Dataset Information

Evaluation of a Newly Developed Vacuum Dried Microtiter Plate for Rapid Biocide Susceptibility Testing of Clinical Enterococcus Faecium Isolates.

ABSTRACT: We investigated the suitability of a newly developed biocide susceptibility test system based on microtiter plates containing vacuum dried biocides as a fast and reliable screening method. The evaluated substances included the cationic biocides benzalkonium chloride (BAC), chlorhexidine dihydrochloride (CHX), cetylpyridinium chloride, didecyldimethylammonium chloride, and octenidine dihydrochloride. Testing a selection of Escherichia coli and enterococci, the biocide microtiter plates provided results comparable to those obtained from broth microdilution according to ISO 20776-1. Broad MIC ranges allowed for testing gram-positive and gram-negative species with the same plate design. In the second part of our study, we applied the established method to analyze the susceptibility of 90 clinical Enterococcus faecium isolates from a German university hospital, as previous studies have indicated a link between reduced susceptibility to substances such as CHX and BAC and vancomycin resistance. We therefore determined MIC and minimum bactericidal concentrations (MBC) for 48 non-clonal vancomycin susceptible and 42 non-clonal vancomycin resistant isolates, but MIC95 and MBC95 were quite similar in both groups. Our easy to handle and ready to use test system enables the routine surveillance of bacterial tolerance towards disinfectants in hospitals. As a result, hygiene measures can be adapted and nosocomial infections controlled despite increasing prevalence of antibiotic-resistant bacteria.

SUBMITTER: Roedel A

PROVIDER: S-EPMC7232460 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature

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Evaluation of a Newly Developed Vacuum Dried Microtiter Plate for Rapid Biocide Susceptibility Testing of Clinical <i>Enterococcus Faecium</i> Isolates.

Roedel Alice A Dieckmann Ralf R Makarewicz Oliwia O Hartung Anita A Noll Matthias M Pletz Mathias W MW Dahouk Sascha Al SA Vincze Szilvia S

Microorganisms 20200411 4

We investigated the suitability of a newly developed biocide susceptibility test system based on microtiter plates containing vacuum dried biocides as a fast and reliable screening method. The evaluated substances included the cationic biocides benzalkonium chloride (BAC), chlorhexidine dihydrochloride (CHX), cetylpyridinium chloride, didecyldimethylammonium chloride, and octenidine dihydrochloride. Testing a selection of <i>Escherichia coli</i> and enterococci, the biocide microtiter plates pro ...[more]

PMID: 32290364

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Project description:Chlorhexidine (CHX) is a broad-spectrum antiseptic widely used in community and clinical contexts for many years that has recently acquired higher relevance in nosocomial infection control worldwide. Despite this, CHX tolerance among Enterococcus faecium bacteria, representing one of the leading agents causing nosocomial infections, has been poorly understood. This study provides new phenotypic and molecular data for better identification of CHX-tolerant E. faecium subpopulations in community and clinical contexts. The chlorhexidine MIC (MICCHX) distribution of 106 E. faecium isolates suggested the occurrence of tolerant subpopulations in diverse sources (human, animal, food, environment) and phylogenomic backgrounds (clades A1/A2/B), with predominance in clade A1. They carried a specific variant of the 2CS-CHXT operon, identified here. It encodes glucose and amino acid-polyamine-organocation family transporters, besides the DNA-binding response regulator ChtR, with a P102H mutation previously described only in CHX-tolerant clade A1 E. faecium, and the ChtS sensor. 2CS-CHXT seems to be associated with three regulons modulating diverse bacterial biological functions. Combined data from normal MIC distribution and 2CS-CHXT operon characterization support a tentative epidemiological cutoff (ECOFF) of 8?mg/liter to CHX, which is useful to detect tolerant E. faecium populations in future surveillance studies. The spread of tolerant E. faecium in diverse epidemiological backgrounds calls for the prudent use of CHX in multiple contexts.IMPORTANCE Chlorhexidine is one of the substances included in the World Health Organization's list of essential medicines, which comprises the safest and most effective medicines needed in global health systems. Although it has been widely applied as a disinfectant and antiseptic in health care (skin, hands, mouthwashes, eye drops) since the 1950s, its use in hospitals to prevent nosocomial infections has increased worldwide in recent years. Here, we provide a comprehensive study on chlorhexidine tolerance among strains of Enterococcus faecium, one of the leading nosocomial agents worldwide, and identify a novel 2CS-CHXT operon as a signature of tolerant strains occurring in diverse phylogenomic groups. Our data allowed for the proposal of a tentative epidemiological cutoff of 8?mg/liter, which is useful to detect tolerant E. faecium populations in surveillance studies in community and clinical contexts. The prediction of 2CS-CHXT regulons will also facilitate the design of future experimental studies to better uncover chlorhexidine tolerance among E. faecium bacteria.

Project description:The objective of this study was to determine the ability of an isolated strain (EGY_NRC1) or commercial (NCIMB 11181) Enterococcus faecium as a probiotic for lactating cows. Two experiments were conducted: In Experiment 1, the effects of three levels (1, 2, and 3 g/kg diet, DM basis) of isolated and commercial E. faecium on in vitro ruminal fermentation kinetics, gas, methane (CH4) and nutrient degradability were determined. In Experiment 2, thirty multiparous Holstein cows (633 ± 25.4 kg body weight) with 7 days in milk, were randomly assigned to 3 treatments in a completely randomized design in a 60-day experiment. Cows were fed without any additives (control treatment) or supplemented with 2 g/kg feed daily of E. faecium EGY_NRC1 (contain 1.1 × 109 CFU/g) or commercial E. faecium NCIMB 11181 (contain 2 × 1012 CFU/g). Diets were prepared to meet cow's nutrient requirements according to NRC recommendations. Probiotic doses were based on the in vitro Experiment 1. Feed intake, digestibility, blood parameters and lactation performance were evaluated. In Experiment 1, the isolated E. faecium linearly and quadratically increased (P < 0.001) in vitro total gas production (TGP), the degradability of dry matter (dDM) and organic matter (dOM) while decreased (P < 0.05) methane (CH4) percent of TGP, NH3CH4 production, and pH. The commercial E. faecium increased TGP and decreased (P < 0.01) CH4 production, pH and increased the dDM and dOM, short chain fatty acids and ruminal NH3-N concentration. In Experiment 2, the isolated E. faecium increased (P < 0.01) total tract digestibility of DM, neutral and acid detergent fiber, daily milk production and feed efficiency compared to the control treatment without affecting feed intake and milk composition. Moreover, the isolated E. faecium increased (P < 0.05) the proportion of C18:1 trans-9, C18:2 cis-9-12 and C18:2 trans-10 cis-12. Both isolated and commercial E. faecium improved (P < 0.01) organic matter, crude protein and nonstructural carbohydrates digestibility, increased serum glucose (P = 0.002) and decreased serum cholesterol (P = 0.002). Additionally, both E. faecium strains decreased C23:0 (P = 0.005) in milk. In conclusion, the use of E. faecium (isolated and commercial) at 2 g/kg DM of feed improved feed efficiency and production performance, with superior effects on animal performance from isolated E. faecium compared to the commercial one.

Dataset Information

Evaluation of a Newly Developed Vacuum Dried Microtiter Plate for Rapid Biocide Susceptibility Testing of Clinical Enterococcus Faecium Isolates.

Publications

Evaluation of a Newly Developed Vacuum Dried Microtiter Plate for Rapid Biocide Susceptibility Testing of Clinical <i>Enterococcus Faecium</i> Isolates.

Similar Datasets

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