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      Ethanol and Isopropyl Alcohol Exposure Increases Biofilm Formation in Staphylococcus aureus and Staphylococcus epidermidis

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          Abstract

          Introduction

          Alcohols, including ethanol and isopropyl alcohol, are used in clinical practice for disinfection and infection prevention. Recent studies, however, demonstrate that alcohols may enhance biofilm production in Staphylococci.

          Methods

          We quantified biofilm formation in the presence of ethanol and isopropyl alcohol in six different, well-characterized strains of Staphylococcus epidermidis and Staphylococcus aureus. After 24 h of biofilm development, each strain was exposed to normal saline (NS), ethanol, or isopropyl alcohol (40%, 60%, 80% and 95%) for additional 24 h incubation. Adherent biofilms were stained and optical density was determined. Viability of strains was also determined after alcohol exposure.

          Results

          Ethanol increased biofilm formation in all six strains compared to normal saline ( p < 0.05). There was increased biofilm formation with increasing ethanol concentration. Isopropyl alcohol also increased biofilm formation with increasing alcohol concentration in all six strains ( p < 0.01 vs NS). The slime-negative, chemical mutant strain of S. epidermidis increased biofilm formation after exposure to both alcohols, likely reverting back its primary phenotype through modulation of the intercellular adhesin repressor. All strains demonstrated viability after exposure to each alcohol concentration, though viability was decreased.

          Conclusion

          Ethanol and isopropyl alcohol exposure increases biofilm formation of S. aureus and S. epidermidis at concentrations used in clinical settings. Ethanol and isopropyl alcohol did not eradicate viable Staphylococci from formed biofilm.

          Electronic supplementary material

          The online version of this article (doi:10.1007/s40121-015-0065-y) contains supplementary material, which is available to authorized users.

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          Most cited references23

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          Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices.

          The adherence of coagulase-negative staphylococci to smooth surfaces was assayed by measuring the optical densities of stained bacterial films adherent to the floors of plastic tissue culture plates. The optical densities correlated with the weight of the adherent bacterial film (r = 0.906; P less than 0.01). The measurements also agreed with visual assessments of bacterial adherence to culture tubes, microtiter plates, and tissue culture plates. Selected clinical strains were passed through a mouse model for foreign body infections and a rat model for catheter-induced endocarditis. The adherence measurements of animal passed strains remained the same as those of the laboratory-maintained parent strain. Spectrophotometric classification of coagulase-negative staphylococci into nonadherent and adherent categories according to these measurements had a sensitivity, specificity, and accuracy of 90.6, 80.8, and 88.4%, respectively. We examined a previously described collection of 127 strains of coagulase-negative staphylococci isolated from an outbreak of intravascular catheter-associated sepsis; strains associated with sepsis were more adherent than blood culture contaminants and cutaneous strains (P less than 0.001). We also examined a collection of 84 strains isolated from pediatric patients with cerebrospinal fluid (CSF) shunts; once again, pathogenic strains were more adherent than were CSF contaminants (P less than 0.01). Finally, we measured the adherence of seven endocarditis strains. As opposed to strains associated with intravascular catheters and CSF shunts, endocarditis strains were less adherent than were saprophytic strains of coagulase-negative staphylococci. The optical densities of bacterial films adherent to plastic tissue culture plates serve as a quantitative model for the study of the adherence of coagulase-negative staphylococci to medical devices, a process which may be important in the pathogenesis of foreign body infections.
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            Biofilm matrix of Candida albicans and Candida tropicalis: chemical composition and role in drug resistance.

            Matrix material was extracted from biofilms of Candida albicans and Candida tropicalis and analysed chemically. Both preparations contained carbohydrate, protein, hexosamine, phosphorus and uronic acid. However, the major component in C. albicans matrix was glucose (32%), whereas in C. tropicalis matrix it was hexosamine (27%). Biofilms of C. albicans were more easily detached from plastic surfaces by treatment with the enzyme lyticase (beta-1,3-glucanase) than were those of C. tropicalis. Biofilms of C. albicans were also partially detached by treatment with proteinase K, chitinase, DNase I, or beta-N-acetylglucosaminidase, whereas C. tropicalis biofilms were only affected by lipase type VII or chitinase. To investigate a possible role for the matrix in biofilm resistance to antifungal agents, biofilms of C. albicans were grown under conditions of continuous flow in a modified Robbins device (MRD). These biofilms produced more matrix material than those grown statically, and were significantly more resistant to amphotericin B. Biofilms of C. tropicalis synthesized large amounts of matrix material even when grown statically, and such biofilms were completely resistant to both amphotericin B and fluconazole. Mixed-species biofilms of C. albicans and a slime-producing strain of Staphylococcus epidermidis (RP62A), when grown statically or in the MRD, were also completely resistant to amphotericin B and fluconazole. Mixed-species biofilms of C. albicans and a slime-negative mutant of S. epidermidis (M7), on the other hand, were completely drug resistant only when grown under flow conditions. These results demonstrate that the matrix can make a significant contribution to drug resistance in Candida biofilms, especially under conditions similar to those found in catheter infections in vivo, and that the composition of the matrix material is an important determinant in resistance.
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              Ethanol tolerance in bacteria.

              The adverse effects of ethanol on bacterial growth, viability, and metabolism are caused primarily by ethanol-induced leakage of the plasma membrane. This increase in membrane leakage is consistent with known biophysical properties of membranes and ethanolic solutions. The primary actions of ethanol result from colligative effects of the high molar concentrations rather than from specific interactions with receptors. The ethanol tolerance of growth in different microorganisms appears to result in large part from adaptive and evolutionary changes in cell membrane composition. Different cellular activities vary in their tolerance to ethanol. Therefore, it is essential that the aspect of cellular function under study be specifically defined and that comparisons of ethanol tolerance among systems share this common definition. Growth is typically one of the most sensitive cellular activities to inhibition by ethanol, followed by survival, or loss of reproductive ability. Glycolysis is the most resistant of these three activities. Since glycolysis is an exergonic process, a cell need not be able to grow or remain viable for glycolysis to occur.
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                Author and article information

                Contributors
                kerrylaplante@uri.edu
                Journal
                Infect Dis Ther
                Infect Dis Ther
                Infectious Diseases and Therapy
                Springer Healthcare (Cheshire )
                2193-8229
                2193-6382
                3 May 2015
                3 May 2015
                June 2015
                : 4
                : 2
                : 219-226
                Affiliations
                [ ]Department of Pharmacy Practice, University of Rhode Island, 7 Greenhouse Road, Kingston, RI 02881 USA
                [ ]Rhode Island Infectious Diseases (RIID) Research Program Laboratory, Veterans Affairs Medical Center, Providence, RI USA
                [ ]Rhode Island College, Providence, RI USA
                [ ]Warren Alpert Medical School of Brown University, Providence, RI USA
                [ ]Rhode Island Hospital, Providence, RI USA
                Article
                65
                10.1007/s40121-015-0065-y
                4471055
                25935134
                ea0ee58b-3e9d-4512-b5bb-cfc7bb877733
                © The Author(s) 2015
                History
                : 25 February 2015
                Categories
                Brief Report
                Custom metadata
                © Springer Healthcare 2015

                alcohol,biofilm,ethyl alcohol,isopropanol,staphylococcus aureus,staphylococcus epidermidis

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