scholarly journals A Novel Biofilm Model System to Visualise Conjugal Transfer of Vancomycin Resistance by Environmental Enterococci

2021 ◽  
Vol 9 (4) ◽  
pp. 789
Author(s):  
Michael Conwell ◽  
James S. G. Dooley ◽  
Patrick J. Naughton
Keyword(s):  
Antibiotic Resistance ◽  
Gene Transfer ◽  
Antibiotic Resistance Gene ◽  
In Situ Hybridisation ◽  
Model System ◽  
Vancomycin Resistance ◽  
Conjugal Transfer ◽  
Clinical Settings ◽  
Resistance Transfer ◽  
Non Destructive

Enterococci and biofilm-associated infections are a growing problem worldwide, given the rise in antibiotic resistance in environmental and clinical settings. The increasing incidence of antibiotic resistance and its propagation potential within enterococcal biofilm is a concern. This requires a deeper understanding of how enterococcal biofilm develops, and how antibiotic resistance transfer takes place in these biofilms. Enterococcal biofilm assays, incorporating the study of antibiotic resistance transfer, require a system which can accommodate non-destructive, real-time experimentation. We adapted a Gene Frame® combined with fluorescence microscopy as a novel non-destructive platform to study the conjugal transfer of vancomycin resistance in an established enterococcal biofilm.A multi-purpose fluorescent in situ hybridisation (FISH) probe, in a novel application, allowed the identification of low copy number mobile elements in the biofilm. Furthermore, a Hoechst stain and ENU 1470 FISH probe identified Enterococcus faecium transconjugants by excluding Enterococcus faecalis MF06036 donors. Biofilm created with a rifampicin resistant E. faecalis (MW01105Rif) recipient had a transfer efficiency of 2.01 × 10−3; double that of the biofilm primarily created by the donor (E. faecalis MF06036). Conjugation in the mixed enterococcal biofilm was triple the efficiency of donor biofilm. Double antibiotic treatment plus lysozyme combined with live/dead imaging provided fluorescent micrographs identifying de novo enterococcal vancomycin resistant transconjugants inside the biofilm. This is a model system for the further study of antibiotic resistance transfer events in enterococci. Biofilms promote the survival of enterococci and reduce the effectiveness of drug treatment in clinical settings, hence giving enterococci an advantage. Enterococci growing in biofilms exchange traits by means of horizontal gene transfer, but currently available models make study difficult. This work goes some way to providing a non-destructive, molecular imaging-based model system for the detection of antibiotic resistance gene transfer in enterococci.

scholarly journals Monitoring horizontal antibiotic resistance gene transfer in a colonic fermentation model

2011 ◽  
Vol 78 (2) ◽  
pp. 210-219 ◽  
Author(s):  
Martina C. Haug ◽  
Sabine A. Tanner ◽  
Christophe Lacroix ◽  
Marc J.A. Stevens ◽  
Leo Meile
Keyword(s):  
Antibiotic Resistance ◽  
Gene Transfer ◽  
Resistance Gene ◽  
Antibiotic Resistance Gene ◽  
Colonic Fermentation ◽  
Fermentation Model

scholarly journals eDNA Inactivation and Biofilm Inhibition by the Polymeric Biocide Polyhexamethylene Guanidine Hydrochloride (PHMG-Cl)

2022 ◽  
Vol 23 (2) ◽  
pp. 731
Author(s):  
Olena V. Moshynets ◽  
Taras P. Baranovskyi ◽  
Olga S. Iungin ◽  
Nadiia P. Kysil ◽  
Larysa O. Metelytsia ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Gene Transfer ◽  
Resistance Gene ◽  
Antibiotic Resistance Gene ◽  
Guanidine Hydrochloride ◽  
Dust Particles ◽  
Biofilm Inhibition ◽  
Water Contaminants ◽  
Polyhexamethylene Guanidine ◽  
Biofilm Development

The choice of effective biocides used for routine hospital practice should consider the role of disinfectants in the maintenance and development of local resistome and how they might affect antibiotic resistance gene transfer within the hospital microbial population. Currently, there is little understanding of how different biocides contribute to eDNA release that may contribute to gene transfer and subsequent environmental retention. Here, we investigated how different biocides affect the release of eDNA from mature biofilms of two opportunistic model strains Pseudomonas aeruginosa ATCC 27853 (PA) and Staphylococcus aureus ATCC 25923 (SA) and contribute to the hospital resistome in the form of surface and water contaminants and dust particles. The effect of four groups of biocides, alcohols, hydrogen peroxide, quaternary ammonium compounds, and the polymeric biocide polyhexamethylene guanidine hydrochloride (PHMG-Cl), was evaluated using PA and SA biofilms. Most biocides, except for PHMG-Cl and 70% ethanol, caused substantial eDNA release, and PHMG-Cl was found to block biofilm development when used at concentrations of 0.5% and 0.1%. This might be associated with the formation of DNA–PHMG-Cl complexes as PHMG-Cl is predicted to bind to AT base pairs by molecular docking assays. PHMG-Cl was found to bind high-molecular DNA and plasmid DNA and continued to inactivate DNA on surfaces even after 4 weeks. PHMG-Cl also effectively inactivated biofilm-associated antibiotic resistance gene eDNA released by a pan-drug-resistant Klebsiella strain, which demonstrates the potential of a polymeric biocide as a new surface-active agent to combat the spread of antibiotic resistance in hospital settings.

scholarly journals eDNA Inactivation and Biofilm Inhibition by the Polymeric Biocide Polyhexamethylene Guanidine Hydrochloride (PHMG-Cl)

2021 ◽  
Author(s):  
Olena V. Moshynets ◽  
Taras P. Baranovskyi ◽  
Olga S. Iungin ◽  
Nadiia P. Kysil ◽  
Larysa O. Metelytsia ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Gene Transfer ◽  
Resistance Gene ◽  
Antibiotic Resistance Gene ◽  
Guanidine Hydrochloride ◽  
Active Agent ◽  
Dust Particles ◽  
Biofilm Inhibition ◽  
Water Contaminants ◽  
Biofilm Development

The choice of effective biocides used for routine hospital practice should consider the role of disinfectants in the maintenance and development of local resistome and how they might affect antibiotic resistance gene transfer within the hospital microbial population. Currently, there is little understanding of how different biocides contribute to eDNA release that may contribute to gene transfer and subsequent environmental retention. Here we investigated how different biocides affected the release of eDNA from mature biofilms of two opportunistic model strains Pseudomonas aeruginosa ATCC 27853 (PA) and Staphylococcus aureus ATCC 25923 (SA) and contribute to the hospital resistome in the form of surface and water contaminants and dust particles. The effect of four groups of biocides including alcohols, hydrogen peroxide, quaternary ammonium compounds, and polymeric guanidines were evaluated using PA and SA biofilms. Most biocides, except for PHMG-Cl and 70% ethanol, caused substantial eDNA release and PHMG-Cl was found to block biofilm development when used at concentrations of 0.5% and 0.1%. This might be associated with the formation of DNA-PHMG-Cl complexes as PHMG-Cl is predicted to bind to AT base pairs by molecular docking assays. PHMG-Cl was found to bind high molecular DNA and plasmid DNA and continued to inactivate DNA on surfaces even after four weeks. PHMG-Cl also effectively inactivated biofilm-associated antibiotic resistance gene eDNA released by a pan-drug-resistant Klebsiella strain which demonstrates the potential of PHMG-Cl as a new surface-active agent to combat the spread of antibiotic resistance in hospital settings.

scholarly journals In vitro evalution of antibiotic resistance of Lactobacillus bulgaricus strains isolated from traditional dairy products

2019 ◽  
Vol 37 (No. 1) ◽  
pp. 36-43
Author(s):  
Huiling Guo ◽  
Bilige Menghe
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Antibiotic Resistance Gene ◽  
Antibiotic Resistance Genes ◽  
Fermented Milk ◽  
Lactobacillus Bulgaricus ◽  
Dilution Method ◽  
Resistance Transfer ◽  
Traditional Dairy Products

The antimicrobial susceptibility of 20 Lb. bulgaricus isolates from traditional fermented milk-originated was assessed and then determined the ability to transfer antibiotic resistance genes to other bacteria. The minimum inhibitory concentration of each strain was determined using a standardized dilution method. All the tested strains were found to be susceptible to gentamicin, erythromycin, clindamycin, neomycin, tetracycline, linezolid, chloramphenicol, rifampicin, and quinupristin/dalfopristin, while their susceptibilities to kanamycin, ciprofloxacin, streptomycin, trimethoprim, ampicillin, and vancomycin varied. Polymerase chain reaction (PCR) was used to check whether specific antibiotic resistance genes were present in these Lb. bulgaricus. We detected the rpoB, erm(B), aadA, bla, cat and vanX. Finally, a filter mating assay was applied to investigate the transferability of these resistance markers; and we observe no antibiotic resistance transfer between bacteria. This work demonstrates a low risk of lateral transfer of the antibiotic resistance gene of Lb. bulgaricus.

scholarly journals Antibiotic-resistance and virulence genes in Enterococcus isolated from tropical recreational waters

2013 ◽  
Vol 11 (3) ◽  
pp. 387-396 ◽  
Author(s):  
Tasha M. Santiago-Rodriguez ◽  
Jessica I. Rivera ◽  
Mariel Coradin ◽  
Gary A. Toranzos
Keyword(s):  
Antibiotic Resistance ◽  
Virulence Genes ◽  
Tetracycline Resistance ◽  
Surface Protein ◽  
Vancomycin Resistance ◽  
Clinical Settings ◽  
Recreational Waters ◽  
Tropical Environments ◽  
Encoding Gene ◽  
Enterococcus Isolate

The prevalence of enterococci harboring tetracycline- and vancomycin-resistance genes, as well as the enterococcal surface protein (esp) has mostly been determined in clinical settings, but their prevalence in tropical recreational waters remains largely unknown. The present study determined the prevalence of tetM (tetracycline-resistance), vanA and vanB (vancomycin-resistance) in the bacterial and viral fractions, enterococci and their induced phages isolated from tropical recreational marine and fresh waters, dry and wet sands. Since lysogenic phages can act as vectors for antibiotic-resistance and virulence factors, the prevalence of the mentioned genes, as well as that of an integrase-encoding gene (int) specific for Enterococcus faecalis phages was determined. Up to 60 and 54% of the bacterial fractions and enterococci, respectively, harbored at least one of the tested genes suggesting that bacteria in tropical environments may be reservoirs of antibiotic-resistance and virulence genes. int was detected in the viral fractions and in one Enterococcus isolate after induction. This study presents the opportunity to determine if the presence of bacteria harboring antibiotic-resistance and virulence genes in tropical recreational waters represents a threat to public health.

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