title: Molecular typing, virulence traits and antimicrobial resistance of diabetic foot staphylococci.
abstract: BACKGROUND
  Diabetes mellitus is a major chronic disease that continues to increase significantly. One of the most important and costly complications of diabetes are foot infections that may be colonized by pathogenic and antimicrobial resistant bacteria, harboring several virulence factors, that could impair its successful treatment. Staphylococcus aureus is one of the most prevalent isolate in diabetic foot infections, together with aerobes and anaerobes.
  
  
  METHODS
  In this study, conducted in the Lisbon area, staphylococci isolated (n = 53) from diabetic foot ulcers were identified, genotyped and screened for virulence and antimicrobial resistance traits. Genetic relationship amongst isolates was evaluated by pulsed-field-gel-electrophoresis with further multilocus sequence typing of the identified pulsotypes. PCR was applied for detection of 12 virulence genes and e-test technique was performed to determine minimal inhibitory concentration of ten antibiotics.
  
  
  RESULTS
  Among the 53 isolates included in this study, 41 Staphylococcus aureus were identified. Staphylococcal isolates were positive for intercellular adhesins icaA and icaD, negative for biofilm associated protein bap and pantone-valentine leucocidin pvl. S. aureus quorum sensing genes agrI and agrII were identified and only one isolate was positive for toxic shock syndrome toxin tst. 36 % of staphylococci tested were multiresistant and higher rates of resistance were obtained for ciprofloxacin and erythromycin. Clonality analysis revealed high genomic diversity and numerous S. aureus sequence types, both community- and hospital-acquired, belonging mostly to clonal complexes CC5 and C22, widely diffused in Portugal nowadays.
  
  
  CONCLUSIONS
  This study shows that diabetic foot ulcer staphylococci are genomically diverse, present resistance to medically important antibiotics and harbour virulence determinants. These properties suggest staphylococci can contribute to persistence and severity of these infections, leading to treatment failure and to the possibility of transmitting these features to other microorganisms sharing the same niche. In this context, diabetic patients may become a transmission vehicle for microorganisms' clones between community and clinical environments.
authors.affiliation: Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinária, Avenida da Universidade Técnica, 1300-477, Lisbon, Portugal. carlamottola@fmv.ulisboa.pt.
authors.forename: Carla
authors.initials: C
authors.lastname: Mottola
authors.affiliation: BioFIG, Centro para a Biodiversidade, Genómica Integrativa e Funcional, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016, Lisbon, Portugal. tmsemedo@fc.ul.pt.
authors.forename: Teresa
authors.initials: T
authors.lastname: Semedo-Lemsaddek
authors.affiliation: Departamento de Medicina Interna, Hospital de Santa Marta/Centro Hospitalar de Lisboa Central, EPE, Rua de Santa Marta, 1169-024, Lisbon, Portugal. joaojoaomendes@hotmail.com.
authors.forename: João J
authors.initials: JJ
authors.lastname: Mendes
authors.affiliation: Faculdade de Medicina, Universidade de Lisboa, Instituto de Microbiologia, Avenida Prof. Egas Moniz, 1649-028, Lisbon, Portugal. melo_cristino@medicina.ulisboa.pt.
authors.forename: José
authors.initials: J
authors.lastname: Melo-Cristino
authors.affiliation: Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinária, Avenida da Universidade Técnica, 1300-477, Lisbon, Portugal. ltavares@fmv.ulisboa.pt.
authors.forename: Luís
authors.initials: L
authors.lastname: Tavares
authors.affiliation: TechnoPhage, S.A., Avenida Prof. Egas Moniz, 1600-190, Lisbon, Portugal. patsilva@netcabo.pt.|Centro de Investigação Interdisciplinar Egas Moniz (CiiEM), Instituto Superior de Ciências da Saúde Egas Moniz, Via Alternativa ao Monte de Caparica, 2829-511, Caparica, Portugal. patsilva@netcabo.pt.
authors.forename: Patrícia
authors.initials: P
authors.lastname: Cavaco-Silva
authors.affiliation: Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinária, Avenida da Universidade Técnica, 1300-477, Lisbon, Portugal. moliveira@fmv.ulisboa.pt.
authors.forename: Manuela
authors.initials: M
authors.lastname: Oliveira
mesh_terms: D017719:Diabetic Foot / Q000382:microbiology*; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D005260:Female; D006801:Humans; D008297:Male; D019655:Quantitative Trait, Heritable; D013211:Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity; D037521:Virulence Factors / Q000235:genetics*
chemical_list: D037521:Virulence Factors