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      Phenotypic and genotypic virulence features of staphylococcal strains isolated from difficult-to-treat skin and soft tissue infections

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          Abstract

          Chronic infections represent an important burden on the healthcare system and have a significant impact on the patients’ quality of life. While Staphylococcus spp. are commensal bacteria, they can become pathogenic, leading to various types of infections. In this study we aimed to characterize the virulence profiles of staphylococcal strains involved in difficult-to-treat skin and soft tissue infections, from both phenotypic and genotypic points of view. Phenotypic ability of the strains to secrete soluble virulence factors was assessed by a culturing dependent assay and their capacity to develop biofilms on inert substrate was screened by an adapted crystal violet microtiter method. We also tested the presence of several virulence genes by PCR. Most of the studied strains were isolated from purulent secretions of acne lesions and frequently secreted two or three soluble virulence factors. Most frequently secreted soluble virulence factors were caseinase (89%), lipase (71%) and lecithinase (67%). Almost half of the strains produced a well-represented biofilm. The molecular characterization showed the presence of the genes cna, hlg, clfA, and clfB. Staphylococcal strains that produce difficult-to-treat skin and soft tissue infections seem to be characterized by an enhanced ability to produce different soluble virulence factors and to develop biofilms in vitro. Further studies need to be developed in other Staphylococcus spp. infections in order to confirm this hypothesis.

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          Virulent combinations of adhesin and toxin genes in natural populations of Staphylococcus aureus.

          Most cases of severe Staphylococcus aureus disease cannot be explained by the action of a single virulence determinant, and it is likely that a number of factors act in combination during the infective process. This study examined the relationship between disease in humans and a large number of putative virulence determinants, both individually and in combination. S. aureus isolates (n = 334) from healthy blood donors and from patients with invasive disease were compared for variation in the presence of 33 putative virulence determinants. After adjusting for the effect of clonality, seven determinants (fnbA, cna, sdrE, sej, eta, hlg, and ica) were significantly more common in invasive isolates. All seven factors contributed independently to virulence. No single factor predominated as the major predictor of virulence, their effects appearing to be cumulative. No combinations of the seven genes were either more or less likely to cause disease than others with the same number of virulence-associated genes. There was evidence of considerable horizontal transfer of genes on a background of clonality. Our findings also suggested that allelic variants of a polymorphic locus can make different contributions to the disease process, further study of which is likely to expand our understanding of staphylococcal disease pathogenesis.
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            Involvement of Panton-Valentine leukocidin-producing Staphylococcus aureus in primary skin infections and pneumonia.

            Panton-Valentine leukocidin (PVL) is a cytotoxin that causes leukocyte destruction and tissue necrosis. It is produced by fewer than 5% of Staphylococcus aureus strains. A collection of 172 S. aureus strains were screened for PVL genes by polymerase chain reaction amplification. PVL genes were detected in 93% of strains associated with furunculosis and in 85% of those associated with severe necrotic hemorrhagic pneumonia (all community-acquired). They were detected in 55% of cellulitis strains, 50% of cutaneous abscess strains, 23% of osteomyelitis strains, and 13% of finger-pulp-infection strains. PVL genes were not detected in strains responsible for other infections, such as infective endocarditis, mediastinitis, hospital-acquired pneumonia, urinary tract infection, and enterocolitis, or in those associated with toxic-shock syndrome. It thus appears that PVL is mainly associated with necrotic lesions involving the skin or mucosa.
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              Nanomaterials for Wound Healing and Infection Control

              Wound healing has been intensely studied in order to develop an “ideal” technique that achieves expeditious recovery and reduces scarring to the minimum, thus ensuring function preservation. The classic approach to wound management is represented by topical treatments, such as antibacterial or colloidal agents, in order to prevent infection and promote a proper wound-healing process. Nanotechnology studies submicroscopic particles (maximum diameter of 100 nm), as well as correlated phenomena. Metal nanoparticles (e.g., silver, gold, zinc) are increasingly being used in dermatology, due to their beneficial effect on accelerating wound healing, as well as treating and preventing bacterial infections. Other benefits include: ease of use, less frequent dressing changes and a constantly moist wound environment. This review highlights recent findings regarding nanoparticle application in wound management.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: Project administrationRole: ValidationRole: VisualizationRole: Writing – original draft
                Role: Formal analysisRole: MethodologyRole: Writing – original draftRole: Writing – review & editing
                Role: Formal analysisRole: InvestigationRole: MethodologyRole: SoftwareRole: Writing – original draft
                Role: Formal analysisRole: MethodologyRole: SoftwareRole: Supervision
                Role: Formal analysisRole: MethodologyRole: ValidationRole: Writing – review & editing
                Role: Formal analysisRole: Visualization
                Role: Formal analysis
                Role: InvestigationRole: Writing – review & editing
                Role: Data curationRole: Formal analysisRole: Software
                Role: Formal analysisRole: Funding acquisitionRole: ResourcesRole: SupervisionRole: Validation
                Role: Formal analysisRole: Funding acquisitionRole: ResourcesRole: Supervision
                Role: Editor
                Journal
                PLoS One
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                2 February 2021
                2021
                : 16
                : 2
                : e0246478
                Affiliations
                [1 ] Department of Microbiology, Parasitology and Virology, Faculty of Midwives and Nursing, ‘Carol Davila’ University of Medicine and Pharmacy, Bucharest, Romania
                [2 ] ‘Cantacuzino’ National Medico-Military Research and Development Institute, Bucharest, Romania
                [3 ] Department of Oncologic Dermatology, ‘Carol Davila’ University of Medicine and Pharmacy, Bucharest, Romania
                [4 ] Department of Dermatology, ‘Elias’ University Emergency Hospital, Bucharest, Romania
                [5 ] Department of Bioinformatics, The National Institute of Research and Development for Biological Sciences, Bucharest, Romania
                [6 ] Research Institute of the University of Bucharest (ICUB), Bucharest, Romania
                [7 ] Department of Microbiology, Faculty of Biology, University of Bucharest, Bucharest, Romania
                [8 ] Department of Microbiology, Faculty of Dental Medicine, ‘Carol Davila’ University of Medicine and Pharmacy, Bucharest, Romania
                [9 ] Department of Microbiology, Faculty of Medicine, ‘Carol Davila’ University of Medicine and Pharmacy, Bucharest, Romania
                Mississippi State University, UNITED STATES
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                https://orcid.org/0000-0002-1436-0088
                Article
                PONE-D-20-22067
                10.1371/journal.pone.0246478
                7853507
                33529240
                ddc19c76-6e69-4fb2-9c71-d422c206dc1a
                © 2021 Preda et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 16 July 2020
                : 19 January 2021
                Page count
                Figures: 4, Tables: 4, Pages: 17
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100007700, Universitatea de Medicină şi Farmacie "Carol Davila" Bucureşti;
                Award ID: 12.659/20.05.2019
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100006730, Ministerul Educației și Cercetării Științifice;
                Award ID: PN-III-P4-ID-PCCF2016-0114
                Award Recipient :
                M.C.C. ”Selection and dissemination of antibiotic resistance genes from wastewater treatment plants into the aquatic environment and clinical reservoirs”; Grant number: PN-III-P4-ID-PCCF2016-0114. Funder: Ministry of Education and Research, https://uefiscdi.gov.ro/ M.P. Grant number: 12.659/20.05.2019; Funder: University of Medicine and Pharmacy "Carol Davila", Bucharest Romania; https://umfcd.ro/ The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
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                Biology and Life Sciences
                Organisms
                Bacteria
                Staphylococcus
                Staphylococcus Aureus
                Biology and Life Sciences
                Microbiology
                Medical Microbiology
                Microbial Pathogens
                Bacterial Pathogens
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