Extended-spectrum beta-lactamase, AmpC, and carbapenemase-producing Gram-negative wastewater isolates from Aotearoa New Zealand.

Extended-spectrum beta-lactamase, AmpC, and carbapenemase-producing bacteria were isolated from raw sewage, effluent, oxidation pond water, and sediment from a wastewater treatment plant in Aotearoa New Zealand. Here, we report the assemblies of 17 isolates belonging to the species Aeromonas veronii, Aeromonas hydrophila, Enterobacter cloacae, Enterobacter soli, Lelliottia amnigena, Aeromonas caviae, Escherichia coli, Pseudomonas moraviensis, Pseudomonas putida, and Kluyvera ascorbata.

Responding to and managing multijurisdictional outbreaks of COVID-19 in Canadian industrial worksite/work camp settings.

SETTING: Early in the COVID-19 pandemic, the Public Health Agency of Canada (PHAC) and provincial/territorial (P/T) public health identified the need for a coordinated response to complex multijurisdictional COVID-19 outbreaks. The first large multijurisdictional industrial worksite COVID-19 outbreak highlighted the risk of transmission within these congregate work settings, the risk of transmission to the broader community(ies), and the need to develop setting-specific outbreak response frameworks. INTERVENTION: PHAC assembled a team to provide national outbreak support for multijurisdictional COVID-19 outbreaks in May 2020. The COVID-19 Outbreak Response Unit (ORU) worked with P/T partners to develop guiding principles for outbreak response and outbreak investigation processes, guidance documents, and investigation tools (e.g., minimum data elements and questionnaires). OUTCOMES: The ORU, P/T partners, and onsite industrial worksite health and safety staff leveraged outbreak investigation guidelines, industrial worksite outbreak process documents (including minimum data elements), and enhanced case questionnaires to respond to multiple COVID-19 outbreak investigations in industrial worksites. Clear roles/responsibilities and processes, along with standardized data, allowed for more efficient outbreak investigations and earlier implementation of mitigation measures. IMPLICATIONS: Multijurisdictional COVID-19 outbreaks highlighted the importance of public health collaboration with industry partners onsite. The assembly of a national outbreak response team was important to facilitate information sharing and provide technical support. Lessons learned and recommendations on outbreak preparation, detection, management, and communication are included to enhance a response framework applicable to future emerging or re-emerging pathogens with epidemic and/or pandemic potential.

RéSUMé: CONTEXTE: Au début de la pandémie de COVID-19, l'Agence de la santé publique du Canada (ASPC) et les autorités provinciales/territoriales de santé publique ont reconnu la nécessité d'une réponse coordonnée en cas d'éclosions complexes multi-juridictionnelles de COVID-19. La première grande éclosion multi-juridictionnelle de COVID-19 dans un chantier industriel a mis en évidence le risque de transmission dans ces milieux de travail collectifs, le risque de transmission à l'ensemble de la (des) communauté(s) et la nécessité d'élaborer des cadres d'intervention en cas d'éclosion spécifiques aux types de milieux. INTERVENTION: L'ASPC a formé une équipe chargée de soutenir la réponse nationale contre les éclosions multi-juridictionnelles de COVID-19 en mai 2020. L'Unité d'intervention en cas d'éclosion (UIE) de COVID-19 a collaboré avec des partenaires provinciaux et territoriaux pour élaborer des principes de référence pour la lutte contre les éclosions de COVID-19 et des processus d'enquête sur les éclosions, des documents d'orientation et des outils d'enquête (p.ex. des éléments de données minimales et des questionnaires). RéSULTATS: L'UIE, les provinces et territoires et le personnel chargé de la santé et sécurité du travail sur le site se sont appuyés sur des principes de référence aux enquêtes sur les éclosions, les documents de processus d'enquête sur les éclosions dans les sites industriels, y compris les éléments de données minimales et le questionnaire détaillé sur les cas, pour répondre à multiples enquêtes d'éclosions de COVID-19 dans les sites industriels. Des rôles/responsabilités et des processus clairs, ainsi que des données standardisées, ont permis de mener des enquêtes plus efficaces sur les éclosions et de mettre en œuvre plus rapidement des mesures d'atténuation. IMPLICATIONS: Les éclosions multi-juridictionnelles de COVID-19 ont mis en évidence l'importance de la collaboration entre les autorités de santé publique et les partenaires industriels sur site. La constitution d'une équipe nationale d'intervention en cas d'éclosion a été importante pour faciliter le partage des informations et fournir un soutien technique. Les connaissances acquises et les recommandations sur la préparation, la détection, la gestion et la communication des éclosions sont incluses afin d'améliorer le cadre de réponse aux futurs agents pathogènes émergents ou ré-émergents ayant un potentiel épidémique et/ou pandémique.

Draft genome sequences of 12 triclosan tolerant bacteria isolated from returned activated sewage sludge.

Herein we report the whole genome sequences of 12 highly triclosan tolerant bacteria isolated from returned activated sludge spiked with triclosan.

Comparative reductions of norovirus, echovirus, adenovirus, Campylobacter jejuni and process indicator organisms during water filtration in alluvial sand.

Sand filtration is a cost-effective means of reducing microbial pathogens in drinking-water treatment. Our understanding of pathogen removal by sand filtration relies largely on studies of process microbial indicators, and comparative data from pathogens are sparse. In this study, we examined the reductions of norovirus, echovirus, adenovirus, bacteriophage MS2 and PRD1, Campylobacter jejuni, and Escherichia coli during water filtration through alluvial sand. Duplicate experiments were conducted using 2 sand columns (50 cm long, 10 cm diameter) and municipal tap water sourced from chlorine-free untreated groundwater (pH 8.0, 1.47 mM) at filtration rates of 1.1-1.3 m/day. The results were analysed using colloid filtration theory and the HYDRUS-1D 2-site attachment-detachment model. The average log10 reduction values (LRVs) of the normalised dimensionless peak concentrations (Cmax/C0) over 0.5 m were: MS2: 0.28; E. coli: 0.76; C. jejuni: 0.78; PRD1: 2.00; echovirus: 2.20; norovirus: 2.35; and adenovirus: 2.79. The relative reductions largely corresponded to the organisms' isoelectric points rather than their particle sizes or hydrophobicities. MS2 underestimated virus reductions by 1.7-2.5 log, and the LRVs, mass recoveries relative to bromide, collision efficiencies, and attachment and detachment rates differed mostly by ∼1 order of magnitude. Conversely, PRD1 reductions were comparable with those of all 3 viruses tested, and its parameter values were mostly within the same orders of magnitude. E. coli seemed an adequate process indicator for C. jejuni with similar reductions. Comparative data describing pathogen and indicator reductions in alluvial sand have important implications for sand filter design, risk assessments of drinking-water supplies from riverbank filtration and the determination of safe setback distances for drinking-water supply wells.

Nonbacterial Microflora in Wastewater Treatment Plants: an Underappreciated Potential Source of Pathogens.

Wastewater treatment plants (WWTPs) receive and treat large volumes of domestic, industrial, and urban wastewater containing pathogenic and nonpathogenic microorganisms, chemical compounds, heavy metals, and other potentially hazardous substances. WWTPs play an essential role in preserving human, animal, and environmental health by removing many of these toxic and infectious agents, particularly biological hazards. Wastewater contains complex consortiums of bacterial, viral, archaeal, and eukaryotic species, and while bacteria in WWTP have been extensively studied, the temporal and spatial distribution of nonbacterial microflora (viruses, archaea, and eukaryotes) is less understood. In this study, we analyzed the viral, archaeal, and eukaryotic microflora in wastewater throughout a treatment plant (raw influent, effluent, oxidation pond water, and oxidation pond sediment) in Aotearoa (New Zealand) using Illumina shotgun metagenomic sequencing. Our results suggest a similar trend across many taxa, with an increase in relative abundance in oxidation pond samples compared to influent and effluent samples, except for archaea, which had the opposite trend. Additionally, some microbial families, such as Podoviridae bacteriophages and Apicomplexa alveolates, appeared largely unaffected by the treatment process, with their relative abundance remaining stable throughout. Several groups encompassing pathogenic species, such as Leishmania, Plasmodium, Toxoplasma, Apicomplexa, Cryptococcus, Botrytis, and Ustilago, were identified. If present, these potentially pathogenic species could be a threat to human and animal health and agricultural productivity; therefore, further investigation is warranted. These nonbacterial pathogens should be considered when assessing the potential for vector transmission, distribution of biosolids to land, and discharge of treated wastewater to waterways or land. IMPORTANCE Nonbacterial microflora in wastewater remain understudied compared to their bacterial counterparts despite their importance in the wastewater treatment process. In this study, we report the temporal and spatial distributions of DNA viruses, archaea, protozoa, and fungi in raw wastewater influent, effluent, oxidation pond water, and oxidation pond sediments by using shotgun metagenomic sequencing. Our study indicated the presence of groups of nonbacterial taxa which encompass pathogenic species that may have potential to cause disease in humans, animals, and agricultural crops. We also observed higher alpha diversity in viruses, archaea, and fungi in effluent samples than in influent samples. This suggests that the resident microflora in the wastewater treatment plant may be making a greater contribution to the diversity of taxa observed in wastewater effluent than previously thought. This study provides important insights to better understand the potential human, animal, and environmental health impacts of discharged treated wastewater.

Canadian COVID-19 Outbreak Surveillance System: implementation of national surveillance during a global pandemic.

SETTING: Early in the SARS-CoV-2 pandemic, the need to develop systematic outbreak surveillance at the national level to monitor trends in SARS-CoV-2 outbreaks was identified as a priority for the Public Health Agency of Canada (PHAC). The Canadian COVID-19 Outbreak Surveillance System (CCOSS) was established to monitor the frequency and severity of SARS-CoV-2 outbreaks across various community settings. INTERVENTION: PHAC engaged with provincial/territorial partners in May 2020 to develop goals and key data elements for CCOSS. In January 2021, provincial/territorial partners began submitting cumulative outbreak line lists on a weekly basis. OUTCOMES: Eight provincial and territorial partners, representing 93% of the population, submit outbreak data on the number of cases and severity indicators (hospitalizations and deaths) for 24 outbreak settings to CCOSS. Outbreak data can be integrated with national case data to supply information on case demographics, clinical outcomes, vaccination status, and variant lineages. Data aggregated to the national level are used to conduct analyses and report on outbreak trends. Evidence from CCOSS analyses has been useful in supporting provincial/territorial outbreak investigations, informing policy recommendations, and monitoring the impact of public health measures (vaccination, closures) in specific outbreak settings. IMPLICATIONS: The development of a SARS-CoV-2 outbreak surveillance system complemented case-based surveillance and furthered the understanding of epidemiological trends. Further efforts are required to better understand SARS-CoV-2 outbreaks for Indigenous populations and other priority populations, as well as create linkages between genomic and epidemiological data. As SARS-CoV-2 outbreak surveillance enhanced case surveillance, outbreak surveillance should be a priority for emerging public health threats.

RéSUMé: CONTEXTE: Au début de la pandémie de SRAS-CoV-2, l'Agence de la santé publique du Canada (ASPC) a déterminé comme priorité la nécessité de développer un système de surveillance systématique des éclosions à l'échelle nationale afin de suivre les tendances des éclosions de SRAS-CoV-2. Le système canadien de surveillance des éclosions de COVID-19 (SCSEC) a été établi pour surveiller la fréquence et la gravité des éclosions de SRAS-CoV-2 dans différents milieux communautaires. INTERVENTION: L'ASPC s'est engagée avec les partenaires provinciaux et territoriaux en mai 2020 pour élaborer des objectifs et des éléments de données clés pour le SCSEC. En janvier 2021, les partenaires provinciaux et territoriaux ont commencé à transmettre des listes d'éclosions cumulatives hebdomadaires. RéSULTATS: Huit partenaires provinciaux et territoriaux, représentant 93 % de la population, transmettent au SCSEC des données sur les éclosions sur le nombre de cas et les indicateurs de gravité (les hospitalisations et les décès) pour 24 types de milieux. Les données sur les éclosions peuvent être intégrées avec les données nationales sur les cas pour obtenir des informations sur la démographie des cas, les résultats cliniques, le statut vaccinal et les lignées de variants. Les données agrégées à l'échelle nationale sont utilisées pour effectuer des analyses et faire rapport des tendances sur les éclosions. Les résultats des analyses du SCSEC ont été utiles pour soutenir les enquêtes provinciales/territoriales sur les éclosions, informer les recommandations politiques et surveiller l'impact des mesures de santé publique (la vaccination, les fermetures) dans des milieux d'éclosions spécifiques. IMPLICATIONS: Le développement d'un système de surveillance des éclosions de SRAS-CoV-2 a permis de complémenter la surveillance des cas et d'approfondir notre compréhension des tendances épidémiologiques. Des efforts supplémentaires sont nécessaires pour mieux comprendre les éclosions de SRAS-CoV-2 chez les populations autochtones et d'autres populations minoritaires, ainsi que pour créer des liens entre les données génomiques et les données épidémiologiques. Comme la surveillance des éclosions de SRAS-CoV-2 a enrichi la surveillance des cas, la surveillance des éclosions devrait être une priorité pour les menaces émergentes pour la santé publique.

COVID-19 outbreak trends in Canada, 2021.

Background: In January 2021, the Public Health Agency of Canada launched an outbreak surveillance system, the Canadian COVID-19 Outbreak Surveillance System (CCOSS), with the goal of monitoring incidence and severity of coronavirus disease 2019 (COVID-19) outbreaks across various community settings and complementing case surveillance. Methods: Seven provinces were included in this report; these provinces submitted weekly cumulative COVID-19 outbreak line lists to CCOSS in 2021. Data includes administrative variables (e.g. date outbreak declared, date outbreak declared over, outbreak identifier), 24 outbreak settings, and number of confirmed cases and outcomes (hospitalization, death). Descriptive analyses for COVID-19 outbreaks across Canada from January 3, 2021, to January 1, 2022, were performed examining trends over time, severity, and outbreak size. Results: Incidence of outbreaks followed similar trends to case incidence. Outbreaks were most common in school and childcare settings (39%) and industrial/agricultural settings (21%). Outbreak size ranged from 2 to 639 cases per outbreak; the median size was four cases per outbreak. Correctional facilities had the largest median outbreak size with 18 cases per outbreak, followed by long-term care facilities with 10 cases per outbreak. During periods of high case incidence, outbreaks may be under-ascertained due to limited public health capacity, or reporting may be biased towards high-risk settings prioritized for testing. Outbreaks reported to CCOSS were dominated by jurisdictions with the largest populations. Conclusion: The trends illustrate that COVID-19 outbreaks in 2021 were reported most frequently in community settings such as schools; however, the largest outbreaks occurred in congregate living settings. The information gathered from outbreak surveillance complemented case incidence trends and furthered understanding of COVID-19 in Canada.

Multijurisdictional outbreak of COVID-19 associated with a wake/funeral event in a northern Saskatchewan First Nations community.

Background: Sixty-eight laboratory-confirmed cases of the coronavirus disease 2019 (COVID-19) (12 in Alberta [AB], 56 in Saskatchewan [SK]) were linked to a gathering at a hospital in Alberta on June 1-4, 2020, and a wake/funeral in a First Nations community in northern Saskatchewan on June 9-11, 2020. Objective: The objectives were to provide a comprehensive description of the epidemiology of the outbreak and describe the chains of transmission to inform the hypothesis that there were multiple introductions of COVID-19 at the wake/funeral. Methods: Case investigation and contact tracing was conducted by local public health in AB and SK. The Public Health Agency of Canada conducted a centralized case analysis. An epidemic curve and a Gantt chart for period of communicability were created to support or refute whether there had been multiple introductions of COVID-19 at the wake/funeral. Results: Illness onset dates ranged from May 31 to July 1, 2020. Ages ranged from 2 to 80 years (median age=43 years). Five cases were hospitalized; there were no deaths. The available case exposure information supports the hypothesis that there had been multiple introductions of COVID-19 at the wake/funeral. Public health authorities in AB and SK declared the outbreak over on July 20, 2020; based on two incubation periods (i.e. 28 days) following the illness onset of the last primary case. Conclusion: During multijurisdictional outbreaks, data sharing, coordination across health authorities and centralized analysis is essential to understanding the events that lead to the outbreak and possible hypotheses around chains of transmission.

Reductions of human enteric viruses in 10 commonly used activated carbon, polypropylene and polyester household drinking-water filters.

Drinking-water treatment in non-networked rural communities relies on the use of point-of-use (PoU) household filters. Source waters treated by PoU filters are often microbially contaminated, but information about human enteric virus reductions in these filters is limited. This study evaluated human rotavirus, adenovirus and norovirus reductions in 10 commonly used, new PoU carbon, polypropylene and polyester microfilters. The viruses were spiked into chlorine-free tap water (pH 8.0, ionic strength 1.22 mM), and 3 sequential challenge tests were conducted in each filter under a constant flow rate of 1 L/min. In most of the filters investigated, the norovirus and adenovirus reductions were similar (P > 0.49). Compared with the norovirus and adenovirus reductions, the rotavirus reductions were significantly lower in the carbon filters (P ≤ 0.009), which may relate to rotavirus's higher zeta potential and lower hydrophobicity. Virus reductions appeared to be dictated by the filter media type through electrostatic and hydrophobic interactions; the effects of filter media pore sizes on virus reductions via physical size-exclusion were very limited. The virus reductions in the carbon filters were significantly greater than those in the polypropylene and polyester filters (P ≤ 0.0001), and they did not differ significantly between the polypropylene and polyester filters (P > 0.24). None of the filters met the "protective" rotavirus reduction level (≥3 log10) required for household drinking-water treatment. Our study's findings highlight a critical need for additional water treatment when using PoU microfilters, for example, water boiling or ultraviolet radiation, or the use of effective surface-modified filter media to prevent drinking-waterborne infections from enteric viruses.

Comparison of faecal indicator and viral pathogen light and dark disinfection mechanisms in wastewater treatment pond mesocosms.

This study compared light and dark disinfection of faecal bacteria/viral indicator organisms (E. coli and MS2 (fRNA) bacteriophage) and human viruses (Echovirus and Norovirus) in Wastewater Treatment Pond (WTP) mesocosms. Stirred pond mesocosms were operated in either outdoor sunlight-exposed or laboratory dark conditions in two experiments during the austral summer. To investigate wavelength-dependence of sunlight disinfection, three optical filters were used: (1) polyethylene film (light control: transmitting all solar UV and visible wavelengths), (2) acrylic (removing most UVB <315 nm), and (3) polycarbonate (removing both UVB and UVA <400 nm). To assess different dark disinfection processes WTP effluent was treated before spiking with target microbes, by (a) 0.22 µm filtration to remove all but colloidal particles, (b) 0.22 µm filtration followed by heat treatment to destroy enzymes, and (c) addition of Cytochalasin B to supress protozoan grazing. Microbiological stocks containing E. coli, MS2 phage, Echovirus, and Norovirus were spiked into each mesocosm 10 min before the experiments commenced. The light control exposed to all sunlight wavelengths achieved >5-log E. coli and MS2 phage removal (from ~1.0 × 106 to <1 PFU/mL) within 3 h compared with up to 6 h in UV-filtered mesocosms. This result confirms that UVB contributes to inactivation of E. coli and viruses by direct sunlight inactivation. However, the very high attenuation with depth of UVB in WTP water (99% removal in the top 8 cm) suggests that UVB disinfection may be less important than other removal processes averaged over time and full-scale pond depth. Dark removal was appreciably slower than sunlight-mediated inactivation. The dark control typically achieved higher removal of E. coli and viruses than the 0.22 µm filtered (dark) mesocosms. This result suggests that adsorption of E. coli and viruses to WTP particles (e.g., algae and bacteria bio-flocs) is an important mechanism of dark disinfection, while bacteria and virus characteristics (e.g. surface charge) and environmental conditions can influence dark disinfection processes.

Heartworm infection in domestic dogs in Canada, 1977-2016: Prevalence, time trend, and efficacy of prophylaxis.

Dirofilaria immitis (heartworm) is a mosquito-borne parasite that primarily infects domestic and wild canids. The objectives of this study were to i) determine if there has been a temporal change in prevalence of heartworm infection among domestic dogs in Canada from 1977 to 2016; ii) explore the spatial extension of heartworm across Canada using choropleth maps; and iii) assess the efficacy of preventive drugs using the estimated "Attributable Fraction Exposed." Heartworm surveys that collected data from 1977 to 2010 and serological laboratory data from 2007 to 2016 were analyzed. The data depicted a decrease in heartworm prevalence, both nationally and provincially, from 1977 to the early 2000s. However, an increase in prevalence was identified for tested dog populations in Manitoba and Quebec from 2007 to 2016. Chemoprophylaxis was associated with an estimated 93.0% [95% confidence interval (CI): 92.85, 93.3] reduction of heartworm infections in domestic dogs in Canada from 1977 to 2010.

Infection par le ver du coeur chez les chiens domestiques au Canada, 1977­2016 : prévalence, tendance dans le temps et efficacité de la prophylaxie. Dirofilaria immitis (ver du coeur) est un parasite transporté par des moustiques qui infecte principalement les canidés domestiques et sauvages. Les objectifs de cette étude consistaient : i) à déterminer s'il y avait eu un changement temporel de la prévalence de l'infection parmi les chiens domestiques au Canada de 1977 à 2016; ii) à explorer le prolongement spatial du ver du coeur au Canada à l'aide de cartes choroplèthe; et iii) à évaluer l'efficacité des médicaments préventifs en utilisant l'estimation de la «fraction attribuable exposée¼. Les sondages sur le ver du coeur qui ont recueilli des données de 1977 à 2010 et les données sérologiques de laboratoire de 2007 à 2016 ont été analysés. Les données illustraient une baisse de la prévalence du ver du coeur, tant à l'échelle nationale que provinciale, de 1977 au début des années 2000. Cependant, une hausse de la prévalence a été identifiée pour les populations de chiens testées au Manitoba et au Québec de 2007 à 2016. La chimioprophylaxie était associée à une réduction estimée de 93,0 % [intervalle de confiance (IC) de 95 % : 92,85, 93,3] des infections du ver du coeur chez les chiens domestiques au Canada de 1977 à 2010.(Traduit par Isabelle Vallières).

Epidemiology of canine heartworm (Dirofilaria immitis) infection in domestic dogs in Ontario, Canada: Geographic distribution, risk factors and effects of climate.

Dirofilaria immitis is the causal agent of heartworm, a mosquito-borne parasite that primarily infects domestic and wild canids. The infection is endemic in parts of Canada, and Ontario has been identified as the province where the majority of heartworm infections occur. Test results for blood samples submitted by veterinary clinics for the years 2007-2016 were used to conduct a spatial risk analysis of heartworm among domestic dogs in Ontario. The geographic extent of the apparent heartworm prevalence was examined through smoothed choropleth maps for all 49 census division regions. Furthermore, the regions were assessed for local clusters in apparent prevalence using the flexible spatial scan statistic. Three clusters were found and located in western, southern and eastern Ontario, respectively. A spatial Poisson regression model for heartworm prevalence among pet dog populations in southern Ontario census divisions was fit to determine the association between human population size, heartworm development units (HDUs), climate moisture index (CMI), precipitation and directions, east or north, with heartworm infection. The model identified the spatial distribution of HDUs and CMI as positively associated with heartworm infection and therefore important predictors of the infection. In contrast, human population size, increasing northern latitude and drier areas were negatively associated with heartworm infection. The east direction and precipitation were not significant.

Tracking effluent discharges in undisturbed stony soil and alluvial gravel aquifer using synthetic DNA tracers.

With the intensification of human activities, fresh water resources are increasingly being exposed to contamination from effluent disposal to land. Thus, there is a greater need to identify the sources and pathways of water contamination to enable the development of better mitigation strategies. To track discharges of domestic effluent into soil and groundwater, 10 synthetic double-stranded DNA (dsDNA)3 tracers were developed in this study. Laboratory column experiment and field groundwater and soil lysimeter studies were carried out spiking DNA with oxidation-pond domestic effluent. The selected DNA tracers were compared with a non-reactive bromide (Br) tracer with respect to their relative mass recoveries, speeds of travel and dispersions using the method of temporal moments. In intact stony soil and gravel aquifer media, the dsDNA tracers typically showed earlier breakthrough and less dispersion than the Br tracer, and underwent mass reduction. This suggests that the dsDNA tracers were predominantly transported through the network of larger pores or preferential flow paths. Effluent tracking experiments in soil and groundwater demonstrated that the dsDNA tracers were readily detectable in effluent-contaminated soil and groundwater using quantitative polymerase chain reaction. DNA tracer spiked in the effluent at quantities of 36µg was detected in groundwater 37m down-gradient at a concentration 3-orders of magnitude above the detection limit. It is anticipated it could be detected at far greater distances. Our findings suggest that synthetic dsDNA tracers are promising for tracking effluent discharges in soils and groundwater but further studies are needed to investigate DNA-effluent interaction and the impact of subsurface environmental conditions on DNA attenuation. With further validation, synthetic dsDNA tracers, especially when multiple DNA tracers are used concurrently, can be an effective new tool to track effluent discharge in soils and groundwater, providing spatial estimation on the presence or absence of contamination sources and pathways.

Influence of colloids on the attenuation and transport of phosphorus in alluvial gravel aquifer and vadose zone media.

Phosphorous (P) leaching (e.g., from effluents, fertilizers) and transport in highly permeable subsurface media can be an important pathway that contributes to eutrophication of receiving surface waters as groundwater recharges the base-flow of surface waters. Here we investigated attenuation and transport of orthophosphate-P in gravel aquifer and vadose zone media in the presence and absence of model colloids (Escherichia coli, kaolinite, goethite). Experiments were conducted using repacked aquifer media in a large column (2m long, 0.19m in diameter) and intact cores (0.4m long, 0.24m in diameter) of vadose zone media under typical field flow rates. In the absence of the model colloids, P was readily traveled through the aquifer media with little attenuation (up to 100% recovery) and retardation, and P adsorption was highly reversible. Conversely, addition of the model colloids generally resulted in reduced P concentration and mass recovery (down to 28% recovery), and increased retardation and adsorption irreversibility in both aquifer and vadose zone media. The degree of colloid-assisted P attenuation was most significant in the presence of fine material and Fe-containing colloids at low flow rate but was least significant in the presence of coarse gravels and E. coli at high flow rate. Based on the experimental results, setback distances of 49-53m were estimated to allow a reduction of P concentrations in groundwater to acceptable levels in the receiving water. These estimates were consistent with field observations in the same aquifer media. Colloid-assisted P attenuation can be utilized to develop mitigation strategies to better manage effluent applications in gravelly soils. To efficiently retain P within soil matrix and reduce P leaching to groundwater, it is recommended to select soils that are rich in iron oxides, to periodically disturb soil preferential flow paths by tillage, and to apply a low irrigation rate.

Size exclusion-based purification and PCR-based quantitation of MS2 bacteriophage particles for environmental applications.

MS2 bacteriophage is the most commonly used surrogate for pathogenic viruses in laboratory and field studies. In order to determine the number of infectious viral particles in samples, the use of accurate quantitation methods is essential. We have optimised a size exclusion chromatography-based method for MS2 purification and a SYBR Green-based single-step quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) assay for the quantitation of MS2. The qRT-PCR enabled accurate quantitation of viral RNA of the purified stock with a detection limit of 2 genome copy equivalents/µl. Detection inhibition, if any, was eliminated by reducing sample volume added to the qRT-PCR reaction mix when MS2 was detected in environmental water samples. The purification method eliminated the impurities and the purified stock yielded a high concentration of infectious MS2 particles. The qRT-PCR assay enabled the accurate quantitation of the viral particles thus providing an alternative to the traditional plaque assays. A combined use of purified MS2 stock and PCR-based quantitation gives the opportunity to explore virus characteristics, behaviour and interactions in the environment.