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Aims: Rat‐associated zoonotic pathogen transmission at the human–wildlife interface is a public health concern in urban environments where Norway rats (Rattus norvegicus) thrive on abundant anthropogenic resources and live in close contact with humans and other animal species. To identify potential factors influencing zoonotic pathogen occurrence in rats, we investigated associations between environmental and sociodemographic factors and Leptospira interrogans and Bartonella spp. infections in rats from Windsor, Ontario, Canada, while controlling for the potential confounding effects of animal characteristics (i.e., sexual maturity and body condition). Methods and Results: Between November 2018 and June 2021, 252 rats were submitted by collaborating pest control professionals. Kidney and spleen samples were collected for L. interrogans and Bartonella spp. PCR and sequencing, respectively. Of the rats tested by PCR, 12.7% (32/252) were positive for L. interrogans and 16.3% (37/227) were positive for Bartonella species. Associations between infection status and environmental and sociodemographic variables of interest were assessed via mixed multivariable logistic regression models with a random intercept for social group and fixed effects to control for sexual maturity and body condition in each model. The odds of L. interrogans infection were significantly higher in rats from areas with high building density (odds ratio [OR]: 3.76; 95% CI: 1.31–10.79; p = 0.014), high human population density (OR: 3.31; 95% CI: 1.20–9.11; p = 0.021), high proportion of buildings built in 1960 or before (OR: 11.21; 95% CI: 2.06–60.89; p = 0.005), and a moderate number of reports of uncollected garbage compared to a low number of reports (OR: 4.88; 95% CI: 1.01–23.63; p = 0.049). A negative association was observed between median household income and Bartonella spp. infection in rats (OR: 0.26; 95% CI: 0.08–0.89; p = 0.031). Conclusions: Due to the complexity of the ecology of rat‐associated zoonoses, consideration of environmental and sociodemographic factors is of critical importance to better understand the nuances of host–pathogen systems and inform how urban rat surveillance and intervention efforts should be distributed within cities. [ABSTRACT FROM AUTHOR]

Lyme disease risk areas have increased across Canada in recent decades with the ongoing range expansion of Ixodes scapularis and Borrelia burgdorferi. Different methodologies are used by federal and provincial governments to determine local Lyme disease risk, which can make comparisons between regions challenging. In this study, seven Canadian Lyme disease risk classification methodologies were compared with each other to highlight the strengths and limitations of how each definition measured I. scapularis and B. burgdorferi risk. Each methodology was applied to active surveillance data from Ontario, and per cent agreement and kappa statistics were calculated. The methodologies varied in their measurements of the risk of exposure to I. scapularis and B. burgdorferi based on their use of active surveillance techniques, multiple types of collected surveillance data and laboratory confirmation of B. burgdorferi. Most initial Lyme disease risk site classifications were maintained over time. Kappa and per cent agreement statistics highlighted large differences between 8 of the 15 methodology pairings, indicating the presence of inconsistencies between most methodologies. Accurate, consistent surveillance and assessment of the spread of I. scapularis and its pathogens will aid with communicating Lyme disease risk to the public and preventing tick‐borne pathogen transmission. [ABSTRACT FROM AUTHOR]

Norway rats (Rattus norvegicus) inhabit cities worldwide and live in close association with humans. Studies of urban rat zoonoses often rely on live‐trapping, with fewer studies using rats sourced through lethal pest control interventions. Our objectives were to evaluate the utility of rats collected by pest control professionals for zoonotic pathogen surveillance and determine whether we could detect Leptospira interrogans and Streptobacillus moniliformis in pest control sourced rats. Rat carcasses were submitted from Windsor, Canada by pest control professionals between November 2018 and March 2020. Submissions were categorized by season and land use. Necropsies were performed to classify carcass quality, collect tissue samples, and record demographic data. The association between carcass quality and the ability to collect tissue samples for pathogen surveillance was assessed via an exact logistic regression model. Using PCR, a subset of kidney and spleen samples were tested for L. interrogans and S. moniliformis, respectively. Our sample of pest control sourced rats had similar sex and age distributions to those of live‐trapping studies. Rats were primarily submitted from residential and industrial locations during fall, winter, and spring, which may reflect pest control service areas and peak business periods, rather than rat distribution. Of 124 submissions, 98 (79.0%) of rats showed only mild decomposition. The odds of collecting all tissue samples were reduced for fair compared to good‐quality carcasses (OR: 0.029; 95% CI: 0–0.25; p =.0009) and for poor compared to fair‐quality carcasses (OR: 0.048; 95% CI: 0.00085–0.53; p =.0065). Leptospira interrogans and S. moniliformis were detected in 9.1% (4/44) and 27.3% (15/55) of a subset of rats tested, respectively. Our results suggest that pest control sourced rats are suitable for surveillance for multiple zoonotic pathogens in urban environments. This method of rat collection may provide preliminary information to guide more detailed ecological studies. [ABSTRACT FROM AUTHOR]

Prior to 2012, Echinococcus multilocularis was not known to occur in any host in Ontario, Canada. However, since that year, five cases of alveolar echinococcosis have been diagnosed in dogs that resided at the western end of Lake Ontario. In addition, E. multilocularis has been shown to be a common infection in wild canids (i.e. coyotes and foxes) across southern Ontario with a high‐risk infection cluster in the area surrounding the western shores of Lake Ontario and northern shores of Lake Erie. In regions endemic for E. multilocularis, dog ownership is considered a risk factor for human alveolar echinococcosis. A study was therefore carried out to determine the prevalence of E. multilocularis intestinal infections in dogs within the high‐risk infection cluster. From May to November 2018, faecal samples were collected from 477 dogs aged ≥6 months that visited 12 off‐leash dog parks in the Halton, Hamilton and Niagara public health units. Faecal samples were analysed via a magnetic capture probe DNA extraction and real‐time PCR method for E. multilocularis DNA. Overall, 0% (97.5% CI: 0%–0.80%) of samples tested positive. This result informs preventive recommendations for E. multilocularis infections in dogs in this region. [ABSTRACT FROM AUTHOR]

Campylobacter is a leading cause of foodborne illness in humans worldwide. Sources of infection are often difficult to identify, and are, generally, poorly understood. Recent work suggests that wildlife may represent a source of Campylobacter for human infections. Using a repeated cross‐sectional study design, raccoons were trapped on five swine farms and five conservation areas in southern Ontario from 2011 to 2013. Our objectives were to: (a) assess the impact of seasonal, climatic, location, annual and raccoon demographic factors on the occurrence of Campylobacter jejuni in these animals; and (b) identify clusters of C. jejuni in space, time and space‐time using spatial scan statistics. Multi‐level multivariable logistic regression was used to examine the odds of isolating C. jejuni, with site and animal modelled as random intercepts. The following independent variables were examined: raccoon age and sex, year, location type, season, temperature and rainfall. A total of 1,096 samples were obtained from 627 raccoons; 46.3% were positive for C. jejuni. The following interactions and their main effects were significant (p <.05) and retained in the final model: season × temperature, year × rainfall, year × temperature. Based on the results from our multivariable model and spatial scan statistics, climatic variables (i.e. rainfall, temperature and season) were associated with the carriage of C. jejuni by raccoons, but the effects were not consistent, and varied by location and year. Although raccoons may pose a zoonotic risk due to their carriage of Campylobacter, further work is required to characterize the transmission and movement of this microorganism within the ecosystem. [ABSTRACT FROM AUTHOR]

Echinococcus multilocularis was recently reported in wild canids across southern Ontario, a newly recognized endemic area in Canada. In such areas, a comprehensive understanding of factors associated with infection in definitive hosts (wild canids) is critical for mitigating risk of transmission to humans. However, little is known about the transmission dynamics of the parasite in definitive hosts for this region. A study was therefore carried out to investigate the association of host‐level (sex, body condition), environmental (southern Ontario region, land cover), temporal (season, hunting season, calendar year) and extraneous factors (submitter type) with E. multilocularis infection in coyotes in southern Ontario. Between November 2015 and March 2017, 416 coyotes were collected from across the region as part of a study that investigated the prevalence and distribution of the parasite in wild canids; approximately 24% of coyotes were positive for E. multilocularis. Associations between infection and factors of interest were assessed via a mixed‐effects logistic regression model with a random intercept for submitter to account for clustering. Coyotes with poor body condition were at greater odds of E. multilocularis infection than those in good condition (odds ratio [OR] 2.14; 95% CI: 1.08–4.26; p =.030). A negative association was observed between infection in coyotes and the proportion of natural land in a coyote's estimated home range (OR: 0.67; 95% CI: 0.52–0.85; p =.001). Coyotes from the western region of southern Ontario had lower odds of infection compared to coyotes from the central region (OR: 0.26; 95% CI: 0.12–0.55; p <.001). These results can be used to help guide future public health prevention strategies for human alveolar echinococcosis. [ABSTRACT FROM AUTHOR]

Traditionally, zoonotic pathogen ecology studies in wildlife have focused on the interplay among hosts, their demographic characteristics and their pathogens. But pathogen ecology is also influenced by factors that traverse the hierarchical scale of biological organization, ranging from within‐host factors at the molecular, cellular and organ levels, all the way to the host population within a larger environment. The influence of host disease and co‐infections on zoonotic pathogen carriage in hosts is important because these factors may be key to a more holistic understanding of pathogen ecology in wildlife hosts, which are a major source of emerging infectious diseases in humans. Using wild Norway rats (Rattus norvegicus) as a model species, the purpose of this study was to investigate how host disease and co‐infections impact the carriage of zoonotic pathogens. Following a systematic trap and removal study, we tested the rats for the presence of two potentially zoonotic bacterial pathogens (Bartonella tribocorum and Leptospira interrogans) and assessed them for host disease not attributable to these bacteria (i.e., nematode parasites, and macroscopic and microscopic lesions). We fitted multilevel multivariable logistic regression models with pathogen status as the outcome, lesions and parasites as predictor variables and city block as a random effect. Rats had significantly increased odds of being infected with B. tribocorum if they had a concurrent nematode infection in one or more organ systems. Rats with bite wounds, any macroscopic lesion, cardiomyopathy or tracheitis had significantly increased odds of being infected with L. interrogans. These results suggest that host disease may have an important role in the ecology and epidemiology of rat‐associated zoonotic pathogens. Our multiscale approach to assessing complex intrahost factors in relation to zoonotic pathogen carriage may be applicable to future studies in rats and other wildlife hosts. [ABSTRACT FROM AUTHOR]

The role of free‐ranging wildlife in the epidemiology of enteropathogens causing clinical illness in humans and domestic animals is unclear. Salmonella enterica and anti‐microbial resistant bacteria have been detected in the faeces of raccoons (Procyon lotor), but little is known about the carriage of these bacteria in other sympatric meso‐mammals. Our objectives were to: (a) report the prevalence of Salmonella and associated anti‐microbial resistance, Campylobacter spp, Clostridium difficile, and anti‐microbial resistant Escherichia coli in the faeces of striped skunks (Mephitis mephitis) and Virginia opossums (Didelphis virginiana) in southern Ontario; and (b) compare the prevalence of these bacteria in the faeces of these meso‐mammal hosts with raccoons from a previously reported study. Faecal swabs were collected from striped skunks and Virginia opossums on five swine farms and five conservation areas from 2011 to 2013. Salmonella was detected in 41% (9/22) and 5% (5/95) of faecal swabs from Virginia opossums and striped skunks, respectively. None of the Salmonella serovars carried resistance to anti‐microbials. The prevalence of Campylobacter spp., C. difficile, and anti‐microbial resistant E. coli ranged from 6% to 22% in striped skunk and Virginia opossums. Using exact logistic regression, Salmonella was significantly more likely to be detected in faecal swabs of Virginia opossums than skunks and significantly less likely in faecal swabs from skunks than raccoons from a previously reported study. In addition, Campylobacter spp. was significantly more likely to be detected in raccoons than opossums. Salmonella Give was detected in 8/9 (89%) of Salmonella‐positive Virginia opossum faecal swabs. Our results suggest that striped skunks and Virginia opossums have the potential to carry pathogenic enteric bacteria in their faeces. The high prevalence of Salmonella Give in Virginia opossum faecal swabs in this study as well as its common occurrence in other Virginia opossum studies throughout North America suggests Virginia opossums may be reservoirs of this serovar. [ABSTRACT FROM AUTHOR]

From May through October 2016, we conducted a repeated cross‐sectional study examining the effects of temporal, spatial, flock and demographic factors (i.e. juvenile vs. adult) on the prevalence of Campylobacter and antimicrobial resistant Enterobacteriaceae among 344 fresh faecal samples collected from Canada geese (Branta canadensis) from four locations where birds nested in Guelph, Ontario, Canada. The overall prevalence of Campylobacter among all fresh faecal samples was 9.3% and was greatest in the fall when these birds became more mobile following the nesting season. Based on 40 gene comparative genomic fingerprinting (CGF40), the increase in prevalence noted in the fall was matched by an increase in the number of unique CGF40 subtypes identified. Resistance to colistin was detected most commonly, in 6% of Escherichia coli isolates, and was highest in the late summer months. All colistin‐resistant isolates were negative for the mcr‐1 to mcr‐5 genes; a chromosomal resistance mechanism (PmrB) was identified in all of these isolates. The prevalence of samples with E. coli exhibiting multi‐class resistance or extended spectrum beta‐lactamase was low (i.e. <2% of samples). The intra‐class correlation coefficients, estimated from the variance components of multilevel logistic regression models, indicated that the shedding of Campylobacter and antimicrobial resistant E. coli among geese within a flock (i.e. birds collected from the same site on the same day) was moderately correlated. Spatial, temporal, and spatiotemporal clusters identified using the spatial scan statistic, largely supported the findings from our multi‐level models. Salmonella was not isolated from any of the fresh faecal samples collected suggesting that its prevalence in this population of birds was very low. [ABSTRACT FROM AUTHOR]

Abstract: Antimicrobial resistant bacteria and zoonotic pathogens have previously been isolated from Canada geese. We examined the prevalence of three enteric bacteria (i.e. Campylobacter, Salmonella, Escherichia coli) among Canada geese from three sampling sources in southern Ontario from 2013 through 2015. Samples were obtained by convenience from hunting groups, diagnostic birds submitted for post‐mortem, and fresh faeces from live birds in parks. Escherichia coli isolates were isolated and tested for susceptibility to 15 antimicrobials using the Canadian Integrated Program for Antimicrobial Resistance Surveillance test panel. The prevalences of Salmonella, Campylobacter and E. coli were 0%, 11.2% and 72.6%, respectively. Among E. coli isolates, 7.9% were resistant to ≥1 class of antimicrobials and 5.6% were resistant to ≥2 classes of antimicrobials, with some including resistance to antimicrobials of highest importance in human medicine. A significant association between season and E. coli resistance among samples from live birds was noted; summer samples had no resistant E. coli isolates, whereas spring samples demonstrated the highest prevalence of E. coli resistant to ≥1 class of antimicrobials (20.0%) among all sources. In addition, Campylobacter coli were only isolated from the spring faecal samples. Flock‐level clustering was an important statistical consideration, as flock was a significant random effect in all but two of our models. Detection of Campylobacter and antimicrobial resistant E. coli from Canada geese suggests that these birds may play a role in disseminating these organisms within the environment. [ABSTRACT FROM AUTHOR]