Characterization of third-generation cephalosporin-resistant Escherichia coli from slaughter calves and fattening pigs: A pilot study for monitoring antimicrobial resistance by whole genome sequencing in Switzerland.

INTRODUCTION: Whole genome sequencing (WGS) was introduced into Swiss antimicrobial resistance monitoring in 2022 as an additional method to phenotypic antimicrobial susceptibility testing by broth microdilution to characterize presumptive third-generation cephalosporin-resistant (3GC-R) Escherichia coli. Caecal samples from Swiss slaughter calves and fattening pigs, as well as beef and pork meat from Swiss retail taken in 2021, were analyzed for the presence of 3GC-R E. coli according to European harmonized protocols. In 2021, 3GC-R E. coli was detected in 23,8 % of slaughter calves, 5,9 % of fattening pigs, and 0 % of meat. Comparative analysis of the antimicrobial resistance results obtained by phenotypic measurement and those obtained by the detection of corresponding underlying molecular mechanisms by WGS showed very high agreement (99 %). Resistance to third-generation cephalosporins (3GCs) was mainly associated with the presence of blaCTX-M-15 in E. coli isolates from calves and blaCTX-M-1 in E. coli isolates from pigs and mutations in the ampC-promoter (g.-42 C>T) in E. coli isolates from both animal species. Moreover, WGS data were used for phylogenetic analysis based on multi locus sequence types (MLST) and core genome MLST(cgMLST) revealing that 3GC-R E. coli isolated from Swiss slaughter calves and fattening pigs were genetically diverse. In this study, it was shown that using WGS alone to monitor antimicrobial resistance could detect trends in known molecular antimicrobial resistance mechanisms while also providing other valuable information about the isolates, such as genetic relatedness. However, only by combining phenotypic susceptibility testing and WGS early detection of previously unknown resistance mechanisms will be possible.

INTRODUCTION: Le séquençage du génome entier (Whole Genome Sequencing, WGS) a été introduit dans la surveillance suisse de la résistance aux antibiotiques en 2022 en tant que méthode supplémentaire aux tests phénotypiques de sensibilité aux antibiotiques pour caractériser les Escherichia coli résistants aux céphalosporines de troisième génération (3GC-R). Des échantillons de cæcum pris en 2021 à l'abattoir de veaux et de porcs suisses, ainsi que de viande de bœuf et de porc provenant de détaillants suisses ont été analysés pour détecter la présence d'E. coli 3GC-R conformément aux protocoles européens harmonisés. En 2021, les E. coli 3GC-R ont été détectés dans 23,8 % des veaux d'abattage, 5,9 % des porcs d'engraissement et 0 % dans la viande. Les résultats de résistance aux antibiotiques obtenus par mesure phénotypique et ceux obtenus par la détection des mécanismes moléculaires sous-jacents concordaient à 99 %. La résistance aux céphalosporines de troisième génération était principalement associée à la pré-sence de blaCTX-M-15 dans les isolats d'E. coli provenant de veaux et de blaCTX-M-1 dans les isolats d'E. coli provenant de porcs et à des mutations dans le promoteur ampC (g.-42 C>T) dans les isolats d'E. coli provenant des deux espèces animales. Les données WGS ont également été utilisées pour une analyse phylogénétique basée sur les types de séquences multilocus (MLST) et MLST du génome de base (cgMLST) révélant que les E. coli 3GC-R isolés des veaux et des porcs suisses étaient génétiquement divers. Dans cette étude, il a été démontré que l'utilisation du WGS seul pour surveiller la résistance aux antibiotiques pouvait détecter des tendances dans les mécanismes moléculaires connus de la résistance aux antibiotiques tout en fournissant d'autres informations précieuses sur les isolats, comme la parenté génétique. Cependant, ce n'est qu'en combinant les tests de sensibilité phénotypique avec le WGS que la détection pré-coce de mécanismes de résistance inconnus sera possible.

Prevalence and characterization of -methicillin-resistant Macrococcus spp. in food producing animals and meat in Switzerland in 2019.

INTRODUCTION: The prevalence of methicillin-resistant Macrococcus spp. in calves and pigs at slaughterhouses and in retail beef and pork meat was determined using samples taken in 2019 within the framework of the national monitoring of methicillin-resistant Staphylococcus aureus in food producing animals in Switzerland. The isolates were submitted to antimicrobial susceptibility testing of 19 antibiotics and to molecular techniques (e.g. PCR, microarray, WGS) for the identification of resistance genes, elements containing the methicillin resistance genes mec and sequence type (ST). Methicillin-resistant Macrococcus spp. (M. caseolyticus (n=38), M. bohemicus (n=4) and Macrococcus spp. (n=2)) were isolated in 40 of 299 nasal swabs from calves representing a prevalence of 13,38 % (95 % CI, 9,98 % - 17,70 %), and in four of 303 nasal swabs from pigs [1,32 % (95 % CI, 0,36 % - 3,35 %)]. One of 311 samples of Swiss pork meat contained a Macrococcus sp. [0,32 % (95 % CI, 0,01 % - 1,78 %)], and four of 309 beef meat samples (260 domestic and 49 imported) contained M. caseolyticus [1,29 % (95 % CI, 0,35 % - 3,28 %)]. The M. caseolyticus strains belonged to diverse STs, with ST21 being the most common in both pigs and calves. The mecD gene was located on Macrococcus resistance island mecD (McRImecD) in 42 strains and on staphylococcal cassette chromosome mec (SCCmecD) in three strains, while mecB was found on plasmids in four strains. In addition to resistance to ß-lactams, the strains also exhibited resistance to tetracycline (n=17; tet(L), tet(K), tet(M)), streptomycin (n=13; str, ant(6)-Ia, rpsL mutation [K56R in ribosomal protein S12]), kanamycin (n=10; aac(6')-Ie - aph(2'')-Ia, aph(2')-Ib, aph(2')-Ic, ant(4')-Ia), clindamycin (n=9; erm(B), erm(45)), erythromycin (n=9; erm(B), msr(G), erm(45)), fusidic acid (n=9; fusC) and gentamicin (n=1; aac(6')-Ie - aph(2'')-Ia). This study represents the first national prevalence study of methicillin-resistant Macrococcus spp. in pigs, calves, pork and beef meat in Switzerland and revealed a reservoir of genetically diverse strains carrying several resistance traits.

INTRODUCTION: La prévalence des macrococques résistants à la méthicilline chez les veaux et les porcs à l'abattoir et dans la viande de bœuf et de porc au détail a été déterminée à partir d'échantillons prélevés en 2019 dans le cadre du monitoring national des Staphylococcus aureus résistants à la méthicilline chez les animaux de rente en Suisse. Les isolats ont été soumis à des tests de sensibilité à 19 antibiotiques et à des techniques moléculaires (par exemple PCR, microarray, WGS) pour l'identification des gènes de résistance, des éléments contenant les gènes mec responsible de la résistance à la méthicilline, ainsi que du type de séquence (ST). Des macocoques résistants à la méthicilline (M. caseolyticus (n=38), M. bohemicus (n=4) et Macrococcus spp. (n=2)) ont été isolés dans 40 des 299 écouvillons nasaux de veaux, ce qui représente une prévalence de 13,38 % (IC 95 %, 9,98 % ­ 17,70 %), et dans quatre des 303 écouvillons nasaux de porcs [1,32 % (IC 95 %, 0,36 % ­ 3,35 %)]. Un des 311 échantillons de viande de porc suisse contenait un Macrococcus sp. [0,32 % (IC 95 %, 0,01 % ­ 1,78 %)], et quatre des 309 échantillons de viande de bœuf (260 domestiques et 49 importés) contenaient M. caseolyticus [1,29 % (IC 95 %, 0,35 % ­ 3,28 %)]. Les souches de M. caseolyticus appartenaient à divers ST, le ST21 étant le plus fréquent chez les porcs et les veaux. Le gène mecD a été localisé sur des éléments du chromosome McRImecD dans 42 souches et SCCmecD dans trois souches, tandis que le gène mecB se trouvait sur des plasmides dans quatre souches. En plus de la résistance aux ß-lactamines, les souches étaient également résistantes à la tétracycline (n=17 ; tet(L), tet(K), tet(M)), à la streptomycine (n=13 ; str, ant(6)-Ia, mutation rpsL [K56R dans la protéine ribosomale S12]), à la kanamycine (n=10 ; aac(6')-Ie ­ aph(2'')-Ia, aph(2')-Ib, aph(2')-Ic, ant(4')-Ia), clindamycine (n=9 ; erm(B), erm(45)), érythromycine (n=9 ; erm(B), msr(G), erm(45)), acide fusidique (n=9 ; fusC) et gentamicine (n=1 ; aac(6')-Ie ­ aph(2'')-Ia). Cette étude est la première à déterminer prévalence des Macrococcus spp. résistants à la méthicilline chez les porcs, les veaux, la viande de porc et de bœuf en Suisse et a révélé un réservoir de souches génétiquement diverses et portant plusieurs traits de résistance.

[Laboratory diagnostic examinations as part of «Pathocalf¼ in case of herd problems on Swiss calf rearing and fattening farms]. / Labordiagnostische Untersuchungen im Rahmen von «Pathocalf¼ bei ­Bestandesproblemen auf Schweizer ­Kälberaufzucht und -mastbetrieben.

INTRODUCTION: «PathoCalf¼ represents a project promoted by the Swiss Federal Food Safety and Veterinary Office. It was the aim of this project to use laboratory diagnostic services for farms raising calves (dairy farms, fattening units, beef cow operations) and dealing with a herd health issue in order to obtain an overview about the spectrum of infectious agents and bacterial resistance patterns in Switzerland. From January 2015 to March 2018, the Bovine Health Service and the farm veterinarian straightened out 148 stock problems on 125 farms. For this, samples were collected from 342 animals. In addition, 98 necropsies were performed. The service related to «PathoCalf¼ was utilized most frequently related to stock problems with respiratory disease (40,5%; 60/148), gastrointestinal disease (37,2%, 55/148) and fatalities with unknown causation (8,8%, 13/148). The majority of all investigated animals (71,8%) were younger than 10 weeks of age. In calves suffering from respiratory disease, most frequently Pasteurella (P.) multocida was found (40,6%, 67/165), followed by mycoplasmen (35,8%, 59/165) and Mannheimia (M.) haemolytica (13,3%, 22/165). The proportion of tested P. multocida and M. haemolytica strains being resistant against oxytetracyline was 67,7% (42/62), 21,1% (4/19) for tilmicosine and 11,3% (7/62) for danofloxacine. For calves suffering from gastrointestinal problems, most frequently Rotavirus (57,6%, 19/33), E. coli (47,7%, 74/155) and Campylobacter spp. (27,7%, 43/155) were found. A resistance against tetracycline was evident for 95,0% (19/20) of all isolated E. coli, and for 80,0% (16/20) in respect to sulfonamides and for 57,1% (12/21) against enrofloxacine. For fatalities with unknown causation, a perforated abomasal ulcer was the most important finding (33,3%, 7/21), followed by diarrhea (28,6%, 6/21) and indigestion related to ruminal acidosis (19,1%, 4/21). The results indicate that aetiological investigations facilitate the evaluation of stock problems. The systematic assessment of abiotic risk factors remains, however, indispensable for the factorial diseases most frequently found on farms raising calves.

INTRODUCTION: «PathoCalf¼ est un projet promu par l'Office fédéral suisse de la sécurité alimentaire et des affaires vétérinaires. L'objectif de ce projet était d'utiliser les services de diagnostic de laboratoire pour les exploitations élevant des veaux (fermes laitières, unités d'engraissement, exploitations de vaches à viande) et confrontées à un problème de santé de troupeau afin d'obtenir une vue d'ensemble du spectre des agents infectieux et de la résistance bactérienne en Suisse. De janvier 2015 à mars 2018, le Service Sanitaire Bovin et/ou le vétérinaire d'exploitation ont traité 148 problèmes dans 125 exploitations. Pour cela, des échantillons ont été prélevés sur 342 animaux. De plus, 98 autopsies ont été réalisées. Le service lié à «PathoCalf¼ a été utilisé le plus souvent en raison de problèmes d'exploitation avec une maladie respiratoire (40,5% ; 60/148), une maladie gastro-intestinale (37,2%, 55/148) ou des décès de cause inconnue (8,8%, 13/148). La majorité de tous les animaux étudiés (71,8%) avaient moins de 10 semaines d'âge. Chez les veaux souffrant de maladies respiratoires, c'est le plus souvent Pasteurella (P.) multocida qui a été trouvé (40,6%, 67/165), suivi par les mycoplasmes (35,8%, 59/165) et Mannheimia (M.) haemolytica (13,3%, 22/165). La proportion des souches de P. multocida et de M. haemolytica testées résistantes à l'oxytétracycline était de 67,7% (42/62), de 21,1% (4/19) pour la tilmicosine et de 11,3% (7/62) pour la danofloxacine. Pour les veaux souffrant de troubles gastro-intestinaux, on a trouvé le plus souvent des Rotavirus (57,6%, 19/33), des E. coli (47,7%, 74/155) et des Campylobacter spp. (27,7%, 43/155). Une résistance à la tétracycline était évidente pour 95,0% 50 (19/20) de tous les E. coli isolés, pour 80,0% (16/20) en ce qui concerne les sulfamides et pour 57,1% (12/21) contre l'enrofloxacine. Pour les décès de cause inconnue, un ulcère de la caillette perforé était la cause la plus importante (33,3%, 7/21), suivie de la diarrhée (28,6%, 6/21) et des indigestions liées à une acidose ruminale (19,1%, 4/21). Les résultats indiquent que les investigations étiologiques facilitent l'évaluation des problèmes de d'exploitation. L'évaluation systématique des facteurs de risque abiotiques reste cependant indispensable pour les maladies factorielles les plus fréquemment recensées sur les exploitations élevant des veaux.

[Survey on the use of antibiotics in exotic pets among Swiss veterinarians]. / Umfrage bei Schweizer Tierärzte/-innen über den Einsatz von Antibiotika bei exotischen Heimtieren.

INTRODUCTION: Monitoring programs and guidelines on the use of antibiotics and antibiotic resistance in human and veterinary medicine have been developed worldwide to promote the prudent use of antibiotics in recent years. However, such information on the use of antibiotics in exotic pets is absent. This fact must be taken into account, since the number of exotic pet patients is constantly increasing and the administration of antibiotics is particular challenging due to the diversity of species and the differences in physiology. The present study reports the results of a survey of frequently used antibiotics and the criteria of antibiotic usage in exotic pets (rabbits, rodents, birds and reptiles) among Swiss veterinarians. These data should form the basis for the development of antibiotic usage guidelines in exotic pets. A total of 61 veterinarians returned the questionnaire. The most important decision-making criteria for antibiotic treatment and antibiotic selection were clinical symptoms (55/59, 93% for antibiotic treatment and 40/59, 68% for antibiotic selection), experience (41/59, 69% and 36/59, 61%, respectively) and textbooks (39/59, 66% and 40/59, 68%). The most important decision-making criteria for dosage and duration of therapy were textbooks (59/59, 100%) and experience (31/59, 53%). The use of a microbial culture was used as a decision criterion for both antibiotic treatment and antibiotic selection by 39% of the participants, sensitivity testing was chosen as a decision criterion by 37% for antibiotic treatment and by 46% for antibiotic selection. Fluoroquinolones were used most frequently, 46/56 (82% of the participants) for rabbits, 49/57 (86%) for rodents, 36/37 (97%) for reptiles and 38/46 (83%) for birds. The vast majority of veterinarians (57/58, 98%) would consult a guide for the use of antibiotics. The frequent use of critical antibiotics in exotic pets underscores the need for a guide to the prudent use of antibiotics. The positive influence of such guidelines has already been proven in dogs and cats.

INTRODUCTION: Pour promouvoir une utilisation prudente des antibiotiques, des programmes de surveillance et des lignes directrices sur l'utilisation des antibiotiques et la résistance aux antibiotiques en médecine humaine et vétérinaire ont été élaborés dans le monde entier ces dernières années. Cependant, il n'existe actuellement aucune information de ce type en ce qui concerne les nouveaux animaux de compagnie ainsi que des informations exhaustives sur l'utilisation d'antibiotiques dans la pratique vétérinaire. Ce fait doit être pris en compte, d'autant plus que le nombre d'nouveaux animaux de compagnie est en constante augmentation dans la patientèle et que l'administration d'antibiotiques est un défi particulier en raison de la diversité des espèces et des différences de physiologie. Dans la présente étude, une enquête a été menée pour avoir un aperçu des antibiotiques fréquemment utilisés et de leurs critères d'utilisation chez les nouveaux animaux de compagnie (lapins, rongeurs, oiseaux et reptiles) auprès de vétérinaires suisses. Ces données sont destinées à servir de base à l'élaboration d'un guide sur les antibiotiques pour les animaux exotiques. Au total 61 vétérinaires ont répondu au questionnaire. Les critères de décision les plus importants pour le traitement antibiotique et la sélection d'un antibiotique étaient les symptômes (55/59, 93% pour le traitement antibiotique et 40/59, 68% pour la sélection de l'antibiotique), l'expérience (41/59, 69% et 36/59, 61%, respectivement) et la littérature spécialisée (39/59, 66% et 40/59, 68%). Les critères de décision les plus importants pour e dosage et la durée du traitement étaient la littérature spécialisée (59/59, 100%) et l'expérience (31/59, 53%). L'utilisation d'une culture bactériologique a été donnée par 39% des participants comme critère de décision à la fois pour le traitement antibiotique et le choix de l'antibiotique et un antibiogramme a été choisi par 37% comme critère de décision pour le traitement antibiotique et 46% pour le choix de l'antibiotique. Il a été constaté que les fluoroquinolones sont utilisées le plus fréquemment, chez 46/56 (82% des participants) pour les lapins, 49/57 (86%) pour les rongeurs, 36/37 (97%) pour les reptiles et 38/46 (83%) pour les oiseaux. La grande majorité des vétérinaires (57/58, 98%) a déclaré qu'ils consulteraient un guide pour l'utilisation des antibiotiques. L'utilisation fréquente d'antibiotiques critiques chez les nouveaux animaux de compagnie souligne la nécessité d'un guide sur l'utilisation prudente des antibiotiques. L'influence positive de ces recommandations a déjà été prouvée pour les chiens et les chats.

Prevalence and antimicrobial resistance of opportunistic pathogens associated with bovine respiratory disease isolated from nasopharyngeal swabs of veal calves in Switzerland.

The composition of the bacterial flora in the calf nasopharynx might influence the risk of bovine respiratory disease (BRD). The aims of the present study were, firstly, to investigate the prevalence of bacteria potentially involved in BRD in the nasopharynx of veal calves and to identify associated risk factors for their presence, and, secondly, to provide data on antimicrobial resistance levels in these bacteria. Deep nasopharyngeal swabs were collected from veal calves on 12 Swiss farms over a period of one year by non-random, but systematic sampling for isolation of Pasteurellaceae and Mycoplasma (M.) bovis and dispar. Associations of potential risk factors with occurrence of these bacteria were tested in multivariable mixed logistic regression analyses, based on information gained from extensive questionnaires completed with the farmers. Antimicrobial susceptibility testing was performed for Pasteurellaceae by broth microdilution method to obtain minimal inhibitory concentrations (MIC). Pasteurellaceae, including Pasteurella (P.) multocida, Mannheimia (M.) haemolytica, Bisgaard Taxon 39 and Histophilus (H.) somni, were almost twice as prevalent as M. bovis and dispar in this study. Continuous stocking was a risk factor for the presence of Pasteurellaceae, especially when calves originated from more than six suppliers. In young calves (≤ 91 days), feeding of California Mastitis Test (CMT) positive milk was an additional risk factor for the presence of Pasteurellaceae whereas transport of calves by farmers and livestock traders (as opposed to transport only by farmers) increased the risk in older calves (> 91 days). Risk factors for the presence of M. bovis/dispar were higher number of calves per drinking nipple in young calves, and no access to an outside pen and feeding of CMT positive milk in older calves, respectively. While further research will have to investigate the observed associations in more detail, this suggests that management can play an important role in the prevalence of nasopharyngeal bacteria with a potential subsequent involvement in BRD. Antimicrobial resistance differed between the three bacterial species tested in this study and was highest to oxytetracycline and spectinomycin in P. multocida, oxytetracycline and penicillin in M. haemolytica, and ampicillin and penicillin in H. somni. Only two European VetCAST breakpoints (for florfenicol in P. multocida and M. haemolytica) have been published to date, matching the MIC distribution of the present isolate populations well, in contrast to certain commonly applied American Clinical and Laboratory Institute interpretive criteria. This highlights the potential for further refinement of clinical breakpoints in veterinary medicine.

Associations between antimicrobial treatment modalities and antimicrobial susceptibility in Pasteurellaceae and E. coli isolated from veal calves under field conditions.

Antimicrobial consumption, with bovine respiratory disease as main indication, is higher in the veal calf industry compared to other livestock production branches. The aim of the present study was to investigate possible associations between antimicrobial drug use and resistance in Pasteurellaceae and indicator Escherichia (E.) coli from veal calves under field conditions in a prospective trial. Over a period of one year, nasopharyngeal and rectal swabs were collected from 2587 animals on 12 and 43 farms, respectively. Antimicrobial susceptibility testing was performed on 346 Mannheimia (M.) haemolytica, 1162 Pasteurella (P.) multocida and 2138 E. coli. Drug use was quantified as treatment incidence for each farm based on the used daily dose methodology (TIUDD), separately for group and individual treatments, and for antimicrobial classes. In multivariable mixed logistic regression analyses, risk factors could be identified for reduced susceptibility to certain antimicrobial classes. Group treatment was generally associated with higher rates of not susceptible (NS) M. haemolytica and P. multocida and non-wildtype (non-WT) E. coli. Individual treatment was associated with less NS and non-WT isolates. Age and entry protocol were important confounders with younger animals showing higher rates of NS and non-WT strains. The present findings suggest that, under field conditions, targeted individual treatment of calves can reduce the development of antimicrobial resistance compared to oral group treatment. For the different microorganisms, risk factors for resistance were partially different. This demonstrates that indicator organisms like E. coli do not necessarily reflect the associations observed in respiratory pathogens.

A dangerous hobby? Erysipelothrix rhusiopathiae bacteremia most probably acquired from freshwater aquarium fish handling.

Erysipelothrix rhusiopathiae is a facultative anaerobic Gram-positive rod that occurs widely in nature and is best known in veterinary medicine for causing swine erysipelas. In humans, infections are rare and mainly considered as occupationally acquired zoonosis. A case of E. rhusiopathiae bacteremia most likely associated with home freshwater aquarium handling is reported. The route of transmission was probably a cut with the dorsal fin of a dead pet fish. A short review of clinical presentations, therapeutic considerations and pitfalls of E. rhusiopathiae infections in humans is presented.

Virulence-associated gene pattern of porcine and human Yersinia enterocolitica biotype 4 isolates.

Yersinia enterocolitica 4/O:3 is the most important human pathogenic bioserotype in Europe and the predominant pathogenic bioserotype in slaughter pigs. Although many studies on the virulence of Y. enterocolitica strains have showed a broad spectrum of detectable factors in pigs and humans, an analysis based on a strict comparative approach and serving to verify the virulence capability of porcine Y. enterocolitica as a source for human yersiniosis is lacking. Therefore, in the present study, strains of biotype (BT) 4 isolated from Swiss slaughter pig tonsils and feces and isolates from human clinical cases were compared in terms of their spectrum of virulence-associated genes (yadA, virF, ail, inv, rovA, ymoA, ystA, ystB and myfA). An analysis of the associated antimicrobial susceptibility pattern completed the characterization. All analyzed BT 4 strains showed a nearly similar pattern, comprising the known fundamental virulence-associated genes yadA, virF, ail, inv, rovA, ymoA, ystA and myfA. Only ystB was not detectable among all analyzed isolates. Importantly, neither the source of the isolates (porcine tonsils and feces, humans) nor the serotype (ST) had any influence on the gene pattern. From these findings, it can be concluded that the presence of the full complement of virulence genes necessary for human infection is common among porcine BT 4 strains. Swiss porcine BT 4 strains not only showed antimicrobial susceptibility to chloramphenicol, cefotaxime, ceftazidime, ciprofloxacin, colistin, florfenicol, gentamicin, kanamycin, nalidixic acid, sulfamethoxazole, streptomycin, tetracycline and trimethoprim but also showed 100% antibiotic resistance to ampicillin. The human BT 4 strains revealed comparable results. However, in addition to 100% antibiotic resistance to ampicillin, 2 strains were resistant to chloramphenicol and nalidixic acid. Additionally, 1 of these strains was resistant to sulfamethoxazole. The results demonstrated that Y. enterocolitica BT 4 isolates from porcine tonsils, as well as from feces, show the same virulence-associated gene pattern and antibiotic resistance properties as human isolates from clinical cases, consistent with the etiological role of porcine BT 4 in human yersiniosis. Thus, cross-contamination of carcasses and organs at slaughter with porcine Y. enterocolitica BT 4 strains, either from tonsils or feces, must be prevented to reduce human yersiniosis.

[Lung and pleural lesions before and after implementation of a national eradication program against enzootic pneumonia and actinobacillosis as well as changes of slaughter carcass organs in slaughter pigs in Switzerland]. / Lungen- und Pleuraläsionen vor und nach der Flächensanierung gegen Enzootische Pneumonie und Actinobazillose sowie Organ- und Schlachtkörperveränderungen bei Schlachtschweinen in der Schweiz.

In a representative cross-sectional study during 12 months of the years 2008/2009 in four abattoirs in Switzerland, lung and pleura lesions as well as lesions of slaughter carcasses and organs of 34 706 pigs were studied for frequency and type of macroscopic lesions. Of the 24276 examined pigs, 91.2% of the lungs, 94.4% of the heart and 95.5% of the livers showed no macroscopically visible lesions. Pigs that were produced for a label program had significantly less bronchopneumonia and pneumonia residuals, pleuritis and liver lesions due to echinococcosis. Pigs supervised by the Swiss Pig Health Service (SGD), showed significantly less bronchopneumonia and pneumonia residuals, diffuse pleuritis, pleuritis/pericarditis and milkspots compared to the non-SGD supervised farms. Thanks to the national eradication program for enzootic pneumonia (EP) and actinobacillosis, the health-status of lungs has been considerably improved and the prevalence of pleurisy decreased considerably. The results of this study indicate a good herd health in Swiss pig production.

Genotypes and antibiotic resistance of bovine Campylobacter and their contribution to human campylobacteriosis.

Campylobacter jejuni and Campylobacter coli are the most important bacterial causes of human gastroenteritis. Chicken has been recognized as a major source for human infection, whereas cattle might also contribute to a lesser extent. However, there is a paucity of information available regarding Campylobacter in Swiss cattle and their role for human campylobacteriosis. To gain more information on genotypes and antibiotic resistance of bovine C. jejuni and C. coli and on their contribution to human disease, 97 cattle isolates were analysed. Multilocus sequence typing (MLST) and flaB typing were applied and the gyrA and 23S rRNA genes were screened for point mutations responsible for quinolone and macrolide resistance, respectively. A total of 37 sequence types (STs) and 44 flaB types were identified, including two sequence types and five flaB types not previously described. Most common sequence types were ST21 (21%), ST61 (12%) and ST48 (11%). Only one isolate was macrolide resistant while 31% (n = 30) were quinolone resistant. Source attribution indicated chicken as the main source of human infection with cattle being second. In conclusion, cattle should not be underestimated as a potential source of human campylobacteriosis.

Nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) among Swiss veterinary health care providers: detection of livestock- and healthcare-associated clones.

We screened a total of 340 veterinarians (including general practitioners, small animal practitioners, large animal practitioners, veterinarians working in different veterinary services or industry), and 29 veterinary assistants for nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus pseudintermedius (MRSP) at the 2012 Swiss veterinary annual meeting. MRSA isolates (n = 14) were detected in 3.8 % (95 % CI 2.1 - 6.3 %) of the participants whereas MRSP was not detected. Large animal practitioners were carriers of livestock-associated MRSA (LA-MRSA) ST398-t011-V (n = 2), ST398-t011-IV (n = 4), and ST398-t034-V (n = 1). On the other hand, participants working with small animals harbored human healthcare-associated MRSA (HCA-MRSA) which belonged to epidemic lineages ST225-t003-II (n = 2), ST225-t014-II (n = 1), ST5-t002-II (n = 2), ST5-t283-IV (n = 1), and ST88-t186-IV (n = 1). HCA-MRSA harbored virulence factors such as enterotoxins, ß-hemolysin converting phage and leukocidins. None of the MRSA isolates carried Panton-Valentine leukocidin (PVL). In addition to the methicillin resistance gene mecA, LA-MRSA ST398 isolates generally contained additional antibiotic resistance genes conferring resistance to tetracycline [tet(M) and tet(K)], trimethoprim [dfrK, dfrG], and the aminoglycosides gentamicin and kanamycin [aac(6')-Ie - aph(2')-Ia]. On the other hand, HCA-MRSA ST5 and ST225 mainly contained genes conferring resistance to the macrolide, lincosamide and streptogramin B antibiotics [erm(A)], to spectinomycin [ant(9)-Ia], amikacin and tobramycin [ant(4')-Ia], and to fluoroquinolones [amino acid substitutions in GrlA (S84L) and GyrA (S80F and S81P)]. MRSA carriage may represent an occupational risk and veterinarians should be aware of possible MRSA colonization and potential for developing infection or for transmitting these strains. Professional exposure to animals should be reported upon hospitalization and before medical intervention to allow for preventive measures. Infection prevention measures are also indicated in veterinary medicine to avoid MRSA transmission between humans and animals, and to limit the spread of MRSA both in the community, and to animal and human hospitals.

Antimicrobial susceptibility of gram-positive udder pathogens from bovine mastitis milk in Switzerland.

We evaluated the susceptibility of the gram-positive mastitis pathogens S. aureus, Str. uberis, Str. dysgalactiae, E. faecalis and L. garviae to antibiotics that are of epidemiological interest or are critically important for mastitis therapy and human medicine. Penicillin resistance was found to be most frequent in S. aureus, and nearly 5 % of the Str. uberis strains displayed a decreased susceptibility to this antibiotic. Resistance to aminoglycosides and macrolides was also detected in the strains tested. The detection of methicillin-resistant S. aureus (MRSA) and of a ciprofloxacin-resistant Str. dysgalactiae isolate corroborated the emergence of mastitis pathogens resistant to critically important antibiotics and underscores the importance of susceptibility testing prior to antibiotic therapy. The monitoring of antibiotic susceptibility patterns and antibiogram analyses are strongly recommended for targeted antimicrobial treatment and to avoid the unnecessary use of the latest generation of antibiotics.

Dans cette étude, on étudie la sensibilité des germes gram positifs causant des mammites, tels que S. aureus, Str. uberis, Str. dysgalactiae, E. faecalis und L. garviae, vis à vis d'une part d'antibiotiques à signification thérapeutique et, dautre part, vis à vis d'antibiotiques d'importance épidémiologique et d'antibiotiques de réserve en médecine humaine. On constate le plus souvent des résistances face à la pénicilline chez S. aureus mais environ 5 % des souches de Str. uberis présentaient également une sensibilité réduite à cet antibiotique. On a en outre constaté des résistances face aux aminoglycosides et aux macrolides. L'identification de S.aureus résistants à la méthicilline (MRSA) et d'un Str. dysgalactiae résistant à la ciprofloxacine démontre que les germes de mammites développent des résistances face aux antibiotiques de réserve importants ainsi que l'importance de la réalisation d'un antibiogramme avant un traitement antibiotique. Une surveillance de la situation de résistance et une interprétation des résultats des antibiogrammes sont indispensables pour effectuer un traitement antibactérien ciblé pour éviter l'utilisation inutile d'antibiotiques de nouvelle génération.

Genotypes and antibiotic resistance of Campylobacter coli in fattening pigs.

Campylobacter coli is a food-borne zoonotic pathogen causing human gastroenteritis worldwide. The organism is a commensal in the intestine of many food production animals including fattening pigs. The role of the pig as a potential reservoir for C. coli affecting human either directly or via poultry has hardly been investigated and genetic characterization of porcine strains is needed to address this question. For this aim multilocus sequence typing (MLST) and flaB typing was applied to 256 C. coli isolates from faeces of fattening pig collected during 2009 at different slaughterhouses in Switzerland. In addition genotypic resistances towards macrolides and quinolones based on point mutations in the 23S rRNA and gyrA genes, respectively, were determined. Of the 67 sequence types (STs) obtained by MLST, 37 were found for the first time. flaB typing revealed 46 different types with 14 of them being novel and was useful to further differentiate strains with an identical ST. Quinolone resistance was detected in 33.6% and macrolide resistance was found in 10.6% of isolates. Comparison with 99 C. coli pig isolates from 2001 revealed a significant decrease in antibiotic resistance towards both groups of antibiotics and there was high overlap between genotypes of 2001 and 2009. Little overlap of porcine genotypes was found with 97 C. coli isolates from poultry collected 2008, however, macrolide resistance was significantly higher in pig isolates. In conclusion, C. coli from Swiss pig are heterogeneous containing many novel STs, findings that could reflect the partitioned Swiss pig production with almost no international breed exchange. The antibiotic resistance echoes the use of corresponding drugs in the Swiss livestock production and indicates the efficacy of restrictive application of antibiotics in order to reduce resistances.

Seroprevalence of Leptospira spp. in clinically healthy horses in Switzerland.

A retrospective, cross-sectional study was conducted to determine the leptospiral seroprevalence in clinically healthy horses in Switzerland. A representative sample of 615 horse sera was examined by microscopic agglutination test for the presence of antibodies against 15 Leptospira spp. serovars. In total, 58.5 % (n = 360) of the horses were positive for one or more of the antigens analysed, with 20.3 % of them showing titres >= 400. The most prevalent serovar was Pyrogenes (22.6 %), followed by serovars Canicola (22.1 %) and Australis (19.2 %). Older horses, mares, ponies and animals spending increased time on pasture exhibited significantly higher prevalence rates (p < 0.05). Moreover, the prevalence was higher in summer and autumn (p = 0.003). The high seroprevalence in healthy horses indicates that they are often exposed to or infected with Leptospira spp. without developing signs of disease. Therefore, other laboratory and clinical data should always be taken into consideration when interpreting serological test results for Leptospira spp.

Genotyping of Mycoplasma hyopneumoniae in wild boar lung samples.

Mycoplasma hyopneumoniae is the etiologic agent of enzootic pneumonia (EP), an important cause of disease-associated losses in swine production and a role of wild boar in recurrent infections can be supposed. Genotypes of M. hyopneumoniae from wild boar are unknown but could indicate its role as a potential reservoir. Therefore, 34 lung samples being PCR-positive for M. hyopneumoniae from wild boar from the Geneva region in Switzerland were assayed by genotyping using the p146 and multi-locus sequence typing (MLST) approaches and compared to data from outbreak cases from domestic swine in Switzerland. Successful genotyping was dependent on a sufficiently high concentration of M. hyopneumoniae DNA in the samples as assessed by different real-time PCR assays. The p146 genotyping was more successful with 24 samples (70.5%) being typeable whereas only 6 samples (17.6%) could be genotyped using the MLST approach. Variability of genotypes was high but identical types were found in geographically related animals. Genotypes from wild boar showed phylogenetic relatedness to those from domestic pigs but no matching types could be identified. Results show that direct genotyping from wild boar lung samples is possible and provides a promising approach to investigate future EP outbreak related samples from wild boar.

Comparison of real-time PCR assays for detection, quantification, and differentiation of campylobacter jejuni and campylobacter coli in broiler neck skin samples.

We tested the use of multiplex real-time PCR for detection and quantification of Campylobacter jejuni and Campylobacter coli on broiler carcass neck skin samples collected during 2008 from slaughterhouses in Switzerland. Results from an established TaqMan assay based on two different targets (hipO and ceuE for C. jejuni and C. coli, respectively) were corroborated with data from a newly developed assay based on a single-nucleotide polymorphism in the fusA gene, which allows differentiation between C. jejuni and C. coli. Both multiplex real-time PCRs were applied simultaneously for direct detection, differentiation, and quantification of Campylobacter from 351 neck skin samples and compared with culture methods. There was good correlation in detection and enumeration between real-time PCR results and quantitative culture, with real-time PCR being more sensitive. Overall, 251 (71.5%) of the samples were PCR positive for Campylobacter, with 211 (60.1%) in the hipO-ceuE assays, 244 (69.5%) in the fusA assay, and 204 (58.1%) of them being positive in both PCR assays. Thus, the fusA assay was similarly sensitive to the enrichment culture (72.4% positive); however, it is faster and allows for quantification. In addition, real-time PCR allowed for species differentiation; roughly 60% of positive samples contained C. jejuni, less than 10% C. coli, and more than 30% contained both species. Real-time PCR proved to be a suitable method for direct detection, quantification, and differentiation of Campylobacter from carcasses, and could permit time-efficient surveillance of these zoonotic agents.

Organization of the equine immunoglobulin heavy chain constant region genes; III. Alignment of c mu, c gamma, c epsilon and c alpha genes.

Previous restriction analysis of cloned equine DNA and genomic DNA of equine peripheral blood mononuclear cells had indicated the existence of one c epsilon, one c alpha and up to six c gamma genes in the haploid equine genome. The c epsilon and c alpha genes have been aligned on a 30 kb DNA fragment in the order 5' c epsilon-c alpha 3'. Here we describe the alignment of the equine c mu and c gamma genes by deletion analysis of one IgM, four IgG and two equine light chain expressing heterohybridomas. This analysis establishes the existence of six c gamma genes per haploid genome. The genomic alignment of the cH-genes is 5' c mu/(/) c gamma 1/(/) c gamma 2/(/) c gamma 3/(/) c gamma 4/(/) c gamma 5/(/) c gamma 6/(/) c epsilon-c alpha 3', naming the c gamma genes according to their position relative to c mu. For three of the c gamma genes the corresponding IgG isotypes could be identified as IgGa for c gamma 1, IgG(T) for c gamma 3 and IgGb for c gamma 4.

Organisation of the equine immunoglobulin constant heavy chain genes. II. Equine cgamma genes.

The number of immunoglobulin G constant heavy chain genes (cgamma genes) varies broadly among mammalian species, reflecting structural and functional differences between expressed immunoglobulin G (IgG) isotypes and allotypes. Up to now equine IgG isotypes have been defined only at the biochemical and serological level. It is still not clear how many IgG isotypes exist in horses and whether there are any allotypes. Here, we describe the isolation and characterisation of equine cgamma genes. An equine genomic lambda phage library was screened with a human cgamma4 probe. Cross-hybridising equine cgamma sequences were cloned twice and characterised by restriction mapping with the human cgamma4 and a murine sgamma1 probe. Genomic equine DNA probes for both, cgamma genes and corresponding switch regions (sgamma), were isolated and used for a more detailed BamHI restriction analysis, comparing genomic DNA of various horses. This analysis reveals the existence of at least five, or probably six cgamma genes in the equine haploid genome. Beside the porcine system, this is the highest number of cgamma genes described for any mammalian species. Moreover, for two of these cgamma genes, BamHI restriction fragment length polymorphism became evident.