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      Analysis of the Adaptation Capacity of Staphylococcus aureus to Commonly Used Antiseptics by Microplate Laser Nephelometry

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          Abstract

          Background: Bacterial colonization and infection are important factors in compromised wound healing. Antiseptics have become an alternative for antimicrobial applications as antibiotic resistance is increasing; they have multiple targets with a broad spectrum of activity. Hence, the risk for developing resistance should be low. However, concerns have been raised that their growing use may result in bacteria that are less susceptible. Methods: The capacity of common antiseptics such as silver nitrate, polihexanide, octenidine, chlorhexidine and polyvinylpyrrolidone (PVP)-iodine to induce adaptation in a Staphylococcus aureus strain was analyzed in vitro using microplate laser nephelometry. S. aureus was repeatedly incubated with the respective half maximal inhibitory concentration (IC<sub>50</sub>) over a time period of 100 days. The influence of the continued treatment was determined by in situ monitoring of changes in the dose-response curves and calculation of the current IC<sub>50</sub> values for the substances tested. Results: During the experiment, S. aureus quickly adapted to high concentrations of the antibiotic mupirocin during repeated treatment. Moreover, a significant increase of the IC<sub>50</sub> for silver nitrate was observed over time. On the other hand, no significant difference was observed for polihexanide or chlorhexidine. While the IC<sub>50</sub> for octenidine was also found to increase significantly, although the change was only marginal, reiterated incubation with PVP-iodine led to a decrease in the IC<sub>50</sub>. Conclusion: Repeated treatment of S. aureus with polihexanide, chlorhexidine, octenidine and PVP-iodine did not trigger bacterial adaptation to these substances.

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          Most cited references31

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          Bacterial silver resistance: molecular biology and uses and misuses of silver compounds.

          Resistance to silver compounds as determined by bacterial plasmids and genes has been defined by molecular genetics. Silver resistance conferred by the Salmonella plasmid pMGH100 involves nine genes in three transcription units. A sensor/responder (SilRS) two-component transcriptional regulatory system governs synthesis of a periplasmic Ag(I)-binding protein (SilE) and two efflux pumps (a P-type ATPase (SilP) plus a three-protein chemiosmotic RND Ag(I)/H+ exchange system (SilCBA)). The same genes were identified on five of 19 additional IncH incompatibility class plasmids but thus far not on other plasmids. Of 70 random enteric isolates from a local hospital, isolates from catheters and other Ag-exposed sites, and total genomes of enteric bacteria, 10 have recognizable sil genes. The centrally located six genes are found and functional in the chromosome of Escherichia coli K-12, and also occur on the genome of E. coli O157:H7. The use of molecular epidemiological tools will establish the range and diversity of such resistance systems in clinical and non-clinical sources. Silver compounds are used widely as effective antimicrobial agents to combat pathogens (bacteria, viruses and eukaryotic microorganisms) in the clinic and for public health hygiene. Silver cations (Ag+) are microcidal at low concentrations and used to treat burns, wounds and ulcers. Ag is used to coat catheters to retard microbial biofilm development. Ag is used in hygiene products including face creams, "alternative medicine" health supplements, supermarket products for washing vegetables, and water filtration cartridges. Ag is generally without adverse effects for humans, and argyria (irreversible discoloration of the skin resulting from subepithelial silver deposits) is rare and mostly of cosmetic concern.
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            Distribution, organization, and ecology of bacteria in chronic wounds.

            Between 1 and 2% of the population in the developed world experiences a nonhealing or chronic wound characterized by an apparent arrest in a stage dominated by inflammatory processes. Lately, research groups have proposed that bacteria might be involved in and contribute to the lack of healing of these wounds. To investigate this, we collected and examined samples from chronic wounds obtained from 22 different patients, all selected because of suspicion of Pseudomonas aeruginosa colonization. These wound samples were investigated by standard culturing methods and peptide nucleic acid-based fluorescence in situ hybridization (PNA FISH) for direct identification of bacteria. By means of the culturing methods, Staphylococcus aureus was detected in the majority of the wounds, whereas P. aeruginosa was observed less frequently. In contrast, using PNA FISH, we found that a large fraction of the wounds contained P. aeruginosa. Furthermore, PNA FISH revealed the structural organization of bacteria in the samples. It appeared that P. aeruginosa aggregated as microcolonies imbedded in the matrix component alginate, which is a characteristic hallmark of the biofilm mode of growth. The present investigation suggests that bacteria present within these wounds tend to be aggregated in microcolonies imbedded in a self-produced matrix, characteristic of the biofilm mode of growth. Additionally, we must conclude that there exists no good correlation between bacteria detected by standard culturing methods and those detected by direct detection methods such as PNA FISH. This strongly supports the development of new diagnostic and treatment strategies for chronic wounds.
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              Antimicrobial activities of silver dressings: an in vitro comparison.

              A range of silver-coated or -impregnated dressings are now commercially available for use but comparative data on their antimicrobial efficacies are limited. The antibacterial activities of five commercially available silver-coated/impregnated dressings were compared against nine common burn-wound pathogens, namely methicillin-sensitive and -resistant Staphylococcus aureus (MRSA), Enterococcus faecalis, Pseudomonas aeruginosa, Escherichia coli, Enterobacter cloacae, Proteus vulgaris, Acinetobacter baumannii and a multi-drug-efflux-positive Acinetobacter baumannii (BM4454), using a broth culture method. The rapidity and extent of killing of these pathogens under in vitro conditions were evaluated. All five silver-impregnated dressings investigated exerted bactericidal activity, particularly against Gram-negative bacteria, including Enterobacter species, Proteus species and E. coli. The spectrum and rapidity of action, however, ranged widely for different dressings. Acticoat and Contreet had a broad spectrum of bactericidal activities against both Gram-positive and -negative bacteria. Contreet was characterized by a very rapid bactericidal action and achieved a reduction of > or =10,000 c.f.u. ml(-1) in the first 30 min for Enterobacter cloacae, Proteus vulgaris, Pseudomonas aeruginosa and Acinetobacter baumanii. Other dressings demonstrated a narrower range of bactericidal activities. Understanding the characteristics of these dressings may enable them to be targeted more appropriately according to the specific requirements for use of a particular dressing, as in for prophylaxis in skin grafting or for an infected wound with MRSA.
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                Author and article information

                Journal
                SPP
                Skin Pharmacol Physiol
                10.1159/issn.1660-5527
                Skin Pharmacology and Physiology
                S. Karger AG
                1660-5527
                1660-5535
                2012
                October 2012
                09 August 2012
                : 25
                : 6
                : 288-297
                Affiliations
                aDepartment of Dermatology, University Medical Center Jena, Jena, bLohmann &amp; Rauscher GmbH and Co. KG, Rengsdorf, and cLohmann &amp; Rauscher GmbH and Co. KG, Neuwied, Germany
                Author notes
                *Cornelia Wiegand, Department of Dermatology, University Medical Center Jena, Erfurter Str. 35, DE–07740 Jena (Germany), Tel. +49 364 193 7584, E-Mail C.Wiegand@med.uni-jena.de
                Article
                341222 Skin Pharmacol Physiol 2012;25:288–297
                10.1159/000341222
                22890487
                878ebfb0-aeb0-4077-83b9-920fcbc2477c
                © 2012 S. Karger AG, Basel

                Open Access License: This is an Open Access article licensed under the terms of the Creative Commons Attribution-NonCommercial 3.0 Unported license (CC BY-NC) ( http://www.karger.com/OA-license), applicable to the online version of the article only. Distribution permitted for non-commercial purposes only. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

                History
                : 16 February 2012
                : 19 June 2012
                Page count
                Figures: 5, Tables: 1, Pages: 10
                Categories
                Original Paper

                Oncology & Radiotherapy,Pathology,Surgery,Dermatology,Pharmacology & Pharmaceutical medicine
                Bacterial adaptation,Microplate laser nephelometry,Antiseptics, Staphylococcus aureus ,Mupirocin

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