title: Antimicrobial-resistant Escherichia coli and Enterococcus spp. isolated from Miranda donkey (Equus asinus): an old problem from a new source with a different approach.
abstract: OBJECTIVE
  The Miranda donkey (Equus asinus) is an endangeredasinine from Miranda do Douro region, located in the north east of Portugal. We studied the antimicrobial resistance and virulence genes in Escherichia coli and Enterococcus spp. isolates from these animals.
  
  
  METHODS
  In March 2014, a total of 66 faecal samples were recovered from independent animals. Antibiotic resistance was determined by the disc diffusion method. Carriage of genes coding for antibiotic-resistant and virulent factors was analysed by PCR.
  
  
  RESULTS
  A total of 66 E. coli and 41 enterococcal isolates were detected, with Enterococcus faecium (61 %) and Enterococcus hirae (24 %) being the most prevalent species. For enterococcal isolates, high percentages of resistance rates to tetracycline (68.3 %), quinupristin/dalfopristin (51.2 %) and ciprofloxacin (48.8 %) were observed. The genes erm(A) and/or erm(B), tet(M) and/or tet(L), vat(D) and/or vat(E) and aph(3')-IIIa were also found. The most frequent virulence gene detected was gel(E), followed by ace, cpd and hyl. Escherichia coli isolates were highly resistant to streptomycin (78 %), whereas 39 % of them exhibited resistance to aminoglycosides and tetracycline. Genes sul1 and/or sul2 were detected in 66.7 % of trimethoprim/sulfamethoxazole-resistant isolates. The virulence genes detected were fim(A) (46 %) and cnf1 (27 %).
  
  
  CONCLUSIONS
  To the best of our knowledge, this is the first report showing antibiotic resistance among Escherichiacoli and Enterococcus spp. isolates from the Miranda donkey in Portugal, indicating possible antibiotic-resistant bacterial reservoirs. However, the detection of these resistances presents a low risk for other animals and human beings in that rural area.
authors.affiliation: Functional Genomics and Proteomics Unit, Vila Real, Portugal.|Department of Genetics and Biotechnology, Vila Real, Portugal.|Department of Veterinary Sciences, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal.
authors.forename: Isabel
authors.initials: I
authors.lastname: Carvalho
authors.affiliation: Microbiology Department, University Hospital Ramón y Cajal, and Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Madrid, and Red Española de Investigación en Patología Infecciosa (REIPI); Spain.
authors.forename: Rosa Del
authors.initials: RD
authors.lastname: Campo
authors.affiliation: Functional Genomics and Proteomics Unit, Vila Real, Portugal.|Department of Genetics and Biotechnology, Vila Real, Portugal.|Department of Veterinary Sciences, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal.
authors.forename: Margarida
authors.initials: M
authors.lastname: Sousa
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Penicuik, UK.
authors.forename: Nuno
authors.initials: N
authors.lastname: Silva
authors.affiliation: Environmental and Biological Department and CITAB, University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal.
authors.forename: João
authors.initials: J
authors.lastname: Carrola
authors.affiliation: Functional Genomics and Proteomics Unit, Vila Real, Portugal.
authors.forename: Catarina
authors.initials: C
authors.lastname: Marinho
authors.affiliation: Functional Genomics and Proteomics Unit, Vila Real, Portugal.
authors.forename: Tiago
authors.initials: T
authors.lastname: Santos
authors.affiliation: Department of Genetics and Biotechnology, Vila Real, Portugal.|Department of Veterinary Sciences, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal.
authors.forename: Sílvia
authors.initials: S
authors.lastname: Carvalho
authors.affiliation: Association for the Study and Protection of Asinine Cattle (AEPGA), Sendim, Portugal.
authors.forename: Miguel
authors.initials: M
authors.lastname: Nóvoa
authors.affiliation: Department of Veterinary Sciences, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal.
authors.forename: Miguel
authors.initials: M
authors.lastname: Quaresma
authors.affiliation: Animal and Veterinary Research Centre (CECAV), Vila Real, Portugal.|Department of Veterinary Sciences, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal.
authors.forename: José Eduardo
authors.initials: JE
authors.lastname: Pereira
authors.affiliation: Microbiology Department, University Hospital Ramón y Cajal, and Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Madrid, and Red Española de Investigación en Patología Infecciosa (REIPI); Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Cobo
authors.affiliation: UCIBIO-REQUIMTE, Faculty of Science and Technology, New University of Lisbon, Monte da Caparica, Portugal.|Functional Genomics and Proteomics Unit, Vila Real, Portugal.|Department of Genetics and Biotechnology, Vila Real, Portugal.
authors.forename: Gilberto
authors.initials: G
authors.lastname: Igrejas
authors.affiliation: Department of Veterinary Sciences, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal.|UCIBIO-REQUIMTE, Faculty of Science and Technology, New University of Lisbon, Monte da Caparica, Portugal.
authors.forename: Patrícia
authors.initials: P
authors.lastname: Poeta
mesh_terms: D000617:Aminoglycosides / Q000494:pharmacology; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics; D002939:Ciprofloxacin / Q000494:pharmacology; D004269:DNA, Bacterial / Q000235:genetics; D024901:Drug Resistance, Multiple, Bacterial; D016983:Enterococcus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D018397:Equidae / Q000382:microbiology*; D004926:Escherichia coli / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D019868:Kanamycin Kinase / Q000235:genetics; D008780:Methyltransferases / Q000235:genetics; D008826:Microbial Sensitivity Tests; D011174:Portugal; D013420:Sulfamethoxazole / Q000494:pharmacology; D013752:Tetracycline / Q000494:pharmacology; D014295:Trimethoprim / Q000494:pharmacology; D014769:Virginiamycin / Q000494:pharmacology
chemical_list: D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D014769:Virginiamycin; C062940:quinupristin-dalfopristin; D002939:Ciprofloxacin; D014295:Trimethoprim; D008780:Methyltransferases; C431709:ErmA protein, Bacteria; D019868:Kanamycin Kinase; D013752:Tetracycline; D013420:Sulfamethoxazole