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      Virulence Factors Found in Nasal Colonization and Infection of Methicillin-Resistant Staphylococcus aureus (MRSA) Isolates and Their Ability to Form a Biofilm

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          Abstract

          Hospitalizations related to Methicillin-resistant Staphylococcus aureus (MRSA) are frequent, increasing mortality and health costs. In this way, this study aimed to compare the genotypic and phenotypic characteristics of MRSA isolates that colonize and infect patients seen at two hospitals in the city of Niterói—Rio de Janeiro, Brazil. A total of 147 samples collected between March 2013 and December 2015 were phenotyped and genotyped to identify the protein A (SPA) gene, the mec staphylococcal chromosomal cassette (SCC mec), mecA, Panton-Valentine Leucocidin (PVL), icaC, icaR, ACME, and hla virulence genes. The strength of biofilm formation has also been exploited. The prevalence of SCC mec type IV (77.1%) was observed in the colonization group; however, in the invasive infection group, SCC mec type II was prevalent (62.9%). The Multilocus Sequence Typing (MLST), ST5/ST30, and ST5/ST239 analyses were the most frequent clones in colonization, and invasive infection isolates, respectively. Among the isolates selected to assess the ability to form a biofilm, 51.06% were classified as strong biofilm builders. Surprisingly, we observed that isolates other than the Brazilian Epidemic Clone (BEC) have appeared in Brazilian hospitals. The virulence profile has changed among these isolates since the ACME type I and II genes were also identified in this collection.

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          Microbial biofilms.

          Direct observations have clearly shown that biofilm bacteria predominate, numerically and metabolically, in virtually all nutrient-sufficient ecosystems. Therefore, these sessile organisms predominate in most of the environmental, industrial, and medical problems and processes of interest to microbiologists. If biofilm bacteria were simply planktonic cells that had adhered to a surface, this revelation would be unimportant, but they are demonstrably and profoundly different. We first noted that biofilm cells are at least 500 times more resistant to antibacterial agents. Now we have discovered that adhesion triggers the expression of a sigma factor that derepresses a large number of genes so that biofilm cells are clearly phenotypically distinct from their planktonic counterparts. Each biofilm bacterium lives in a customized microniche in a complex microbial community that has primitive homeostasis, a primitive circulatory system, and metabolic cooperativity, and each of these sessile cells reacts to its special environment so that it differs fundamentally from a planktonic cell of the same species.
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            Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci.

            The details of all steps involved in the quantification of biofilm formation in microtiter plates are described. The presented protocol incorporates information on assessment of biofilm production by staphylococci, gained both by direct experience as well as by analysis of methods for assaying biofilm production. The obtained results should simplify quantification of biofilm formation in microtiter plates, and make it more reliable and comparable among different laboratories.
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              Emerging Strategies to Combat ESKAPE Pathogens in the Era of Antimicrobial Resistance: A Review

              The acronym ESKAPE includes six nosocomial pathogens that exhibit multidrug resistance and virulence: Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp. Persistent use of antibiotics has provoked the emergence of multidrug resistant (MDR) and extensively drug resistant (XDR) bacteria, which render even the most effective drugs ineffective. Extended spectrum β-lactamase (ESBL) and carbapenemase producing Gram negative bacteria have emerged as an important therapeutic challenge. Development of novel therapeutics to treat drug resistant infections, especially those caused by ESKAPE pathogens is the need of the hour. Alternative therapies such as use of antibiotics in combination or with adjuvants, bacteriophages, antimicrobial peptides, nanoparticles, and photodynamic light therapy are widely reported. Many reviews published till date describe these therapies with respect to the various agents used, their dosage details and mechanism of action against MDR pathogens but very few have focused specifically on ESKAPE. The objective of this review is to describe the alternative therapies reported to treat ESKAPE infections, their advantages and limitations, potential application in vivo, and status in clinical trials. The review further highlights the importance of a combinatorial approach, wherein two or more therapies are used in combination in order to overcome their individual limitations, additional studies on which are warranted, before translating them into clinical practice. These advances could possibly give an alternate solution or extend the lifetime of current antimicrobials.
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                Author and article information

                Journal
                Toxins (Basel)
                Toxins (Basel)
                toxins
                Toxins
                MDPI
                2072-6651
                25 December 2020
                January 2021
                : 13
                : 1
                : 14
                Affiliations
                [1 ]Biotechnology and Molecular Epidemiology Laboratory, Fluminense Federal University, Niteroi, 24220-900 Rio de Jeneiro, Brazil; thamiris.santana.machado@ 123456gmail.com (T.S.M.); felipepinheiro@ 123456id.uff.br (F.R.P.); lialyz@ 123456live.com (L.S.P.A.); renatafreireapereira@ 123456gmail.com (R.F.A.P.); gabrielac.mello@ 123456hotmail.com (G.C.d.M.)
                [2 ]Pathology Postgraduate Program, Fluminense Federal University, Niteroi, 24220-900 Rio de Janeiro, Brazil
                [3 ]Applied Microbiology and Parasitology Postgraduate Program, Fluminense Federal University, Niteroi, 24220-900 Rio de Janeiro, Brazil
                [4 ]Santa Martha Hospital, Department of Clinical Pathology, Niterói, 24241-265 Rio de Janeiro, Brazil; correaregi@ 123456hotmail.com
                [5 ]Microbiological Testing Laboratory Associated with Materials and Drugs of the Center for Studies, Research and Innovation in Biofunctional Materials and Biotechnology, Federal University of Itajubá, 37500-903 Itajubá, Brazil; tainara.ribeiro@ 123456unifei.edu.br (T.A.N.R.); danisachs@ 123456gmail.com (D.S.)
                [6 ]Gram-Positive Cocci Laboratory, Biomedical Institute, Fluminense Federal University, Niteroi, 24220-900 Rio de Janeiro, Brazil; bpenna@ 123456id.uff.br
                Author notes
                [* ]Correspondence: faalves@ 123456gmail.com ; Tel.: +55-(21)-9852-5189
                Author information
                https://orcid.org/0000-0002-2776-1331
                https://orcid.org/0000-0002-7917-901X
                https://orcid.org/0000-0001-6235-9367
                Article
                toxins-13-00014
                10.3390/toxins13010014
                7823648
                33375552
                80c5ad06-b66e-49f0-bd7e-382d8075c3df
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 21 August 2020
                : 27 October 2020
                Categories
                Article

                Molecular medicine
                mrsa,resistance profile,virulence factors,biofilm,genotyping
                Molecular medicine
                mrsa, resistance profile, virulence factors, biofilm, genotyping

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