. With TD applications (37 and 56 m(3) ha(-1)), nutrients and bacteria did not move to tiles at the time of application, but with rains that fell within 3 d after application, concentrations increased (e.g., 0.1 mg DRP L(-1)), although less than with INJ. Overall, sidedress injection rates that supplied adequate crop nutrients did not compromise drainage water quality.

Brominated flame retardants and perfluoroalkyl acids in groundwater, tile drainage, soil, and crop grain following a high application of municipal biosolids to a field.

Dewatered municipal biosolids (DMB) were applied at a rate of 22Mgdwha-1 to an agricultural field in fall 2008. Concentrations of polybrominated diphenyl ethers (PBDEs; BDE-47, -99, -100, -153, -154, -183, -197, -207, -209), other brominated flame retardants (BFRs; HBB, PBEB, DBDPE, BTBPE) and perfluoroalkyl acids (PFAAs; PFHxS, PFOS, PFDS, PFOSA, PFHpA, PFOA, PFNA, PFDA, PFUnA, PFDoA, PFTA) were monitored in tile drainage, groundwater (2m, 4m and 6m depth), soil cores (0-0.3m) pre- and post-application, DMB aggregates incorporated into the soil post-application, and in wheat (Triticum spp.) planted post-application. Several compounds were detected in soil and water pre-application and on a reference field plot. PBDEs, other BFRs and PFAAs were detected in tile drainage and 2m groundwater throughout the post-application study period; a few PBDEs were also detected sporadically at lower depths in groundwater. Some of these compounds had not been detected pre-application, while some exceeded reference field plot/pre-application levels (some significantly (p<0.05) in tile drainage); both cases indicating biosolid-based water contamination. In DMB aggregates, several PBDE congeners were found to have dissipated exponentially, with reductions >90% in many of them within 1year post-application. Exponential dissipation of other BFRs and PFAAs in DMB aggregates were not significant. No PBDEs, other BFRs, or PFAAs were detected in wheat grain.

Towards a more accurate quantitative assessment of seasonal Cryptosporidium infection risks in surface waters using species and genotype information.

Many Cryptosporidium species/genotypes are not considered infectious to humans, and more realistic estimations of seasonal infection risks could be made using human infectious species/genotype information to inform quantitative microbial risk assessments (QMRA). Cryptosporidium oocyst concentration and species/genotype data were collected from three surface water surveillance programs in two river basins [South Nation River, SN (2004-09) and Grand River, GR (2005-13)] in Ontario, Canada to evaluate seasonal infection risks. Main river stems, tributaries, agricultural drainage streams, water treatment plant intakes, and waste water treatment plant effluent impacted sites were sampled. The QMRA employed two sets of exposure data to compute risk: one assuming all observed oocysts were infectious to humans, and the other based on the fraction of oocysts that were C. hominis and/or C. parvum (dominant human infectious forms of the parasite). Viability was not considered and relative infection risk was evaluated using a single hypothetical recreational exposure. Many sample site groupings for both river systems, had significant seasonality in Cryptosporidium occurrence and concentrations (p ≤ 0.05); occurrence and concentrations were generally highest in autumn for SN, and autumn and summer for GR. Mean risk values (probability of infection per exposure) for all sites combined, for each river system, were roughly an order of magnitude lower (avg. of SN and GR 5.3 × 10-5) when considering just C. parvum and C. hominis oocysts, in relation to mean infection risk (per exposure) assuming all oocysts were infectious to humans (5.5 × 10-4). Seasonality in mean risk (targeted human infectious oocysts only) was most strongly evident in SN (e.g., 7.9 × 10-6 in spring and 8.1 × 10-5 in summer). Such differences are important if QMRA is used to quantify effects of water safety/quality management practices where inputs from a vast array of fecal pollution sources can readily occur. Cryptosporidium seasonality in water appears to match the seasonality of human infections from Cryptosporidium in the study regions. This study highlights the importance of Cryptosporidium species/genotype data to help determine surface water pollution sources and seasonality, as well as to help more accurately quantify human infection risks by the parasite.