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      Is heparinized 40% ethanol lock solution efficient for reducing bacterial and fungal biofilms in an in vitro model?

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          Abstract

          Background

          We applied an in vitro model to evaluate the efficacy of a heparinized 40% ethanol-based lock solution in a wide variety of clinical isolates causing C-RBSI.

          Methods

          A total of 100 clinical strains were collected retrospectively from the blood of patients with C-RBSI. The reduction in biomass and metabolic activity of biofilms was measured using the crystal violet (CV) assay and XTT assay, respectively. Regrowth inhibition (RI) was measured within 24 hours and 72 hours of ethanol lock therapy. Percentage reduction of ≥ 85% in RI was considered to be successful.

          Results

          Ethanol lock was more effective in reducing metabolic activity than in reducing biomass (83% vs. 50%, respectively). Percentages of RI diminished as regrowth was prolonged (57% for 24 hours and 17% for 72 hours of regrowth). No statistically significant intraspecies differences were found in biofilm reduction or in RI (p>0.05).

          Conclusions

          The use of heparinized 40% ethanol lock solution for 72 hours significantly reduced biomass and metabolic activity in clinical isolates from patients with C-RBSI. However, as biofilm has an important regrowth rate, 40% ethanol solution was not able to fully eradicate biofilm in vitro.

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          Most cited references29

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          The importance of the viable but non-culturable state in human bacterial pathogens

          Many bacterial species have been found to exist in a viable but non-culturable (VBNC) state since its discovery in 1982. VBNC cells are characterized by a loss of culturability on routine agar, which impairs their detection by conventional plate count techniques. This leads to an underestimation of total viable cells in environmental or clinical samples, and thus poses a risk to public health. In this review, we present recent findings on the VBNC state of human bacterial pathogens. The characteristics of VBNC cells, including the similarities and differences to viable, culturable cells and dead cells, and different detection methods are discussed. Exposure to various stresses can induce the VBNC state, and VBNC cells may be resuscitated back to culturable cells under suitable stimuli. The conditions that trigger the induction of the VBNC state and resuscitation from it are summarized and the mechanisms underlying these two processes are discussed. Last but not least, the significance of VBNC cells and their potential influence on human health are also reviewed.
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            Current Perspectives on Viable but Non-Culturable (VBNC) Pathogenic Bacteria

            Under stress conditions, many species of bacteria enter into starvation mode of metabolism or a physiologically viable but non-culturable (VBNC) state. Several human pathogenic bacteria have been reported to enter into the VBNC state under these conditions. The pathogenic VBNC bacteria cannot be grown using conventional culture media, although they continue to retain their viability and express their virulence. Though there have been debates on the VBNC concept in the past, several molecular studies have shown that not only can the VBNC state be induced under in vitro conditions but also that resuscitation from this state is possible under appropriate conditions. The most notable advance in resuscitating VBNC bacteria is the discovery of resuscitation-promoting factor (Rpf), which is a bacterial cytokines found in both Gram-positive and Gram-negative organisms. VBNC state is a survival strategy adopted by the bacteria, which has important implication in several fields, including environmental monitoring, food technology, and infectious disease management; and hence it is important to investigate the association of bacterial pathogens under VBNC state and the water/foodborne outbreaks. In this review, we describe various aspects of VBNC bacteria, which include their proteomic and genetic profiles under the VBNC state, conditions of resuscitation, methods of detection, antibiotic resistance, and observations on Rpf.
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              Adverse effects associated with ethanol catheter lock solutions: a systematic review.

              Antimicrobial lock therapy has been widely utilized internationally for the prevention and management of intravascular catheter-related bloodstream infections. One of the agents commonly utilized for lock therapy is ethanol. However, a systematic review of adverse events associated with ethanol locks has not been published.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Project administrationRole: SupervisionRole: ValidationRole: Writing – original draft
                Role: ConceptualizationRole: Writing – review & editing
                Role: Methodology
                Role: Resources
                Role: Resources
                Role: Funding acquisitionRole: Supervision
                Role: ConceptualizationRole: Funding acquisitionRole: SupervisionRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                8 July 2019
                2019
                : 14
                : 7
                : e0219098
                Affiliations
                [1 ] Instituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain
                [2 ] Department of Clinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, Madrid, Spain
                [3 ] Cardiac Surgery Postoperative Care Unit, Hospital General Universitario Gregorio Marañón, Madrid, Spain
                [4 ] CIBER Enfermedades Respiratorias-CIBERES (CB06/06/0058), Madrid, Spain
                [5 ] Pharmacy Department, Hospital General Universitario Gregorio Marañón, Madrid, Spain
                [6 ] Medicine Department, School of Medicine, Universidad Complutense de Madrid, Madrid, Spain
                Karl-Franzens-Universitat Graz, AUSTRIA
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0002-0062-2999
                Article
                PONE-D-19-08734
                10.1371/journal.pone.0219098
                6613681
                31283786
                beb077a2-e385-415d-9541-74487f059be4
                © 2019 Alonso et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 27 March 2019
                : 16 June 2019
                Page count
                Figures: 1, Tables: 2, Pages: 9
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100004587, Instituto de Salud Carlos III;
                Award ID: MS13/00268
                Award Recipient :
                M. Guembe is supported by the Miguel Servet Program (ISCIIIMICINN, MS13/00268) from the Health Research Fund (FIS) of the Carlos III Health Institute (ISCIII), Madrid, Spain. The study has been partially financed by the European Regional Development Fund (FEDER) “A way of making Europe”.
                Categories
                Research Article
                Physical Sciences
                Chemistry
                Chemical Compounds
                Organic Compounds
                Alcohols
                Ethanol
                Physical Sciences
                Chemistry
                Organic Chemistry
                Organic Compounds
                Alcohols
                Ethanol
                Biology and Life Sciences
                Microbiology
                Biofilms
                Biology and Life Sciences
                Bioengineering
                Biotechnology
                Medical Devices and Equipment
                Catheters
                Engineering and Technology
                Bioengineering
                Biotechnology
                Medical Devices and Equipment
                Catheters
                Medicine and Health Sciences
                Medical Devices and Equipment
                Catheters
                Medicine and Health Sciences
                Pharmacology
                Drugs
                Heparin
                Medicine and Health Sciences
                Pharmacology
                Drugs
                Antimicrobials
                Antibiotics
                Biology and Life Sciences
                Microbiology
                Microbial Control
                Antimicrobials
                Antibiotics
                Biology and Life Sciences
                Organisms
                Bacteria
                Staphylococcus
                Staphylococcus Aureus
                Biology and Life Sciences
                Microbiology
                Medical Microbiology
                Microbial Pathogens
                Bacterial Pathogens
                Staphylococcus
                Staphylococcus Aureus
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Pathogens
                Microbial Pathogens
                Bacterial Pathogens
                Staphylococcus
                Staphylococcus Aureus
                Biology and Life Sciences
                Organisms
                Eukaryota
                Fungi
                Yeast
                Candida
                Candida Albicans
                Biology and Life Sciences
                Microbiology
                Medical Microbiology
                Microbial Pathogens
                Fungal Pathogens
                Candida Albicans
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Pathogens
                Microbial Pathogens
                Fungal Pathogens
                Candida Albicans
                Biology and Life Sciences
                Mycology
                Fungal Pathogens
                Candida Albicans
                Research and Analysis Methods
                Animal Studies
                Experimental Organism Systems
                Yeast and Fungal Models
                Candida Albicans
                Biology and Life Sciences
                Microbiology
                Bacteriology
                Bacterial Biofilms
                Biology and Life Sciences
                Microbiology
                Biofilms
                Bacterial Biofilms
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

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