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Summary The Asian longhorned tick (ALHT) ( Haemaphysalis longicornis Neumann, 1901) is a vector for many pathogenic micro-organisms of veterinary and public health importance. The ALHT is native to east Asia but was accidentally introduced into New Zealand and Australia in the early 1990s/late 1880s, respectively. It is now well established in these countries and it is a competent vector for Theileria orientalis Ikeda, which can cause significant livestock morbidity and mortality. Recently, this tick was introduced into North America. It was first formally detected on sheep in New Jersey in 2017, although it is now thought that it may have been present in the USA since 2010 but was initially misidentified as the morphologically similar rabbit tick H. leporispalustris (Packard, 1869). As of August 2022, this tick has been reported in 17 US eastern states. In this case study, we illustrate how we engaged an interdisciplinary team to examine the habitat suitability for the ALHT across North America under current and future climatic conditions. Using a transdisciplinary approach, we also considered potential routes of entry into Canada from the north-eastern states of the USA. Using published and current surveillance data, we generated habitat suitability models to predict the most likely areas in North America for the geographic expansion of the ALHT. Following discussions with climate change experts and entomologists, models were also developed to examine future habitat suitability under selected climate change scenarios. Interviews with experts and government colleagues suggest that likely routes of entry into Canada include cross-border movement of infested terrestrial wildlife and livestock, dog importations and the movement of migratory birds. Enhanced tick surveillance, including the use of citizen science, is required to gain a better understanding of how this tick might spread in the future. The logo of T C C dash 3 W. A map is depicted and the bottom-left part of the map is highlighted. The logo of the University of Calgary Faculty of Veterinary Medicine. The logo of the Canadian Animal Health Surveillance System or C A H S S.

Salmonella infections are a major human health concern. In the elderly and immunocompromised, infections can be life-threatening and may require antibiotic therapy. Where antibiotic therapy is required, antimicrobials of choice include fluoroquinolones and extended-spectrum cephalosporins (ESC). The aim of this study is to utilize data from the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) to compare the prevalence of Salmonella serovars between broiler chicken and turkey flocks across Canada, and to gain an understanding of the prevalence of resistance to antimicrobials categorized as important to human health. There were 1,596 Salmonella isolates obtained from 514 broiler chicken flocks, and 659 Salmonella isolates obtained from 217 turkey flocks (2013-2018). All isolates were obtained from pooled faecal samples. Among broiler chicken flocks, the top three serovars were Kentucky (n = 573, 36%), Enteritidis (n = 314, 20%) and Heidelberg (n = 127, 8%). Resistance to ceftriaxone among Salmonella ser. Kentucky decreased from 27% in 2013 to 22% in 2018. There was no resistance among Salmonella ser. Enteritidis reported until 2018 when one isolate from British Columbia was resistant to ampicillin, streptomycin, sulphisoxazole and tetracycline. Salmonella ser. Heidelberg resistance to ceftriaxone decreased from 19% in 2013 to 14% in 2018. Among turkey flocks the top three serovars were Uganda (n = 109, 16.5%), Hadar (n = 85, 12%) and Muenchen (n = 66, 10%). No isolates of Salmonella ser. Uganda or Salmonella ser. Muenchen were resistant to any β-lactams. Salmonella ser. Hadar (34/81, 42%) exhibited resistance to ampicillin. There was no resistance to quinolones among turkey isolates. Emerging resistance among Salmonella ser. Enteritidis, and resistance to β-lactams and fluoroquinolones among Salmonella ser. Kentucky from broilers are cause for concern as these classes of antimicrobials are important for treatment of salmonellosis.

Approximately 238,000 to 450,000 Albertans rely on private water wells for their water needs. In Canada, private well owners are responsible for monitoring and maintaining the quality of their wate...

Shiga-toxin-producing Escherichia coli (STEC) represent a major concern for waterborne disease outbreaks associated with consumption of contaminated groundwater. Over 4 million people rely on private groundwater systems as their primary drinking water source in Canada; many of these systems do not meet current standards for water quality. This manuscript provides a scoping overview of studies examining STEC prevalence and occurrence in groundwater, and it includes a synopsis of the environmental variables affecting survival, transport, persistence, and overall occurrence of these important pathogenic microbes in private groundwater wells used for drinking purposes.

Enterococcus spp. have arisen as important nosocomial pathogens and are ubiquitous in the gastrointestinal tracts of animals and the environment. They carry many intrinsic and acquired antimicrobial resistance genes. Because of this, surveillance of Enterococcus spp. has become important with whole genome sequencing emerging as the preferred method for the characterization of enterococci. A scoping review was designed to determine how the use of whole genome sequencing in the surveillance of Enterococcus spp. adds to our knowledge of antimicrobial resistance in Enterococcus spp. Scoping review design was guided by the PRISMA extension and checklist and JBI Reviewer's Guide for scoping reviews. A total of 72 articles were included in the review. Of the 72 articles included, 48.6% did not state an association with a surveillance program and 87.5% of articles identified Enterococcus faecium. The majority of articles included isolates from human clinical or screening samples. Significant findings from the articles included novel sequence types, the increasing prevalence of vancomycin-resistant enterococci in hospitals, and the importance of surveillance or screening for enterococci. The ability of enterococci to adapt and persist within a wide range of environments was also a key finding. These studies emphasize the importance of ongoing surveillance of enterococci from a One Health perspective. More studies are needed to compare the whole genome sequences of human enterococcal isolates to those from food animals, food products, the environment, and companion animals.

We previously demonstrated the existence of naturalized strains of E. coli in wastewater and herein perform an in-depth comparative whole genome analysis of these strains (n = 17). Fourteen of the Canadian E. coli strains, isolated from geographically separated wastewater treatment plants, were virtually identical at the core genome and were ≥96% similar at the whole genome level, suggesting clonal-relatedness among these isolates. Remarkably, these strains were shown to be extremely similar to the genome of an E. coli isolated from wastewater in Switzerland, suggesting a global distribution of these strains. The genomes of three other Canadian wastewater strains were more diverse but very similar to the genomes of E. coli isolates collected from U.S. wastewater samples. Based on maximum likelihood phylogenetic analysis, wastewater strains from Canada, the U.S. and Switzerland formed a clade separate from other known enteric phylogroups (i.e., A, B1, B2, D, E) and the cryptic clades. All Canadian, Swiss and U.S. wastewater strains possessed a common SNP biomarker pattern across their genomes, and a sub-population (i.e., 14 Canadian and 1 Swiss strain) also possessed a previously identified wastewater-specific marker known as uspC-IS30-flhDC element. Biochemical heat mapping of 518 categories of genes recapitulated phylogeny, with wastewater strains phenotypically clustering separately from enteric and cryptic clades. Wastewater strains were enriched for stress-response genes (i.e., nutrient acquisition/deprivation, DNA repair, oxidative stress, and UV resistance) - elements reflective of their environmental survival challenges. Wastewater strains were shown to carry a plethora of known antibiotic resistance (AR) genes, the patterns of which were remarkably similar among all Canadian, U.S. and Swiss wastewater strains. Virulence gene composition was also similar among all the wastewater strains, with an abundant representation of virulence genes commonly associated with urinary pathogenic E. coli (UPEC) as well as enterohemorrhagic (EHEC) E. coli. The remarkable degree of similarity between all wastewater strains from Canada, Switzerland and the U.S. suggests the evolution and global-dissemination of water treatment-resistant clone of E. coli. These finding, along with others, raise some important concerns about the potential for emergence of E. coli pathotypes resistant to water-treatment.

Muskoxen (Ovibos moschatus) are an integral component of Arctic biodiversity. Given low genetic diversity, their ability to respond to future and rapid Arctic change is unknown, although paleontological history demonstrates adaptability within limits. We discuss status and limitations of current monitoring, and summarize circumpolar status and recent variations, delineating all 55 endemic or translocated populations. Acknowledging uncertainties, global abundance is ca 170 000 muskoxen. Not all populations are thriving. Six populations are in decline, and as recently as the turn of the century, one of these was the largest population in the world, equaling ca 41% of today’s total abundance. Climate, diseases, and anthropogenic changes are likely the principal drivers of muskox population change and result in multiple stressors that vary temporally and spatially. Impacts to muskoxen are precipitated by habitat loss/degradation, altered vegetation and species associations, pollution, and harvest. Which elements are relevant for a specific population will vary, as will their cumulative interactions. Our summaries highlight the importance of harmonizing existing data, intensifying long-term monitoring efforts including demographics and health assessments, standardizing and implementing monitoring protocols, and increasing stakeholder engagement/contributions. Electronic supplementary material The online version of this article (10.1007/s13280-019-01205-x) contains supplementary material, which is available to authorized users.

Water wells are important in providing water for rural populations that are far removed from municipal water supplies. Over 400,000 rural Albertans use water wells for domestic purposes. Th...

Risk assessments are utilized to understand the risks associated with bacterial foodborne pathogens with increased antimicrobial resistance (AMR). This review aimed at identifying and describing the literature on risk assessments for the study of antimicrobial resistance of foodborne bacteria in humans, with focus only on products intended for the treatment of bacterial infections. The definition of what constituted a risk assessment from a methodological perspective is based on the Codex Guidelines for Risk Analysis of Foodborne Antimicrobial Resistance. Thirty-eight articles produced between 1989–2018 were identified following a comprehensive search strategy. Most articles (25/38) were quantitative risk assessments. The most frequently identified hazard of concern was the consumption of retail meat. Knowledge gaps were evident throughout the four components of the risk assessments, particularly in the hazard characterization component. There appears to be a lack of evidence available to inform risk assessments regarding the additional effects of disease with an AMR organism. The adverse health effects attributable to AMR were addressed in less than half of the articles identified. Additionally, vulnerable populations were not generally included in the risk assessment. There is the potential for risk assessments to consider a broader scope for sources of information to identify and characterize the hazard of concern. Codex guidelines provide a structured framework by which to conduct a risk assessment relating to the microbiological hazards and AMR and should be addressed in every risk assessment on these topics.

Infectious laryngotracheitis (ILT) is an infectious upper respiratory tract disease that impacts the poultry industry worldwide. ILT is caused by an alphaherpesvirus commonly referred to as infectious laryngotracheitis virus (ILTV). Vaccination with live attenuated vaccines is practiced regularly for the control of ILT. However, extensive and improper use of live attenuated vaccines is related to vaccine viruses reverting to virulence. An increase in mortality and pathogenicity has been attributed to these vaccine revertant viruses. Recent studies characterized Canadian ILTV strains originating from ILT outbreaks as related to live attenuated vaccine virus revertants. However, information is scarce on the pathogenicity and transmission potential of these Canadian isolates. Hence, in this study, the pathogenicity and transmission potential of two wildtype ILTVs and a chicken embryo origin (CEO) vaccine revertant ILTV of Canadian origin were evaluated. To this end, 3-week-old specific pathogen-free chickens were experimentally infected with each of the ILTV isolates and compared to uninfected controls. Additionally, naïve chickens were exposed to the experimentally infected chickens to mimic naturally occurring infection. Pathogenicity of each of these ILTV isolates was evaluated by the severity of clinical signs, weight loss, mortality, and lesions observed at the necropsy. The transmission potential was evaluated by quantification of ILTV genome loads in oropharyngeal and cloacal swabs and tissue samples of the experimentally infected and contact-exposed chickens, as well as in the capacity to produce ILT in contact-exposed chickens. We observed that the CEO vaccine revertant ILTV isolate induced severe disease in comparison to the two wildtype ILTV isolates used in this study. According to ILTV genome load data, CEO vaccine revertant ILTV isolate was successfully transmitted to naïve contact-exposed chickens in comparison to the tested wildtype ILTV isolates. Overall, the Canadian origin CEO vaccine revertant ILTV isolate possesses higher virulence, and dissemination potential, when compared to the wildtype ILTV isolates used in this study. These findings have serious implications in ILT control in chickens.

Horizontal gene transfer is an important mechanism which facilitates bacterial populations in overcoming antimicrobial treatment. In this study, a total of 120 Escherichia coli and 62 Salmonella enterica subsp. enterica isolates were isolated from broiler chicken farms in Alberta. Fourteen serovars were identified among Salmonella isolates. Thirty one percent of E. coli isolates (37/120) were multiclass drug resistant (resistant to ≥ 3 drug classes), while only about 16% of Salmonella isolates (10/62) were multiclass drug resistant. Among those, eight E. coli isolates had an AmpC-type phenotype, and one Salmonella isolate had an extended-spectrum beta-lactamase (ESBL)-type beta-lactamase phenotype. We identified both AmpC-type (blaCMY-2) and ESBL-type (blaTEM) genes in both E. coli and Salmonella isolates. Plasmids from eight of nine E. coli and Salmonella isolates were transferred to recipient strain E. coli J53 through conjugation. Transferable plasmids in the eight E. coli and Salmonella isolates were also transferred into a lab-made sodium azide-resistant Salmonella recipient through conjugation. The class 1 integrase gene, int1, was detected on plasmids from two E. coli isolates. Further investigation of class 1 integron cassette regions revealed the presence of an aadA gene encoding streptomycin 3’’-adenylyltransferase, an aadA1a/aadA2 gene encoding aminoglycoside 3’’-O-adenyltransferase, and a putative adenylyltransferase gene. This study provides some insight into potential horizontal gene transfer events of antimicrobial resistance genes between E. coli and Salmonella in broiler chicken production.

Tick infestation is the most reported parasitological problem in cattle in Bhutan. In May and June 2019, we collected ticks from 240 cattle in two districts of Eastern Bhutan. Tick presence, diversity, and infestation prevalence were examined by morphological identification of 3600 live adult ticks. The relationships between cattle, geographic factors, and infestation prevalence were assessed using logistic regression analyses. Habitat suitability for the tick species identified was determined using MaxEnt. Four genera and six species of ticks were found. These were Rhipicephalus microplus (Canestrini) (70.2% (95% confidence interval (CI): 68.7–71.7)), Rhipicephalus haemaphysaloides Supino (18.8% (95% CI: 17.5–20.1)), Haemaphysalis bispinosa Neumann (8.2% (95% CI: 7.3–9.1)), Haemaphysalis spinigera Neumann (2.5% (95% CI: 2–3)), Amblyomma testudinarium Koch (0.19% (95% CI: 0.07–0.4)), and a single unidentified Ixodes sp. Logistic regression indicated that the variables associated with infestation were: longitude and cattle age for R. microplus; latitude for R. haemaphysaloides; and altitude and cattle breed for H. bispinosa and H. spinigera. MaxEnt models showed land cover to be an important predictor for the occurrence of all tick species examined. These findings provide information that can be used to initiate and plan enhanced tick surveillance and subsequent prevention and control programs for ticks and tick-borne diseases in cattle in Bhutan.

Livestock farming plays an important role in supporting the livelihood of resource-poor subsistence farmers in Bhutan. However, ticks and tick-borne diseases (TBDs) are one of the major constraints to livestock farming due to their negative effect on health and production. To date, no study has been conducted in Bhutan to assess farmers’ knowledge, attitude, and practices (KAP) about ticks and TBDs in cattle, although such information is essential in ensuring the development and adoption of effective prevention and control measures. Therefore, a KAP survey was conducted among 246 cattle owners in the Samkhar sub-district of eastern Bhutan in June 2019, using a structured questionnaire. Based on our scoring criteria, 52% [95%CI: 45.5–58.4] had adequate knowledge about ticks as potential vectors of diseases. Logistic regression analysis showed that the individuals who practiced a stall-feeding system of cattle rearing were 2.8 times [OR = 2.8 (95%CI: 1.66–4.78)] more likely to have adequate knowledge than others. Sixty-eight percent [95%CI: 62.5–74.4] had a favorable attitude toward tick prevention and control programs. Men were 1.95 times [OR = 1.95 (95%CI: 1.09–3.55)] more likely to have a favorable attitude than women, and the individuals who practiced a stall-feeding system were 2.59 times [OR = 2.59 95%CI: 1.45–4.78)] more likely to have a favorable attitude than others, after adjusting for the effect of other variables in the model. Overall, only 38% [95%CI 32.5–45] of the respondents reported tick infestation as one of the most important animal health problems, but 100% reported using acaricides to control ticks in cattle. Despite a high level of acaricide usage, the level of knowledge was low among the farmers interviewed. Findings from this study underline the importance of considering identified knowledge gaps and initiating education efforts to improve the adoption of effective tick prevention and control measures among farmers.

Erysipelothrix rhusiopathiae was detected immunohistochemically in contagious ecthyma (orf virus) dermatitis in two muskoxen (Ovibos moschatus), harvested and found dead in 2014 and 2015, respectively, on Victoria Island, Canada. This may help target further research on E. rhusiopathiae epidemiology and mechanisms of infection in muskoxen, recently associated with widespread mortalities in Canada's Arctic.

Infectious laryngotracheitis virus (ILTV) is a herpes virus that causes an acute respiratory disease of poultry known as infectious laryngotracheitis (ILT). Chicken embryo origin (CEO) and tissue culture origin (TCO) live attenuated vaccines are routinely used for the control of ILT. However, vaccine virus is known to revert to virulence, and it has been recently shown that ILT field viral strains can undergo recombination with vaccinal ILTV and such recombinant ILT viruses possess greater transmission and pathogenicity potential. Based on complete or partial genes of the ILTV genome, few studies genotyped ILTV strains circulating in Canada, and so far, information is scarce on whole-genome sequencing or the presence of recombination in Canadian ILTV isolates. The objective of this study was to genetically characterize the 14 ILTV isolates that originated from three provinces in Canada (Alberta, British Columbia and Quebec). To this end, a phylogenetic analysis of 50 ILTV complete genome sequences, including 14 sequences of Canadian origin, was carried out. Additional phylogenetic analysis of the unique long, unique short and inverted repeat regions of the ILTV genome was also performed. We observed that 71%, 21% and 7% of the ILTV isolates were categorized as CEO revertant, wild-type and TCO vaccine-related, respectively. The sequences were also analyzed for potential recombination events, which included evidence in the British Columbia ILTV isolate. This event involved two ILTV vaccine (CEO) strains as parental strains. Recombination analysis also identified that one ILTV isolate from Alberta as a potential parental strain for a United States origin ILTV isolate. The positions of the possible recombination breakpoints were identified. These results indicate that the ILTV wild-type strains can recombine with vaccinal strains complicating vaccine-mediated control of ILT. Further studies on the pathogenicity of these ILTV strains, including the recombinant ILTV isolate are currently ongoing.

Horizontal gene transfer is an important mechanism which facilitates bacterial populations in overcoming antimicrobial treatment. In this study, a total of 120 Escherichia coli and 62 Salmonella enterica subsp. enterica isolates were isolated from broiler chicken farms in Alberta. Fourteen serovars were identified among Salmonella isolates. Thirty one percent of E. coli isolates (37/120) were multiclass drug resistant (resistant to ≥ 3 drug classes), while only about 16% of Salmonella isolates (10/62) were multiclass drug resistant. Among those, eight E. coli isolates had an AmpC-type phenotype, and one Salmonella isolate had an extended-spectrum beta-lactamase (ESBL)-type beta-lactamase phenotype. We identified both AmpC-type (bla CMY-2) and ESBL-type (bla TEM) genes in both E. coli and Salmonella isolates. Plasmids from eight of nine E. coli and Salmonella isolates were transferred to recipient strain E. coli J53 through conjugation. Transferable plasmids in the eight E. coli and Salmonella isolates were also transferred into a lab-made sodium azide-resistant Salmonella recipient through conjugation. The class 1 integrase gene, int1, was detected on plasmids from two E. coli isolates. Further investigation of class 1 integron cassette regions revealed the presence of an aadA gene encoding streptomycin 3’’-adenylyltransferase, an aadA1a/aadA2 gene encoding aminoglycoside 3’’-O-adenyltransferase, and a putative adenylyltransferase gene. This study provides some insight into potential horizontal gene transfer events of antimicrobial resistance genes between E. coli and Salmonella in broiler chicken production.

Muskoxen (Ovibos moschatus), a taxonomically unique Arctic species, are increasingly exposed to climate and other anthropogenic changes. It is critical to develop and validate reliable tools to monitor their physiological stress response in order to assess the impacts of these changes. Here, we measured fecal glucocorticoid metabolite (FGM) levels in response to the administration of adrenocorticotropic hormone (ACTH) in the winter (1 IU/kg) and summer (2 IU/kg) using two enzyme immunoassays, one targeting primarily cortisol and the other targeting primarily corticosterone. Fecal cortisol levels varied substantially within and among individuals, and none of the animals in either challenge showed an increase in fecal cortisol following the injection of ACTH. By contrast, two of six (winter) and two of five (summer) muskoxen showed a clear response in fecal corticosterone levels (i.e., maximal percentage increase as compared to time 0 levels > 100%). Increases in fecal corticosterone post-ACTH injection occurred earlier and were of shorter duration in the summer than in the winter and fecal corticosterone levels were, in general, lower during the summer. These seasonal differences in FGM responses may be related to the use of different individuals (i.e., influence of sex, age, social status, etc.) and to seasonal variations in the metabolism and excretion of glucocorticoids, intestinal transit time, voluntary food intake, and fecal output and moisture content. Results from this study support using FGMs as a biomarker of hypothalamic–pituitary–adrenal axis activity in muskoxen, advance our understanding of the physiological adaptations of mammals living in highly seasonal and extreme environments such as the Arctic, and emphasize the importance of considering seasonality in other species when interpreting FGM levels.

Horizontal gene transfer is an important mechanism which facilitates bacterial populations in overcoming antimicrobial treatment. In this study, a total of 120 Escherichia coli and 62 Salmonella enterica subsp. enterica isolates were isolated from poultry farms in Alberta. Fourteen serovars were identified among Salmonella isolates. Thirty one percent of E. coli isolates were multiclass drug resistant (resistant to ≥ 3 drug classes), while only about 16% of Salmonella isolates were multiclass drug resistant. Among those, eight E. coli isolates had an AmpC-type phenotype, and one Salmonella isolate had an extended-spectrum beta-lactamase (ESBL)-type β-lactamase phenotype. We identified both AmpC-type (blaCMY-2) and ESBL-type (blaTEM) genes in both E. coli and Salmonella isolates. Plasmids from eight of nine E. coli and Salmonella isolates were transferred to recipient strain E. coli J53 through conjugation. Transferable plasmids in above total eight E. coli and Salmonella isolates were also transferred into a lab-made sodium azide-resistant Salmonella recipient through conjugation. The class 1 integrase gene, int1, was detected on plasmids from two E. coli isolates. Further investigation of class 1 integron cassette regions revealed the presence of an aadA gene encoding streptomycin 3”-adenylyltransferase, an aadA1a/aadA2 gene encoding aminoglycoside 3”-O-adenyltransferase, and a putative adenylyltransferase gene. This study provides some insight into potential horizontal gene transfer events of antimicrobial resistance genes between E. coli and Salmonella in poultry production.

BackgroundMuskoxen are a key species of Arctic ecosystems and are important for food security and socio-economic well-being of many Indigenous communities in the Arctic and Subarctic. Between 2009 and 2014, the bacterium Erysipelothrix rhusiopathiae was isolated for the first time in this species in association with multiple mortality events in Canada and Alaska, raising questions regarding the spatiotemporal occurrence of the pathogen and its potential impact on muskox populations.Materials and methodsWe adapted a commercial porcine E. rhusiopathiae enzyme-linked immunosorbent assay to test 958 blood samples that were collected from muskoxen from seven regions in Alaska and the Canadian Arctic between 1976 and 2017. The cut-off between negative and positive results was established using mixture-distribution analysis, a data-driven approach. Based on 818 samples for which a serological status could be determined and with complete information, we calculated trends in sample seroprevalences in population time-series and compared them with population trends in the investigated regions.ResultsOverall, 219/818 (27.8%, 95% Confidence Interval: 24.7-31.0) samples were classified as positive for exposure to E. rhusiopathiae. There were large variations between years and regions. Seropositive animals were found among the earliest serum samples tested; 1976 in Alaska and 1991 in Canada. In Alaskan muskoxen, sample seroprevalence increased after 2000 and, in two regions, peak seroprevalences occurred simultaneously with population declines. In one of these regions, concurrent unusual mortalities were observed and E. rhusiopathiae was isolated from muskox carcasses. In Canada, there was an increase in sample seroprevalence in two muskox populations following known mortality events that had been attributed to E. rhusiopathiae.ConclusionOur results indicate widespread exposure of muskoxen to E. rhusiopathiae in western Canada and Alaska. Although not new to the Arctic, we documented an increased exposure to the pathogen in several regions concurrent with population declines. Understanding causes for the apparent increased occurrence of this pathogen and its association with large scale mortality events for muskoxen is critical to evaluate the implications for wildlife and wildlife-dependent human populations in the Arctic.

Emergence and dissemination of antimicrobial resistance (AMR) in food-borne bacteria is a public health issue. Retail meat is considered an important carrier for antimicrobial-resistant Escherichia coli. Currently, resistance of E. coli strains to third generation cephalosporins are of particular concern, with significant potential animal and public health consequences. Resistance to tetracyclines is also a concern, due to high prevalence of this resistance and important co-resistance patterns. However, the actual likelihood of exposure to antimicrobial-resistant bacteria via the consumption of retail meats, beyond the simple frequencies of resistance found in samples of meat at the grocery store, in Canada remains to be investigated. This study was conducted to estimate the potential human exposure to ceftriaxone- and tetracycline-resistant E.coli (CREC and TREC; the hazards of interest) through the consumption of ground beef in Western Canada. Our exposure assessment simulation model included five modules: 1) estimation of prevalence and concentration of the hazards of interest in retail ground beef samples collected by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS), 2011–2014; 2) potential growth of the hazards during transportation from retail to home; 3) potential growth or reduction of the hazards during home storage; 4) thermal inactivation of the hazards due to cooking; and 5) consumption. The outputs of the model were presented as the probability of consumers' exposure to various threshold levels of the hazards [10 to 106 colony forming units (CFU)] in one serving of cooked ground beef. Overall, the probabilities of exposure to high threshold levels of the hazards (>4 log10 CFU/serving) were estimated to be lower than 0.12% in the study population (2011–2014). The mean probabilities of exposure to at least 1 log10 CFU CREC and TREC in a single meal containing cooked ground beef were 0.067% (SD: 0.001%) and 1.20% (SD: 0.02%), respectively. This probability substantially decreased as the threshold level of exposure increased to ≥6 log10 CFU. The probability of exposure to TREC was consistently greater than that for CREC. Cooking led to a prominent drop in the mean concentration of the hazards (4.7 log10 CFU/g). The findings from this research could inform the policy-making process and provide suggestions for adjustments in future retail surveillance plans. In addition, important knowledge gaps in this area have been highlighted.

Monitoring and surveillance of wildlife populations, including demographics and health, is often challenging, particularly in resource-constrained and remote settings. However, in areas characterized by subsistence oriented societies, the users of renewable resources hold a vast and holistic ecological knowledge about the natural environment. This knowledge can be instrumental for understanding and early detection of changes in wildlife populations. Using qualitative research methods and participatory epidemiology techniques we documented the local knowledge from resource users of the community of Iqaluktutiaq (Nunavut, Canada) to assess the health and population status and trends for muskoxen and caribou in the area. Semi-structured individual interviews, followed by group interviews, were implemented with 38 participants, and research findings were summarized and then verified with 31 interviewees. Local knowledge identified major declines in the number of muskoxen and caribou in the study area that were corroborated by subsequent aerial population estimates for both species. Observations made by participants allowed inference of possible mechanisms for the recent population declines, including poor recruitment, poor body condition, and increased morbidity and mortality (including endemic and emerging diseases). Engaging resource users in the process of knowledge generation was useful to identify further research priorities and fostered trust among parties that facilitated the subsequent collaborative development of management plans for these species. We use our experience to illustrate that local knowledge contributes to a holistic understanding of wildlife health and can serve as an early warning system to detect changes in wildlife populations. These participatory approaches are portable to other species and settings and can enhance conservation and co-management efforts for wildlife species worldwide.

As shrimp farming can be an important means of income generation, particularly among Sri Lanka’s rural communities, it is important that this industry grows in a sustainable manner, starting at the farm level. The objectives of this study were to; 1) create baseline farm-level sustainability indices for smallholder shrimp farms in Sri Lanka using both content-based and system-based frameworks adapted from agriculture; 2) determine whether arranging indicators within both content- and system-based sustainability frameworks would provide additional insight into relative farm sustainability (rather than using just one framework); and 3) seek differences between the North Western Province (NWP) and the Eastern Province (EP) in sustainability index, sub-index, or indicator scores that might be used to create province-specific policies and education programs designed to potentially improve sustainable practices at the farm level. Since little has been published on the development of practical indicators and evaluation of farm-level sustainability in aquaculture, this study adapted two types of frameworks used for measuring sustainability in agriculture: content- and system-based. Using both frameworks, along with expert opinion, indicators of farm level sustainability were developed into a questionnaire and measured on 225 farms in two provinces of Sri Lanka. Indicators primarily included modifiable practices that farmers could influence to improve the chances of their farms’ survival. The farm indicators were normalized using “min-max” normalization, where scores of zero and one were assigned for the least and most sustainable options, respectively. Farm indicators in each province were then aggregated into sub-indices as well an overall aggregated sustainability index score. Similar themes were gleaned from both the content-based and system-based sustainability frameworks, and there was no significant difference between mean overall sustainability scores for the two different frameworks. Farms in the NWP scored significantly higher on the overall sustainability indices of both models despite the NWP’s history of severe disease outbreaks while the EP farms had no disease. Possible reasons for this difference were explored with an in-depth analysis of sub-indices as well as individual indicators. An overall aggregated sustainability index score was a useful baseline measure for monitoring changes in sustainability over time; however, its use was limited when attempting to identify gaps in sustainable management practices and provide practical information for farmers and stakeholders to improve farm-level sustainability. It was important to evaluate individual indicators in addition to sub-indices and overall index scores when making recommendations or identifying gaps in sustainable practices at the farm level.

The prevalence and levels of enteric viruses in untreated groundwater of private wells used for drinking and/or agricultural practices in rural Alberta were studied using the qPCR panel assay, integrated cell culture with qPCR and cell culture in the volume of 500 liters per sample through serial sampling. Seven viruses were assessed including adenovirus, rotavirus, norovirus, astrovirus, sapovirus, reovirus and JC virus. Five viruses were detected with an overall positive detection rate of 6.33 % (45 of 711 samples). The most frequently detected virus was adenovirus (48.9%, 22/45) followed by rotavirus (44.4%, 20/45), reovirus (20%, 9/45), JC virus (6.7%, 3/45) and norovirus (6.7%, 3/45). There was no significant difference in the positive detection rates, ranging from 1.1% to 3.4% by various well settings used for broiler farms, cow/calf farms, feedlots and rural acreages. Effects of well characteristics (aquifer type, well depth, static level of water, well seal) and well completion lithology on potential viral contamination of groundwater of private wells were also analyzed upon available data. The findings demonstrate that occurrence of enteric viruses is low and viral contamination is sporadic in groundwater of private wells in rural Alberta. Conventional fecal bacterial indicators (coliform and/or E. coli) were not a representative marker for viral contamination in groundwater wells in rural Alberta.

Muskoxen (Ovibos moschatus) are increasingly exposed to a broad diversity of stressors in their rapidly changing Arctic environment. There is an urgent need to develop validated tools to monitor the impact of these stressors on the hypothalamic-pituitary-adrenal (HPA) axis activity of muskoxen to help inform conservation actions. Here, we evaluated whether muskox qiviut (dense wooly undercoat) cortisol accurately reflects changes in HPA axis activity. Two repeated pharmacological challenges, involving weekly administrations of saline (control group) or adrenocorticotropic hormone (ACTH) during five consecutive weeks, were done on captive muskoxen, in winter (no hair growth) and summer (maximum hair growth). Pre-challenge qiviut cortisol levels were significantly higher in the shoulder than in the neck, but neither differed from rump concentrations. Qiviut cortisol levels significantly increased (p

A devastating flood occurred in southern Alberta on June 19, 2013, from greater than normal snowfalls in the Rocky Mountains and excess precipitation during the early spring that left soils saturated and unable to absorb any additional precipitation. This flood was Canada's most costly natural disaster, with five to six billion Canadian dollars in damages. The first objective of this study was to determine if the flood caused an increase in private drinking water well contamination in the Calgary Health Zone by comparing contamination rates to previous years. The second objective was to determine which environmental factors were associated with contamination during this flood event. Test results of total coliforms (TC) and Escherichia coli (EC) of private water wells were used to determine contamination. A geographically weighted Poisson regression analysis suggested that TC contamination was not associated with this flood. The EC contamination is positively associated with floodways, flood fringe, farms, and negatively associated with intermittent water (sloughs). These results suggest that for the 2013 flood, individual well characteristics are more important than surrounding geographic features. Thus, it is recommended that homeowners who live in a high-risk area ensure their wells are properly maintained to reduce risk of water well contamination.

Brucella serostatus was evaluated in 3189 muskoxen sampled between 1989 and 2016 from various locations of the Canadian Arctic archipelago and mainland, near the communities of Sachs Harbour and Ulukhaktok, Northwest Territories, and Cambridge Bay and Kugluktuk, Nunavut. Brucella antibodies were found only in muskoxen sampled around Cambridge Bay, both on southern Victoria Island and on the adjacent mainland (Kent Peninsula). Consistent with participatory epidemiology data documented from local harvesters describing increased Brucella-like syndromes (swollen joints and lameness) and a decreased proportion of juveniles, the apparent Brucella seroprevalence in the sampled muskoxen of the Cambridge Bay area increased from 0.9% (95% CI 0.3–2.1) in the period of 1989–2001 to 5.6% (95% CI 3.3–8.9) in 2010–2016. The zoonotic bacteria Brucella suis biovar 4 was also cultured from tissues of muskoxen sampled on Victoria Island near Ulukhaktok in 1996 (n = 1) and Cambridge Bay in 1998, 2014, and 2016 (n = 3). Overall, our data demonstrate that B. suis biovar 4 is found in muskoxen that are harvested for food and by guided hunts on Victoria Island and Kent Peninsula, adding an important public health dimension to this study. Robust participatory epidemiology data on muskox health and diseases greatly enhanced the interpretation of our Cambridge Bay data and, combined with the serological and microbiological data, provide compelling evidence that the prevalence of B. suis biovar 4 has increased in this area since the late 1990s. This study enhances the available knowledge on Brucella exposure and infection in muskoxen and provides an example of how scientific knowledge and local knowledge can work together to better understand disease status in wildlife.

BackgroundWe have previously reported, in a systematic review of 181 studies, that restriction of antibiotic use in food-producing animals is associated with a reduction in antibiotic-resistant bacterial isolates. While informative, that report did not concretely specify whether different types of restriction are associated with differential effectiveness in reducing resistance. We undertook a sub-analysis of the systematic review to address this question.MethodsWe created a classification scheme of different approaches to antibiotic restriction: (1) complete restriction; (2) single antibiotic-class restriction; (3) single antibiotic restriction; (4) all non-therapeutic use restriction; (5) growth promoter and prophylaxis restriction; (6) growth promoter restriction and (7) other/undetermined. All studies in the original systematic review that were amenable to meta-analysis were included into this substudy and coded by intervention type. Meta-analyses were conducted using random effects models, stratified by intervention type.ResultsA total of 127 studies were included. The most frequently studied intervention type was complete restriction (n=51), followed by restriction of non-therapeutic (n=33) and growth promoter (n=19) indications. None examined growth promoter and prophylaxis restrictions together. Three and seven studies examined single antibiotic-class and single antibiotic restrictions, respectively; these two intervention types were not significantly associated with reductions in antibiotic resistance. Though complete restrictions were associated with a 15% reduction in antibiotic resistance, less prohibitive approaches also demonstrated reduction in antibiotic resistance of 9%–30%.ConclusionBroad interventions that restrict global antibiotic use appear to be more effective in reducing antibiotic resistance compared with restrictions that narrowly target one specific antibiotic or antibiotic class. Importantly, interventions that allow for therapeutic antibiotic use appear similarly effective compared with those that restrict all uses of antibiotics, suggesting that complete bans are not necessary. These findings directly inform the creation of specific policies to restrict antibiotic use in food-producing animals.

Dog bites are a public health concern that also implicates animal welfare, with negative outcomes such as rehoming or euthanasia for the animals responsible. Previous research has shown that the severity of dog-bite injuries reflects multiple factors, including the degree of inhibition exhibited by dogs and how people behave towards dogs. This study utilizes an objective dog bite injury assessment tool: The Dunbar aggression scale. Trained officers employed by The City of Calgary systematically use the Dunbar scale whenever investigating dog-bite complaints. We analyzed The City of Calgary&rsquo, s administrative data on confirmed dog-bite injuries in people, 2012&ndash, 2017, with a multivariable generalized ordered logistic regression model. Severe dog-bite injuries occurred more frequently in the family home than in any other setting. Young children, youths and older adults were at higher risk of more serious bites than adults. There has been a decreasing trend in the probability of a high or medium severity bite, and an increasing trend in the probability of a low severity bite since 2012. These results indicate that greater public awareness regarding dog-bite injuries is needed. Consideration should be given to campaigns targeted towards different demographics, including older adults, to provide an understanding of dog behaviour and to emphasize the need to supervise children closely in the presence of all dogs at all times, including family dogs in the home environment. Given that dog-bite injuries are not just a public health issue, but also an animal welfare issue, we endorse One Health responses in educational campaigns, policy development, and professional practice.

Waterborne outbreaks of infectious disease continue to be a public health risk, particularly those in areas where testing of private and public small system groundwater systems is left to the owner...

Campylobacter infections in humans are usually self-limiting; however, antibiotic intervention may be necessary in the case of severe infection. Fluoroquinolones are often the drug of choice for treatment of campylobacteriosis; however, resistance to these drugs can develop rapidly, complicating treatment protocols. Increasing resistance to fluoroquinolones in human infections has coincided with approval of use of fluoroquinolones in animals, therefore, isolation of fluoroquinolone resistant (FQr) Campylobacter in broiler flocks is concerning. This cross-sectional study utilized data collected from 2013-2018 by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) on-farm surveillance program to investigate prevalence factors associated with the isolation of FQr C. jejuni from broiler faecal samples. Mixed effects logistic regression models accounting for clustering of flocks within hatcheries, with and without a fixed effect for the presence of flock level tetracycline resistance were used to assess prevalence factors among 536 C. jejuni isolates from 158 flocks. Both models indicated that the type of bird used (Ross versus Cobb or mixed), the use of virginiamycin as a feed additive, the use of traps to control rodent populations in the barn, and the total number of birds in the barn were significant prevalence factors for increased FQr C. jejuni in a flock. In the model where flock level tetracycline resistance was included as a fixed effect, the odds of FQr C. jejuni increased by 16 (95% CI: 3.74, 68), and the magnitude of the effect of each of the identified prevalence factors was larger. Both models indicated that methods of disinfection of water lines between production cycles is important, with the use of chlorine being protective in the model where tetracycline resistance was included as a fixed effect, and the use of hydrogen peroxide being a risk factor in the model where tetracycline resistance was not included as a fixed effect. The use of hot water to wash the barn between production cycles was also a significant protective factor in the model where tetracycline resistance was not included as a fixed effect. These results indicate that biosecurity and sanitation procedures play a role in the dissemination of FQr C. jejuni in broiler flocks. Future analysis should seek to understand the effect of different disinfectant products on the isolation of FQr C. jejuni. Gaining a better understanding of the management of these critical practices may allow for the reduction of this enteric pathogen in broiler flocks in Canada.

A growing body of evidence has demonstrated that extraintestinal pathogenic E. coli (ExPEC), such as the urinary pathogenic E. coli (UPEC), are common constituents of treated wastewater, and therefore represent a potential public health risk. However, no single virulence gene, or set of virulence genes, can be used to conclusively identify this genetically diverse pathotype. As such we sought to identify and characterize the public health relevance of potential UPEC found in treated sewage/wastewater using a comparative genomics approach. Presumptive wastewater UPEC (W-UPEC) were initially identified by virulence gene screening against 5 virulence genes, and for which isolates containing ≥3 virulence genes were whole genome sequenced (n = 24). Single nucleotide polymorphic (SNP) spanning tree analysis demonstrated that many of these wastewater UPEC (WUPEC) were virtually identical at the core genome (0.4 Mbp) when compared to clinical UPEC (C-UPEC) sequences obtained from NCBI, varying by as little as 1 SNP. Remarkably, at the whole genome level, W-UPEC isolates displayed >96% whole genome similarity to C-UPEC counterparts in NCBI, with one strain demonstrating 99.5% genome similarity to a particular C-UPEC strain. The W-UPEC populations were represented by sequence types (ST) known to be clinically important, including ST131, ST95, ST127 and ST640. Many of the W-UPEC carried the exact same complement of virulence genes as their most closely related C-UPEC strains. For example, O25b-ST131 W-UPEC strains possessed the same 80 virulence genes as their most closely related C-UPEC counterparts. Concerningly, W-UPEC strains also carried a plethora of antibiotic resistance genes, and O25b-ST131strains were designated as extended spectrum beta-lactamase (ESBL) producing E. coli by both genome profiling and phenotypic resistance testing. W-UPEC ST131 strains were found in the effluents of a single treatment plant at different times, as well as different wastewater treatment plants, suggesting a differentially ability to survive wastewater treatment. Indeed, in sewage samples treated with chlorine doses sufficient for inducing a ∼99.99% reduction in total E. coli levels, UPEC represented a significant proportion of the chlorine-resistant population. By contrast, no Shiga toxin-producing E. coli were observed in these chlorinated sewage libraries. Our results suggest that clinically-relevant UPEC exist in treated wastewater effluents and that they appear to be specifically adapted to survive wastewater treatment processes.

Selective pressure exerted by the widespread use of antibacterial drugs is accelerating the development of resistant bacterial populations. The purpose of this scoping review was to summarise the range of studies that use dynamic models to analyse the problem of bacterial resistance in relation to antibacterial use in human and animal populations. A comprehensive search of the peer-reviewed literature was performed and non-duplicate articles (n = 1486) were screened in several stages. Charting questions were used to extract information from the articles included in the final subset (n = 81). Most studies (86%) represent the system of interest with an aggregate model; individual-based models are constructed in only seven articles. There are few examples of inter-host models outside of human healthcare (41%) and community settings (38%). Resistance is modelled for a non-specific bacterial organism and/or antibiotic in 40% and 74% of the included articles, respectively. Interventions with implications for antibacterial use were investigated in 67 articles and included changes to total antibiotic consumption, strategies for drug management and shifts in category/class use. The quality of documentation related to model assumptions and uncertainty varies considerably across this subset of articles. There is substantial room to improve the transparency of reporting in the antibacterial resistance modelling literature as is recommended by best practice guidelines.

The extra-label use of ceftiofur in Canadian hatcheries was cause for concern due to an increased prevalence of ceftiofur resistant Salmonella Heidelberg in chickens and humans in Quebec. Due to on-going concerns related to human health the use of ceftiofur was eventually phased out of the poultry production industry in 2014-2015. Simultaneous resistance to amoxicillin-clavulanic acid, ceftiofur and cefoxitin, a pattern known as A2C, caused by the presence of blaCMY-2 has become increasingly prevalent in broiler flocks worldwide. This study used data from the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) broiler farm surveillance program to examine the risk factors for A2C resistant Escherichia coli isolates (A2C-REI) from 1482 samples taken from 371 broiler flocks in Canada, between 2013 and 2015. Results of a mixed effects Poisson model with fixed effects at the flock level and random effects at the hatchery level indicated that the in ovo use of ceftiofur (Prevalence Ratio (PR): 1.91, 95% CI: 1.43-2.57), and the use of hydrogen peroxide to disinfect water lines during the growing period (PR: 1.52, 95% CI: 1.11-2.07), were risk factors for increased expected prevalence of A2C-REI, compared to farms not using these strategies. Controlling for wild birds (PR: 0.64, 95% CI: 0.48-0.84), use of avilamycin in feed (PR: 0.63, 95% CI: 0.41-0.98), and storage of manure on the farm (PR: 0.67, 95% CI: 0.51-0.89) were associated with a lower expected prevalence of A2C-REI, than farms that did not use these strategies. This study identified important management factors that were associated with the prevalence of A2C-REI on Canadian broiler farms. The findings of this study can be used as a baseline for the monitoring of the effects of the removal of ceftiofur from poultry production in Canada.

With increasing stress on our water resources and recent waterborne disease outbreaks, understanding the epidemiology of waterborne pathogens is crucial to build surveillance systems. The purpose of this study was to explore techniques for describing microbial water quality in rural drinking water wells, based on spatiotemporal analysis, time series analysis and relative risk mapping. Tests results for Escherichia coli and coliforms from private and small public well water samples, collected between 2004 and 2012 in Alberta, Canada, were used for the analysis. Overall, 14.6 and 1.5% of the wells were total coliform and E. coli-positive, respectively. Private well samples were more often total coliform or E. coli-positive compared with untreated public well samples. Using relative risk mapping we were able to identify areas of higher risk for bacterial contamination of groundwater in the province not previously identified. Incorporation of time series analysis demonstrated peak contamination occurring for E. coli in July and a later peak for total coliforms in September, suggesting a temporal dissociation between these indicators in terms of groundwater quality, and highlighting the potential need to increase monitoring during certain periods of the year.

Antimicrobial resistance is considered one of the greatest threats to global and public health today. The World Health Organization, the Food and Agriculture Organization, and the World Organisation for Animal Health, known as the Tripartite Collaboration, have called for urgent action. We have previously published a systematic review of 181 studies, demonstrating that interventions that restrict antibiotic use in food-producing animals are associated with a reduction in antibiotic resistant bacterial isolates in both animals and humans. What remains unknown, however, are whether (and what) unintended consequences may arise from such interventions. We therefore undertook a sub-analysis of the original review to address this research question. A total of 47 studies described potential consequences of antibiotic restrictions. There were no consistent trends to suggest clear harm. There may be increased bacterial contamination of food products, the clinical significance of which remains unclear. There is a need for rigorous evaluation of the unintended consequences of antibiotic restrictions in human health, food availability, and economics, given their possible widespread implications. Keywords: Antimicrobial resistance, One health, Antimicrobial use

This study determined the prevalence of Salmonella serovars, antimicrobial resistance (AMR) and resistance genes in Salmonella isolated from retail meats purchased in Alberta, Canada. Samples were collected during one year period (May 2007-April 2008) on weekly basis from 19 census divisions in Alberta. A total of 564 samples including chicken (n = 206), turkey (n = 91), beef (n = 134) and pork (n = 133) were purchased. Salmonella were recovered from chicken (40%), turkey (27%) and pork (2%) samples and was not found in ground beef. A total of 21, 8, and 3 different serovars were recovered from chicken, turkey and pork meats, respectively. Salmonella Hadar was most common in chicken whereas S. Heidelberg was common in turkey meat. Overall 29% (32/110) of isolates were susceptible to tested antimicrobials and resistance to ciprofloxacin, amikacin and nalidixic acid was not found in any isolate. Multiresistance (≥2 antimicrobials) was found in 56% of isolates. Resistance to amoxicillin-clavulanic acid (AMC), ceftiofur (TIO), and ceftriaxone (CRO) was found in about 21% of chicken and 25% of turkey isolates. Resistance to either of tetracycline (TET), streptomycin (STR) or ampicillin (AMP) was unconditionally associated with S. Hadar but resistance to either of TET, AMP, AMC, TIO, CRO or cefoxitin was associated with S. Heidelberg. The strA/B (42% isolates), tet(A) (28% isolates), bla(CMY-2) (21% isolates) and bla(TEM) (17% isolates) were the most common resistance genes found. The bla(CMY-2) and bla(TEM) genes were unconditionally associated with S. Heidelberg; tet(A) and strA/B with S. Hadar and tet(B) gene with S. Kentucky. The strA/B genes were not associated with S. Heidelberg. Our data suggests that the prevalence of Salmonella serovars varied by the meat type and that AMR and resistance genes varied by the Salmonella serovars.

An adult male muskox ( Ovibos moschatus ), harvested on 26 August 2014 on Victoria Island, Nunavut, in the Canadian Arctic, had proliferative dermatitis on the muzzle and fetlocks suggestive of contagious ecthyma or orf (Parapoxvirus). Histopathologic features of the lesions were consistent with this diagnosis. Orf virus DNA, phylogenetically similar to an isolate from a captive muskox of the Minnesota Zoo, US, was detected in the lesions by PCR using Parapoxvirus primers. Additionally, there was a metaphyseal abscess with a cortical fistula in the right metacarpus from which Brucella suis biovar 4 was isolated and identification supported by PCR. Brucella spp. antibodies were detected in serum. Finally, 212 nodules were dissected from the lungs. Fecal analysis and lung examination demonstrated co-infection with the lungworms Umingmakstrongylus pallikuukensis and Varestrongylus eleguneniensis. The zoonotic potential of orf and rangiferine brucellosis adds an important public health dimension to this case, particularly given that muskoxen are a valuable source of food for Arctic residents. Careful examination of these pathogens at a population level is needed as they may contribute to muskox population decline and potentially constitute a driver of food insecurity for local communities. This case underscores the importance of wildlife health surveillance as a management tool to conserve wildlife populations and maintain food security in subsistence-oriented communities.

Campylobacter spp. are the leading cause of bacterial gastroenteritis worldwide, and water is increasingly seen as a risk factor in transmission. Here we describe a most-probable-number (MPN)–quantitative PCR (qPCR) assay in which water samples are centrifuged and aliquoted into microtiter plates and the bacteria are enumerated by qPCR. We observed that commonly used Campylobacter molecular assays produced vastly different detection rates. In irrigation water samples, detection rates varied depending upon the PCR assay and culture method used, as follows: 0% by the de Boer Lv1-16S qPCR assay, 2.5% by the Van Dyke 16S and Jensen glyA qPCR assays, and 75% by the Linton 16S endpoint PCR when cultured at 37°C. Primer/probe specificity was the major confounder, with Arcobacter spp. routinely yielding false-positive results. The primers and PCR conditions described by Van Dyke et al. (M. I. Van Dyke, V. K. Morton, N. L. McLellan, and P. M. Huck, J Appl Microbiol 109:1053–1066, 2010, http://dx.doi.org/10.1111/j.1365-2672.2010.04730.x ) proved to be the most sensitive and specific for Campylobacter detection in water. Campylobacter occurrence in irrigation water was found to be very low (Campylobacter -specific qPCR was used, with the most commonly detected species being C. jejuni , C. coli , and C. lari . Campylobacters in raw sewage were present at ∼10 2 /100 ml, with incubation at 42°C required for reducing microbial growth competition from arcobacters. Overall, when Campylobacter prevalence and/or concentration in water is reported using molecular methods, considerable validation is recommended when adapting methods largely developed for clinical applications. Furthermore, combining MPN methods with molecular biology-based detection algorithms allows for the detection and quantification of Campylobacter spp. in environmental samples and is potentially suited to quantitative microbial risk assessment for improved public health disease prevention related to food and water exposures. IMPORTANCE The results of this study demonstrate the importance of assay validation upon data interpretation of environmental monitoring for Campylobacter when using molecular biology-based assays. Previous studies describing Campylobacter prevalence in Canada utilized primers that we have determined to be nonspecific due to their cross-amplification of Arcobacter spp. As such, Campylobacter prevalence may have been vastly overestimated in other studies. Additionally, the development of a quantitative assay described in this study will allow accurate determination of Campylobacter concentrations in environmental water samples, allowing more informed decisions to be made about water usage based on quantitative microbial risk assessment.

We studied the presence of linear enamel hypoplasias (LEHs; tooth defects associated with physiological stress) in caribou (Rangifer tarandus). A timeline of tooth enamel development was determined by radiographic examination of 48 mandibles from caribou aged 3–24 months old. We examined mandibles from the Bluenose East (n = 56) and Bluenose West (n = 15) caribou herds in the Northwest Territories and Nunavut, Canada, for LEHs and 21.1% (15/71) were affected. We concluded that LEHs do occur in caribou and tracking these over time may provide a tool to track population dynamics in extant wildlife. © 2012 The Wildlife Society.

This study analyzed antimicrobial resistance (AMR) and resistance genes in generic Escherichia coli isolated from retail meat samples purchased (2007-2008) in Alberta, Canada, and determined potential associations between resistance phenotypes and resistance genes with relation to the meat types. A total of 422 E. coli isolates from retail chicken, turkey, beef, and pork meats were tested for antimicrobial susceptibility. Multiplex PCRs were used to detect major resistance genes for tetracyclines [tet(A), tet(B), tet(C)], sulfonamides (sul1, sul2, sul3), aminoglycosides (strA/B, aadA, aadB, aac(3)IV, aphA1, aphA2), and β-lactamase (bla(CMY-2), bla(TEM), bla(SHV), bla(PSE-1)). Resistance to ciprofloxacin was not found in any isolate. Overall resistances to clinically important antimicrobials amoxicillin-clavulanic acid (16.8% of isolates) and ceftriaxone (12.6% isolates) were observed. These resistances were observed more frequently (p0.0001) in chicken-derived E. coli than those from the other meat types. Resistance to multiple antimicrobials (≥ 5) was found in more chicken derived E. coli (32%) than E. coli from other meat types. The β-lactamase genes of clinical importance, including bla(CMY-2) and bla(TEM), were found in about 18% of poultry-derived E. coli and in only 5% of ground beef. The bla(CMY-2) gene was more likely to be found in E. coli from chicken than turkey, beef, or pork meats. The tet(A) gene was associated with bla(CMY-2), whereas bla(CMY-2) and bla(TEM) genes were associated with strA/B genes. Resistance genes for tetracycline, sulfonamides, and aminoglycosides were associated with the phenotypic expression of resistance to unrelated classes of antimicrobials. These data suggest the prevalence of AMR and select resistance genes were higher in poultry-derived E. coli. The multiple associations found between AMR phenotypes and resistance genes suggest a complex nature of resistance in E. coli from retail meat, and hence the use of a single antimicrobial could result in the selection of resistant E. coli not only to the drug being used but to other unrelated classes of antimicrobials as well.