Redefining piscine lactococcosis.

Piscine lactococcosis is a significant threat to cultured and wild fish populations worldwide. The disease typically presents as a per-acute to acute hemorrhagic septicemia causing high morbidity and mortality, recalcitrant to antimicrobial treatment or management interventions. Historically, the disease was attributed to the gram-positive pathogen Lactococcus garvieae. However, recent work has revealed three distinct lactococcosis-causing bacteria (LCB)-L. garvieae, L. petauri, and L. formosensis-which are phenotypically and genetically similar, leading to widespread misidentification. An update on our understanding of lactococcosis and improved methods for identification are urgently needed. To this end, we used representative isolates from each of the three LCB species to compare currently available and recently developed molecular and phenotypic typing assays, including whole-genome sequencing (WGS), end-point and quantitative PCR (qPCR) assays, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), API 20 Strep and Biolog systems, fatty acid methyl ester analysis (FAME), and Sensititre antimicrobial profiling. Apart from WGS, sequencing of the gyrB gene was the only method capable of consistent and accurate identification to the species and strain level. A qPCR assay based on a putative glycosyltransferase gene was also able to distinguish L. petauri from L. garvieae/formosensis. Biochemical tests and MALDI-TOF MS showed some species-specific patterns in sugar and fatty acid metabolism or protein profiles but should be complemented by additional analyses. The LCB demonstrated overlap in host and geographic range, but there were relevant differences in host specificity, regional prevalence, and antimicrobial susceptibility impacting disease treatment and prevention. IMPORTANCE: Lactococcosis affects a broad range of host species, including fish from cold, temperate, and warm freshwater or marine environments, as well as several terrestrial animals, including humans. As such, lactococcosis is a disease of concern for animal and ecosystem health. The disease is endemic in European and Asian aquaculture but is rapidly encroaching on ecologically and economically important fish populations across the Americas. Piscine lactococcosis is difficult to manage, with issues of vaccine escape, ineffective antimicrobial treatment, and the development of carrier fish or biofilms leading to recurrent outbreaks. Our understanding of the disease is also widely outdated. The accepted etiologic agent of lactococcosis is Lactococcus garvieae. However, historical misidentification has masked contributions from two additional species, L. petauri and L. formosensis, which are indistinguishable from L. garvieae by common diagnostic methods. This work is the first comprehensive characterization of all three agents and provides direct recommendations for species-specific diagnosis and management.

Observational study on antimicrobial resistance in Escherichia coli and Salmonella isolates from Ontario calf samples submitted to a diagnostic laboratory from 2007 to 2020.

The objectives of this study were to i) describe Escherichia coli and Salmonella isolates; ii) investigate the temporal trends in antimicrobial resistance (AMR) profiles; and iii) evaluate the impact of season and age on these AMR profiles from diagnostic and post-mortem samples in Ontario calves ≤ 2-months-old submitted from 2007 to 2020 to the Animal Health Laboratory in Guelph, Ontario, Canada. Antimicrobial susceptibility testing results were measured by the Kirby-Bauer disk diffusion method. A total of 1291 isolates with AMR profiles were obtained from calves, with E. coli (n = 434) and Salmonella (n = 378) being the most common bacteria characterized for AMR. For E. coli, 79% of isolates tested showed a positive result in F5/K99, whereas for Salmonella isolates, S. Typhimurium (33%) and S. Dublin (22%) were the 2 most common serotypes identified. Multivariable logistic regression models were built to evaluate AMR profiles for E. coli (n = 414) and Salmonella (n = 357) to each antimicrobial tested. Most E. coli isolates (91%) and Salmonella isolates (97%) were resistant to at least one of the antimicrobials tested. In general, E. coli and Salmonella had higher odds of resistance in calves aged ≥ 2 wk compared to 1-week-old calves, and little difference was seen in the level of resistance over the years observed or between seasons in most of the antimicrobials tested. Prospective research should investigate potential risk factors for the development of AMR in calves examples being antimicrobial use and farm management practices.

Étude observationnelle sur la résistance aux antimicrobiens d'isolats d'Escherichia coli et de Salmonella provenant d'échantillons de veaux de l'Ontario soumis à un laboratoire de diagnostic de 2007 à 2020. Les objectifs de cette étude étaient de i) décrire les isolats d'Escherichia coli et de Salmonella; ii) étudier les tendances temporelles des profils de résistance aux antimicrobiens (RAM); et iii) évaluer l'impact de la saison et de l'âge sur ces profils de RAM à partir d'échantillons diagnostiques et post-mortem de veaux de l'Ontario âgés de ≤ 2 mois soumis de 2007 à 2020 au Laboratoire de santé animale de Guelph, Ontario, Canada. Les résultats des tests de sensibilité aux antimicrobiens ont été mesurés par la méthode de diffusion en disque de Kirby-Bauer. Un total de 1291 isolats avec des profils de RAM ont été obtenus à partir de veaux, E. coli (n = 434) et Salmonella (n = 378) étant les bactéries les plus courantes caractérisées pour la RAM. Pour E. coli, 79 % des isolats testés ont montré un résultat positif en F5/K99, alors que pour les isolats de Salmonella, S. Typhimurium (33 %) et S. Dublin (22 %) étaient les deux sérotypes les plus fréquemment identifiés. Des modèles de régression logistique multivariable ont été construits pour évaluer les profils de RAM pour E. coli (n = 414) et Salmonella (n = 357) pour chaque antimicrobien testé. La plupart des isolats d'E. coli (91 %) et des isolats de Salmonella (97 %) étaient résistants à au moins un des antimicrobiens testés. En général, E. coli et Salmonella présentaient un risque de résistance plus élevé chez les veaux âgés de ≥ 2 semaines par rapport aux veaux âgés d'une semaine, et peu de différence a été observée dans le niveau de résistance au cours des années observées ou entre les saisons pour la plupart des antimicrobiens testés. La recherche prospective devrait étudier les facteurs de risque potentiels pour le développement de la RAM chez les veaux, par exemple l'utilisation d'antimicrobiens et les pratiques de gestion de la ferme.(Traduit par Dr Serge Messier).

Laboratory investigation of cases of fatal bacterial pneumonia in dairy cows.

Objective: Bacterial bronchopneumonia occurs in mature dairy cows but much of the information is extrapolated from knowledge of the disease in calves. The study was prompted by perceptions of an increasing occurrence and a paucity of information on fatal Mannheimia haemolytica pneumonia in dairy cows in Ontario. The study objectives were to describe the seasonality, main pathogens involved, and suggested predisposing factors for cases of fatal bacterial bronchopneumonia in mature dairy cows submitted for postmortem examination to a diagnostic laboratory, and to evaluate if the frequency of such submissions has increased over time. Animals: Mature dairy cows. Procedure: Retrospective study of cases submitted for postmortem examination to a diagnostic laboratory from 2007-2020 that were diagnosed as bacterial bronchopneumonia. Results: Most of the postmortem cases of bacterial bronchopneumonia in dairy cows were submitted from November to February (54% of cases). Mannheimia haemolytica was isolated from lung of 61/101 cases. Viruses were only identified in 8/55 cases tested. A minority (29/92) of bacterial isolates had in vitro resistance to antimicrobials used to treat pneumonia. Frequently suggested predisposing factors included recent introductions or movement of animals, recent or imminent calving, inclement weather, concurrent diseases, and poor ventilation in barns. Conclusion and clinical relevance: This study describes seasonal and annual trends, major pathogens, antimicrobial resistance profiles, and suggested predisposing factors in Ontario dairy cows submitted to a diagnostic laboratory for postmortem investigation of pneumonia and provides insights for understanding why outbreaks occur.

Objectif: La bronchopneumonie bactérienne survient chez les vaches laitières matures, mais une grande partie de l'information est extrapolée à partir de la connaissance de la maladie chez les veaux. L'étude a été motivée par la perception d'une occurrence croissante et d'un manque d'information sur la pneumonie mortelle à Mannheimia haemolytica chez les vaches laitières en Ontario. Les objectifs de l'étude étaient de décrire la saisonnalité, les principaux agents pathogènes impliqués et les facteurs prédisposants suggérés pour les cas de bronchopneumonie bactérienne mortelle chez les vaches laitières matures soumises à un examen post-mortem à un laboratoire de diagnostic, et d'évaluer si la fréquence de telles soumissions a augmenté au fil du temps. Animaux: Vaches laitières matures. Procédure: Étude rétrospective des cas soumis pour examen post-mortem à un laboratoire de diagnostic, entre 2007 et 2020, qui ont été diagnostiqués comme une bronchopneumonie bactérienne. Résultats: La plupart des cas post-mortem de bronchopneumonie bactérienne chez les vaches laitières ont été soumis de novembre à février (54 % des cas). Mannheimia haemolytica a été isolée du poumon de 61/101 cas. Des virus n'ont été identifiés que dans 8/55 cas testés. Une minorité (29/92) d'isolats bactériens présentaient une résistance in vitro aux antimicrobiens utilisés pour traiter la pneumonie. Les facteurs prédisposants fréquemment suggérés comprenaient des introductions ou des déplacements récents d'animaux, un vêlage récent ou imminent, des conditions météorologiques défavorables, des maladies concomitantes et une mauvaise ventilation dans les étables. Conclusion et pertinence clinique: Cette étude décrit les tendances saisonnières et annuelles, les principaux agents pathogènes, les profils de résistance aux antimicrobiens et les facteurs prédisposants suggérés chez les vaches laitières de l'Ontario soumises à un laboratoire de diagnostic pour une enquête post-mortem sur la pneumonie et fournit des informations pour comprendre pourquoi les épidémies se produisent.(Traduit par Dr Serge Messier).

Antimicrobial resistance in fecal Escherichia coli and Salmonella enterica isolates: a two-year prospective study of small poultry flocks in Ontario, Canada.

BACKGROUND: Although keeping small poultry flocks is increasingly popular in Ontario, information on the antimicrobial susceptibility of enteric bacteria of such flocks is lacking. The current study was conducted on small poultry flocks in Ontario between October 2015 and September 2017, and samples were submitted on a voluntary basis to Ontario's Animal Health Laboratory. From each submission, a pooled cecal sample was obtained from all the birds of the same species from the same flock and tested for the presence of two common enteric pathogens, E. coli and Salmonella. Three different isolates from each E. coli-positive sample and one isolate from each Salmonella-positive sample were selected and tested for susceptibility to 14 antimicrobials using a broth microdilution technique. RESULTS: A total of 433 fecal E. coli isolates (358 chicken, 27 turkey, 24 duck, and 24 game bird) and 5 Salmonella isolates (3 chicken, 1 turkey, and 1 duck) were recovered. One hundred and sixty-seven chicken, 5 turkey, 14 duck, and 15 game bird E. coli isolates were pan-susceptible. For E. coli, a moderate to high proportion of isolates were resistant to tetracycline (43% chicken, 81% turkey, 42% duck, and 38% game bird isolates), streptomycin (29% chicken, 37% turkey, and 33% game bird isolates), sulfonamides (17% chicken, 37% turkey, and 21% duck isolates), and ampicillin (16% chicken and 41% turkey isolates). Multidrug resistance was found in 37% of turkey, 20% of chicken, 13% of duck, and 8% of game bird E. coli isolates. Salmonella isolates were most frequently resistant to streptomycin, tetracycline, and sulfonamides. Resistance to cephalosporins, carbapenems, macrolides, and quinolones was infrequent in both E. coli and Salmonella isolates. Cluster and correlation analyses identified streptomycin-tetracycline-sulfisoxazole-trimethoprim-sulfamethoxazole as the most common resistance pattern in chicken E. coli isolates. Turkey E. coli isolates compared to all the other poultry species had higher odds of resistance to tetracycline and ampicillin, and a higher multidrug resistance rate. CONCLUSIONS: Escherichia coli isolates were frequently resistant to antimicrobials commonly used to treat poultry bacterial infections, which highlights the necessity of judicious antimicrobial use to limit the emergence of multidrug resistant bacteria.

Enhancing the one health initiative by using whole genome sequencing to monitor antimicrobial resistance of animal pathogens: Vet-LIRN collaborative project with veterinary diagnostic laboratories in United States and Canada.

BACKGROUND: Antimicrobial resistance (AMR) of bacterial pathogens is an emerging public health threat. This threat extends to pets as it also compromises our ability to treat their infections. Surveillance programs in the United States have traditionally focused on collecting data from food animals, foods, and people. The Veterinary Laboratory Investigation and Response Network (Vet-LIRN), a national network of 45 veterinary diagnostic laboratories, tested the antimicrobial susceptibility of clinically relevant bacterial isolates from animals, with companion animal species represented for the first time in a monitoring program. During 2017, we systematically collected and tested 1968 isolates. To identify genetic determinants associated with AMR and the potential genetic relatedness of animal and human strains, whole genome sequencing (WGS) was performed on 192 isolates: 69 Salmonella enterica (all animal sources), 63 Escherichia coli (dogs), and 60 Staphylococcus pseudintermedius (dogs). RESULTS: We found that most Salmonella isolates (46/69, 67%) had no known resistance genes. Several isolates from both food and companion animals, however, showed genetic relatedness to isolates from humans. For pathogenic E. coli, no resistance genes were identified in 60% (38/63) of the isolates. Diverse resistance patterns were observed, and one of the isolates had predicted resistance to fluoroquinolones and cephalosporins, important antibiotics in human and veterinary medicine. For S. pseudintermedius, we observed a bimodal distribution of resistance genes, with some isolates having a diverse array of resistance mechanisms, including the mecA gene (19/60, 32%). CONCLUSION: The findings from this study highlight the critical importance of veterinary diagnostic laboratory data as part of any national antimicrobial resistance surveillance program. The finding of some highly resistant bacteria from companion animals, and the observation of isolates related to those isolated from humans demonstrates the public health significance of incorporating companion animal data into surveillance systems. Vet-LIRN will continue to build the infrastructure to collect the data necessary to perform surveillance of resistant bacteria as part of fulfilling its mission to advance human and animal health. A One Health approach to AMR surveillance programs is crucial and must include data from humans, animals, and environmental sources to be effective.

Short communication: Detection of antibiotic resistance, mecA, and virulence genes in coagulase-negative Staphylococcus spp. from buffalo milk and the milking environment.

The aim of this study was to determinate whether coagulase-negative staphylococci (CNS) from buffalo milk or the milking environment possess virulence factors that are associated with intramammary infections or antimicrobial resistance. Milk samples (n = 320) from 80 lactating buffalo were evaluated for clinical and subclinical mastitis by physical examination, the strip cup test, California Mastitis Test (CMT), and somatic cell count (SCC) over a 4-mo period. In addition, swabs were obtained from the hands of consenting milkers (16), liners (64), and from the mouths (15) and nostrils (15) of buffalo calves. No clinical cases of mastitis were observed; however, CMT together with SCC results indicated that 8 animals had subclinical mastitis. Eighty-four CNS isolates were identified by MALDI-TOF MS and cydB real-time PCR (qPCR) and then evaluated by qPCR for presence of the eta, etb, sea, sec, cna, seb, sei, seq, sem, seg, see, and tst toxin genes, adhesion- and biofilm-associated genes (eno, ebps, fib, fnbA, coa), and the methicillin resistance gene (mecA). Resistance to antibiotics commonly used for mastitis treatment in Brazil was determined using the Kirby-Bauer test. Two strains were positive for the see and eta toxin genes; and mecA (1), eno (27), ebps (10), fnbA (10), and coa (5) genes were also detected. A notable number of isolates were resistant to erythromycin (30), penicillin (26), and cotrimoxazole (18); importantly, 10 vancomycin-resistant isolates were also detected. A smaller number of isolates were resistant to rifampicin (8), oxacillin (7), clindamycin (5), cefepime (4), tetracycline (3), ciprofloxacin (2), and chloramphenicol (1), and none were resistant to gentamicin or ciprofloxacin. Isolates with resistance to 2 (13 isolates), 3 (3), 4 (3), 5 (1), and 6 (1) antibiotics were detected. Taken together, our findings suggest that CNS isolates may not be a significant cause of clinical or even subclinical mastitis in buffaloes, but they may be a reservoir of virulence and antibiotic resistance genes.

Resistance to antimicrobials used at hatchery level among clinical Escherichia coli isolates from Ontario broilers.

Antimicrobials are used for prophylactic purposes in some flocks because chicks are susceptible to pathogenic bacterial infection at the hatchery stage. The objectives of this study were to assess temporal changes in resistance to ceftiofur, gentamicin, and spectinomycin in Escherichia coli clinical isolates from Ontario broiler chickens between 2008 and 2015, to determine whether stage of production, year and season are predictors of resistance, and whether published data on antimicrobial use help to interpret the temporal patterns in resistance. Logistic regression revealed that stage of production, year, and season were significant predictors of resistance to all 3 antimicrobials. Resistance to ceftiofur fluctuated over time, with a trend towards decreasing resistance between 2013 to 2015; resistance to gentamicin and spectinomycin increased over the study period, with significantly high resistance clusters identified from 2013 to 2015. Comparisons with published hatchery-level antimicrobial use data suggest that these trends may reflect changes in use of ceftiofur and spectinomycin in Ontario.

Résistance aux antimicrobiens utilisés dans les couvoirs chez des isolats d' Escherichia coli provenant de poulets à griller de l'Ontario. Les antimicrobiens sont utilisés à des fins préventives dans certains troupeaux car les oiseaux sont susceptibles à des infections causées par des bactéries pathogènes lorsqu'ils sont au couvoir. Les objectifs de la présente étude étaient d'évaluer les changements temporaux dans la résistance au ceftiofur, à la gentamicine, et à la spectinomycine chez des isolats cliniques d'Escherichia coli provenant de poulets à griller de l'Ontario entre 2008 et 2015, afin de déterminer si le stade de production, l'année et la saison sont des prédicteurs de résistance, et si les données publiées sur l'utilisation des antimicrobiens aide à interpréter les patrons temporaux de résistance. Une régression logistique a révélé que le stade de production, l'année, et la saison étaient des prédicteurs significatifs de résistance pour les trois antibiotiques. La résistance au ceftiofur fluctuait dans le temps, avec une tendance vers une diminution de résistance entre 2013 et 2015; la résistance à la gentamicine et à la spectinomycine a augmenté durant la période d'étude, avec des agrégats de résistance élevée significative entre 2013 et 2015. Des comparaisons avec les données publiées sur l'utilisation des antimicrobiens dans les couvoirs suggèrent que ces tendances pourraient refléter des changements dans l'utilisation du ceftiofur et de la spectinomycine en Ontario.(Traduit par Dr Serge Messier).

An epidemiological study of Streptococcus suis serotypes of pigs in Ontario determined by a multiplex polymerase chain reaction.

The purpose of the study was to identify the serotypes of Streptococcus suis from tonsil swabs in clinically ill and healthy pigs in Ontario using a multiplex polymerase chain reaction (PCR) method. Although 22 different serotypes were identified, most isolates were S. suis-like bacteria or untypable.

Étude épidémiologique des sérotypes Streptococcus suisdes porcs en Ontario déterminés par amplification en chaîne par polymérase multiplexe. Le but de cette étude consistait à identifier les sérotypes de Streptococcus suis provenant d'écouvillons des amygdales chez des porcs cliniquement malades et en santé en Ontario en utilisant une méthode multiplexe d'amplification en chaîne par polymérase (PCR). Même si 22 sérotypes différents ont été identifiés, la plupart des isolats étaient des bactéries de type S. suis ou non typables.(Traduit par Isabelle Vallières).

Flock-level prevalence, geographical distribution, and seasonal variation of avian reovirus among broiler flocks in Ontario.

Avian reovirus (ARV) is an economically significant pathogen of broiler chickens. Our objective was to determine the prevalence, geographical distribution, and seasonal variation of ARV infection among commercial broiler flocks in Ontario, Canada during grow-out. A cross-sectional study of 231 randomly selected flocks was conducted from July 2010 to January 2012. Fifteen blood samples, 15 whole intestines, and 15 cloacal swabs per flock were collected at slaughter; ELISA and PCR were used to determine a flock's ARV exposure status. Avian reovirus prevalence was 91% (95% CI: 87 to 94). District alone did not significantly explain the overall variation in the prevalence of ARV (univariable logistic regression; P = 0.073), although geographical differences were identified. The odds of ARV presence were significantly lower in the summer/autumn compared to the winter/spring (univariable exact logistic regression; P < 0.001). There was no association between flock mortality and flock ELISA mean titer or PCR status.

Prévalence au niveau du troupeau, distribution géographique et variations saisonnières du réovirus aviaire parmi les troupeaux de poulets à griller en Ontario. Le réovirus aviaire (RVA) est un agent pathogène important sur le plan économique pour les poulets à griller. Notre objectif consistait à déterminer la prévalence, la distribution géographique et les variations saisonnières de l'infection par le RVA parmi les troupeaux de poulets à griller commerciaux en Ontario, au Canada, durant la phase d'engraissement. Une étude transversale de 231 troupeaux choisis au hasard a été réalisée de juillet 2010 à janvier 2012. Quinze échantillons sanguins, 15 intestins complets et 15 écouvillons cloacaux ont été prélevés à l'abattage; des tests ELISA et ACP ont été utilisés pour déterminer l'état d'exposition du troupeau au RVA. La prévalence du réovirus aviaire était de 91 % (IC de 95 % : de 87 à 94). Le district à lui seul ne peut pas expliquer significativement la variation générale dans la prévalence du RVA (régression logistique univariable; P = 0,073), quoique des différences géographiques ont été identifiées. Les risques de la présence du RVA étaient significativement inférieurs à l'été/automne comparativement à l'hiver/printemps (régression logistique exacte univariable; P < 0,001). Il n'y avait aucune association entre la mortalité du troupeau et le titre moyen ELISA du troupeau ou de son état d'ACP.(Traduit par Isabelle Vallières).

Biosecurity practices and causes of enteritis on Ontario meat rabbit farms.

Infectious enterocolitis is a significant cause of mortality in meat rabbits. Disease risk is enhanced by intensive rearing practices and poor on-farm biosecurity. This investigation was undertaken in farmed meat rabbits during an Ontario-wide outbreak of enteritis with high mortality to determine the prevalence of causative agents. A survey evaluating on-farm biosecurity practices was also conducted to identify potential means of pathogen contamination and zoonotic risks. Gross and microscopic pathology evaluations combined with microbiologic testing were conducted on 95 rabbits over spring and winter months. Escherichia coli and Clostridium spiroforme were most commonly associated with enteritis in rabbits regardless of age or season and lesions were significantly more severe in mature does (P < 0.0001). The survey results demonstrated a lack of consistent on-farm biosecurity practices. The infectious nature of enteric disease of rabbits combined with poor biosecurity practices may contribute to disease transmission within and between farms.

Pratiques de biosécurité et causes d'entérite dans des fermes d'élevage de lapins de l'Ontario. L'entérocolite infectieuse est une cause importante de mortalité chez les lapins d'élevage. Le risque de maladie est accru par des pratiques d'élevage intensives et une mauvaise biosécurité à la ferme. Cette enquête a été entreprise chez des lapins d'élevage durant une éclosion d'entérite à l'échelle de l'Ontario qui présentait un taux de mortalité élevé afin de déterminer la prévalence des agents étiologiques. On a aussi réalisé un sondage évaluant les pratiques de biosécurité à la ferme afin d'identifier les modes potentiels de contamination des agents pathogènes et les risques zoonotiques. Des évaluations pathologiques macroscopiques et microscopiques combinées à des tests microbiologiques ont été réalisés sur 95 lapins au cours des mois d'été et d'hiver. Escherichia coli et Clostridium spiroforme étaient le plus communément associés à l'entérite chez les lapins sans égard à l'âge ou à la saison et les lésions étaient significativement plus graves chez les lapines adultes (P < 0,0001). Les résultats du sondage ont démontré l'absence de pratiques de biosécurité uniformes à la ferme. La nature infectieuse de la maladie entérique des lapins et de mauvaises pratiques de biosécurité peuvent contribuer à la transmission de la maladie dans les fermes et entre ces dernières.(Traduit par Isabelle Vallières).

The first case of porcine epidemic diarrhea in Canada.

In January, 2014, increased mortality was reported in piglets with acute diarrhea on an Ontario farm. Villus atrophy in affected piglets was confined to the small intestine. Samples of colon content were PCR-positive for porcine epidemic diarrhea virus (PEDV). Other laboratory tests did not detect significant pathogens, confirming this was the first case of PED in Canada.

Premier cas de diarrhée épidémique porcine au Canada. En janvier 2014, une mortalité accrue a été signalée chez des porcelets atteints de diarrhée aiguë dans une ferme de l'Ontario. L'atrophie des villosités chez les porcelets touchés a été confinée au petit intestin. Des échantillons du contenu du côlon étaient positifs par RCP pour le virus de la diarrhée épidémique porcine (VDEP). D'autres tests de laboratoire n'ont pas détecté d'agents pathogènes importants, ce qui confirme qu'il s'agit du premier cas de DEP au Canada.(Traduit par Isabelle Vallières).

Antimicrobial susceptibility of Escherichia coli F4, Pasteurella multocida, and Streptococcus suis isolates from a diagnostic veterinary laboratory and recommendations for a surveillance system.

Antimicrobial susceptibility data on Escherichia coli F4, Pasteurella multocida, and Streptococcus suis isolates from Ontario swine (January 1998 to October 2010) were acquired from a comprehensive diagnostic veterinary laboratory in Ontario, Canada. In relation to the possible development of a surveillance system for antimicrobial resistance, data were assessed for ease of management, completeness, consistency, and applicability for temporal and spatial statistical analyses. Limited farm location data precluded spatial analyses and missing demographic data limited their use as predictors within multivariable statistical models. Changes in the standard panel of antimicrobials used for susceptibility testing reduced the number of antimicrobials available for temporal analyses. Data consistency and quality could improve over time in this and similar diagnostic laboratory settings by encouraging complete reporting with sample submission and by modifying database systems to limit free-text data entry. These changes could make more statistical methods available for disease surveillance and cluster detection.

Sensibilité antimicrobienne des isolats d'Escherichia coliF4, dePasteurella multocidaet deStreptococcus suistransmise par un laboratoire de diagnostic vétérinaire et recommandations pour un système de surveillance. Les données de sensibilité antimicrobienne sur les isolats d'Escherichia coli F4, de Pasteurella multocida et de Streptococcus suis provenant des porcs de l'Ontario (de janvier 1998 à octobre 2010) ont été acquises auprès d'un laboratoire de diagnostic vétérinaire complet situé en Ontario, au Canada. En relation avec la création éventuelle d'un système de surveillance pour l'antibiorésistance, des données ont été évaluées pour déterminer la facilité de gestion, l'intégralité, la cohérence et l'applicabilité des analyses temporelles et spatiales. Des données limitées sur l'emplacement de la ferme empêchaient des analyses spatiales et des données démographiques manquantes limitaient leur utilisation comme prédicteurs au sein de modèles statistiques multivariables. Les changements du groupe standard d'antimicrobiens utilisés pour les tests de sensibilité ont réduit le nombre d'antimicrobiens disponibles pour des analyses temporelles. La cohérence et la qualité des données pourraient être améliorées au fil du temps dans ce laboratoire de diagnostic et d'autres installations semblables en encourageant la production de rapports complets avec la soumission d'échantillons et en modifiant les systèmes des bases de données afin de limiter l'entrée de données en forme libre. Ces changements pourraient rendre d'autres méthodes statistiques disponibles pour la surveillance des maladies et la détection de grappes.(Traduit par Isabelle Vallières).

An investigation of ear necrosis in pigs.

Porcine ear necrosis was investigated in 23 conveniently chosen farms, consisting of 14 case farms and 9 control farms. Biopsies of lesions and oral swabs from pigs on 11 case farms were examined by histology and bacterial culture. All farms were visited for observations and a survey on management, housing, and the presence of other clinical signs or behavioral vices. Histological examination revealed that the lesions began on the surface and progressed to deeper layers, and that vascular damage did not appear to be the initiating cause. Spirochetes were only rarely observed in histological examination and were not cultured from biopsies and oral swabs. Staphylococcus aureus and Staphylococcus hyicus were cultured from 91% and 66% of samples, respectively. Ear biting and a humid environment were associated with ear necrosis. On some farms large numbers of pigs were affected and lesions were sometimes extensive. The condition appears to be an infectious disease beginning on the surface of the skin; contributing environmental and management factors are likely.

Enquête sur la nécrose des oreilles chez les porcs. La nécrose des oreilles porcines a fait l'objet d'une enquête dans un échantillonnage de convenance de 23 fermes, composé de 14 fermes de cas et de 9 fermes témoins. Les biopsies de lésions et des prélèvements oraux faits chez des porcs dans 11 fermes de cas ont été examinés par histologie et culture bactérienne. Toutes les fermes ont été visitées pour recueillir des observations et effectuer un examen de la gestion, du logement et de la présence d'autres signes cliniques ou vices de comportement. L'examen histologique a révélé que les lésions commençaient sur la surface et progressaient vers des couches plus profondes et que les dommages vasculaires ne semblaient pas être la cause initiale. Des spirochètes ont été rarement observés à l'examen histologique et n'ont pas été cultivés dans les biopsies et les prélèvements oraux. Staphylococcus aureus et Staphylococcus hyicus ont été cultivés dans 91 % et 66 % des échantillons, respectivement. Les morsures d'oreilles et un environnement humide étaient associés à la nécrose des oreilles. Dans certaines fermes, un grand nombre de porcs étaient affectés et les lésions étaient parfois importantes. L'affection semble être une maladie infectieuse qui commence à la surface de la peau; des facteurs contributifs liés à l'environnement et à la gestion sont probables.(Traduit par Isabelle Vallières).

Bacterial diskospondylitis in juvenile mink from 2 Ontario mink farms.

Nine juvenile mink with hind-limb paresis/paralysis from 2 Ontario farms were submitted for necropsy. Diagnostic tests revealed spinal compression and severe thoracic diskospondylitis with intralesional Gram-positive coccoid bacterial colonies. Streptococcus canis, Streptococcus dysgalactiae subsp. equisimilis, and hemolytic Staphylococcus spp. were isolated from vertebral lesions.

Discospondylite bactérienne chez des jeunes visons provenant de 2 fermes de visons de l'Ontario. Neuf jeunes visons atteints d'une parésie/paralysie des membres postérieurs provenant de 2 fermes de l'Ontario ont été soumis à une nécropsie. Les tests diagnostiques ont révélé une compression médullaire et une discospondylite thoracique grave avec des colonies de bactéries coccoïdes à Gram positif. Les bactéries Streptococcus canis, Streptococcus dysgalactiae subsp. equisimilis, et Staphylococcus spp. hémolytiques ont été isolés des lésions vertébrales.(Traduit par Isabelle Vallières).

Rapid identification of Salmonella serovars Enteritidis and Typhimurium using whole cell matrix assisted laser desorption ionization - Time of flight mass spectrometry (MALDI-TOF MS) coupled with multivariate analysis and artificial intelligence.

Salmonella is a common food-borne pathogen with Enteritidis and Typhimurium being among the most important serovars causing numerous outbreaks. A rapid method was investigated to identify these serovars using whole-cell MALDI-TOF MS coupled with multivariate analysis and artificial intelligence and 113 Salmonella strains, including 38 Enteritidis (SE), 38 Typhimurium (ST) and 37 strains from 32 other Salmonella serovars (SG). Datasets of ions (presence/absence) with high discriminative power were created using newly developed criteria and subject to multivariate analyses and eight artificial intelligence (AI) tools. Principal Component Analysis based on 55 or 88 selected ions separated SE, ST and SG without overlap on the first three principal components. Datasets were partitioned using five partitioning methods with 70% of samples for AI model training and 30% for validation. Of the eight AI models evaluated, high performance (HP) SVM and HP Neural were the top performers, identified three serovar groups 97% correctly on average (range 82%-100%) according to the validation results. Selection of serovar specific ions facilitated differentiation of serotypes using unsupervised model PCA and improved the accuracy of classification using AI significantly (p < 0.01). MALDI-TOF MS incorporated with advanced data processing and classification tools is a promising method to allow rapid identification of Salmonella serovars of concern in routine diagnostic laboratories.

AAV Vectors Pseudotyped with Capsids from Porcine and Bovine Species Mediate In Vitro and In Vivo Gene Delivery.

Adeno-associated virus (AAV) vectors are among the most widely used delivery vehicles for in vivo gene therapy as they mediate robust and sustained transgene expression with limited toxicity. However, a significant impediment to the broad clinical success of AAV-based therapies is the widespread presence of pre-existing humoral immunity to AAVs in the human population. This immunity arises from the circulation of non-pathogenic endemic human AAV serotypes. One possible solution is to use non-human AAV capsids to pseudotype transgene-containing AAV vector genomes of interest. Due to the low probability of human exposure to animal AAVs, pre-existing immunity to animal-derived AAV capsids should be low. Here, we characterize two novel AAV capsid sequences: one derived from porcine colon tissue and the other from a caprine adenovirus stock. Both AAV capsids proved to be effective transducers of HeLa and HEK293T cells in vitro. In vivo, both capsids were able to transduce the murine nose, lung, and liver after either intranasal or intraperitoneal administration. In addition, we demonstrate that the porcine AAV capsid likely arose from multiple recombination events involving human- and animal-derived AAV sequences. We hypothesize that recurrent recombination events with similar and distantly related AAV sequences represent an effective mechanism for enhancing the fitness of wildtype AAV populations.

Methicillin-resistant Staphylococcus aureus in a neonatal alpaca.

A 6-hour-old alpaca was presented for evaluation of respiratory difficulty. As part of routine surveillance, methicillin-resistant Staphylococcus aureus (MRSA) was identified from a nasal swab taken upon admission to the hospital. No signs of MRSA infection were noted. The MRSA strain recovered was a human epidemic clone that has been associated with horses. Methicillin-resistant S. aureus colonization can occur in camelids, and the potential animal and public health risks require consideration.

Clostridium perfringens type A fatal acute hemorrhagic gastroenteritis in a dog.

The morning after participating in a dog show, a 2-year-old Pomeranian dog was found dead in a pool of bloody feces. Necropsy revealed hemorrhagic gastroenteritis of the entire gastrointestinal tract, with many Gram-positive bacilli on the surface and in the lumen and crypts of the intestine. Enterotoxin-positive type A Clostridium perfringens were isolated in large numbers. This dramatic case of fatal C. perfringens gastroenteritis highlights the need to better understand the role of this bacterium in enteric disease of dogs.

Diagnostic and public health investigation of Mycobacterium tuberculosis infection in a dog in Ontario, Canada.

A 4-y-old, female mixed-breed dog was presented to the Ontario Veterinary College for further evaluation of multiple pulmonary and hepatic masses, intrathoracic lymphadenitis, and recent development of a pyogranulomatous pleural effusion. Along with other comprehensive tests, a thoracic lymph node biopsy was performed, and Mycobacterium tuberculosis complex infection was confirmed by real-time PCR. The dog's condition declined post-operatively, and euthanasia was elected. Postmortem examination confirmed severe granulomatous pneumonia, hepatitis, intrathoracic and intraabdominal lymphadenitis, omentitis, and nephritis. Line-probe assays performed on samples collected postmortem confirmed the species as M. tuberculosis. 24-loci MIRU-VNTR genotyping, spoligotyping, and whole-genome sequencing revealed relations to known human isolates, but no epidemiologic link to these cases was investigated. Given the concern for potential human exposure during this animal's disease course, a public health investigation was initiated; 45 individuals were tested for M. tuberculosis exposure, and no subsequent human infections related to this animal were identified. Our case highlights the need for more readily available, minimally invasive testing for the diagnosis of canine mycobacteriosis, and highlights the ability of canid species to act as potential contributors to the epidemiology of M. tuberculosis infections.

The lipopolysaccharide core of Actinobacillus suis and its relationship to those of Actinobacillus pleuropneumoniae.

The Gram-negative bacteria Actinobacillus suis colonizes the upper respiratory and genital tracts of swine. Along with capsular polysaccharides, lipopolysaccharides (O-chain→core→lipid A~cell) are a main cell-surface component of A. suis. In this study, we determined that A. suis lipopolysaccharide incorporates a conserved core that shares some structural features with several core types of A. pleuropneumoniae . These common core structural features likely account for the observed serological cross-reactivity between A. suis and A. pleuropneumoniae, and the data suggest that the structural epitopes responsible for immunogenicity are those in the outer core domain.