Bacterial genomics reveal the complex epidemiology of an emerging pathogen in Arctic and boreal ungulates

Northern ecosystems are currently experiencing unprecedented ecological change, largely driven by a rapidly changing climate. Pathogen range expansion, and emergence and altered patterns of infectious disease, are increasingly reported in wildlife at high latitudes. Understanding the causes and consequences of shifting pathogen diversity and host-pathogen interactions in these ecosystems is important for wildlife conservation, and for indigenous populations that depend on wildlife. Among the key questions are whether disease events are associated with endemic or recently introduced pathogens, and whether emerging strains are spreading throughout the region. In this study, we used a phylogenomic approach to address these questions of pathogen endemicity and spread for Erysipelothrix rhusiopathiae, an opportunistic multi-host bacterial pathogen associated with recent mortalities in arctic and boreal ungulate populations in North America. We isolated E. rhusiopathiae from carcasses associated with large-scale die-offs of muskoxen in the Canadian Arctic Archipelago, and from contemporaneous mortality events and/or population declines among muskoxen in northwestern Alaska and caribou and moose in western Canada. Bacterial genomic diversity differed markedly among these locations; minimal divergence was present among isolates from muskoxen in the Canadian Arctic, while in caribou and moose populations, strains from highly divergent clades were isolated from the same location, or even from within a single carcass. These results indicate that mortalities among northern ungulates are not associated with a single emerging strain of E. rhusiopathiae, and that alternate hypotheses need to be explored. Our study illustrates the value and limitations of bacterial genomic data for discriminating between ecological hypotheses of disease emergence, and highlights the importance of studying emerging pathogens within the broader context of environmental and host factors

A global assessment of Echinococcus multilocularis infections in domestic dogs: proposing a framework to overcome past methodological heterogeneity

Echinococcus multilocularis , the aetiological agent of human Alveolar Echinococcosis (AE), is transmitted between small mammals and wild or domestic canids. Dogs infected with E. multilocularis can transmit this infection to humans and infect themselves with canine AE as dead-end hosts. Whereas E. multilocularis infections in wild hosts and humans have been well-studied in the last decades, infections in domestic dogs are sparsely reported. This literature review and meta-analysis highlights gaps in the available data and provided a re-assessment of the global distribution of domestic dog E. multilocularis infections. We found 46 published articles documenting the prevalence of E. multilocularis in domestic dogs from 21 countries across Europe, Asia, and North America. Apparent prevalence estimates ranged from 0.00% (0.00-0.33%) in Germany to 55.50% (26.67-81.12%) in China. Most studies were conducted in areas of high human AE. By accounting for reassessed diagnostic sensitivity and specificity, we estimated true prevalence in a sub-set of studies, which varied between 0.00% (0.00-12.42%) and 41.09% (21.12- 65.81%) as these true prevalence estimates were seldom reported in the articles themselves. Articles also showed a heavy emphasis on rural dogs dismissing urban ones, which is concerning due to the role urbanization plays in the transmission of zoonotic diseases, especially those utilizing pets as definitive hosts. Lastly, population studies on canine AE were absent, highlighting the relative focus on human rather than animal health. We thus developed a framework for investigating domestic dog E. multilocularis infections and performing risk assessment of dog-associated transmission to fill the gaps found in these literatures

Analysis of whole-genome sequences of infectious laryngotracheitis virus isolates from poultry flocks in Canada : evidence of recombination

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

Pathogenic and transmission potential of wildtype and chicken embryo origin (CEO) vaccine revertant infectious laryngotracheitis virus

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

Factors associated with antimicrobial resistant enterococci in Canadian beef cattle: A scoping review

IntroductionAntimicrobial resistance (AMR) is a global health concern, occurring when bacteria evolve to render antimicrobials no longer effective. Antimicrobials have important roles in beef production; however, the potential to introduce AMR to people through beef products is a concern. This scoping review identifies factors associated with changes in the prevalence of antimicrobial-resistant Enterococcus spp. applicable to the Canadian farm-to-fork beef continuum.MethodsFive databases (MEDLINE, BIOSIS, Web of Science, Embase, and CAB Abstracts) were searched for articles published from January 1984 to March 2022, using a priori inclusion criteria. Peer-reviewed articles were included if they met all the following criteria: written in English, applicable to the Canadian beef production context, primary research, in vivo research, describing an intervention or exposure, and specific to Enterococcus spp.ResultsOut of 804 screened articles, 26 were selected for inclusion. The included articles discussed 37 factors potentially associated with AMR in enterococci, with multiple articles discussing at least two of the same factors. Factors discussed included antimicrobial administration (n = 16), raised without antimicrobials (n = 6), metal supplementation (n = 4), probiotics supplementation (n = 3), pen environment (n = 2), essential oil supplementation (n = 1), grass feeding (n = 1), therapeutic versus subtherapeutic antimicrobial use (n = 1), feeding wet distiller grains with solubles (n = 1), nutritional supplementation (n = 1) and processing plant type (n = 1). Results were included irrespective of their quality of evidence.DiscussionComparability issues arising throughout the review process were related to data aggregation, hierarchical structures, study design, and inconsistent data reporting. Findings from articles were often temporally specific in that resistance was associated with AMR outcomes at sampling times closer to exposure compared to studies that sampled at longer intervals after exposure. Resistance was often nuanced to unique gene and phenotypic resistance patterns that varied with species of enterococci. Intrinsic resistance and interpretation of minimum inhibitory concentration varied greatly among enterococcal species, highlighting the importance of caution when comparing articles and generalizing findings.Systematic Review Registration[http://hdl.handle.net/1880/113592

Comparative pathogenicity of infectious bronchitis virus Massachusetts and Delmarva (DMV/1639) genotypes in laying hens

Infectious bronchitis (IB) is a highly contagious and acute viral disease of chicken caused by the infectious bronchitis virus (IBV) of the family Coronaviridae. Even with extensive vaccination against IB by the poultry industry, the occurrence of new IBV genotypes is a continuous challenge encountered by the global poultry industry. This experiment was designed to compare the pathogenicity of two IBV strains belonging to Massachusetts (Mass) and Delmarva DMV/1639 genotypes. Specific pathogen-free laying hens were challenged during the peak of production (30 weeks), keeping a mock-infected control group. During 21 days of observation following infection, a significant drop in egg production with miss-shaped and soft shells was observed in the DMV/1639 IBV-infected hens only. The DMV/1639 IBV infected group showed prolonged and higher cloacal viral shedding compared with the Mass IBV-infected group. At the end of the study (21 days post-infection), the viral genome loads in the respiratory, urogenital, and immune tissues were significantly higher in the DMV/1639 IBV-infected group compared with the Mass IBV-infected group. Macroscopic lesions such as distorted ova leading to egg peritonitis were observed only in the DMV/1639 IBV-infected group. Moreover, microscopic lesion scores were significantly higher in the lung, kidney, cecal tonsils, and oviduct of the DMV/1639 IBV-infected group compared with the Mass IBV-infected group. Finally, the apoptosis index in the kidney, ovary, magnum, isthmus, and shell gland was significantly higher in the DMV/1639 IBV-infected group compared with the control and Mass-infected groups. This study examined the pathogenicity of two IBV genotypes that are impacting the layer industry in North America

The epidemiology of antimicrobial resistance in fecal escherichia coli isolates of feedlot cattle in western Canada

Prevalence of antimicrobial resistance in fecal Escherichia coli isolates from feedlot cattle was characterized. Tetracycline resistance in isolates from newly weaned, auction market derived calves on arrival at the feedlot in a clinical trial was 9.8% and resistance to three or more antimicrobials was 2.1% compared to 17.6 % and 5.9% in a cohort study. The prevalence of tetracycline resistance at 78.3% and resistance to three or more antimicrobials at 52.5 % in isolates from spring calves submitted to a regional diagnostic laboratory were higher than those found on arrival at the feedlot. Of isolates from composite feedyard pen samples late in the feeding period, 39.4% were tetracycline resistant and 7.6% were resistant to three or more antimicrobials, somewhat higher than on arrival. Use of oxytetracycline in the feed for disease prophylaxis and the metaphylactic use of long-acting injectable oxytetracycline were associated with increased proportions of cattle with one or more resistant E. coli isolates early in the feeding period, while the use of individual animal treatments was not. The proportion of animals with one or more tetracycline resistant E. coli isolates was not different between the control, metaphylactic treatment and prophylactic treatment groups preslaughter; however, there were significantly more resistant animals in all groups preslaughter than at arrival. There were also no associations found between the total volume of parenteral antimicrobials used for disease treatment in individual animals and antimicrobial resistance in the cohort study. In addition, no strong associations were found between pen-level prevalence of antimicrobial resistance antimicrobial use or other variables. There was no significant difference between the proportion of isolates per pen resistant to tetracycline, one or more, two or more antimicrobials, or three or more antimicrobials when using 20, 15, 10 or 5 isolates from composite pen-level fecal samples. Variance for isolates resistant to three or more antimicrobials was partitioned as 12.7% at the feedyard-level and 28.7% at the pen-level. The use of diagnostic laboratory data for AMR surveillance was also discussed, and alternatives to antimicrobial treatment in the feedlot were also investigated. Overall a significant contribution to our understanding of antimicrobial resistance in feedlot cattle was achieved

Rainfall and microbial contamination in Alberta well water

Spatial and seasonal patterns in the positive rates of total coliforms and Escherichia coli in Alberta well water were investigated to gain insight into well water microbial contamination. Analysis was conducted in the presence of total coliforms (77  135 tests) and E. coli (77  132 tests) in well water from 2004 to 2009 along with monthly estimates of precipitation, all of which were aggregated to 13 zones across Alberta by using Voronoi tessellation. Regression combined with autocorrelation analysis was employed to develop wave functions for data assembled in each zone. Precipitation was found to peak in June or July in all 13 regions. The positive E. coli rate was found to peak in June, July or August, but the positive total coliform rate peaked in August, September or October. Spatial statistical analysis revealed a potential association of total coliforms and E. coli with precipitation in two heavily populated basins. Spatial density analysis revealed a cluster of positive tests of total coliforms and E. coli in a narrow spatial extent in June and July of 2005.FacultyReviewe