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      Antibacterial Efficacy Testing of a Bioelectric Wound Dressing Against Clinical Wound Pathogens

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          Abstract

          Silver-based wound dressings have been developed for the control of bioburden in wounds. However, the popularity and extensive use of silver-based dressings has been associated with emerging microbial resistances to silver. In this study we examined in vitro antibacterial efficacy of a bioelectric dressing containing silver and zinc against various wound pathogens. Antibiotic-sensitive clinical wound isolates showed a 100% reduction in bacterial growth, except that Enterococcus faecalis isolate was shown to survive with a bacterial log 10 reduction rate of less than 10 2 CFU. We also investigated antibacterial efficacy against the extended spectrum β-lactamase (ESBL) bacteria, multidrug-resistant (MDR) bacteria, and methicillin-resistant Staphylococcus aureus (MRSA). The bioelectric dressing was effective in killing wound pathogens including ESBL, MDR, and MRSA in vitro. Furthermore, based on the primary results against E. faecalis, we carried out extensive studies against several nosocomial Enterococcus species including vancomycin-resistant species. Overall, the vancomycin-sensitive or -resistant Enterococcus species were resistant to this dressing at up to 48 h, except for the vancomycin-resistant Enterococcus raffinosus isolate only showing a 100% bacterial reduction at 48 h, but not at 24 h. The results demonstrated the effective bactericidal activity of a bioelectric dressing against antibiotic-sensitive and MDR strains, but Enterococcus species are bacteriostatic.

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          Most cited references28

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          Antibacterial activity and mechanism of action of zinc oxide nanoparticles against Campylobacter jejuni.

          The antibacterial effect of zinc oxide (ZnO) nanoparticles on Campylobacter jejuni was investigated for inhibition and inactivation of cell growth. The results showed that C. jejuni was extremely sensitive to treatment with ZnO nanoparticles. The MIC of ZnO nanoparticles for C. jejuni was determined to be 0.05 to 0.025 mg/ml, which is 8- to 16-fold lower than that for Salmonella enterica serovar Enteritidis and Escherichia coli O157:H7 (0.4 mg/ml). The action of ZnO nanoparticles against C. jejuni was determined to be bactericidal, not bacteriostatic. Scanning electron microscopy examination revealed that the majority of the cells transformed from spiral shapes into coccoid forms after exposure to 0.5 mg/ml of ZnO nanoparticles for 16 h, which is consistent with the morphological changes of C. jejuni under other stress conditions. These coccoid cells were found by ethidium monoazide-quantitative PCR (EMA-qPCR) to have a certain level of membrane leakage. To address the molecular basis of ZnO nanoparticle action, a large set of genes involved in cell stress response, motility, pathogenesis, and toxin production were selected for a gene expression study. Reverse transcription-quantitative PCR (RT-qPCR) showed that in response to treatment with ZnO nanoparticles, the expression levels of two oxidative stress genes (katA and ahpC) and a general stress response gene (dnaK) were increased 52-, 7-, and 17-fold, respectively. These results suggest that the antibacterial mechanism of ZnO nanoparticles is most likely due to disruption of the cell membrane and oxidative stress in Campylobacter.
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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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              Bacterial heavy metal resistance: new surprises.

              Bacterial plasmids encode resistance systems for toxic metal ions including Ag+, AsO2-, AsO4(3-), Cd2+, CO2+, CrO4(2-), Cu2+, Hg2+, Ni2+, Pb2+, Sb3+, TeO3(2-), Tl+, and Zn2+. In addition to understanding of the molecular genetics and environmental roles of these resistances, studies during the last few years have provided surprises and new biochemical mechanisms. Chromosomal determinants of toxic metal resistances are known, and the distinction between plasmid resistances and those from chromosomal genes has blurred, because for some metals (notably mercury and arsenic), the plasmid and chromosomal determinants are basically the same. Other systems, such as copper transport ATPases and metallothionein cation-binding proteins, are only known from chromosomal genes. The largest group of metal resistance systems function by energy-dependent efflux of toxic ions. Some of the efflux systems are ATPases and others are chemiosmotic cation/proton antiporters. The CadA cadmium resistance ATPase of gram-positive bacteria and the CopB copper efflux system of Enterococcus hirae are homologous to P-type ATPases of animals and plants. The CadA ATPase protein has been labeled with 32P from gamma-32P-ATP and drives ATP-dependent Cd2+ uptake by inside-out membrane vesicles. Recently isolated genes defective in the human hereditary diseases of copper metabolism, Menkes syndrome and Wilson's disease, encode P-type ATPases that are more similar to the bacterial CadA and CopB ATPases than to eukaryote ATPases that pump different cations. The arsenic resistance efflux system transports arsenite, using alternatively either a two-component (ArsA and ArsB) ATPase or a single polypeptide (ArsB) functioning as a chemiosmotic transporter. The third gene in the arsenic resistance system, arsC, encodes an enzyme that converts intracellular arsenate [As (V)] to arsenite [As (III)], the substrate of the efflux system. The three-component Czc (Cd2+, Zn2+, and CO2+) chemiosmotic efflux pump of soil microbes consists of inner membrane (CzcA), outer membrane (CzcC), and membrane-spanning (CzcB) proteins that together transport cations from the cytoplasm across the periplasmic space to the outside of the cell. Finally, the first bacterial metallothionein (which by definition is a small protein that binds metal cations by means of numerous cysteine thiolates) has been characterized in cyanobacteria.
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                Author and article information

                Journal
                Open Microbiol J
                Open Microbiol J
                TOMICROJ
                The Open Microbiology Journal
                Bentham Open
                1874-2858
                21 February 2014
                2014
                : 8
                : 15-21
                Affiliations
                [1 ]Diagnostics and Translational Research Center, Henry M Jackson Foundation for the Advancement of Military Medicine, Gaithersburg, MD, USA
                [2 ]Combat Wound Initiative Program, Bethesda, MD, USA
                [3 ]Vomaris, Inc. Vomaris Innovations Inc., Chandler, AZ, USA
                [4 ]United States Army Medical Materiel Agency (USAMMA), Fort Detrick, MD, USA
                [5 ]Walter Reed National Military Medical Center, Bethesda, MD, USA
                Author notes
                [* ] Address correspondence to this author at the Diagnostics and Translational Research Center, Henry M Jackson Foundation for the Advancement of Military Medicine, 401 Professional Drive, Suite 210, Gaithersburg, MD, USA; Tel: 240-833-4971; Fax: 240-833-4940; E-mail: mizadjoo@ 123456dtrc-hjfresearch.org.
                Article
                TOMICROJ-8-15
                10.2174/1874285801408010015
                3950956
                24627730
                5d73d5c8-5ea7-497c-bd88-76aa7c468f4d
                © Kim et al.; Licensee Bentham Open.

                This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

                History
                : 17 December 2013
                : 17 January 2014
                : 3 February 2014
                Categories
                Article

                Microbiology & Virology
                antibacterial efficacy testing,bioelectric dressing,clinical wound isolates.

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