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      MINIMUM BACTERICIDAL CONCENTRATION OF COMMERCIAL DISINFECTANTS ON STAPHYLOCOCCUS SPP. ISOLATED FROM MASTITIS IN GOATS AND DETECTION OF THE icaD GENE Translated title: CONCENTRAÇÃO BACTERICIDA MÍNIMA DE DESINFETANTES COMERCIAIS FRENTE À STAPHYLOCOCCUS SPP. ISOLADOS DE MASTITE EM CAPRINOS E DETECÇÃO DO GENE icaD

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          Abstract

          Abstract This study aimed to evaluate the in vitro efficacy of commercial disinfectants on Staphylococcus spp., isolated from mastitis cases in goats, and to associate the observed resistance with the presence of the icaD gene. Broth microdilution was employed to evaluate the in vitro antimicrobial activity of the disinfectants, whereas the Congo Red technique was used for the evaluation of biofilm production and amplification of the icaD gene. All evaluated samples were sensitive to disinfectants, with the following ranges of activity: quaternary ammonium (0.13 - 21.33 µg/ml), chlorhexidine (4.00 - 313.00 µg/ml) and iodine (190.00 - 12500.00 µg/ml); however, the sodium hypochlorite-based disinfectant showed no bactericidal activity in the concentration range from 15.0 to 0.03 µg/ml. The icaD gene presented a frequency of 14.7% in the isolate samples. Fisher’s exact test showed a significant effect of the relation between the minimum bactericidal concentration value of the quaternary ammonium-based disinfectant and the presence/absence of the icaD gene (P <0.01). Commercial disinfectants with quaternary ammonium, chlorhexidine and iodine active ingredients presented in vitro activity even at concentrations lower than those recommended by the manufacturers. Therefore, the periodic evaluation of the sensitivity profile of the disinfectants must be performed.

          Translated abstract

          Resumo O presente trabalho objetivou avaliar a eficácia in vitro de desinfetantes comerciais frente à Staphylococcus spp. isolados de casos de mastite em cabras e, relacionar a resistência observada com a presença do gene icaD. A microdiluição em caldo foi utilizada para avaliar a atividade antimicrobiana in vitro dos desinfetantes, enquanto a técnica de vermelho congo para a avaliação da produção de biofilme e amplificação do gene icaD. Todas as amostras avaliadas foram sensíveis aos desinfetantes, com as seguintes faixas de atividade: amônia quaternária (0,13 - 21,33 µg/ml), clorexidina (4,00 - 313,00 µg/ml) e iodo (190,00 - 12500,00 µg/ml), no entanto o desinfetante a base de hipoclorito de sódio não apresentou atividade bactericida na faixa de concentração 15,0 a 0,03 µg/ml. O gene icaD apresentou frequência de 14,7% nos isolados. O teste exato de Fisher apontou efeito significativo da relação entre o valor da concentração bactericida mínima do desinfetante à base de amônia quaternária e a presença/ausência do gene icaD (P<0,01). Os desinfetantes comerciais com os princípios ativos amônia quaternária, clorexidina e iodo, apresentaram atividade in vitro até mesmo em concentrações inferiores às preconizadas pelos fabricantes o que torna necessária a realização periódica da avaliação do perfil de sensibilidade dos desinfetantes.

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          Biofilm formation in Staphylococcus implant infections. A review of molecular mechanisms and implications for biofilm-resistant materials.

          Implant infections in orthopaedics, as well as in many other medical fields, are chiefly caused by staphylococci. The ability of growing within a biofilm enhances the chances of staphylococci to protect themselves from host defences, antibiotic therapies, and biocides. Advances in scientific knowledge on structural molecules (exopolysaccharide, proteins, teichoic acids, and the most recently described extracellular DNA), on the synthesis and genetics of staphylococcal biofilms, and on the complex network of signal factors that intervene in their control are here presented, also reporting on the emerging strategies to disrupt or inhibit them. The attitude of polymorphonuclear neutrophils and macrophages to infiltrate and phagocytise biofilms, as well as the ambiguous behaviour exhibited by these innate immune cells in biofilm-related implant infections, are here discussed. Research on anti-biofilm biomaterials is focused, reviewing materials loaded with antibacterial substances, or coated with anti-adhesive/anti-bacterial immobilized agents, or surfaced with nanostructures. Latter approaches appear promising, since they avoid the spread of antibacterial substances in the neighbouring tissues with the consequent risk of inducing bacterial resistance. Copyright © 2012 Elsevier Ltd. All rights reserved.
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            Presence of icaA and icaD genes and slime production in a collection of staphylococcal strains from catheter-associated infections.

            Both Staphylococcus epidermidis and Staphylococcus aureus are important causes of infections associated with catheters and other medical devices. It has recently been shown that not only S. epidermidis but also S. aureus can produce slime and carries the ica operon responsible for slime production. In the operon, coexpression of icaA and icaD is required for full slime synthesis. In this study, the presence of icaA and icaD was determined in a collection of 91 staphylococcal (68 S. epidermidis and 23 S. aureus) strains from intravenous catheter-associated infections, in 10 strains from the skin and mucosa of healthy volunteers, and in two reference strains by a PCR method. Slime-forming ability was tested on Congo red agar plates; 49% of S. epidermidis strains from catheters and, surprisingly, 61% of S. aureus strains were icaA and icaD positive and slime forming. All the saprophytic strains turned out to be negative for both icaA and icaD and also non-slime forming. Two S. aureus and one S. epidermidis strain from catheters, detected as icaA and icaD positive by PCR analysis and as slime forming (black colonies) at 24 h on Congo red agar, at 48 h exhibited tiny red spikes at the center of black colonies. The onset of these variants could not be ascribed to a mutagenic potential of Congo red, which, in the Ames test, was devoid of mutagenicity. PCR analysis showed that these red variants were negative for both icaA and icaD and even lacking the entire icaADBC operon. The data reported indicate an important role of ica genes as a virulence marker in staphylococcal infections from intravenous catheters.
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              Characterization of the N-acetylglucosaminyltransferase activity involved in the biosynthesis of the Staphylococcus epidermidis polysaccharide intercellular adhesin.

              The polysaccharide intercellular adhesin (PIA) is an important factor in the colonization of medical devices by Staphylococcus epidermidis. The genes encoding PIA production are organized in the icaADBC (intercellular adhesion) operon. To study the function of the individual genes, we have established an in vitro assay with UDP-N-acetylglucosamine, the substrate for PIA biosynthesis, and analyzed the products by thin-layer chromatography and mass spectrometry. IcaA alone exhibited a low N-acetylglucosaminyltransferase activity and represents the catalytic enzyme. Coexpression of icaA with icaD led to a significant increase in activity. The newly identified icaD gene is located between icaA and icaB and overlaps both genes. N-Acetylglucosamine oligomers produced by IcaAD reached a maximal length of 20 residues. Only when icaA and icaD were expressed together with icaC were oligomer chains that react with PIA-specific antiserum synthesized. IcaA and IcaD are located in the cytoplasmic membrane, and IcaC also has all the structural features of an integral membrane protein. These results indicate a close interaction between IcaA, IcaD, and IcaC. Tunicamycin and bacitracin did not affect the in vitro synthesis of PIA intermediates or the complete PIA biosynthesis in vivo, suggesting that a undecaprenyl phosphate carrier is not involved. IcaAD represents a novel protein combination among beta-glycosyltransferases.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                cab
                Ciência Animal Brasileira
                Ciênc. anim. bras.
                Universidade Federal de Goiás (Goiânia, GO, Brazil )
                1518-2797
                1809-6891
                2019
                : 20
                : e53193
                Affiliations
                [3] Petrolina PE orgnameInstituto Federal do Sertão Pernambucano Brazil
                [2] Petrolina orgnameUniversidade Federal do Vale do São Francisco Brazil
                [1] Araripina Pernambuco orgnameFaculdade de Ciências Agrárias de Araripina Brazil
                Article
                S1809-68912019000100318
                10.1590/1089-6891v20e-53193
                861d00c3-4017-4f3f-8890-3c264adb862a

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 26 May 2018
                : 20 March 2019
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 22, Pages: 0
                Product

                SciELO Brazil

                Categories
                Veterinary Medicine

                ordenha,caprinos leiteiros,biofilme,atividade antimicrobiana,chemical substances,milking,dairy goat,biofilm,antimicrobial activity,produtos químicos

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