title: Effect of Photodynamic Therapy on the Virulence Factors of Staphylococcus aureus.
abstract: Staphylococcus aureus is a Gram-positive bacterium that is present in the human microbiota. Nevertheless, these bacteria can be pathogenic to the humans. Due to the increasing occurrence of antibiotic-resistant S. aureus strains, new approaches to control this pathogen are necessary. The antimicrobial photodynamic inactivation (PDI) process is based in the combined use of light, oxygen, and an intermediary agent (a photosensitizer). These three components interact to generate cytotoxic reactive oxygen species that irreversibly damage vital constituents of the microbial cells and ultimately lead to cell death. Although PDI is being shown to be a promising alternative to the antibiotic approach for the inactivation of pathogenic microorganisms, information on effects of photosensitization on particular virulence factors is strikingly scarce. The objective of this work was to evaluate the effect of PDI on virulence factors of S. aureus and to assess the potential development of resistance of this bacterium as well as the recovery of the expression of the virulence factors after successive PDI cycles. For this, the photosensitizer 5,10,15,20-tetrakis(1-methylpyridinium-4-yl)porphyrin tetra-iodide (Tetra-Py(+)-Me) and six strains of S. aureus [one reference strain, one strain with one enterotoxin, two strains with three enterotoxins and two methicillin resistant strains (MRSA) - one with five enterotoxins and the other without enterotoxins] were used. The effect of photosensitization on catalase activity, beta hemolysis, lipases, thermonuclease, enterotoxins, coagulase production, and resistance/susceptibility to methicillin was tested. To assess the development of resistance after successive cycles of treatment, three strains of S. aureus (ATCC 6538, 2065 MA, and SA 3 MRSA) were used. The surviving colonies of a first cycle of PDI were collected from the solid medium and subjected to further nine consecutive cycles of PDI. The results indicate that the expression of some external virulence factors is affected by PDI and enterotoxin producing strains were more susceptible to PDI than non-toxigenic strains. The surviving bacteria did not develop resistance. PDI, contrarily to traditional antibiotics, inhibited the expression of virulence factors, efficiently inactivating either highly virulent strains and low virulent S. aureus strains, inactivating also antibiotic susceptible and resistant strains, without development of photoresistance after at least 10 consecutive cycles of treatment, and so this therapy may become a strong promising alternative to antibiotics to control pathogenic microorganisms.
authors.affiliation: Departamento de Biologia and Centro de Estudos do Ambiente e do Mar, University of Aveiro Aveiro, Portugal.
authors.forename: Maria
authors.initials: M
authors.lastname: Bartolomeu
authors.affiliation: Departamento de Biologia and Centro de Estudos do Ambiente e do Mar, University of Aveiro Aveiro, Portugal.
authors.forename: Sónia
authors.initials: S
authors.lastname: Rocha
authors.affiliation: Departamento de Biologia and Centro de Estudos do Ambiente e do Mar, University of Aveiro Aveiro, Portugal.
authors.forename: Ângela
authors.initials: Â
authors.lastname: Cunha
authors.affiliation: Departamento de Química and Unidade de Investigação em Química Orgânica, Produtos Naturais e Agroalimentares, University of Aveiro Aveiro, Portugal.
authors.forename: M G P M S
authors.initials: MG
authors.lastname: Neves
authors.affiliation: Departamento de Química and Unidade de Investigação em Química Orgânica, Produtos Naturais e Agroalimentares, University of Aveiro Aveiro, Portugal.
authors.forename: Maria A F
authors.initials: MA
authors.lastname: Faustino
authors.affiliation: Departamento de Biologia and Centro de Estudos do Ambiente e do Mar, University of Aveiro Aveiro, Portugal.
authors.forename: Adelaide
authors.initials: A
authors.lastname: Almeida
keywords: Staphylococcus aureus; antibiotic/methicillin resistance; coagulase; enterotoxins; photodynamic inactivation (PDI); virulence factors