Development of a "big data" groundwater microbial contamination index and spatial comparisons with enteric infection rates in southern Ontario.

Big data have become increasingly important for policymakers and scientists but have yet to be employed for the development of spatially specific groundwater contamination indices or protecting human and environmental health. The current study sought to develop a series of indices via analyses of three variables: Non-E. coli coliform (NEC) concentration, E. coli concentration, and the calculated NEC:E. coli concentration ratio. A large microbial water quality dataset comprising 1,104,094 samples collected from 292,638 Ontarian wells between 2010 and 2021 was used. Getis-Ord Gi* (Gi*), Local Moran's I (LMI), and space-time scanning were employed for index development based on identified cluster recurrence. Gi* and LMI identify hot and cold spots, i.e., spatially proximal subregions with similarly high or low contamination magnitudes. Indices were statistically compared with mapped well density and age-adjusted enteric infection rates (i.e., campylobacteriosis, cryptosporidiosis, giardiasis, verotoxigenic E. coli (VTEC) enteritis) at a subregional (N = 298) resolution for evaluation and final index selection. Findings suggest that index development via Gi* represented the most efficacious approach. Developed Gi* indices exhibited no correlation with well density, implying that indices are not biased by rural population density. Gi* indices exhibited positive correlations with mapped infection rates, and were particularly associated with higher bacterial (Campylobacter, VTEC) infection rates among younger sub-populations (p < 0.05). Conversely, no association was found between developed indices and giardiasis rates, an infection not typically associated with private groundwater contamination. Findings suggest that a notable proportion of bacterial infections are associated with groundwater and that the developed Gi* index represents an appropriate spatiotemporal reflection of long-term groundwater quality. Bacterial infection correlations with the NEC:E. coli ratio index (p < 0.001) were markedly different compared to correlations with the E. coli index, implying that the ratio may supplement E. coli monitoring as a groundwater assessment metric capable of elucidating contamination mechanisms. This study may serve as a methodological blueprint for the development of big data-based groundwater contamination indices across the globe.

An outbreak of Salmonella Infantis linked to shredded pork products from an unlicensed source in multiple health districts, Ontario, Canada, 2021.

Background: An outbreak of Salmonella Infantis was associated with the consumption of shredded pork products at multiple restaurants in Ontario between July 2021 and October 2021. The outbreak involved 36 case-patients from six public health units. The implicated shredded pork products were obtained from an unlicensed source. This is the largest reported outbreak of Salmonella Infantis linked to restaurant food exposures in Ontario, with complexities related to the investigation of unlicensed foods. This article aims to describe the epidemiological, food safety and laboratory investigations that led to the identification and removal of the source of the outbreak from implicated restaurants, including the challenges encountered while investigating an outbreak related to an unlicensed source of food. Methods: Epidemiological and laboratory analyses were conducted to identify the source of the outbreak. Food safety investigations were conducted to ascertain the origin and distribution of the implicated food. Results: Whole-genome sequencing identified the outbreak strain from the isolates of 36 case-patients across six public health units in Ontario. Seven case-patients (19%) were hospitalized. No deaths were reported. The outbreak was linked to shredded pork products (i.e., rinds or skins) that were distributed by an unlicensed meat processor and consumed at various restaurants that served Southeast Asian fusion cuisine concentrated in the Greater Toronto Area. The product was removed from implicated restaurants. Conclusion: Historically, foods from unlicensed sources have been implicated in multiple large outbreaks and continue to be of significant public health risk. The outbreak investigation emphasized the threat of food from unlicensed sources to the public's health and the importance of additional public health interventions to prevent outbreaks linked to unlicensed sources.

Salmonella Vitkin Outbreak Associated with Bearded Dragons, Canada and United States, 2020-2022.

We identified 2 cases of Salmonella enterica serovar Vitkin infection linked by whole-genome sequencing in infants in Ontario, Canada, during 2022. Both households of the infants reported having bearded dragons as pets. The outbreak strain was also isolated from an environmental sample collected from a patient's bearded dragon enclosure. Twelve cases were detected in the United States, and onset dates occurred during March 2021-September 2022 (isolates related to isolates from Canada within 0-9 allele differences by core-genome multilocus sequence typing). Most US patients (66.7%) were <1 year of age, and most (72.7%) had reported bearded dragon exposure. Hospitalization was reported for 5 (38.5%) of 13 patients. Traceback of bearded dragons identified at least 1 potential common supplier in Southeast Asia. Sharing rare serovar information and whole-genome sequencing data between Canada and the United States can assist in timely identification of outbreaks, including those that might not be detected through routine surveillance.

Two Listeria monocytogenes outbreaks in a cancer centre: onsite food premises and their potential health risk to patients.

BACKGROUND: This report describes two L. monocytogenes outbreak investigations that occurred in March and September of 2018 and that linked illness to a food premises located in an Ontario cancer centre. The cancer centre serves patients from across the province. METHODS: In Ontario, local public health agencies follow up with all reported laboratory-confirmed cases of listeriosis to identify possible sources of disease acquisition and to carry out investigations, including at suspected food premises. The Canadian Food Inspection Agency (CFIA) is notified of any Listeria-positive food product collected in relation to a case. The CFIA traces Listeria-positive product through the food distribution system to identify the contamination source and ensure the implicated manufacturing facility implements corrective measures. RESULTS: Outbreaks one and two each involved three outbreak-confirmed listeriosis cases. All six cases were considered genetically related by whole genome sequencing (WGS). In both outbreaks, outbreak-confirmed cases reported consuming meals at a food premises located in a cancer centre (food premises A) before illness onset. Various open deli meat samples and, in outbreak two, environmental swabs (primarily from the meat slicer) collected from food premises A were genetically related to the outbreak-confirmed cases. Food premises A closed as a result of the investigations. CONCLUSIONS: When procuring on-site food premises, healthcare facilities and institutions serving individuals with immuno-compromising conditions should consider the potential health risk of foods available to their patient population.

Integrating consumer risk perception and awareness with simulation-based quantitative microbial risk assessment using a coupled systems framework: A case study of private groundwater users in Ontario.

Private well users in Ontario are responsible for ensuring the potability of their own private drinking water source through protective actions (i.e., water treatment, well maintenance, and regular water quality testing). In the absence of regulation and limited surveillance, quantitative microbial risk assessment (QMRA) represents the most practical and robust approach to estimating the human health burden attributable to private wells. For an increasingly accurate estimation, QMRA of private well water should be represented by a coupled model, which includes both the socio-cognitive and physical aspects of private well water contamination and microbial exposure. The objective of the current study was to determine levels of waterborne exposure via well water consumption among three sub-groups (i.e., clusters) of private well users in Ontario and quantify the risk of waterborne acute gastrointestinal illness (AGI) attributed to Giardia, shiga-toxin producing E. coli (STEC) and norovirus from private drinking water sources in Ontario. Baseline simulations were utilized to explore the effect of varying socio-cognitive scenarios on model inputs (i.e., increased awareness, protective actions, aging population). The current study uses a large spatio-temporal groundwater quality dataset and cross-sectional province-wide survey to create socio-cognitive-specific QMRA simulations to estimate the risk of waterborne AGI attributed to three enteric pathogens in private drinking waters source in Ontario. Findings suggest significant differences in the level of exposure among sub-groups of private well users. Private well users within Cluster 3 are characterised by higher levels of exposure and annual illness attributable to STEC, Giardia and norovirus than Clusters 1 and 2. Provincial incidence rates of 520.9 (1522 illness per year), 532.1 (2211 illness per year) and 605.5 (5345 illness per year) cases/100,000 private well users per year were predicted for private well users associated with Clusters 1 through 3. Established models will enable development of necessary tools tailored to specific groups of at-risk well users, allowing for preventative public health management of private groundwater sources.

Classification of sub-populations for quantitative risk assessment based on awareness and perception: A cross-sectional population study of private well users in Ontario.

Private well users in Ontario are responsible for protective actions, including source maintenance, treatment, and submitting samples for laboratory testing. However, low participation rates are reported, thus constituting a public health concern, as risk mitigation behaviours can directly reduce exposure to waterborne pathogens. The current study examined the combined effects of socio-demographic profile, experience(s), and "risk domains" (i.e., awareness, attitudes, risk perceptions and beliefs) on behaviours, and subsequently classified private well users in Ontario based on cognitive factors. A province-wide online survey (n = 1228) was employed to quantify Ontario well owners' awareness, perceptions, and behaviours in relation to their personal groundwater supply and local contamination sources. A scoring protocol for four risk domains was developed. Two-step cluster analysis was used to classify respondents based on individual risk domain scores. Logistic regression was employed to identify key variables associated with cluster membership (i.e., profile analysis). Overall, 1140 survey respondents were included for analyses. Three distinct clusters were identified based on two risk domains; groundwater awareness and source risk perception. Profile analyses indicate "low awareness and source risk perception" (Low A/SRP) members were more likely male, while "low awareness and moderate source risk perception" (Low A/Mod SRP) members were more likely female and bottled water users. Well users characterised as "high awareness and source risk perception" (High A/SRP) were more likely to report higher educational attainment and previous well water testing. Findings illustrate that socio-cognitive clusters and their components (i.e., demographics, awareness, attitudes, perceptions, experiences, and protective actions) are distinct based on the likelihood, frequency, and magnitude of waterborne pathogen exposures (i.e., risk-based). Risk-based clustering, when incorporated into quantitative microbial risk assessment, enables the development of effective risk management and communication initiatives that are demographically focused and tailored to specific sub-groups.

Assessing the risk of acute gastrointestinal illness attributable to three enteric pathogens from contaminated private water wells in Ontario.

The province of Ontario compromises the largest groundwater reliant population in Canada serving approximately 1.6 million individuals. Unlike municipal water systems, private well water is not required to meet water quality regulatory standards and thus source maintenance, treatment and testing remains the responsibility of the well owner. Infections associated with private drinking water systems are rarely documented given their typically sporadic nature, thus the human health effects (e.g., acute gastrointestinal illness (AGI)) on consumers remains relatively unknown, representing a significant gap in water safety management. The current study sought to quantify the risk of waterborne AGI attributed to Giardia, shiga-toxin producing E. coli (STEC) and norovirus from private drinking water sources in Ontario using Monte Carlo simulation-based quantitative microbial risk assessment (QMRA). Findings suggest that consumption of contaminated private well water in Ontario is responsible for approximately 4823 AGI cases annually, with 3464 (71.8%) and 1359 (28.1%) AGI cases predicted to occur in consolidated and unconsolidated aquifers, respectively. By pathogen, waterborne AGI was attributed to norovirus (62%; 2991/4823), Giardia (24.6%; 1186/4823) and STEC (13.4%; 646/4823). The developed QMRA framework was used to assess the potential health impacts of partial and total well water treatment system failure. In the unlikely event of total treatment failure, total mean annual illnesses are predicted to almost double (4217 to 7064 cases per year), highlighting the importance of effective water treatment and comprehensive testing programs in reducing infectious health risks attributable to private well water in Ontario. Study findings indicate significant underreporting of waterborne AGI rates at the provincial level likely biasing public health interventions and programs that are effective in monitoring and minimizing the health risk associated with private well water.

Community Legionella outbreak linked to a cooling tower, 2022.

Background: Thirty-five laboratory-confirmed legionellosis cases were reported to the Simcoe Muskoka District Health Unit (Ontario, Canada) between September 27, 2022, and October 15, 2022, resulting in one death and 29 hospitalizations. This article describes the Legionella outbreak and highlights activities for managing the outbreak, including various environmental and infrastructural controls associated with the public health response and some of the unique challenges and potential solutions to mitigate future outbreaks. Methods: All cases of legionellosis were reported to and investigated by the local provincial health unit. Within a 6 km radius around the community, 27 cooling towers (CTs) were identified as potential sources of Legionella. Environmental samples were collected from 19 CTs and a long-term care home. Outcome: Of the 35 cases, 29 (83%) were hospitalized (including three long-term care residents) with two requiring intubation/ventilation. Of the five sputa (clinical isolates) collected from confirmed cases, four tested positive for Legionella pneumophila (one was positive for L. pneumophila serogroup 1-with the same sequence type as one of the CT isolates). Education and recommendations were provided by the local provincial health unit to operators to improve CT operation. Conclusion: Detection and management of community legionellosis outbreaks associated with CTs involve resources and time to properly identify and control risks. Measures for community risk mitigation included coordinating with provincial and community partners, developing methods to rapidly identify CTs as a likely source of infection and applying operational/maintenance/testing standards for CTs to control bacterial growth and minimize the dispersion of contaminated aerosols.

Monkeypox Virus Detection in Different Clinical Specimen Types.

A global monkeypox outbreak began in May 2022. Limited data exist on specimen type performance in associated molecular diagnostics. Consequently, a diverse range of specimen sources were collected in the initial weeks of the outbreak in Ontario, Canada. Our clinical evaluation identified skin lesions as the optimal diagnostic specimen source.

An assessment of total coliforms and associated thresholds as water quality indicators using a large Ontario private drinking water well dataset.

A spatiotemporally static total coliform (TC) concentration threshold of five colony-forming units (CFU) per 100 mL is used in Ontario to determine whether well water is of acceptable quality for drinking. The current study sought to assess the role of TC and associated thresholds as microbial water quality parameters as the authors hypothesized that, since static TC thresholds are not evidence-based, they may not be appropriate for all well water consumers. A dataset containing the microbial water quality information of 795,023 samples (including TC and Escherichia coli (E. coli) counts) collected from 253,136 private wells in Ontario between 2010 and 2017 was used. To accurately assess the relationship between E. coli and non-E. coli TC, "non-E. coli coliform" (NEC) counts were calculated from microbial water quality data and replaced TC throughout analyses. This study analysed NEC and E. coli detection rates to determine differences between the two, and NEC:E. coli concentration ratios to assess links, if any, between NEC and E. coli contamination. Study findings suggest that spatiotemporally static NEC thresholds are not appropriate because seasonal, spatial, and well-specific susceptibility factors are associated with distinct contamination trends. For example, NEC detection rates exhibited bimodality, with summer (29.4 %) and autumn (30.2 %) detection rates being significantly higher (p < 0.05) than winter (21.9 %) and spring (19.9 %). E. coli detection rates also varied seasonally, but peaked in summer rather than autumn. As such, it is recommended that these factors be considered during the development of private well water guidelines and that static thresholds be avoided. Furthermore, the authors propose that, because NEC:E. coli concentration ratios change in the context of the aforementioned factors, they may have a role in inferring groundwater contamination mechanisms, with high ratios being associated with generalized aquifer contamination mechanisms and low ratios with localized contamination mechanisms.

Estimating the duration and overlap of Escherichia coli contamination events in private groundwater supplies for quantitative risk assessment using a multiannual (2010-2017) provincial dataset.

Approximately 1.6 million individuals in Ontario rely on private water wells. Private well water quality in Ontario remains the responsibility of the well owner, and due to the absence of regulation, quantitative microbial risk assessment (QMRA) likely represents the most effective approach to estimating and mitigating waterborne infection risk(s) from these supplies. Annual contamination duration (i.e., contaminated days per annum) represents a central input for waterborne QMRA; however, it is typically based on laboratory studies or meta-analyses, thus representing an important limitation for risk assessment, as groundwater mesocosms cannot accurately replicate subsurface conditions. The present study sought to address these limitations using a large spatio-temporal in-situ groundwater quality dataset (>700,000 samples) to evaluate aquifer-specific E. coli die-off rates (CFU/100 mL per day decline), subsequent contamination sequence duration(s) and the likelihood of overlapping contamination events. Findings indicate median E. coli die-off rates of 0.38 CFU/100 mL per day and 0.64 CFU/100 mL per day, for private wells located in unconsolidated and consolidated aquifers, respectlvely, with mean calculated contamination sequence durations of 18 days (unconsolidated) and 11 days (consolidated). Study findings support and permit development of increasingly evidence-based, regionally- and temporally-specific quantitative waterborne risk assessment.

Rural Raccoons (Procyon lotor) Not Likely to Be a Major Driver of Antimicrobial Resistant Human Salmonella Cases in Southern Ontario, Canada: A One Health Epidemiological Assessment Using Whole-Genome Sequence Data.

Non-typhoidal Salmonella infections represent a substantial burden of illness in humans, and the increasing prevalence of antimicrobial resistance among these infections is a growing concern. Using a combination of Salmonella isolate short-read whole-genome sequence data from select human cases, raccoons, livestock and environmental sources, and an epidemiological framework, our objective was to determine if there was evidence for potential transmission of Salmonella and associated antimicrobial resistance determinants between these different sources in the Grand River watershed in Ontario, Canada. Logistic regression models were used to assess the potential associations between source type and the presence of select resistance genes and plasmid incompatibility types. A total of 608 isolates were obtained from the following sources: humans (n = 58), raccoons (n = 92), livestock (n = 329), and environmental samples (n = 129). Resistance genes of public health importance, including bla CMY-2, were identified in humans, livestock, and environmental sources, but not in raccoons. Most resistance genes analyzed were significantly more likely to be identified in livestock and/or human isolates than in raccoon isolates. Based on a 3,002-loci core genome multi-locus sequence typing (cgMLST) scheme, human Salmonella isolates were often more similar to isolates from livestock and environmental sources, than with those from raccoons. Rare instances of serovars S. Heidelberg and S. Enteritidis in raccoons likely represent incidental infections and highlight possible acquisition and dissemination of predominantly poultry-associated Salmonella by raccoons within these ecosystems. Raccoon-predominant serovars were either not identified among human isolates (S. Agona, S. Thompson) or differed by more than 350 cgMLST loci (S. Newport). Collectively, our findings suggest that the rural population of raccoons on swine farms in the Grand River watershed are unlikely to be major contributors to antimicrobial resistant human Salmonella cases in this region.

Sources of microbial contamination in the watershed and coastal zone of Soufriere, St. Lucia.

The Sustainable Development Goal 6 calls for global progress by 2030 in treating domestic wastewater and providing access to adequate sanitation facilities. However, meeting these goals will be a challenge for most Small Island Developing States, including Caribbean island nations. In the nearshore zone of the Soufriere region on the Caribbean island of St. Lucia, there is a history of high levels of bacteria of fecal origin. Possible land-based sources of microbial contamination in the Soufriere Bay include discharges from the Soufriere River and transport of wastewater, including fecal material from the town of Soufriere. This area is an important tourist destination and supports a local fishery. To identify the sources of microbial contamination in Soufriere Bay, a range of monitoring methods were employed in this study. In grab samples of surface water collected from the Soufriere River, counts of total coliforms and Escherichia coli were elevated above water quality guidelines. However, the spikes in concentrations of these indicator organisms in the river did not necessarily coincide with the spikes in the levels of total coliforms and E. coli detected in samples collected on the same dates in Soufriere Bay, indicating that there are other sources of pollution in the Bay besides discharges from the river. Monitoring for chemical indicators of wastewater (i.e., caffeine, sucralose, fluconazole) in the Soufriere River indicated that there are inputs of sewage or human fecal material throughout the watershed. However, analysis of Bacteroidales 16S rRNA genetic markers for fecal bacteria originating from humans, bovine ruminants, or other warm-blooded animals indicated that the majority of microbial contamination in the river was not from humans. Monitoring for chemical indicators of wastewater using passive samplers deployed in Soufriere Bay indicated that there are two "hot spots" of contamination located offshore of economically depressed areas of the town of Soufriere. This study indicates that efforts to control contamination of Soufriere Bay by fecal microorganisms must include management of pollution originating from both sewage and domestic animals in the watershed.

Real-Time RT-PCR Allelic Discrimination Assay for Detection of N501Y Mutation in the Spike Protein of SARS-CoV-2 Associated with B.1.1.7 Variant of Concern.

The N501Y amino acid mutation caused by a single point substitution A23063T in the spike gene of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is possessed by three variants of concern (VOCs), B.1.1.7, B.1.351, and P.1. A rapid screening tool using this mutation is important for surveillance during the coronavirus disease 2019 (COVID-19) pandemic. We developed and validated a single nucleotide polymorphism real-time reverse transcription PCR assay using allelic discrimination of the spike gene N501Y mutation to screen for potential variants of concern and differentiate them from SARS-CoV-2 lineages without the N501Y mutation. A total of 160 clinical specimens positive for SARS-CoV-2 were characterized as mutant (N501Y) or N501 wild type by Sanger sequencing and were subsequently tested with the N501Y single nucleotide polymorphism real-time reverse transcriptase PCR assay. Our assay, compared to Sanger sequencing for single nucleotide polymorphism detection, demonstrated positive percent agreement of 100% for all 57 specimens displaying the N501Y mutation, which were confirmed by Sanger sequencing to be typed as A23063T, including one specimen with mixed signal for wild type and mutant. Negative percent agreement was 100% in all 103 specimens typed as N501 wild type, with A23063 identified as wild type by Sanger sequencing. The identification of circulating SARS-CoV-2 lineages carrying an N501Y mutation is critical for surveillance purposes. Current identification methods rely primarily on Sanger sequencing or whole-genome sequencing, which are time consuming, labor intensive, and costly. The assay described herein is an efficient tool for high-volume specimen screening for SARS-CoV-2 VOCs and for selecting specimens for confirmatory Sanger or whole-genome sequencing. IMPORTANCE During the coronavirus disease 2019 (COVID-19) pandemic, several variants of concern (VOCs) have been detected, for example, B.1.1.7, B.1.351, P.1, and B.1.617.2. The VOCs pose a threat to public health efforts to control the spread of the virus. As such, surveillance and monitoring of these VOCs is of the utmost importance. Our real-time RT-PCR assay helps with surveillance by providing an easy method to quickly survey SARS-CoV-2 specimens for VOCs carrying the N501Y single nucleotide polymorphism (SNP). Samples that test positive for the N501Y mutation in the spike gene with our assay can be sequenced to identify the lineage. Thus, our assay helps to focus surveillance efforts and decrease turnaround times.

Impact of coronavirus disease 2019 (COVID-19) pre-test probability on positive predictive value of high cycle threshold severe acute respiratory coronavirus virus 2 (SARS-CoV-2) real-time reverse transcription polymerase chain reaction (RT-PCR) test results.

OBJECTIVES: Performance characteristics of SARS-CoV-2 nucleic acid detection assays are understudied within contexts of low pre-test probability, including screening asymptomatic persons without epidemiological links to confirmed cases, or asymptomatic surveillance testing. SARS-CoV-2 detection without symptoms may represent presymptomatic or asymptomatic infection, resolved infection with persistent RNA shedding, or a false-positive test. This study assessed the positive predictive value of SARS-CoV-2 real-time reverse transcription polymerase chain reaction (rRT-PCR) assays by retesting positive specimens from 5 pre-test probability groups ranging from high to low with an alternate assay. METHODS: In total, 122 rRT-PCR positive specimens collected from unique patients between March and July 2020 were retested using a laboratory-developed nested RT-PCR assay targeting the RNA-dependent RNA polymerase (RdRp) gene followed by Sanger sequencing. RESULTS: Significantly fewer (15.6%) positive results in the lowest pre-test probability group (facilities with institution-wide screening having ≤3 positive asymptomatic cases) were reproduced with the nested RdRp gene RT-PCR assay than in each of the 4 groups with higher pre-test probability (individual group range, 50.0%-85.0%). CONCLUSIONS: Large-scale SARS-CoV-2 screening testing initiatives among low pre-test probability populations should be evaluated thoroughly prior to implementation given the risk of false-positive results and consequent potential for harm at the individual and population level.

Characteristics of SARS-CoV-2 testing for rapid diagnosis of COVID-19 during the initial stages of a global pandemic.

Accurate SARS-CoV-2 diagnosis is essential to guide prevention and control of COVID-19. Here we examine SARS-CoV-2 molecular-based test performance characteristics and summarize case-level data related to COVID-19 diagnosis. From January 11 through April 22, 2020, Public Health Ontario conducted SARS-CoV-2 testing of 86,942 specimens collected from 80,354 individuals, primarily using real-time reverse-transcription polymerase chain reaction (rRT-PCR) methods. We analyzed test results across specimen types and for individuals with multiple same-day and multi-day collected specimens. Nasopharyngeal compared to throat swabs had a higher positivity (8.8% vs. 4.8%) and an adjusted estimate 2.9 Ct lower (SE = 0.5, p<0.001). Same-day specimens showed high concordance (98.8%), and the median Ct of multi-day specimens increased over time. Symptomatic cases had rRT-PCR results with an adjusted estimate 3.0 Ct (SE = 0.5, p<0.001) lower than asymptomatic/pre-symptomatic cases. Overall test sensitivity was 84.6%, with a negative predictive value of 95.5%. Molecular testing is the mainstay of SARS-CoV-2 diagnosis and testing protocols will continue to be dynamic and iteratively modified as more is learned about this emerging pathogen.

Duration of SARS-CoV-2 shedding: A population-based, Canadian study.

INTRODUCTION: There is an evidence gap regarding the duration of SARS-CoV-2 shedding and of its variability across different care settings and by age, sex, income, and co-morbidities. Such evidence is part of understanding of infectivity and reinfection. We examine direct measures of viral shedding using a linked population-based health administrative dataset. METHODS: Laboratory and sociodemographic databases for Ontario, Canada were linked to identify those testing positive (RT-PCR) between Jan. 15 and April 30, 2020 who underwent subsequent testing by May 31, 2020. To maximise use of available data, we computed two shedding duration estimates defined as the time between initial positive and most recent positive (documented shedding) or second of two negative tests (documented resolution). We also report multivariable results using quantile regression to examine subgroup differences. RESULTS: In Ontario, of the 16,595 who tested positive before April 30, 2020, 6604 had sufficient subsequent testing to allow shedding duration calculation. Documented shedding median duration calculated in 4,889 (29% of 16,595) patients was 19 days (IQR 12-28). Documented resolution median duration calculated in 3,219 (19% of the 16,595) patients was 25 days (IQR 18-34). Long-term care residents had 3-5 day longer shedding durations using both definitions. Shorter documented shedding durations of 2-4 days were observed in those living in higher income neighbourhoods. Shorter documented resolution durations of 2-3 days were observed at the 25th% of the distribution in those aged 20-49. Only 11.5% of those with definitive negative test results reverted to negative status by day 14. CONCLUSIONS: Viral shedding continued well beyond 14 days among this large subset of a population-based group with COVID-19, and longer still for long-term care residents and those living in less affluent neighborhoods. Our findings do not speak to duration of infectivity but are useful for understanding the expected duration of RT-PCR positivity and for identifying reinfection.

Environmental adaptation of E. coli within private groundwater sources in southeastern Ontario: Implications for groundwater quality monitoring and human health.

Groundwater quality monitoring typically employs testing for the presence of E. coli as a fecal indicator of recent ingress of human or animal fecal material. The efficacy of fecal indicator organisms is based on the primary criteria that the organism does not reproduce in the aquatic environment. However, recent studies have reported that E. coli may proliferate (i.e., has adapted to) in the external environment, including soil and surface water. To date, the presence of environmentally-adapted E. coli in groundwater has not been examined. The current study employed Clermont phylotyping and the presence of six accessory genes to identify the likely presence of adapted E. coli in private groundwater sources. E. coli isolates (n = 325) from 76 contaminated private water wells located in a southeastern Ontario watershed were compared with geographically analogous human and animal fecal E. coli isolates (n = 234). Cryptic clades III-V, a well-described environmentally-adapted Escherichia population, were identified in three separate groundwater wells, one of which exclusively comprised this adapted population. Dimensionality reduction (via Principal Component Analysis) was used to develop an "E. coli adaptation model", comprising three distinct components (groundwater, animal feces, human feces) and suggests adaptation occurs frequently in the groundwater environment. Model findings indicate that 23/76 (30.3%) wells had an entirely adapted community. Accordingly, the use of E. coli as a FIO returned a false positive result in these instances, while an additional 23/76 (30.3%) wells exhibited some evidence of adaptation (i.e., not all isolates were adapted) representing an over-estimate of the magnitude (concentration) of contamination. Study findings highlight the need to further characterize environmentally-adapted E. coli in the groundwater environment and the potential implications with respect to water quality policy, legislation and determinants of human health risk both regionally and internationally.

Exploration of E. coli contamination drivers in private drinking water wells: An application of machine learning to a large, multivariable, geo-spatio-temporal dataset.

Groundwater resources are under increasing threats from contamination and overuse, posing direct threats to human and environmental health. The purpose of this study is to better understand drivers of, and relationships between, well and aquifer characteristics, sampling frequencies, and microbiological contamination indicators (specifically E. coli) as a precursor for improving knowledge and tools to assess aquifer vulnerability and well contamination within Ontario, Canada. A dataset with 795, 023 microbiological testing observations over an eight-year period (2010 to 2017) from 253,136 unique wells across Ontario was employed. Variables in this dataset include date and location of test, test results (E. coli concentration), well characteristics (well depth, location), and hydrogeological characteristics (bottom of well stratigraphy, specific capacity). Association rule analysis, univariate and bivariate analyses, regression analyses, and variable discretization techniques were utilized to identify relationships between E. coli concentration and the other variables in the dataset. These relationships can be used to identify drivers of contamination, their relative importance, and therefore potential public health risks associated with the use of private wells in Ontario. Key findings are that: i) bedrock wells completed in sedimentary or igneous rock are more susceptible to contamination events; ii) while shallow wells pose a greater risk to consumers, deep wells are also subject to contamination events and pose a potentially unanticipated risk to health of well users; and, iii) well testing practices are influenced by results of previous tests. Further, while there is a general correlation between months with the greatest testing frequencies and concentrations of E. coli occurring in samples, an offset in this timing is observed in recent years. Testing remains highest in July while peaks in adverse results occur up to three months later. The realization of these trends prompts a need to further explore the bases for such occurrences.

Drinking Water Consumption Patterns among Private Well Users in Ontario: Implications for Exposure Assessment of Waterborne Infection.

Understanding the water consumption patterns within a specific population informs development of increasingly accurate, spatially specific exposure and/or risk assessment of waterborne infection. The current study examined the consumption patterns of private well users in Ontario while considering potentially influential underlying sociodemographics, household characteristics, and experiential factors. A province-wide online survey was circulated between May and August 2018 (n = 1,162). Overall, 81.5% of respondents reported daily well water consumption (i.e., tap water). Results indicate a mean daily well water consumption rate of 1,132 mL/day (SD = 649 mL/day) among well water consumers. Gender was significantly associated with well water consumption, with higher consumption rates found among female respondents. The experience of acute gastrointestinal illness (AGI) symptoms or diagnosis in the past 12 months did not impact the volume of water consumed, suggesting that experiencing previous AGI does not decrease consumption volumes, and therefore exposure over time. Significantly higher rates of well water consumption were found among respondents who reported previous testing or ongoing water treatment. Approximately 45.5% of survey respondents who stated that they do not consume well water selected bottled water as their primary household drinking water supply. Bottled water consumption was also not associated with previous AGI experiences. Findings will inform future quantitative microbial risk assessments associated with private well water use by providing spatially and demographically specific estimates of well water consumption.