75 results on '"Brodard I"'
Search Results
2. Discovery of insertion element ISCfe1: a new tool for Campylobacter fetus subspecies differentiation
- Author
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Abril, C., Vilei, E.M., Brodard, I., Burnens, A., Frey, J., and Miserez, R.
- Published
- 2007
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3. Impact of the early-life skin microbiota on the development of canine atopic dermatitis in a high-risk breed birth cohort
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Rodriguez-Campos, S, Rostaher, Ana; https://orcid.org/0000-0002-1940-0341, Zwickl, L, Fischer, Nina M; https://orcid.org/0000-0002-0142-226X, Brodard, I, Vidal, S, Brandt, B W, Favrot, Claude; https://orcid.org/0000-0002-0821-0956, Perreten, Vincent; https://orcid.org/0000-0001-5722-9445, Rodriguez-Campos, S, Rostaher, Ana; https://orcid.org/0000-0002-1940-0341, Zwickl, L, Fischer, Nina M; https://orcid.org/0000-0002-0142-226X, Brodard, I, Vidal, S, Brandt, B W, Favrot, Claude; https://orcid.org/0000-0002-0821-0956, and Perreten, Vincent; https://orcid.org/0000-0001-5722-9445
- Abstract
Canine atopic dermatitis (CAD) is a prevalent inflammatory skin disease of dogs worldwide. Certain breeds such as the West Highland White Terriers (WHWT) are predisposed to suffer from CAD. Microbial dysbiosis is known to play a significant role in the pathogenesis of the disease, which is similar to its human counterpart, atopic dermatitis (AD). To date, no large cohort-study has been conducted in a predisposed dog breed to study the impact of the early-life microbiota on the development of CAD, as well as the possible implication of factors such as hygiene and access to the outdoors. In this study skin samples of 143 WHWT, including 109 puppies up to three weeks old and 34 parent dogs, from 17 breeders, were subjected to 16S rRNA gene and ITS2 amplicon sequencing to disclose the bacterial and fungal oral and skin microbiota, respectively. The oral samples served as a control group to confirm differences between haired and mucosal surfaces. The cutaneous microbiota differed between sample sites and age of the dogs. The season of sampling, geographical origin as well as hygiene status of the household and the access to the outdoors shaped the skin microbiota of the puppies significantly. However, we found that the individual early-life microbiota did not predispose for the later development of CAD.
- Published
- 2020
4. Detection of specific Treponema species and Dichelobacter nodosus from digital dermatitis (Mortellaro’s disease) lesions in Swiss cattle
- Author
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Alsaaod, M, primary, Locher, I, additional, Jores, J, additional, Grimm, P, additional, Brodard, I, additional, Steiner, A, additional, and Kuhnert, P, additional
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- 2019
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5. Bacterial, fungal, parasitological and pathological analyses of abortions in small ruminants from 2012-2016
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Schnydrig, P, primary, Vidal, S, additional, Brodard, I, additional, Frey, C, additional, Posthaus, H, additional, Perreten, V, additional, and Rodriguez-Campos, S, additional
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- 2017
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6. Virulence-associated gene pattern of porcine and human Yersinia enterocolitica biotype 4 isolates
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Schneeberger, M., primary, Brodard, I., additional, and Overesch, G., additional
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- 2015
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7. Nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) among Swiss veterinary health care providers: Detection of livestock- and healthcare-associated clones
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Wettstein Rosenkranz, K., primary, Rothenanger, E., additional, Brodard, I., additional, Collaud, A., additional, Overesch, G., additional, Bigler, B., additional, Marschall, J., additional, and Perreten, V., additional
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- 2014
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8. Real-time multiplex PCR assays for reliable detection of Clostridium perfringens toxin genes in animal isolates
- Author
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Albini, S., Brodard, I., Jaussi, A., Wollschlaeger, N., Frey, J., Miserez, R., and Abril, C.
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- 2008
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9. Campylobacter fetus subspecies venerealis transport medium for enrichment and PCR
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Harwood, L. J., primary, Thomann, A., additional, Brodard, I., additional, Perreten, V., additional, and Makaya, P. V., additional
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- 2009
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10. Vorkommen von Clostridium perfringens Typ A und Typ C bei Saugferkeln in der schweizerischen Schweinepopulation
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Wollschläger, N., primary, Zimmermann, W., additional, Brodard, I., additional, Albini, S., additional, Doherr, M., additional, Posthaus, H., additional, and Miserez, R., additional
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- 2009
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11. Novel identification and differentiation of Brucella melitensis, B. abortus, B. suis, B. ovis, B. canis, and B. neotomae suitable for both conventional and real-time PCR systems
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Hinić, V., primary, Brodard, I., additional, Thomann, A., additional, Cvetnić, Ž., additional, Makaya, P.V., additional, Frey, J., additional, and Abril, C., additional
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- 2008
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12. Antimicrobial resistance profile of Actinobacillus pleuropneumoniae and Actinobacillus porcitonsillarum
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MATTER, D, primary, ROSSANO, A, additional, LIMAT, S, additional, VORLETFAWER, L, additional, BRODARD, I, additional, and PERRETEN, V, additional
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- 2007
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13. IS711-based real-time PCR assay as a tool for detection of Brucella spp. in wild boars and comparison with bacterial isolation and serology
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Miserez Raymond, Holub Milena, Thomann Andreas, Brodard Isabelle, Hinić Vladimira, and Abril Carlos
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Veterinary medicine ,SF600-1100 - Abstract
Abstract Background Control of brucellosis in livestock, wildlife and humans depends on the reliability of the methods used for detection and identification of bacteria. In the present study, we describe the evaluation of the recently established real-time PCR assay based on the Brucella-specific insertion sequence IS711 with blood samples from 199 wild boars (first group of animals) and tissue samples from 53 wild boars (second group of animals) collected in Switzerland. Results from IS711 real-time PCR were compared to those obtained by bacterial isolation, Rose Bengal Test (RBT), competitive ELISA (c-ELISA) and indirect ELISA (i-ELISA). Results In the first group of animals, IS711 real-time PCR detected infection in 11.1% (16/144) of wild boars that were serologically negative. Serological tests showed different sensitivities [RBT 15.6%, c-ELISA 7.5% and i-ELISA 5.5%] and only 2% of blood samples were positive with all three tests, which makes interpretation of the serological results very difficult. Regarding the second group of animals, the IS711 real-time PCR detected infection in 26% of animals, while Brucella spp. could be isolated from tissues of only 9.4% of the animals. Conclusion The results presented here indicate that IS711 real-time PCR assay is a specific and sensitive tool for detection of Brucella spp. infections in wild boars. For this reason, we propose the employment of IS711 real-time PCR as a complementary tool in brucellosis screening programs and for confirmation of diagnosis in doubtful cases.
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- 2009
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14. Resistance of zebu cattle (Bos indicus) to colonization by major ruminant hoof pathogens.
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Kuhnert P, Loosli N, Brodard I, Lindtke D, and Jores J
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- Animals, Cattle, Switzerland epidemiology, Treponema genetics, Treponema isolation & purification, Treponema classification, Foot Diseases veterinary, Foot Diseases microbiology, Prevalence, Disease Resistance, Fusobacterium Infections veterinary, Fusobacterium Infections microbiology, Cattle Diseases microbiology, Hoof and Claw microbiology, Dichelobacter nodosus genetics, Dichelobacter nodosus pathogenicity, Fusobacterium necrophorum genetics, Fusobacterium necrophorum pathogenicity, Fusobacterium necrophorum isolation & purification
- Abstract
Zebu cattle (Bos indicus) is reported to be more resistant towards harmful environmental factors than taurine cattle (Bos taurus). A few hundred zebu cattle are kept in Switzerland and in contrast to the Swiss indigenous breeds, infectious hoof disease in zebu is not observed. Therefore, we compared the prevalence of three ruminant hoof pathogens in zebu and taurine cattle. These included Treponema spp., Fusobacterium necrophorum and Dichelobacter nodosus which are associated with bovine digital dermatitis (BDD), different bovine hoof diseases and ovine footrot, respectively. Interdigital swabs and punch biopsies from hind feet of slaughter animals were tested for the three pathogens by PCR. Sixty zebu from eight farms were compared to a convenience sample of 20 taurine cattle from 17 farms. Treponema spp. associated with BDD were not detected in zebu while 23 % of animals and 50 % of farms were positive for benign D. nodosus, with results indicating environmental contamination rather than colonization. Taurine cattle showed 35 % of animals and 41 % of farms positive for T. phagedenis while 90 % of animals and 94 % of farms were colonized by D. nodosus as indicated by a 500-fold higher bacterial load than in zebu. The difference in prevalence of the two pathogens between zebu and taurine cattle was highly significant. F. necrophorum was as well only detected in taurine cattle with values of 15 % of animals and 17.7 % of farms, being significantly different at the animal level. Furthermore, genetic analysis of Swiss zebu indicates high genomic diversity and clear separation from taurine cattle. This is the first evidence that zebu show resistance towards colonization by bacterial hoof pathogens in contrast to taurine cattle., Competing Interests: Declaration of Competing Interest None, (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2024
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15. Zoonotic bacterial and parasitic intestinal pathogens in foxes, raccoons and other predators from eastern Germany.
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Kittl S, Frey CF, Brodard I, Scalisi N, Vargas Amado ME, Thomann A, Schierack P, and Jores J
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- Animals, Germany, Zoonoses microbiology, Zoonoses parasitology, Whole Genome Sequencing, Foxes microbiology, Foxes parasitology, Raccoons microbiology, Raccoons parasitology, Feces microbiology, Feces parasitology, Bacteria genetics, Bacteria classification, Bacteria isolation & purification, Phylogeny
- Abstract
In this study, we investigated faecal specimens from legally hunted and road-killed red foxes, raccoons, raccoon dogs, badgers and martens in Germany for parasites and selected zoonotic bacteria. We found that Baylisascaris procyonis, a zoonotic parasite of raccoons, had spread to northeastern Germany, an area previously presumed to be free of this parasite. We detected various pathogenic bacterial species from the genera Listeria, Clostridium (including baratii), Yersinia and Salmonella, which were analysed using whole-genome sequencing. One isolate of Yersinia enterocolitica contained a virulence plasmid. The Salmonella Cholerasuis isolate encoded an aminoglycoside resistance gene and a parC point mutation, conferring resistance to ciprofloxacin. We also found tetracycline resistance genes in Paeniclostridium sordellii and Clostridium baratii. Phylogenetic analyses revealed that the isolates were polyclonal, indicating the absence of specific wildlife-adapted clones. Predators, which scavenge from various sources including human settlements, acquire and spread zoonotic pathogens. Therefore, their role should not be overlooked in the One Health context., (© 2024 The Authors. Environmental Microbiology Reports published by John Wiley & Sons Ltd.)
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- 2024
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16. Novel tetracycline resistance gene tet(65) located on a multi-resistance Corynebacterium plasmid.
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Kittl S, Brodard I, Tresch M, and Perreten V
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- Animals, Drug Resistance, Multiple, Bacterial genetics, Phylogeny, Dogs, Tetracycline pharmacology, Cloning, Molecular, Corynebacterium Infections microbiology, Dog Diseases microbiology, Plasmids genetics, Tetracycline Resistance genetics, Microbial Sensitivity Tests, Anti-Bacterial Agents pharmacology, Escherichia coli genetics, Escherichia coli drug effects, Corynebacterium genetics, Corynebacterium drug effects
- Abstract
Background: Corynebacterium (C.) sp. 22KM0430 related to C. oculi and isolated from a dog exhibited resistance to tetracycline, and its WGS analysis revealed a putative resistance gene on a 35 562-bp plasmid also harbouring the MLSB resistance gene erm(X)., Objectives: To characterize the novel tetracycline resistance gene tet(65) and demonstrate its functionality by expression in C. glutamicum and Escherichia coli and plasmid curing of the host strain., Methods: tet(65) was cloned with and without its repressor tetR(65) and expressed in C. glutamicum DSM20300 and E. coli DH5α. Plasmid was cured by non-selective passages. Minimal inhibitory concentrations (MICs) of tetracyclines were determined according to CLSI guidelines. Association of tet(65) with efflux was shown by the addition of reserpine to MIC assays. Phylogenetic position and transmembrane structure of Tet(65) were analysed using MEGA11 and DeepTMHMM., Results: Tet(65) shows 73% amino acid identity with the closest related Tet(Z), contains 12 transmembrane domains and is structurally related to the Major Facilitator Superfamily. The tetracycline MICs decreased in the plasmid-cured strain and increased when tet(65) was expressed in C. glutamicum and in E. coli. The MICs of tetracycline decreased in the presence of reserpine indicating that tet(65) functions as an efflux pump. A GenBank search also identified tet(65) in C. diphtheriae and Brevibacterium (B.) casei and B. luteolum., Conclusions: A novel tetracycline efflux gene tet(65) was identified in a C. oculi related species and was also present in the human pathogen C. diphtheriae and in Brevibacterium species indicating broader potential for dissemination., (© The Author(s) 2024. Published by Oxford University Press on behalf of British Society for Antimicrobial Chemotherapy.)
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- 2024
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17. Canine Staphylococcaceae circulating in a Kenyan animal shelter.
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Akarsu H, Liljander AM, Lacasta A, Ssajjakambwe P, Brodard I, Cherbuin JDR, Torres-Puig S, Perreten V, Kuhnert P, Labroussaa F, and Jores J
- Subjects
- Animals, Dogs, Humans, Staphylococcus aureus genetics, Kenya, Staphylococcaceae, Phylogeny, Anti-Bacterial Agents pharmacology, Microbial Sensitivity Tests, Staphylococcal Infections microbiology, Dog Diseases microbiology
- Abstract
Animal shelters, especially in resource-poor countries, bring together pets from different regions and with different backgrounds. The crowding of such animals often results in infectious diseases, such as respiratory infections. This study characterized Staphylococcaceae from diseased and apparently healthy dogs housed in an animal shelter in Kenya, to determine their antibiotic resistance profiles, their genetic relatedness, and the presence of dominant clones. Therefore, bacteria were collected from all 167 dogs present in the shelter in June 2015 and screened for Staphylococcaceae using standard cultivation techniques. In all, 92 strains were isolated from 85 dogs and subsequently sequenced by PacBio long-read sequencing. Strains encompassed nine validated species, while S. aureus ( n = 47), S. pseudintermedius ( n = 21), and Mammaliicoccus (M.) sciuri ( n = 16) were the three most dominant species. Two S . aureus clones of ST15 (CC15) and ST1292 (CC1) were isolated from 7 and 37 dogs, respectively. All 92 strains isolated were tested for their antimicrobial susceptibility by determining the minimum inhibitory concentrations. In all, 86 strains had resistance-associated minimal inhibitory concentrations to at least one of the following antimicrobials: tetracycline, benzylpenicillin, oxacillin, erythromycin, clindamycin, trimethoprim, kanamycin/gentamicin, or streptomycin. Many virulence-encoding genes were detected in the S. aureus strains, other Staphylococcaceae contained a different set of homologs of such genes. The presence of mobile genetic elements, such as plasmids and prophages, known to facilitate the dissemination of virulence- and resistance-encoding genes, was also assessed. The unsuspected high presence of two S . aureus clones in about 50% of dogs suggests dissemination within the shelter and a human source.IMPORTANCEMicrobiological data from sub-Saharan Africa are scarce compared to data from North America, Europe, or Asia, and data derived from dogs, the man's best friend, kept in sub-Saharan Africa are largely missing. This work presents data on Staphylococcaceae mainly isolated from the nasal cavity of dogs stationed at a Kenyan shelter in 2015. We characterized 92 strains isolated from 85 dogs, diseased and apparently healthy ones. The strains isolated covered nine validated species and we determined their phenotypic resistance and characterized their complete genomes. Interestingly, Staphylococcus aureus of two predominant genetic lineages, likely to be acquired from humans, colonized many dogs. We also detected 15 novel sequence types of Mammaliicoccus sciuri and S. pseudintermedius indicating sub-Saharan-specific phylogenetic lineages. The data presented are baseline data that guide antimicrobial treatment for dogs in the region., Competing Interests: The authors declare no conflict of interest.
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- 2024
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18. Serological and molecular detection as well as typing of Leptospira spp. in foxes, raccoons, and other wild carnivores in North-Eastern Germany, 2021-2022.
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Kuhnert P, Brodard I, Ackermann S, Schierack P, and Jores J
- Abstract
Leptospirosis is a worldwide zoonosis caused by pathogenic Leptospira spp. While the latter are reported from various mammal hosts such as humans, dogs, or rodents, less is known about their presence in wild carnivores. We therefore investigated the presence of Leptospira spp. in foxes, raccoons, badgers, raccoon dogs, and martens in North-Eastern Germany. Kidney, urine, and blood specimens obtained from legally hunted or road-killed animals were tested by real-time PCR and by serogroup specific antibody detection for the presence of Leptospira spp. Additionally, kidney and urine specimens were tested by real-time PCR for the presence of Brucella spp. and Francisella tularensis , with all being negative for these two zoonotic pathogens. Leptospira spp. were detected by PCR in 12.6 % (n = 21/166) and serologically in 26.2 % (n = 53/202) of tissue and serum samples, respectively. Antibodies to 15 different serogroups were identified with Javanica (n = 25) and Bataviae (n = 12) being predominant. A high sero-prevalence of 34.0 % and 18.6 % in foxes and raccoons, respectively, and the presence of ST17 associated with human and animal leptospirosis indicates a reservoir and the zoonotic potential of these wild animals., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)
- Published
- 2023
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19. Field validation of an antibiotic-free hoof spray to effectively treat ovine footrot by eliminating virulent Dichelobacter nodosus.
- Author
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Loosli N, Brodard I, Kittl S, Luyet C, and Kuhnert P
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- Sheep, Animals, Dichelobacter nodosus, Foot Rot prevention & control, Gram-Negative Bacterial Infections drug therapy, Gram-Negative Bacterial Infections prevention & control, Gram-Negative Bacterial Infections veterinary, Hoof and Claw, Sheep Diseases drug therapy, Sheep Diseases prevention & control
- Abstract
Ovine footrot caused by Dichelobacter nodosus is a highly contagious hoof disease negatively impacting animal welfare and causing major economic losses to the sheep industry. Bactericidal footbaths have shown to be an efficient treatment option and will be used in the national footrot control program in Switzerland. However, the application of footbaths is laborious and economically not sound for small flock holders. We therefore tested in a field study the Intra Repiderma spray for its applicability and efficacy to treat ovine footrot. Ten independent flocks fulfilling defined parameters (e.g. clinical signs, positive for D. nodosus, flock size) could be identified and were included in the study. Farms were visited weekly to fortnightly and clinical scores and swabs for D. nodosus real-time (rt)PCR were taken. Treatment with the Intra Repiderma spray was started after initial claw trimming at the very first visit and was carried out three times within a week. Clearly visible clinical improvement was evident after one week of treatment. Virulent D. nodosus amounts on feet declined constantly during treatment which was continued until all sheep of a flock tested rtPCR-negative (1-10 weeks). Results indicate that a highly effective improvement of clinical signs and complete elimination of virulent D. nodosus can be achieved with the spray treatment. Therefore, it is a valuable alternative to cumbersome footbaths especially for small flocks. A sustainable control of footrot and its pathogen in a successfully treated flock can be maintained by strict biosecurity measures and continued treatment as far as necessary., Competing Interests: Declaration of Competing Interest none., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2023
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20. Corynebacterium oculi-related bacterium may act as a pathogen and carrier of antimicrobial resistance genes in dogs: a case report.
- Author
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Tresch M, Watté C, Stengard M, Ritter C, Brodard I, Feyer S, Gohl E, Akdesir E, Perreten V, and Kittl S
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- Animals, Dogs, Anti-Bacterial Agents pharmacology, Corynebacterium genetics, Drug Resistance, Bacterial genetics, Microbial Sensitivity Tests veterinary, Corynebacterium Infections veterinary, Corynebacterium Infections microbiology, Dog Diseases
- Abstract
Background: The genus Corynebacterium comprises well-known animal and human pathogens as well as commensals of skin and mucous membranes. Species formerly regarded as contaminants are increasingly being recognized as opportunistic pathogens. Corynebacterium oculi has recently been described as a human ocular pathogen but has so far not been reported in dogs., Case Presentation: Here we present two cases of infection with a novel Corynebacterium sp., a corneal ulcer and a case of bacteriuria. The two bacterial isolates could not be identified by MALDI-TOF MS. While 16 S rRNA gene (99.3% similarity) and rpoB (96.6% identity) sequencing led to the preliminary identification of the isolates as Corynebacterium (C.) oculi, whole genome sequencing revealed the strains to be closely related to, but in a separate cluster from C. oculi. Antimicrobial susceptibility testing showed high minimal inhibitory concentrations of lincosamides, macrolides, tetracycline, and fluoroquinolones for one of the isolates, which also contained an erm(X) and tet-carrying plasmid as well as a nonsynonymous mutation leading to an S84I substitution in the quinolone resistance determining region of GyrA., Conclusions: While the clinical signs of both dogs were alleviated by antimicrobial treatment, the clinical significance of these isolates remains to be proven. However, considering its close relation with C. oculi, a known pathogen in humans, pathogenic potential of this species is not unlikely. Furthermore, these bacteria may act as reservoir for antimicrobial resistance genes also in a One Health context since one strain carried a multidrug resistance plasmid related to pNG3 of C. diphtheriae., (© 2023. The Author(s).)
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- 2023
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21. Genomic Characterization and Antimicrobial Susceptibility of Dromedary-Associated Staphylococcaceae from the Horn of Africa.
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Akarsu H, Liljander A, Younan M, Brodard I, Overesch G, Glücks I, Labroussaa F, Kuhnert P, Perreten V, Monecke S, Drexler JF, Corman VM, Falquet L, and Jores J
- Subjects
- Animals, Cattle, Humans, Camelus, Staphylococcus aureus, Staphylococcaceae, Microbial Sensitivity Tests, Staphylococcus, Anti-Bacterial Agents pharmacology, Genomics, Kenya, Staphylococcal Infections veterinary, Methicillin-Resistant Staphylococcus aureus genetics
- Abstract
Members of the Staphylococcaceae family, particularly those of the genus Staphylococcus, encompass important human and animal pathogens. We collected and characterized Staphylococcaceae strains from apparently healthy and diseased camels ( n = 84) and cattle ( n = 7) in Somalia and Kenya. We phenotypically characterized the strains, including their antimicrobial inhibitory concentrations. Then, we sequenced their genomes using long-read sequencing, closed their genomes, and subsequently compared and mapped their virulence- and resistance-associated gene pools. Genome-based phylogenetics revealed 13 known Staphylococcaceae and at least two novel species. East African strains of different species encompassed novel sequence types and phylogenetically distant clades. About one-third of the strains had non-wild-type MICs. They were resistant to at least one of the following antimicrobials: tetracycline, benzylpenicillin, oxacillin, erythromycin, clindamycin, trimethoprim, gentamicin, or streptomycin, encoded by tet (K), blaZ / bla
ARL , mecA / mecA1 , msrA / mphC , salA , dfrG , aacA-aphD , and str , respectively. We identified the first methicillin- and multidrug-resistant camel S. epidermidis strain of sequence type (ST) 1136 in East Africa. The pool of virulence-encoding genes was largest in the S. aureus strains, as expected, although other rather commensal strains contained distinct virulence-encoding genes. We identified toxin-antitoxin (TA) systems such as the hicA/hicB and abiEii/abiEi families, reported here for the first time for certain species of Staphylococcaceae . All strains contained at least one intact prophage sequence, mainly belonging to the Siphoviridae family. We pinpointed potential horizontal gene transfers between camel and cattle strains and also across distinct Staphylococcaceae clades and species. IMPORTANCE Camels are a high value and crucial livestock species in arid and semiarid regions of Africa and gain importance giving the impact of climate change on traditional livestock species. Our current knowledge with respect to Staphylococcaceae infecting camels is very limited compared to that for other livestock species. Better knowledge will foster the development of specific diagnostic assays, guide promising antimicrobial treatment options, and inform about potential zoonotic risks. We characterized 84 Staphylococcaceae strains isolated from camels with respect to their antimicrobial resistance and virulence traits. We detected potentially novel Staphylococcus species, resistances to different classes of antimicrobials, and the first camel multidrug-resistant S. epidermidis strain of sequence type 1136.- Published
- 2022
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22. Corynebacterium uberis sp. nov. frequently isolated from bovine mastitis.
- Author
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Kittl S, Studer E, Brodard I, Thomann A, and Jores J
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- Animals, Bacterial Typing Techniques, Cattle, Corynebacterium, DNA, Bacterial genetics, Fatty Acids, Female, Nucleic Acid Hybridization, Phylogeny, RNA, Ribosomal, 16S genetics, Sequence Analysis, DNA, Mastitis, Bovine
- Abstract
Several strains belonging to the genus Corynebacterium, but not to any described species of the genus were isolated from bovine mastitic milk samples over the past five years in the diagnostic unit of the University of Bern. Six of these strains (18M0132
T , 17M2518, 18M0913, 19M0083, 20M1046 and 20M1090) that were phenotypically similar were further characterized genotypically. Gram-positive coryneform rods were catalase positive, facultative anaerobe and CAMP-test negative. Whole genome sequencing and subsequent phylogenetic analysis revealed their genome size to be 2.53 Mb and their G + C content to be between 65.4 and 65.5 mol%. Digital DNA-DNA hybridisation (dDDH) showed the highest similarity of only less than 20% with Corynebacterium mastitidis and Corynebacterium frankenforstense, which indicated that the isolates belong to an undescribed Corynebacterium species. This was confirmed by studying the average nucleotide identity (ANI) where the accepted species boundary is around 95% and which ranged between 70.3% and 74.9% with the most closely related species C. mastitidis. We established MALDI-TOF fingerprints of the species, which allows a clear separation from related species and can be used by other laboratories for diagnostic purposes. Based on our analyses we conclude that the selected strains belong to a previously undescribed species and propose the name Corynebacterium uberis sp. nov. The proposed type strain is 18M0132T (=DSM 111922T , = CCOS 1972T )., (Copyright © 2022 The Author(s). Published by Elsevier GmbH.. All rights reserved.)- Published
- 2022
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23. Wielerella bovis gen. nov., sp. nov. a member of the family Neisseriaceae associated with bovine endocarditis.
- Author
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Kuhnert P, Brodard I, Bock S, Hemphill A, Akarsu H, Engelhardt A, and Kutzer P
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- Animals, Bacterial Typing Techniques, Base Composition, Cattle, DNA, Bacterial genetics, Fatty Acids chemistry, Phospholipids chemistry, Phylogeny, RNA, Ribosomal, 16S genetics, Sequence Analysis, DNA, Endocarditis, Neisseriaceae
- Abstract
Seven bacterial strains isolated from bovine endocarditis in six animals from different geographic regions were investigated in a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA gene sequences placed all seven isolates on a distinct, monophyletic cluster in the family Neisseriaceae with closest similarity to type strains of Alysiella filiformis (97.06 %) and Kingella kingae (96.34 %). Whole genome sequence analysis of isolates confirmed their species status, with an average nucleotide identity >96 % between isolates and <80 % to other type species of genera of Neisseriaceae while digital DNA-DNA hybridization values were >80 % and<18 %, respectively. The DNA G+C content was 42.5-43.0 mol%. Whole genome sequence based phylogeny showed the isolates being monophyletic and separated from established genera, thereby forming a new genus within the family Neisseriaceae . Similarly, analysis of MALDI-TOF MS reference spectra clustered the isolates close together and clearly separated from other genera, making this the method of choice for identification. Biochemical markers based on classical as well as commercial identification schemes allowed separation from closely related Neisseriaceae genera, even though the new taxon is biochemically not very active. Major fatty acids are C
12 : 0 , C14 : 0 and C16 : 0 . The major quinone is ubiquinone Q-8. In the polar lipid profile, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phospholipid were predominant. We propose the novel genus Wielerella with the type species Wielerella bovis gen. nov., sp. nov. The type strain is CCUG 44465T (=DSM 113289T =JF 2483T ) isolated post mortem from a cow with endocarditis in Switzerland.- Published
- 2022
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24. Detection of treponemes in digital dermatitis lesions of captive European bison (Bison bonasus).
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Hoby S, Jensen TK, Brodard I, Gurtner C, Eicher R, Steiner A, Kuhnert P, and Alsaaod M
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- Animals, Female, Male, In Situ Hybridization, Fluorescence, Treponema genetics, Treponema isolation & purification, Bison microbiology, Digital Dermatitis microbiology, Digital Dermatitis pathology, Treponemal Infections veterinary, Treponemal Infections microbiology, Treponemal Infections pathology
- Abstract
A newly-discovered foot disease of unknown origin in captive European Bison (Bison bonasus) was recently detected at Berne Animal Park. Dermatitis of the interdigital cleft of varying degrees of severity was diagnosed in all animals (n = 10). The aim of this study was to describe the gross and histological lesions of the interdigital cleft found in 10 captive European bison and to identify involved potential pathogens in affected feet using molecular-based methods for Treponema spp., Dichelobacter nodosus and Fusobacterium necrophorum. Lesions were scored according to the degree of gross pathology at limb level. In a single animal, the gross lesions were restricted to focal lesions on the dorsal aspect of the digital skin of each foot (score 1), whereas all other animals showed at least one foot with extended lesions including the interdigital cleft (score 2). The presence of viable spirochaetes was observed in all animals using dark field microscopy. Applying fluorescence in situ hybridisation (FISH) on biopsies, Treponema spp. were identified, infiltrating the skin lesions in varying numbers in nine animals. Nested PCRs for Treponema medium, Treponema phagedenis and Treponema pedis of swab samples showed three positive animals out of ten for the latter two, whereas pooled biopsy samples were positive in all ten animals for at least T. phagedenis (9/10) and/or T. pedis (7/10), while all samples were negative for T. medium. However, none of these Treponema species could be isolated and sequence analysis of the amplified products showed 100% match of 365 base pairs (bp) to Treponema phylotype PT3 and almost full match (530 of 532 bp, 99.6%) to Treponema phylotype PT13. The presence of T. phagedenis, PT3 and PT13 phylotypes was confirmed by FISH analyses. The phylotypes of T. phagedenis were present in all hybridized positive biopsies of Treponema spp., and PT13 and PT3 were less abundant. Neither D. nodosus nor F. necrophorum were detected. The histological Treponema score was mostly mild. Digital dermatitis in captive European Bison is contagious and differs from bovine digital dermatitis, concerning associated pathogens as well as gross appearance., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2021
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25. Complete Genome Sequences of the Methicillin-Resistant Strain Staphylococcus aureus 17Gst354 and Its Prophage Staphylococcus Phage vB_StaphS-IVBph354.
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Kittl S, Brodard I, Overesch G, Kuhnert P, Jores J, and Labroussaa F
- Abstract
We report the complete 2,783,931-bp circular genome sequence of the human methicillin-resistant strain Staphylococcus aureus 17Gst354, isolated from a nasal swab. The strain possessed an additional 4,397-bp plasmid. Moreover, we induced and sequenced its temperate phage Staphylococcus phage vB_StaphS-IVBph354, which has a circular genome of 41,970 bp.
- Published
- 2021
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26. A filter-assisted culture method for isolation of Treponema spp. from bovine digital dermatitis and their identification by MALDI-TOF MS.
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Brodard I, Alsaaod M, Gurtner C, Jores J, Steiner A, and Kuhnert P
- Subjects
- Animals, Bacteriological Techniques methods, Cattle, Cattle Diseases microbiology, Digital Dermatitis microbiology, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization veterinary, Treponemal Infections microbiology, Treponemal Infections veterinary, Bacteriological Techniques veterinary, Cattle Diseases diagnosis, Digital Dermatitis diagnosis, Treponema isolation & purification, Treponemal Infections diagnosis
- Abstract
Digital dermatitis (DD) is a major infectious foot disease of cattle worldwide. Some DD stages are associated with lameness, and the disease has significant economic and animal welfare consequences. The pathogenesis of the disease is not yet fully understood, but Treponema spp. have been associated consistently with clinical cases. Isolation of these fastidious bacteria is difficult and cumbersome. We describe an improved method enabling the culturing of the 3 Treponema spp. ( T. pedis , T. phagedenis , and T. medium ) from bovine foot specimens derived from DD lesions, using a combination of membrane filtering and subsequent growth on selective agar media. The entire procedure from sampling to verification of individual Treponema spp. takes up to 24 d. In addition, we established a MALDI-TOF MS-based identification method to be applied for confirmation of the different Treponema spp. This scheme provides an unambiguous, simple, and straightforward identification procedure for DD-associated Treponema spp.
- Published
- 2021
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27. Trueperella pecoris sp. nov. isolated from bovine and porcine specimens.
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Schönecker L, Schnydrig P, Brodard I, Thomann A, Hemphill A, Rodriguez-Campos S, Perreten V, Jores J, and Kittl S
- Subjects
- Actinomycetaceae isolation & purification, Animals, Bacterial Typing Techniques, Base Composition, DNA, Bacterial genetics, Female, Nucleic Acid Hybridization, Pregnancy, RNA, Ribosomal, 16S genetics, Sequence Analysis, DNA, Switzerland, Actinomycetaceae classification, Cattle microbiology, Milk microbiology, Phylogeny, Placenta microbiology, Swine microbiology
- Abstract
A novel Gram-stain-positive bacterium was isolated from a purulent bovine milk sample, the bovine placenta from an abortion, the udder secretion of a heifer and the lung of a pig that had succumbed from suppurative bronchopneumonia in Switzerland from 2015 to 2019. The strains grew best under aerobic conditions with 5 % CO
2 and colonies were non-haemolytic and greyish-white. They were non-motile and negative for catalase and oxidase. The genomes of the four strains 19M2397T , 15A0121, 15IMD0307 and 19OD0592 were obtained by sequencing. The results of phylogenetic analyses based on the 16S rRNA gene grouped them within the genus Trueperella in the family Arcanobacteriaceae . The genomes had DNA G+C contents of 61.2-62.2 mol% and showed digital DNA-DNA hybridization (dDDH) values of 21.4-22.8 % and average nucleotide identity (ANI) values of approximately 77 % to their closest relatives Trueperella pyogenes and Trueperella bernardiae . With respect to the presence in different livestock species we propose the name Trueperella pecoris sp. nov. The type strain is 19M2397T (=CCOS 1952T =DSM 111392T ), isolated from the udder secretion of a heifer diagnosed with summer mastitis in 2019.- Published
- 2021
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28. Prevalence of Dichelobacter nodosus and Ovine Footrot in German Sheep Flocks.
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Storms J, Wirth A, Vasiliadis D, Brodard I, Hamann-Thölken A, Ambros C, Moog U, Jores J, Kuhnert P, and Distl O
- Abstract
The bacterium Dichelobacter nodosus ( D. nodosus ) is the causative agent of ovine footrot. The aim of this field study was to determine the prevalence of D. nodosus in German sheep flocks. The sheep owners participated voluntarily in the study. More than 9000 sheep from 207 flocks were screened for footrot scores using a Footrot Scoring System from 0 to 5 and sampling each sheep using one interdigital swab for all four feet of the sheep. The detection and discrimination between benign and virulent strains was done employing a real-time PCR. Our results showed a mean prevalence of 42.93% of D. nodosus in German sheep on an animal level. Underrunning of hoof horn on at least one foot (Scores 3-5) was detected in 567 sheep (6.13%). Sheep with four clinically healthy feet were found through visual inspection in 47.85% of all animals included in this study. In total, 1117 swabs from sheep with four clinically healthy feet tested positive for D. nodosus . In 90.35% of the positive swabs, virulent D. nodosus were detected. Benign D. nodosus were detected in 4.74% of the D. nodosus -positive swabs while 4.91% tested positive for both, benign and virulent D. nodosus . In 59 flocks D. nodosus were not detected and in 115 flocks only virulent D. nodosus were found while seven flocks tested positive for benign strains.
- Published
- 2021
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29. Mannheimia pernigra sp. nov., isolated from bovine respiratory tract.
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Kuhnert P, Brodard I, Schönecker L, Akarsu H, Christensen H, and Bisgaard M
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- Animals, Bacterial Typing Techniques, Base Composition, DNA, Bacterial genetics, Fatty Acids chemistry, Genes, Bacterial, Mannheimia isolation & purification, RNA, Ribosomal, 16S genetics, Sequence Analysis, DNA, Switzerland, Ubiquinone chemistry, Cattle microbiology, Mannheimia classification, Phylogeny, Respiratory System microbiology
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Over a period of 1 year, 270 isolates identified as Taxon 39 of Bisgaard were obtained from the nasopharynx of veal calves at 11 epidemiologically independent Swiss fattening farms. Two isolates from each farm and the Australian Taxon 39 reference strain BNO311 were further characterized by genetic and phenotypic methods. Phylogenetic analysis of 16S rRNA and recN gene sequences placed the isolates in a single, distinct cluster within the genus Mannheimia . As to the rpoB gene, most isolates clustered together, but four strains formed a separate cluster close to Mannheimia varigena . Genome sequence analysis of isolates from both rpoB clusters confirmed their species status, with an average nucleotide identity (ANI) >98.9 % between isolates and <84 % to the closest species, M. varigena . Based upon whole genome sequences, the G+C content was determined as 39.1 mol%. Similarly, analysis of MALDI-TOF MS reference spectra clustered the isolates clearly separated from the other Mannheimia species, making this the method of choice for identification. In addition, numerous biochemical markers based on classical as well as commercial identification schemes were determined, allowing separation from other Mannheimia species and identification of the new taxon. Major fatty acids for strain 17CN0883
T are C14 : 0 , C16 : 0 , C16 : 1 ω7 c and C18 : 1 ω7 c . Major respiratory quinones are ubiquinone-7 and ubiquinone-8. We propose the name Mannheimia pernigra sp. nov. for former Taxon 39 of Bisgaard. The type strain is 17CN0883T (=CCUG 74657T =DSM 111153T ) isolated from a veal calf in Switzerland.- Published
- 2021
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30. Complete Genome Sequences of Four Brucella suis Strains Isolated from Swiss Wild Boars.
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Akarsu H, Brodard I, Kittl S, Overesch G, and Jores J
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We present the complete genomes of four Brucella suis biovar 2 isolates that were obtained from wild boars in Switzerland in 2008 and 2009. Genomes were sequenced with PacBio technology, contained two chromosomes each, had a genome size of 3.3 Mbp, and contained more than 3,225 genes per genome., (Copyright © 2020 Akarsu et al.)
- Published
- 2020
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31. First European report of Francisella tularensis subsp. holarctica isolation from a domestic cat.
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Kittl S, Francey T, Brodard I, Origgi FC, Borel S, Ryser-Degiorgis MP, Schweighauser A, and Jores J
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- Animals, Cat Diseases microbiology, Cats, Genome, Viral, Male, Phylogeny, Switzerland, Tularemia diagnosis, Tularemia microbiology, Cat Diseases diagnosis, Francisella isolation & purification, Tularemia veterinary
- Abstract
Francisella tularensis subsp. holarctica is a select agent causing life-threatening tularemia. It has been isolated from humans and animals, mainly lagomorphs and rodents, rarely other wild carnivore species. Increasing numbers of human tularemia cases have been reported during the last 5 years in Switzerland. Here we report the first isolation of Francisella tularensis subsp. holarctica from a domestic cat in Europe and compare its genome sequence with other Swiss isolates. The cat isolate shows a close phylogenetic relationship with a contemporary hare isolate from close geographic proximity, indicating a possible epidemiological link.
- Published
- 2020
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32. Treponema phagedenis ( ex Noguchi 1912) Brumpt 1922 sp. nov., nom. rev., isolated from bovine digital dermatitis.
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Kuhnert P, Brodard I, Alsaaod M, Steiner A, Stoffel MH, and Jores J
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- Animals, Bacterial Typing Techniques, Base Composition, Cattle, DNA, Bacterial genetics, Fatty Acids chemistry, Humans, Polymerase Chain Reaction, RNA, Ribosomal, 16S genetics, Sequence Analysis, DNA, Switzerland, Treponema isolation & purification, Cattle Diseases microbiology, Digital Dermatitis microbiology, Phylogeny, Treponema classification
- Abstract
' Treponema phagedenis ' was originally described in 1912 by Noguchi but the name was not validly published and no type strain was designated. The taxon was not included in the Approved Lists of Bacterial Names and hence has no standing in nomenclature. Six Treponema strains positive in a ' T. phagedenis ' phylogroup-specific PCR test were isolated from digital dermatitis (DD) lesions of cattle and further characterized and compared with the human strain ' T. phagedenis ' ATCC 27087. Results of phenotypic and genotypic analyses including API ZYM, VITEK2, MALDI-TOF and electron microscopy, as well as whole genome sequence data, respectively, showed that they form a cluster of species identity. Moreover, this species identity was shared with ' T. phagedenis '-like strains reported in the literature to be regularly isolated from bovine DD. High average nucleotide identity values between the genomes of bovine and human ' T. phagedenis ' were observed. Slight genomic as well as phenotypic variations allowed us to differentiate bovine from human isolates, indicating host adaptation. Based on the fact that this species is regularly isolated from bovine DD and that the name is well dispersed in the literature, we propose the species Treponema phagedenis sp. nov., nom. rev. The species can phenotypically and genetically be identified and is clearly separated from other Treponema species. The valid species designation will allow to further explore its role in bovine DD. The type strain for Treponema phagedenis sp. nov., nom. rev. is B43.1
T (=DSM 110455T =NCTC 14362T ) isolated from a bovine DD lesion in Switzerland.- Published
- 2020
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33. Methicillin-Resistant Staphylococcus aureus Strains in Swiss Pigs and Their Relation to Isolates from Farmers and Veterinarians.
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Kittl S, Brodard I, Heim D, Andina-Pfister P, and Overesch G
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- Animals, Anti-Bacterial Agents pharmacology, Methicillin pharmacology, Methicillin-Resistant Staphylococcus aureus classification, Multilocus Sequence Typing veterinary, Polymorphism, Single Nucleotide, Prevalence, Staphylococcal Infections microbiology, Sus scrofa, Swine, Swine Diseases virology, Switzerland epidemiology, Farmers statistics & numerical data, Methicillin-Resistant Staphylococcus aureus isolation & purification, Staphylococcal Infections epidemiology, Staphylococcal Infections veterinary, Swine Diseases epidemiology, Veterinarians statistics & numerical data
- Abstract
Methicillin-resistant Staphylococcus aureus (MRSA) has emerged over the last few decades as a One Health problem with an increasing prevalence in various animal species. The most notable animals are pigs, as asymptomatic carriers, and horses, where there is often an association with infections. The current study looked at the course of MRSA prevalence in Swiss livestock since 2009, with a special focus on pigs, followed by screening of veterinarians and farmers. Livestock isolates were obtained from the Swiss monitoring program and then characterized by spa typing. Concentrating on the year 2017, we analyzed the prevalence of MRSA in Swiss veterinarians and farmers, followed by whole-genome sequencing of selected human and animal strains. The phylogeny was assessed by applying core-genome multilocus sequence typing (MLST) and single-nucleotide polymorphism (SNP) analyses, followed by screening for resistance genes and virulence factors. The prevalence of MRSA in Swiss pigs showed a dramatic increase from 2% in 2009 to 44% in 2017. Isolates typically belonged to clonal complex 398 (CC398), split between spa t011 and t034. The higher prevalence was mainly due to an increase in t011. spa t034 strains from farmers were found to be closely associated with porcine t034 strains. The same could be shown for spa t011 strains from horses and veterinarians. spa t034 strains had a high number of additional resistance genes, and two strains had acquired the immune evasion cluster. However, all but one of the pig spa t011 strains clustered in a separate group. Thus, the increase in pig spa t011 strains does not directly translate to humans. IMPORTANCE MRSA is an important human pathogen; thus, its increasing prevalence in livestock over the last decade has a potentially large impact on public health. Farmers and veterinarians are especially at risk due to their close contact with animals. Our work demonstrates a dramatic increase in MRSA prevalence in Swiss pigs, from 2% in 2009 to 44% in 2017. Whole-genome sequencing allowed us to show a close association between farmer and pig strains as well as veterinarian and horse strains, indicating that the respective animals are a likely source of human colonization. Furthermore, we could demonstrate that pig spa t011 strains cluster separately and are probably less likely to colonize humans than are pig spa t034 strains. This research may provide a basis for a more substantiated risk assessment and preventive measures., (Copyright © 2020 American Society for Microbiology.)
- Published
- 2020
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34. Detection of specific Treponema species and Dichelobacter nodosus from digital dermatitis (Mortellaro's disease) lesions in Swiss cattle.
- Author
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Alsaaod M, Locher I, Jores J, Grimm P, Brodard I, Steiner A, and Kuhnert P
- Subjects
- Animals, Cattle, Dichelobacter nodosus genetics, Polymerase Chain Reaction, Switzerland epidemiology, Treponema genetics, Treponemal Infections epidemiology, Treponemal Infections microbiology, Cattle Diseases epidemiology, Cattle Diseases microbiology, Digital Dermatitis epidemiology, Digital Dermatitis microbiology, Gram-Negative Bacterial Infections veterinary, Treponemal Infections veterinary
- Abstract
Introduction: The aim of this study was to determine the prevalence of the three Treponema species as well as D. nodosus in Digital dermatitis (DD) and slurry of Swiss cattle using PCR. A total of 86 specimens from 24 farms were enrolled in the study. Slurry samples from 21 DD-affected and one unaffected farm were collected to assess the potential of environmental transmission. Nested and real-time PCR were performed from the specimens to detect Treponema species and D. nodosus, respectively. The DD-stages were positive for at least one or more of the DD-associated Treponema species in 50 of 61 cases (82.0%) and in 9 of 25 cases (36.0%) in unaffected animals. Infected animals with small focal active lesions showed a significantly lower prevalence (14.8%) compared to the other DD stages (67.2%; P=0.011). Most prevalent was T. phagedenis (65.1%). D. nodosus was detected in 51.8% of clinical DD lesions and 24.1% in unaffected cases, but its presence was not significantly associated with the various DD-stages. All samples positive for D. nodosus contained the acid protease gene aprB2 but were negative for aprV2, the latter associated with virulence in sheep foot rot. Control farms were negative for all DD-associated Treponema species while positive for aprB2 and negative for aprV2. The presence of aprB2 suggests it is ubiquitous in the animal environment. With respect to the slurry samples, three out of 21 specimens (14.3%) were positive for one or more of the DD-associated Treponema species and eleven out of 21 specimens (52.4%) were positive for aprB2 and negative for aprV2 of D. nodosus. In conclusion, an association was found between the presence of clinical DD and specific Treponema species, while for D. nodosus no such link with DD lesions could be observed.
- Published
- 2019
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35. Otitis in a cat associated with Corynebacterium provencense.
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Kittl S, Brodard I, Rychener L, Jores J, Roosje P, and Gobeli Brawand S
- Subjects
- Animals, Anti-Bacterial Agents therapeutic use, Cat Diseases drug therapy, Cats, Chloramphenicol therapeutic use, Corynebacterium Infections drug therapy, Corynebacterium Infections microbiology, Genome, Bacterial genetics, Male, Microbial Sensitivity Tests veterinary, Otitis Media drug therapy, Otitis Media microbiology, Phylogeny, RNA, Ribosomal, 16S genetics, Cat Diseases microbiology, Corynebacterium genetics, Corynebacterium Infections veterinary, Otitis Media veterinary
- Abstract
Background: The role of corynebacteria in canine and feline otitis has not been investigated in detail; however, members of this genus are increasingly recognized as pathogens of otitis in both human and veterinary medicine., Case Presentation: Here we report the first case of feline otitis associated with the recently described species Corynebacterium provencense. A seven-month old cat presented with a head tilt and ataxia was diagnosed with peripheral vestibular syndrome associated with an otitis media/interna. This took place 6 weeks after resection of a polyp, having initially shown a full recovery with topical ofloxacin and glucocorticoid treatment. Bacteriology of an ear swab yielded a pure culture of corynebacteria, which could not be identified at the species level using routine methods. However, the 16S rRNA gene sequence was 100% identical to the recently published novel corynebacterium species, Corynebacterium provencense. Whole genome sequencing of the cat isolate and calculation of average nucleotide identity (99.1%) confirmed this finding. The cat isolate was found to contain additional presumptive iron acquisition genes that are likely to encode virulence factors. Furthermore, the strain tested resistant to clindamycin, penicillin and ciprofloxacin. The cat was subsequently treated with chloramphenicol, which lead to clinical improvement., Conclusion: Corynebacteria from otitis cases are not routinely identified at the species level and not tested for antimicrobial susceptibility in veterinary laboratories, as they are not considered major pathogens. This may lead to underreporting of this genus or animals being treated with inappropriate antimicrobials since corynebacteria are often resistant to multiple drugs.
- Published
- 2018
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36. Bacterial, fungal, parasitological and pathological analyses of abortions in small ruminants from 2012-2016.
- Author
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Schnydrig P, Vidal S, Brodard I, Frey C, Posthaus H, Perreten V, and Rodriguez-Campos S
- Subjects
- Abortion, Veterinary microbiology, Abortion, Veterinary parasitology, Abortion, Veterinary pathology, Animals, Bacteria classification, Bacteria genetics, Bacteriological Techniques, Female, Fungi classification, Fungi genetics, Goat Diseases microbiology, Goat Diseases parasitology, Goat Diseases pathology, Goats, Pathology, Molecular, Pregnancy, Real-Time Polymerase Chain Reaction, Sheep, Sheep Diseases microbiology, Sheep Diseases parasitology, Sheep Diseases pathology, Abortion, Veterinary diagnosis, Goat Diseases diagnosis, Sheep Diseases diagnosis
- Abstract
Introduction: Abortion in small ruminants presents a clinical and economic problem with legal implications regarding animal health and zoonotic risk by some of the abortive pathogens. Several bacteria, fungi and parasites can cause abortion, but cost-orientated routine diagnostics only cover the most relevant epizootic agents. To cover a broad-range of common as well as underdiagnosed abortifacients, we studied 41 ovine and 36 caprine abortions by Stamp's modification of the Ziehl-Neelsen stain, culture for classical and opportunistic abortive agents, real-time PCR for C. burnetii, C. abortus, pathogenic Leptospira spp., Toxoplasma gondii and Neospora caninum. When the dam's serum was available detection of antibodies against B. melitensis, C. burnetii, C. abortus and Leptospira spp. was performed. In 37 cases sufficient placental tissue was available for pathological and histopathological examination. From the 77 cases 11 (14.3%) were positive by staining whereas real-time PCR detected C. burnetii and C. abortus in 49.3% and 32.5% of the cases. Antibodies against C. abortus and Leptospira spp. (33.3 and 26.7%) were detected. In 23.4% a bacterial culturable pathogen was isolated. Fungal abortion was confirmed in 1.3% of cases. A single abortive agent was identified in 44.2% of the cases and in 31.2% multiple possible abortifacients were present. Our study shows that the highest clarification rate can only be achieved by a combination of methods and evidences the role that multi-infections play as cause of abortion.
- Published
- 2017
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37. Macrococcus canis sp. nov., a skin bacterium associated with infections in dogs.
- Author
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Gobeli Brawand S, Cotting K, Gómez-Sanz E, Collaud A, Thomann A, Brodard I, Rodriguez-Campos S, Strauss C, and Perreten V
- Subjects
- Animals, Bacterial Typing Techniques, Base Composition, Cell Wall chemistry, DNA, Bacterial genetics, Fatty Acids chemistry, Genes, Bacterial, Nucleic Acid Hybridization, Peptidoglycan chemistry, RNA, Ribosomal, 16S genetics, Sequence Analysis, DNA, Staphylococcaceae genetics, Staphylococcaceae isolation & purification, Vitamin K 2 analogs & derivatives, Vitamin K 2 chemistry, Dog Diseases microbiology, Dogs microbiology, Phylogeny, Skin microbiology, Skin Diseases, Bacterial veterinary, Staphylococcaceae classification
- Abstract
Gram-stain-positive cocci were isolated from miscellaneous sites of the skin of healthy dogs as well as from infection sites in dogs. The closest relative by sequencing of the 16S rRNA gene was Macrococcus caseolyticus with 99.7 % sequence identity, but compared with M. caseolyticus, the novel strains shared only 90.8 to 93.5 % DNA sequence identity with cpn60, dnaJ, rpoB and sodA partial genes, respectively. The novel strains also exhibited differential phenotypic characteristics from M. caseolyticus, and the majority displayed a visible haemolysis on sheep blood agar, while M. caseolyticus did not have any haemolytic activity. They generated different matrix-assisted laser-desorption/ionization time-of-flight (MALDI-TOF) MS spectral profiles compared with the other species of the genus Macrococcus. Furthermore, strain KM 45013
T shared only 53.7 % DNA-DNA relatedness with the type strain of M. caseolyticus, confirming that they do not belong to the same species. The DNA G+C content of strain KM 45013T was 36.9 mol%. The most abundant fatty acids were C14 : 0, C18 : 3ω6c (6, 9, 12) and C16 : 0 n alcohol. MK-6 was the menaquinone type of KM 45013T . Cell-wall structure analysis revealed that the peptidoglycan type was A3α l-Lys-Gly2-l-Ser. Based on genotypic and chemotaxonomic characteristics, we propose to classify these strains within a novel species of the genus Macrococcus for which the name Macrococcus canis sp. nov. is proposed. The type strain is KM 45013T (=DSM 101690T =CCOS 969T =CCUG 68920T ).- Published
- 2017
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38. Clinical and epidemiological analysis of Campylobacter fetus subsp. fetus infections in humans and comparative genetic analysis with strains isolated from cattle.
- Author
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Escher R, Brunner C, von Steiger N, Brodard I, Droz S, Abril C, and Kuhnert P
- Subjects
- Adult, Aged, Aged, 80 and over, Animals, Anti-Bacterial Agents pharmacology, Campylobacter Infections drug therapy, Campylobacter Infections microbiology, Campylobacter Infections veterinary, Campylobacter fetus pathogenicity, Cattle, Drug Resistance, Bacterial drug effects, Electrophoresis, Gel, Pulsed-Field, Female, Genotype, Humans, Male, Microbial Sensitivity Tests, Middle Aged, Multilocus Sequence Typing, Phenotype, Phylogeny, Polymerase Chain Reaction, Streptomycin pharmacology, Switzerland epidemiology, Tetracycline pharmacology, Campylobacter Infections epidemiology, Campylobacter fetus drug effects, Campylobacter fetus genetics, Drug Resistance, Bacterial genetics
- Abstract
Background: Campylobacter fetus subspecies fetus (CFF) is an important pathogen for both cattle and humans. We performed a systematic epidemiological and clinical study of patients and evaluated the genetic relatedness of 17 human and 17 bovine CFF isolates by using different genotyping methods. In addition, the serotype, the dissemination of the genomic island containing a type IV secretion system (T4SS) and resistance determinants for tetracycline and streptomycin were also evaluated., Methods: The isolates from patients diagnosed with CFF infection as well as those from faecal samples of healthy calves were genotyped using pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), as well as single locus sequence typing (SLST) targeting cmp1 and cmp2 genes encoding two major outer membrane proteins in CFF. The presence of the genomic island and identification of serotype was determined by PCRs targeting genes of the T4SS and the sap locus, respectively. Tetracycline and streptomycin resistance phenotypes were determined by minimal inhibitory concentration. Clinical data obtained from medical records and laboratory data were supplemented by data obtained via telephone interviews with the patients and treating physicians., Results: PFGE analysis defined two major clusters; cluster A containing 16 bovine (80 %) isolates and cluster B containing 13 human (92 %) isolates, suggesting a host preference. Further genotypic analysis using MLST, SLST as well as sap and T4SS PCR showed the presence of genotypically identical isolates in cattle and humans. The low diversity observed within the cmp alleles of CFF corroborates the clonal nature of this pathogen. The genomic island containing the tetracycline and streptomycin resistance determinants was found in 55 % of the isolates in cluster A and correlated with phenotypic antibiotic resistance., Conclusions: Most human and bovine isolates were separated on two phylogenetic clusters. However, several human and bovine isolates were identical by diverse genotyping methods, indicating a possible link between strains from these two hosts.
- Published
- 2016
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39. DEVRIESEASIS IN A PLUMED BASILISK (BASILISCUS PLUMIFRONS) AND CHINESE WATER DRAGONS (PHYSIGNATHUS COCINCINUS) IN A ZOOLOGIC COLLECTION.
- Author
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Rossier C, Hoby S, Wenker C, Brawand SG, Thomann A, Brodard I, Jermann T, and Posthaus H
- Subjects
- Animals, Animals, Zoo, Female, Gram-Positive Bacteria isolation & purification, Gram-Positive Bacterial Infections microbiology, Male, Gram-Positive Bacteria classification, Gram-Positive Bacterial Infections veterinary, Lizards
- Abstract
Devriesea agamarum is a Gram-positive bacterium that was first described in 2008 as a causative agent of disease in lizards. Until today, reports from several countries reported the presence of this bacterium in various lizard species, which suggests a wide distribution among lizard collections. Pathologic lesions ranged from proliferative dermatitis and cheilitis to abscesses in multiple organs and septicemia in single animals, as well as entire groups. Until now, disease caused by D. agamarum has been reported in several lizard species. Because the bacterium is only identified by 16S rRNA sequencing and no commercially available identification systems contain the agent in their database, it may be underdiagnosed. This report describes a series of fatal devrieseasis in plumed basilisks (Basiliscus plumifrons) and Chinese water dragons (Physignathus cocincinus) from a zoologic collection and extends the range of susceptible species. In 3 mo, five animals died with pyogranulomatous lesions in the subcutis, the coelomic cavity, or multiple organs. In all cases, diffuse swelling or focal skin elevations of different body parts were observed. Devriesea agamarum could be isolated from lesions in all animals. A subsequent clinical survey of the lizard collection including bacteriologic investigation of oral cavity swabs indicated that bearded dragons (Pogona vitticeps) were carriers of D. agamarum, which suggests that this species could be a source of infection with this pathogen.
- Published
- 2016
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40. Uruburuella testudinis sp. nov., isolated from tortoise (Testudo).
- Author
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Kuhnert P, Thomann A, Brodard I, Haefeli W, and Korczak BM
- Subjects
- Animals, Bacterial Typing Techniques, Base Composition, DNA, Bacterial genetics, Fatty Acids chemistry, Genes, Bacterial, Molecular Sequence Data, Neisseriaceae genetics, Neisseriaceae isolation & purification, Nucleic Acid Hybridization, RNA, Ribosomal, 16S genetics, Sequence Analysis, DNA, Neisseriaceae classification, Phylogeny, Turtles microbiology
- Abstract
A polyphasic taxonomic analysis was carried out on 11 uncommon Gram-stain-negative, non-motile, catalase- and oxidase-positive, but indole-negative, bacterial strains isolated from tortoises. Phenotypically and genetically they represented a homogeneous group of organisms most closely related to, but distinct from, Uruburuella suis. In a reconstructed 16S rRNA gene tree they clustered on a monophyletic branch next to U. suis with gene similarities between strains of 99.5-100%, and of up to 98.2% with U. suis . DNA-DNA hybridization indicated the organisms represented a novel species with only 40% DNA-DNA similarity with U. suis . Partial sequencing of rpoB resulted in two subclusters confirming the 16S rRNA gene phylogeny; both genes allowed clear separation and identification of the novel species. Furthermore, they could be unambiguously identified by matrix-assisted laser desorption ionization time-of-flight MS, where, again, they formed a highly homogeneous cluster separate from U. suis and other members of the family Neisseriaceae . The major fatty acids were C(16 : 0) and summed feature C(16 : 1)ω7c/iso-C(15 : 0) 2-OH. The DNA G+C content was 54.4 mol%. Based on phenotypic and genetic data we propose classifying these organisms as representatives of a novel species named Uruburuella testudinis sp. nov. The type strain is 07_OD624(T) ( = DSM 26510(T) = CCUG 63373(T))., (© 2015 IUMS.)
- Published
- 2015
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41. Arsenicicoccus dermatophilus sp. nov., a hypha-forming bacterium isolated from the skin of greater flamingos (Phoenicopterus roseus) with pododermatitis.
- Author
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Gobeli S, Thomann A, Wyss F, Kuehni-Boghenbor K, Brodard I, and Perreten V
- Subjects
- Actinomycetales genetics, Actinomycetales isolation & purification, Animals, Animals, Zoo microbiology, Bacterial Typing Techniques, Base Composition, Cell Wall chemistry, DNA, Bacterial genetics, Fatty Acids chemistry, Molecular Sequence Data, Nucleic Acid Hybridization, Peptidoglycan chemistry, RNA, Ribosomal, 16S genetics, Sequence Analysis, DNA, Skin Diseases, Bacterial microbiology, Switzerland, Vitamin K 2 analogs & derivatives, Vitamin K 2 chemistry, Actinomycetales classification, Bird Diseases microbiology, Birds microbiology, Phylogeny, Skin microbiology, Skin Diseases, Bacterial veterinary
- Abstract
Dermatophilus-like bacteria were observed in histological examinations of samples of diseased foot skin from greater flamingos (Phoenicopterus roseus) living in zoological gardens in Switzerland. When grown on TSA-SB containing polymyxin B, the bacteria isolated from these skin samples formed hyphae, as is typical for Dermatophilus congolensis, but these bacteria were non-haemolytic. The closest relatives based on 16S rRNA gene sequences were the two members of the genus Arsenicicoccus, Arsenicicoccus bolidensis and Arsenicicoccus piscis. A representative of the isolated strains shared 34.3 % DNA-DNA relatedness with the type strain of A. bolidensis, 32.3 % with the type strain of A. piscis and 34.5 % with the type strain of D. congolensis, demonstrating that these strains do not belong to any of these species. The phenotypic characteristics differed from those of members of the genus Arsenicicoccus as well as from those of D. congolensis. The G+C content of strain KM 894/11(T) was 71.6 mol%. The most abundant fatty acids were iso-C15 : 0, summed feature 3 (including C16 : 1ω7c and/or iso-C15 : 0 2-OH) and C18 : 1ω9c. MK-8(H4) was the predominant menaquinone. Cell-wall structure analysis revealed that the peptidoglycan type was A3γ ll-Dpm-Gly (type A41.1). Based on genotypic and chemotaxonomic characteristics, the isolated strains represent a novel species within the genus Arsenicicoccus, for which the name Arsenicicoccus dermatophilus sp. nov. is proposed. The type strain is KM 894/11(T) ( = DSM 25571(T) = CCUG 62181(T) = CCOS 690(T)), and strain KM 1/12 ( = DSM 25572 = CCUG 62182 = CCOS 691) is a reference strain.
- Published
- 2013
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42. Ruminant rhombencephalitis-associated Listeria monocytogenes alleles linked to a multilocus variable-number tandem-repeat analysis complex.
- Author
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Balandyté L, Brodard I, Frey J, Oevermann A, and Abril C
- Subjects
- Alleles, Animals, Cluster Analysis, DNA, Bacterial chemistry, DNA, Bacterial genetics, Encephalitis pathology, Environmental Microbiology, Food Microbiology, Genotype, Humans, Listeria monocytogenes classification, Listeria monocytogenes isolation & purification, Molecular Sequence Data, Rhombencephalon microbiology, Rhombencephalon pathology, Ruminants, Sequence Analysis, DNA, Virulence Factors genetics, Encephalitis microbiology, Encephalitis veterinary, Listeria monocytogenes genetics, Listeria monocytogenes pathogenicity, Minisatellite Repeats, Polymorphism, Genetic
- Abstract
Listeria monocytogenes is among the most important food-borne pathogens and is well adapted to persist in the environment. To gain insight into the genetic relatedness and potential virulence of L. monocytogenes strains causing central nervous system (CNS) infections, we used multilocus variable-number tandem-repeat analysis (MLVA) to subtype 183 L. monocytogenes isolates, most from ruminant rhombencephalitis and some from human patients, food, and the environment. Allelic-profile-based comparisons grouped L. monocytogenes strains mainly into three clonal complexes and linked single-locus variants (SLVs). Clonal complex A essentially consisted of isolates from human and ruminant brain samples. All but one rhombencephalitis isolate from cattle were located in clonal complex A. In contrast, food and environmental isolates mainly clustered into clonal complex C, and none was classified as clonal complex A. Isolates of the two main clonal complexes (A and C) obtained by MLVA were analyzed by PCR for the presence of 11 virulence-associated genes (prfA, actA, inlA, inlB, inlC, inlD, inlE, inlF, inlG, inlJ, and inlC2H). Virulence gene analysis revealed significant differences in the actA, inlF, inlG, and inlJ allelic profiles between clinical isolates (complex A) and nonclinical isolates (complex C). The association of particular alleles of actA, inlF, and newly described alleles of inlJ with isolates from CNS infections (particularly rhombencephalitis) suggests that these virulence genes participate in neurovirulence of L. monocytogenes. The overall absence of inlG in clinical complex A and its presence in complex C isolates suggests that the InlG protein is more relevant for the survival of L. monocytogenes in the environment.
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- 2011
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43. Free-ranging wild boar: a disease threat to domestic pigs in Switzerland?
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Wu N, Abril C, Hinić V, Brodard I, Thür B, Fattebert J, Hüssy D, and Ryser-Degiorgis MP
- Subjects
- Animals, Animals, Domestic, Animals, Wild, Brucellosis epidemiology, Brucellosis transmission, Female, Male, Porcine Reproductive and Respiratory Syndrome epidemiology, Porcine Reproductive and Respiratory Syndrome transmission, Swine, Swine Diseases transmission, Switzerland epidemiology, Brucella suis, Brucellosis veterinary, Disease Transmission, Infectious veterinary, Sus scrofa, Swine Diseases epidemiology
- Abstract
The risk of transmission of pathogens from free-ranging wild boars (Sus scrofa scrofa) to outdoor domestic pigs (S. scrofa domesticus) is of increasing concern in many European countries. We assess this risk, using Switzerland as an example. We estimated 1) the prevalence of important pathogens in wild boars and 2) the risk of interactions between wild boars and outdoor pigs. First, we tested 252 wild boars from selected areas between 2008 and 2010 for infection with Brucella spp. Bacterial prevalence was estimated to 28.8% (confidence interval [CI] 23.0-34.0) when using bacterial culture (B. suis Biovar 2) and real-time polymerase chain reaction. Antibody prevalence was 35.8% (CI 30.0-42.0), which was significantly higher than in previous studies in Switzerland. We also tested 233 wild boars for porcine reproductive and respiratory syndrome virus (PRRSV). Antibody prevalence was 0.43% (CI 0.01-2.4) for EU-PRRSV and real-time reverse transcription polymerase chain reaction results were negative. These findings suggest that B. suis is increasingly widespread in wild boars and PRRSV is currently not of concern. Second, we documented the spatial overlap between free-ranging wild boars and outdoor piggeries by mapping data on their respective occurrence. Wild boars are most widespread in the mountain range along the western and northern Swiss borders, while most piggeries are located in central lowlands. A risk of interaction is mainly expected at the junction between these two bioregions. This risk may increase if wild boars expand eastward and southward beyond anthropogenic barriers believed to limit their range. Therefore, we evaluated the potential of expansion of the wild boar population. Population trends suggest a continuous increase of wild boars for the past 15 yr. Surveillance of selected wildlife passages using cameras on highways and main roads indicates that these barriers are permeable (average of up to 13 wild boar crossings per 100 days). Thus an increase of wild boar range should be considered. There may be a risk of B. suis spillover from wild boars in Switzerland, which could increase in the future. Data on the occurrence of interactions between pigs and wild boars are needed to assess this risk.
- Published
- 2011
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44. A novel isolation method of Brucella species and molecular tracking of Brucella suis biovar 2 in domestic and wild animals.
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Abril C, Thomann A, Brodard I, Wu N, Ryser-Degiorgis MP, Frey J, and Overesch G
- Subjects
- Animals, Animals, Wild microbiology, Bacterial Typing Techniques, Brucella classification, Brucella genetics, Brucella isolation & purification, Brucella suis genetics, Brucellosis epidemiology, Brucellosis genetics, Brucellosis microbiology, Disease Outbreaks, Minisatellite Repeats, Phylogeny, Sensitivity and Specificity, Swine, Swine Diseases epidemiology, Swine Diseases genetics, Switzerland epidemiology, Bacteriological Techniques, Brucella suis isolation & purification, Brucellosis veterinary, Swine Diseases microbiology
- Abstract
Brucella suis biovar 2 is the most common aetiological agent of porcine brucellosis in Europe. B. suis biovar 2 is considered to have low zoonotic potential, but is a causative agent of reproductive losses in pigs, and it is thus economically important. The multilocus variable-number of tandem repeats genotyping analysis of 16 loci (MLVA-16) has proven to be highly discriminatory and is the most suitable assay for simultaneously identifying B. suis and tracking infections. The aim of this study was to investigate the relatedness between isolates of B. suis biovar 2 obtained during a brucellosis outbreak in domestic pigs and isolates from wild boars and hares collected from proximal or remote geographical areas by MLVA-16. A cluster analysis of the MLVA-16 data revealed that most of the isolates obtained from Switzerland clustered together, with the exception of one isolate. The outbreak isolates constituted a unique subcluster (with a genetic similarity >93.8%) distinct from that of the isolates obtained from wild animals, suggesting that direct transmission of the bacterium from wild boars to domestic pigs did not occur in this outbreak. To obtain a representative number of isolates for MLVA-16, alternative methods of Brucella spp. isolation from tissue samples were compared with conventional direct cultivation on a Brucella-selective agar. We observed an enhanced sensitivity when mechanical homogenisation was followed by host cell lysis prior to cultivation on the Brucella-selective agar. This work demonstrates that MLVA-16 is an excellent tool for both monitoring brucellosis and investigating outbreaks. Additionally, we present efficient alternatives for the isolation of Brucella spp., (Copyright © 2011 Elsevier B.V. All rights reserved.)
- Published
- 2011
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45. Two novel antibiotic resistance genes, tet(44) and ant(6)-Ib, are located within a transferable pathogenicity island in Campylobacter fetus subsp. fetus.
- Author
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Abril C, Brodard I, and Perreten V
- Subjects
- Bacterial Proteins genetics, Microbial Sensitivity Tests, Minocycline pharmacology, Molecular Sequence Data, Nucleotidyltransferases genetics, Phylogeny, Polymerase Chain Reaction, Campylobacter fetus drug effects, Campylobacter fetus genetics, Drug Resistance, Multiple, Bacterial genetics, Genomic Islands genetics, Streptomycin pharmacology, Tetracycline Resistance genetics
- Abstract
New tetracycline and streptomycin resistance genes, tet(44) and ant(6)-Ib, were identified in Campylobacter fetus subsp. fetus within a transferable pathogenicity island that is typically unique to Campylobacter fetus subsp. venerealis. The 640-amino-acid tetracycline resistance determinant, Tet 44, belongs to a class of proteins that confers resistance to tetracycline and minocycline by ribosomal protection. The 286-amino-acid streptomycin resistance determinant, ANT(6)-Ib, belongs to a family of aminoglycoside nucleotidyltransferases. The resistance phenotypes were demonstrated by gene inactivation and expression.
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- 2010
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46. Brucellosis in a dog caused by Brucella melitensis Rev 1.
- Author
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Hinić V, Brodard I, Petridou E, Filioussis G, Contos V, Frey J, and Abril C
- Subjects
- Amoxicillin-Potassium Clavulanate Combination therapeutic use, Animals, Anti-Bacterial Agents pharmacology, Anti-Bacterial Agents therapeutic use, Brucella melitensis drug effects, Brucella melitensis genetics, Brucellosis drug therapy, Dog Diseases drug therapy, Dogs, Fatal Outcome, Genotype, Greece, Male, Microbial Sensitivity Tests, Microsatellite Repeats genetics, Brucella melitensis physiology, Brucellosis veterinary, Dog Diseases microbiology
- Published
- 2010
- Full Text
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47. [Occurrence of Clostridium perfringens type A and type C in piglets of the Swiss swine population].
- Author
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Wollschläger N, Zimmermann W, Brodard I, Albini S, Doherr M, Posthaus H, and Miserez R
- Subjects
- Animals, Animals, Newborn, Bacterial Toxins genetics, Clostridium Infections epidemiology, Clostridium Infections microbiology, Clostridium perfringens isolation & purification, Clostridium perfringens pathogenicity, DNA, Bacterial analysis, Disease Outbreaks veterinary, Enteritis epidemiology, Enteritis microbiology, Feces microbiology, Female, Genotype, Male, Necrosis epidemiology, Necrosis microbiology, Necrosis veterinary, Phylogeny, Prevalence, Retrospective Studies, Risk Factors, Swine, Swine Diseases epidemiology, Switzerland epidemiology, Clostridium Infections veterinary, Clostridium perfringens classification, Enteritis veterinary, Enterotoxins genetics, Swine Diseases microbiology
- Abstract
Necrotizing enteritis (NE) of newborn piglets still represents an economical problem in Swiss pig breeding and production. The aim of our study was to identify risk factors for NE and evaluate the prevalence of C. perfringens with the toxingenes cpb and cpb2 in Swiss pig breeding farms. The prevalence of theses C. perfringens was investigated using fecal swabs followed by bacteriological culturing and genotyping. Close proximity to other breeding farms and large herd sizes were shown to predispose to NE. C. perfringens type C, carrying the genes cpa, cpb and cpb2 were frequently identified in herds with acute outbreaks of NE. Farms not affected by NE or those using prophylactic vaccination against NE were predominantly positive for C. perfringens type A strains with cpb2 and showed much lower prevalence of C. perfringens type C, compared to acutely affected herds. Our results demonstrate that C. perfringens type A strains with cpb2 are not associated with NE. Besides typical necropsy finding, only the identification of cpb can be used for the diagnosis of NE in affected herds.
- Published
- 2009
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48. IS711-based real-time PCR assay as a tool for detection of Brucella spp. in wild boars and comparison with bacterial isolation and serology.
- Author
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Hinić V, Brodard I, Thomann A, Holub M, Miserez R, and Abril C
- Subjects
- Animals, Brucellosis blood, Brucellosis epidemiology, Brucellosis microbiology, Female, Male, Reproducibility of Results, Sensitivity and Specificity, Serologic Tests, Spleen microbiology, Swine, Swine Diseases microbiology, Testis microbiology, Uterus microbiology, Brucella isolation & purification, Brucellosis veterinary, Polymerase Chain Reaction veterinary, Swine Diseases diagnosis
- Abstract
Background: Control of brucellosis in livestock, wildlife and humans depends on the reliability of the methods used for detection and identification of bacteria. In the present study, we describe the evaluation of the recently established real-time PCR assay based on the Brucella-specific insertion sequence IS711 with blood samples from 199 wild boars (first group of animals) and tissue samples from 53 wild boars (second group of animals) collected in Switzerland. Results from IS711 real-time PCR were compared to those obtained by bacterial isolation, Rose Bengal Test (RBT), competitive ELISA (c-ELISA) and indirect ELISA (i-ELISA)., Results: In the first group of animals, IS711 real-time PCR detected infection in 11.1% (16/144) of wild boars that were serologically negative. Serological tests showed different sensitivities [RBT 15.6%, c-ELISA 7.5% and i-ELISA 5.5%] and only 2% of blood samples were positive with all three tests, which makes interpretation of the serological results very difficult. Regarding the second group of animals, the IS711 real-time PCR detected infection in 26% of animals, while Brucella spp. could be isolated from tissues of only 9.4% of the animals., Conclusion: The results presented here indicate that IS711 real-time PCR assay is a specific and sensitive tool for detection of Brucella spp. infections in wild boars. For this reason, we propose the employment of IS711 real-time PCR as a complementary tool in brucellosis screening programs and for confirmation of diagnosis in doubtful cases.
- Published
- 2009
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49. Routine phenotypic identification of bacterial species of the family Pasteurellaceae isolated from animals.
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Dousse F, Thomann A, Brodard I, Korczak BM, Schlatter Y, Kuhnert P, Miserez R, and Frey J
- Subjects
- Actinobacillus genetics, Actinobacillus isolation & purification, Aeromonas genetics, Aeromonas isolation & purification, Animals, Animals, Domestic, Bordetella bronchiseptica genetics, Bordetella bronchiseptica isolation & purification, Escherichia coli genetics, Escherichia coli isolation & purification, Mannheimia genetics, Mannheimia isolation & purification, Pasteurellaceae isolation & purification, Pasteurellaceae Infections veterinary, Phenotype, Pseudomonas aeruginosa genetics, Pseudomonas aeruginosa isolation & purification, Pasteurellaceae genetics, Pasteurellaceae Infections diagnosis
- Abstract
Pasteurellaceae are bacteria with an important role as primary or opportunistic, mainly respiratory, pathogens in domestic and wild animals. Some species of Pasteurellaceae cause severe diseases with high economic losses in commercial animal husbandry and are of great diagnostic concern. Because of new data on the phylogeny of Pasteurellaceae, their taxonomy has recently been revised profoundly, thus requiring an improved phenotypic differentiation procedure to identify the individual species of this family. A new and simplified procedure to identify species of Actinobacillus, Avibacterium, Gallibacterium, Haemophilus, Mannheimia, Nicoletella, and Pasteurella, which are most commonly isolated from clinical samples of diseased animals in veterinary diagnostic laboratories, is presented in the current study. The identification procedure was evaluated with 40 type and reference strains and with 267 strains from routine diagnostic analysis of various animal species, including 28 different bacterial species. Type, reference, and field strains were analyzed by 16S ribosomal RNA (rrs) and rpoB gene sequencing for unambiguous species determination as a basis to evaluate the phenotypic differentiation schema. Primary phenotypic differentiation is based on beta-nicotinamide adenine dinucleotide (beta-NAD) dependence and hemolysis, which are readily determined on the isolation medium. The procedure divides the 28 species into 4 groups for which particular biochemical reactions were chosen to identify the bacterial species. The phenotypic identification procedure allowed researchers to determine the species of 240 out of 267 field strains. The procedure is an easy and cost-effective system for the rapid identification of species of the Pasteurellaceae family isolated from clinical specimens of animals.
- Published
- 2008
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50. Rapid diagnosis and quantification of Francisella tularensis in organs of naturally infected common squirrel monkeys (Saimiri sciureus).
- Author
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Abril C, Nimmervoll H, Pilo P, Brodard I, Korczak B, Markus S, Miserez R, and Frey J
- Subjects
- Animals, Animals, Zoo, Bacterial Outer Membrane Proteins genetics, Bacterial Typing Techniques veterinary, Francisella tularensis classification, Francisella tularensis genetics, Francisella tularensis isolation & purification, Humans, Microbial Sensitivity Tests veterinary, Monkey Diseases microbiology, Polymerase Chain Reaction veterinary, Tularemia diagnosis, Tularemia microbiology, Zoonoses microbiology, Francisella tularensis physiology, Monkey Diseases diagnosis, Saimiri microbiology, Tularemia veterinary
- Abstract
Francisella tularensis, a small Gram-negative facultative intracellular bacterium, is the causative agent of tularaemia, a severe zoonotic disease transmitted to humans mostly by vectors such as ticks, flies and mosquitoes. The disease is endemic in many parts of the northern hemisphere. Among animals, the most affected species belong to rodents and lagomorphs, in particular hares. However, in the recent years, many cases of tularaemia among small monkeys in zoos were reported. We have developed a real-time PCR that allows to quantify F. tularensis in tissue samples. Using this method, we identified the spleen and the kidney as the most heavily infected organ containing up to 400 F. tularensis bacteria per simian host cell in two common squirrel monkeys (Saimiri sciureus) from a zoo that died of tularaemia. In other organs such as the brain, F. tularensis was detected at much lower titres. The strain that caused the infection was identified as F. tularensis subsp. holarctica biovar I, which is susceptible to erythromycin. The high number of F. tularensis present in soft organs such as spleen, liver and kidney represents a high risk for persons handling such carcasses and explains the transmission of the disease to a pathologist during post-mortem analysis. Herein, we show that real-time PCR allows a reliable and rapid diagnosis of F. tularensis directly from tissue samples of infected animals, which is crucial in order to attempt accurate prophylactic measures, especially in cases where humans or other animals have been exposed to this highly contagious pathogen.
- Published
- 2008
- Full Text
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