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- Staphylococcus aureusandEscherichia colidual-species biofilms on nanohydroxyapatite loaded with CHX or ZnO nanoparticlesPublication . Barros, Joana; Grenho, Liliana; Fontenente, Sílvia; Manuel, Cândida M.; Nunes, Olga C.; Melo, Luís F.; Monteiro, Fernando J.; Ferraz, Maria PiaImplant-associated infections are caused by surface-adhering microorganisms persisting as biofilms, resistant to host defense and antimicrobial agents. Given the limited efficacy of traditional antibiotics, novel strategies may rely on the prevention of such infections through the design of new biomaterials. In this work, two antimicrobial agents applied to nanohydroxyapatite materials-namely, chlorhexidine digluconate (CHX) and zinc oxide (ZnO) nanoparticles-were compared concerning their ability to avoid single- or dual-species biofilms of Staphylococcus aureus and Escherichia coli. The resulting biofilms were quantified by the enumeration of colony-forming units and examined by confocal microscopy using both Live/Dead staining and bacterial-specific fluorescent in situ hybridization. The sessile population arrangement was also observed by scanning electron microscopy. Both biomaterials showed to be effective in impairing bacterial adhesion and proliferation for either single- or dual-species biofilms. Furthermore, a competitive interaction was observed for dual-species biofilms wherein E. coli exhibited higher proliferative capacity than S. aureus, an inverse behavior from the one observed in single-species biofilms. Therefore, either nanoHA-CHX or nanoHA-ZnO surfaces appear as promising alternatives to antibiotics for the prevention of devices-related infections avoiding the critical risk of antibiotic-resistant strains emergence. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 491-497, 2017.
- Lytic bacteriophages against multidrug-resistant Staphylococcus aureus, Enterococcus faecalis and Escherichia coli isolates from orthopaedic implant-associated infectionsPublication . Barros, Joana; Melo, Luís D.R.; Poeta, Patrícia; Igrejas, Gilberto; Ferraz, Maria Pia; Azeredo, Joana; Monteiro, Fernando J.Orthopaedic implant-associated infections are a devastating complication of orthopaedic surgery with a significant impact on patients and healthcare systems. The aims of this work were to describe the patterns of antimicrobial resistance, pathogenicity and virulence of clinical bacterial isolates from orthopaedic implant-associated infections and to further isolate and characterise bacteriophages that are efficient in controlling these bacteria. Staphylococcus aureus, Enterococcus faecalis and Escherichia coli isolated from orthopaedic infections showed multiresistance patterns to the most frequently used antibiotics in clinical settings. The presence of mobile genetic elements (mecA, Tn916/Tn1545 and intl1) and virulence determinants (icaB, cna, hlb, cylLs, cylM, agg, gelE, fsr and fimA) highlighted the pathogenicity of these isolates. Moreover, the isolates belonged to clonal complexes associated with the acquisition of pathogenicity islands and antimicrobial resistance genes by recombination and horizontal gene transfer. Bacteriophages vB_SauM_LM12, vB_EfaS_LM99 and vB_EcoM_JB75 were characterised and their ability to infect clinical isolates of S. aureus, E. faecalis and E. coli, respectively, was assessed. Morphological and genomic analyses revealed that vB_EfaS_LM99 and vB_EcoM_JB75 belong to the Siphoviridae and Myoviridae families, respectively, and no genes associated with lysogeny were found. The bacteriophages showed low latent periods, high burst sizes, broad host ranges and tolerance to several environmental conditions. Moreover, they showed high efficiency and specificity to infect and reduce clinical bacteria, including methicillin-resistant S. aureus and vancomycin-resistant enterococci. Therefore, the results obtained suggest that the bacteriophages used in this work are a promising approach to control these pathogens involved in orthopaedic implant-associated infections.