Prévalence, facteurs de risque et mécanismes de dissémination des gènes de résistance aux antibiotiques, l’espèce équine et l’espèce porcine ont été étudiées en insistant particulièrement sur les antibiotiques de haute importance en médecine humaine dans chaque filière (céphalosporines de 3e génération et fluoroquinolones respectivement).

La résistance aux antibiotiques a pris une ampleur considérable du fait de l’utilisation des antibiotiques dans de nombreux domaines. Pour respecter l’approche « OneHealth », il est essentiel d’avoir une image spécifique de chaque situation, afin d’orienter les recommandations et de limiter la dissémination des gènes, des plasmides et des clones. Nos objectifs étaient adaptés à nos populations d’étude (chevaux et porcs) afin d’ajuster les résultats aux besoins des filières. Dans la filière équine, nous avons quantifié les résistances phénotypiques et identifié les gènes de β-lactamases à spectre étendu (BLSE/AmpC) présents dans le microbiome des chevaux sains, et nous avons identifié les facteurs de risque associés à leur portage en France et au Québec. En France, nous avons également caractérisé les mécanismes de dissémination des gènes de BLSE/AmpC. Nous avons mis en évidence qu’en France et au Québec, les E. coli commensaux provenant de fèces de chevaux sains étaient majoritairement non susceptibles à l’ampicilline, l’amoxicilline/acide clavulanique et la streptomycine et que des E. coli multirésistants et porteurs de gènes codant pour des BLSE/AmpC étaient détectés dans respectivement environ 45% et 8% des chevaux. Le blaCTX-M-1 était majoritairement détecté bien qu’en France d’autres BLSE aient été identifiés (blaCTX-M-2 et blaCTX-M-14) ainsi que le gène AmpC blaCMY-2. L’administration d’un traitement médical, le nombre de personnes s’occupant des chevaux, le type d’activité et le fait de participer à un évènement équestre dans les trois derniers mois ont été identifiés comme des facteurs de risque du portage des E. coli multirésistants ou producteurs de gènes BLSE/AmpC, soit en France soit au Québec. En France, le plasmide IncHI1-ST9 était majoritairement associé aux gènes blaCTX-M-1/2 et à l’opéron fos. Pour la filière porcine, nos objectifs étaient de colliger les données de la base de données du laboratoire EcL entre 2008 et 2016, d’évaluer la présence d’un agrégat spatio-temporel pour les isolats ETEC:F4 non susceptibles à l’enrofloxacine et de caractériser ces isolats et les éléments génétiques mobiles qu’ils transportent. En effet, l’enrofloxacine est un antibiotique de haute importance en santé humaine, et doit donc faire l’objet d’une surveillance accrue. Nous avons trouvé que plus de 90% des isolats d’E. coli entérotoxinogènes détectés chez des cas de porcs malades soumis au laboratoire EcL de 2008 à 2016 au Québec, étaient multirésistants. Le virotype LT:STb:F4 prédominait jusqu’en 2014, puis a été dépassé par le virotype LT:STb:STa:F4. Un agrégat spatio-temporel d’isolats LT:STb:STa:F4 non susceptibles à l’enrofloxacine a été détecté entre 04/2015 et 09/2016 au centre de la Montérégie. Nous avons démontré la présence d’un clone ETEC:F4 non susceptible à l’enrofloxacine, à haut risque, qui se dissémine en Amérique du Nord depuis 2013. Les isolats appartenant à ce clone sont ST100, O149:H10. Ils sont multirésistants, et associés à une pathogénicité et une virulence augmentée par rapport aux isolats détectés avant 2000. Ils portent le réplicon IncFII. Les résistances et leur mécanisme de dissémination sont différents selon l’espèce animale. Ces divergences sont fonction de l’usage des antibiotiques, et des échanges possibles avec les différents protagonistes en contact avec les animaux.Antimicrobial resistance has become an essential issue in the last decades because of the extensive use of antimicrobials in numerous sectors. In order to follow the OneHealth approach, it is critical to have a precise picture of each situation, to adjust recommendations and prevent resistance gene dissemination as well as plasmid and clone spread. Our objectives were adapted to the animal populations under study. Therefore, our results were compatible with each sector. In the equine sector, we quantified phenotypic resistance and identified β-lactamase (ESBL/AmpC) genes present in the intestinal microbiome of healthy horses and we identified risk factors associated with their carriage both in France and in Quebec. Then, in France we characterized ESBL/AmpC gene spread mechanisms. We demonstrated that commensal E. coli originating from the feces of healthy horses were mostly non-susceptible to ampicillin, amoxicillin/clavulanic acid and streptomycin. The presence of multidrug resistant E. coli and of E. coli carrying ESBL/AmpC genes was found in around 45% and 8% of horses respectively. The most frequently detected gene was blaCTX-M-1, although blaCTX-M-2 and blaCTX-M-14 were also identified in France. The AmpC gene blaCMY-2 was identified in both localities. Medical treatment, staff number, activity, and participation in an equestrian event within the last three months were identified as risk factors for MDR or ESBL/AmpC E. coli. In France, commensal E. coli from healthy horses most commonly possessed the IncHI1-ST9 plasmid. This plasmid carries blaCTX-M-1/2 genes and the fos operon. For the swine sector in Quebec, our objectives were to gather data provided by the Animal pathogenic and zoonotic E. coli (APZEC) database between 2008 and 2017, to assess the presence of a spatio-temporal cluster for enrofloxacin non-susceptible ETEC:F4 and to characterize these isolates and the mobile genetic elements they carry. Enrofloxacin is an antibiotic classified as highly important in human medicine and as such needs to come under higher scrutiny. For this sector, we demonstrated that more than 90% of enterotoxigenic E. coli (ETEC) isolates from diseased swine submitted to the EcL between 2008 and 2016, were multidrug resistant. The main virotype in 2014 was LT:STb:F4. It was subsequently replaced by the LT:STb:STa:F4 virotype. A spatio-temporal cluster of LT:STb:STa:F4 isolates non-susceptible to enrofloxacin was detected between 04/2015 and 09/2016 in the centre of the Monteregie region. These isolates constituted an ETEC:F4 high risk enrofloxacin non-susceptible clone, which has been spreading in North America since 2013. Isolates belonging to this clone are ST100, O149H10, phylogroup A, and fimH gene negative. These isolates are multidrug resistant and associated with a higher pathogenicity and virulence than isolates detected before 2000. They all carry the incFII replicon. Resistance and mechanisms of dissemination are different according to the animal species being studied. This is likely due to different patterns of antimicrobial use in each industry and possible interactions with different protagonists in contact with the animals. It is essential to understand the situation for each animal species in order to adapt recommendations for efficiently limiting the spread of resistance genes, plasmids and clones

Clonal and plasmidic dissemination of critical antimicrobial resistance genes through clinically relevant ExPEC and APEC-like lineages (ST) in the dairy cattle population of Québec, Canada

Antimicrobial resistance can be effectively limited by improving the judicious use of antimicrobials in food production. However, its effect on the spread of AMR genes in animal populations is not well described. In the province of Québec, Canada, a new legislation implemented in 2019 has led to an unprecedented reduction in the use of critical antimicrobials in dairy production. We aimed to investigate the potential link between ESBL/AmpC E. coli isolated before and after legislation and to determine the presence of plasmids carrying genes responsible for critical AMR. We collected fecal samples from calves, cows, and manure pit from 87 Québec dairy farms approximately 2 years before and 2 years after the legislation came into effect. The whole genomes of 183 presumptive ESBL/AmpC E. coli isolated after cefotaxime enrichment were sequenced. Their phylogenetic characteristics (MLST, serogroup, cgMLST) and the presence of virulence and resistance genes and replicons were examined. A maximum likelihood phylogenetic tree was constructed based on single nucleotide polymorphism (SNPs). We identified 10 clonal lineages (same cgMLST) and 7 clones (SNPs ≤ 52). Isolates belonging to these clones could be found on different farms before and after the legislation, strongly suggesting a clonal spread of AMR genes in the population during this 4-year period. All isolates were multidrug resistant (MDR), with clone 2 being notable for the presence of macrolide, fluoroquinolone, and third-generation cephalosporin resistance genes. We also identified clinically relevant ExPEC (ST10) and APEC-like lineages (ST117, ST58, ST88) associated with the presence of ExPEC and APEC virulence genes, respectively. Our data also suggests the presence of one epidemic plasmid belonging to the IncY incompatibility group and carrying qnrs1 and blaCTX–M–15. We demonstrated that AMR genes spread through farms and can persist over a 4-year period in the dairy cattle population through both plasmids and E. coli clones, despite the restriction of critical antimicrobial use. MDR ExPEC and APEC-like STs are present in the normal microbiota of cattle (more frequently in calves). These data increase our knowledge on gene dissemination dynamics and highlight the fact that biosecurity measures should be enhanced in this industry to limit such dissemination

Prévalence des arythmies cardiaques liées à l'effort chez le trotteur

La contre-performance du cheval de course est un défi diagnostique pour tout vétérinaire équin. La plupart du temps elle est multifactorielle et engage les appareils locomoteur, respiratoire et cardiovasculaire. Les arythmies cardiaques peuvent être responsables de baisse de performances notables chez le cheval. La plus importante, et par conséquent la mieux connue, est la fibrillation atriale. La signification clinique des autres arythmies liées à l effort, en particulier les extrasystoles atriales et ventriculaires, est par contre beaucoup plus floue. Les connaissances actuelles dans ce domaine ont été remises en cause largement par une étude de Ryan en 2005 qui montrait que certaines de ces arythmies pouvaient être présentes pendant un effort chez des pur-sang tout fait performant. Notre étude consistait à déterminer l existence, la prévalence et les éventuels facteurs de risque d apparition de ces arythmies chez le trotteur. On a trouvé que 11% des chevaux présentaient des extrasystoles atriales et 30% des chevaux présentaient des extrasystoles ventriculaires au cours des différentes périodes de travail. Certaines arythmies observées lors de cette étude, telle la fibrillation atriale en plein effort, sont difficiles à expliquer. Aucun facteur de risque n a pu être déterminé du fait de la petitesse de l échantillon. Seule des différences significatives entre les effectifs ont pu être mis pour les extrasystoles atriales en fonction de l âge, des gains, du type d échauffement et de la fréquence cardiaque maximum lors de l effort.MAISONS-ALFORT-Ecole Vétérin (940462302) / SudocSudocFranceF

Prevalence, Risk Factors, and Characterization of Multidrug Resistant and ESBL/AmpC Producing Escherichia coli in Healthy Horses in Quebec, Canada, in 2015–2016

Although antimicrobial resistance is an increasing threat in equine medicine, molecular and epidemiological data remain limited in North America. We assessed the prevalence of, and risk factors for, shedding multidrug-resistant (MDR) and extended-spectrum β-lactamase (ESBL) and/or AmpC β-lactamase-producing E. coli in healthy horses in Quebec, Canada. We collected fecal samples in 225 healthy adult horses from 32 premises. A questionnaire on facility management and horse medical history was completed for each horse. Indicator (without enrichment) and specific (following enrichment with ceftriaxone) E. coli were isolated and tested for antimicrobial susceptibility. The presence of ESBL/AmpC genes was determined by PCR. The prevalence of isolates that were non-susceptible to antimicrobials and to antimicrobial classes were estimated at the horse and the premises level. Multivariable logistic regression was used to assess potential risk factors for MDR and ESBL/AmpC isolates. The shedding of MDR E. coli was detected in 46.3% of horses. Non-susceptibility was most commonly observed to ampicillin, amoxicillin/clavulanic acid or streptomycin. ESBL/AmpC producing isolates were detected in 7.3% of horses. The most commonly identified ESBL/AmpC gene was blaCTX-M-1, although we also identified blaCMY-2. The number of staff and equestrian event participation were identified as risk factors for shedding MDR isolates. The prevalence of healthy horses harboring MDR or ESBL/AmpC genes isolates in their intestinal microbiota is noteworthy. We identified risk factors which could help to develop guidelines to preclude their spread

High Risk Clone: A Proposal of Criteria Adapted to the One Health Context with Application to Enterotoxigenic Escherichia coli in the Pig Population

The definition of a high risk clone for antibiotic resistance dissemination was initially established for human medicine. We propose a revised definition of a high risk clone adapted to the One Health context. Then, we applied our criteria to a cluster of enrofloxacin non susceptible ETEC:F4 isolates which emerged in 2013 in diseased pigs in Quebec. The whole genomes of 183 ETEC:F4 strains isolated in Quebec from 1990 to 2018 were sequenced. The presence of virulence and resistance genes and replicons was examined in 173 isolates. Maximum likelihood phylogenetic trees were constructed based on SNP data and clones were identified using a set of predefined criteria. The strains belonging to the clonal lineage ST100/O149:H10 isolated in Quebec in 2013 or later were compared to ETEC:F4 whole genome sequences available in GenBank. Prior to 2000, ETEC:F4 isolates from pigs in Quebec were mostly ST90 and belonged to several serotypes. After 2000, the isolates were mostly ST100/O149:H10. In this article, we demonstrated the presence of a ETEC:F4 high risk clone. This clone (1) emerged in 2013, (2) is multidrug resistant, (3) has a widespread distribution over North America and was able to persist several months on farms, and (4) possesses specific virulence genes. It is crucial to detect and characterize high risk clones in animal populations to increase our understanding of their emergence and their dissemination

High Risk Clone: A Proposal of Criteria Adapted to the One Health Context with Application to Enterotoxigenic <i>Escherichia coli</i> in the Pig Population

The definition of a high risk clone for antibiotic resistance dissemination was initially established for human medicine. We propose a revised definition of a high risk clone adapted to the One Health context. Then, we applied our criteria to a cluster of enrofloxacin non susceptible ETEC:F4 isolates which emerged in 2013 in diseased pigs in Quebec. The whole genomes of 183 ETEC:F4 strains isolated in Quebec from 1990 to 2018 were sequenced. The presence of virulence and resistance genes and replicons was examined in 173 isolates. Maximum likelihood phylogenetic trees were constructed based on SNP data and clones were identified using a set of predefined criteria. The strains belonging to the clonal lineage ST100/O149:H10 isolated in Quebec in 2013 or later were compared to ETEC:F4 whole genome sequences available in GenBank. Prior to 2000, ETEC:F4 isolates from pigs in Quebec were mostly ST90 and belonged to several serotypes. After 2000, the isolates were mostly ST100/O149:H10. In this article, we demonstrated the presence of a ETEC:F4 high risk clone. This clone (1) emerged in 2013, (2) is multidrug resistant, (3) has a widespread distribution over North America and was able to persist several months on farms, and (4) possesses specific virulence genes. It is crucial to detect and characterize high risk clones in animal populations to increase our understanding of their emergence and their dissemination

Prevalence, risk factors, and characterization of multidrug resistant and extended spectrum β-lactamase/AmpC β-lactamase producing Escherichia coli in healthy horses in France in 2015

International audienceAlthough antimicrobial resistance is increasingly common in equine medicine, molecular and epidemiological data remains scarce. We estimated the prevalence of, and risk factors for, shedding of multidrug resistant (MDR), extended spectrum β-lactamase (ESBL)-producing, and AmpC β-lactamase-producing, or some combination of these in Escherichia coli in horses in France. We characterized ESBL/AmpC isolates for antimicrobial susceptibility and the presence of virulence and ESBL/AmpC-associated resistance genes. Fecal samples from healthy adult horses at 41 premises were collected. A questionnaire was completed by each premises manager. A subset of these samples was tested to build 2 bacterial collections. Indicator (without enrichment) and specific (enrichment with ceftriaxone) E. coli tested for antimicrobial susceptibility. Prevalence of isolates nonsusceptible to antimicrobials was estimated at the horse and the premises level. The ESBL/AmpC and virulence genes were identified by PCR. Multivariable logistic regression was used to investigate risk factors for MDR and ESBL/AmpC isolates at premises. Virulence gene iutA was identified in 1 ESBL/AmpC-producing isolate. Medical treatment, staff numbers, and activity were identified as risk factors for housing horses shedding ESBL/AmpC-producing E. coli isolates. Prevalence of healthy horses harboring ESBL/AmpC genes and MDR isolates in their intestinal microbiota is substantial. Risk factors could be used to elaborate guidelines to prevent their dissemination

Spread of multidrug-resistant IncHI1 plasmids carrying ESBL gene blaCTX-M-1 and metabolism operon of prebiotic oligosaccharides in commensal Escherichia coli from healthy horses, France

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Escherichia coli producteurs de β-lactamases à spectre étendu chez le cheval en France

International audienceObjectif-Introduction : Les animaux de compagnie sont considérés comme un réservoir potentiel de bactéries résistantes aux antibiotiques notamment aux céphalosporines de dernières générations. L’objectif de cette étude était de déterminer le portage intestinal de souches d’Escherichia coli résistantes aux céphalosporines de dernières générations (C3/4G) chez le cheval en France et de caractériser les déterminants génétiques de cette résistance.Matériels et méthodes : Sept-cent-trente-huit chevaux de 41 élevages et centres équestres français ont été échantillonnés pendant l’été 2015. La présence d’E. coli résistants aux céphalosporines de dernières générations a été recherchée dans les crottins. Les gènes de résistance aux C3/4G ont été déterminéss par PCR et séquençage. Les plasmides porteurs de gènes BLSE et/ou céphalosporinase AmpC ont été transférés par conjugaison ou transformation afin d’être caractérisés par typage de réplicon, profil de restriction et multilocus sequence typing. Les souches d’E. coli resistantes aux C3/4G rations ont été caractérisées par champs pulsé, détermination du phylogroupe et du contenu en gènes de virulence.Results: Seize structures équestres hébergeaient au minimum un cheval porteur d’E. coli résistants aux C3/4G, ce qui représentait une prévalence de 7,9% de portage parmi les chevaux échantillonnés. Cinquante-une souches distinctes d’E. coli résistantes aux C3/4G ont été incluses dans l’analyse moléculaire de la résistance. Toutes ces souches présentaient une grande diversité de profil de macrorestriction en champs pulsé XbaI, appartenaient majoritairement au phylogroupe B1 et ne possédaient pas les gènes majeurs de virulence testés pour les animaux suggérant qu’il s’agit de souches commensales non pathogènes. Les BLSE du groupe CTX-M étaient largement prédominante (blaCTX-M-1, n=35; blaCTX-M-2, n=8; blaCTX-M-14, n=2). Les gènes BLSE/AmpC ont été localisés sur différents plasmides appartenant aux groupes d’incompatibilité IncHI1, IncI1, IncN, et IncY. Les gènes BLSE blaCTX-M-1 et blaCTX-M-2 étaient principalement portés par des plasmides IncHI1. Conclusion: Cette étude originale a mis en évidence un portage significatif d’E. coli producteur de BLSE chez le cheval sain en France. Les plasmides porteurs de gènes BLSE sont les acteurs majeurs de la dissémination de la résistance aux C3/4G dans le microbiote intestinal du cheval

Fecal carriage and characterization of extended-spectrum β-lactamase- and AmpC β-lactamase-producing Escherichia coli from healthy horses in France

Fecal carriage and characterization of extended-spectrum β-lactamase- and AmpC β-lactamase-producing [i]Escherichia coli[/i] from healthy horses in France. 7. Symposium on Antimicrobial Résistance in Animals and the Environment (ARAE