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      Biofilm Formation of Helicobacter pylori in Both Static and Microfluidic Conditions Is Associated With Resistance to Clarithromycin

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          Abstract

          It is widely accepted that production of biofilm is a protective mechanism against various type of stressors, including exposure to antibiotics. However, the impact of this structure on the spread of antibiotic resistance in Helicobacter pylori is still poorly understood. Therefore, the aim of the current research was to determine the relationship between biofilm formation and antibiotic resistance of H. pylori. The study was carried out on 24 clinical strains with different resistance profiles (antibiotic-sensitive, mono-resistant, double-resistant and multidrug-resistant) against clarithromycin (CLR), metronidazole (MTZ) and levofloxacin (LEV). Using static conditions and a crystal violet staining method, a strong correlation was observed between biofilm formation and resistance to CLR but not MTZ or LEV. Based on the obtained results, three the strongest and three the weakest biofilm producers were selected and directed for a set of microfluidic experiments performed in the Bioflux system combined with fluorescence microscopy. Under continuous flow conditions, it was observed that strong biofilm producers formed twice as much of biofilm and created significantly more eDNA and in particular proteins within the biofilm matrix when compared to weak biofilm producers. Additionally, it was noticed that strong biofilm producers had higher tendency for autoaggregation and presented morphostructural differences (a greater cellular packing, shorter cells and a higher amount of both OMVs and flagella) in relation to weak biofilm counterparts. In conclusion, resistance to CLR in clinical H. pylori strains was associated with a broad array of phenotypical features translating to the ability of strong biofilm formation.

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          Discovery, research, and development of new antibiotics: the WHO priority list of antibiotic-resistant bacteria and tuberculosis

          The spread of antibiotic-resistant bacteria poses a substantial threat to morbidity and mortality worldwide. Due to its large public health and societal implications, multidrug-resistant tuberculosis has been long regarded by WHO as a global priority for investment in new drugs. In 2016, WHO was requested by member states to create a priority list of other antibiotic-resistant bacteria to support research and development of effective drugs.
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            Global Prevalence of Helicobacter pylori Infection: Systematic Review and Meta-Analysis.

            The epidemiology of Helicobacter pylori infection has changed with improvements in sanitation and methods of eradication. We performed a systematic review and meta-analysis to evaluate changes in the global prevalence of H pylori infection.
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              Molecular mechanisms of biofilm-based antibiotic resistance and tolerance in pathogenic bacteria.

              Biofilms are surface-attached groups of microbial cells encased in an extracellular matrix that are significantly less susceptible to antimicrobial agents than non-adherent, planktonic cells. Biofilm-based infections are, as a result, extremely difficult to cure. A wide range of molecular mechanisms contribute to the high degree of recalcitrance that is characteristic of biofilm communities. These mechanisms include, among others, interaction of antimicrobials with biofilm matrix components, reduced growth rates and the various actions of specific genetic determinants of antibiotic resistance and tolerance. Alone, each of these mechanisms only partially accounts for the increased antimicrobial recalcitrance observed in biofilms. Acting in concert, however, these defences help to ensure the survival of biofilm cells in the face of even the most aggressive antimicrobial treatment regimens. This review summarises both historical and recent scientific data in support of the known biofilm resistance and tolerance mechanisms. Additionally, suggestions for future work in the field are provided.
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                Author and article information

                Contributors
                Journal
                Front Cell Infect Microbiol
                Front Cell Infect Microbiol
                Front. Cell. Infect. Microbiol.
                Frontiers in Cellular and Infection Microbiology
                Frontiers Media S.A.
                2235-2988
                25 March 2022
                2022
                : 12
                : 868905
                Affiliations
                [1] 1 Department of Microbiology, Faculty of Medicine, Wroclaw Medical University , Wroclaw, Poland
                [2] 2 Department of Environment, Hygiene and Animal Welfare, Wroclaw University of Environmental and Life Sciences , Wroclaw, Poland
                [3] 3 Department of Pharmacy, University “G. d’Annunzio” of Chieti-Pescara , Chieti, Italy
                Author notes

                Edited by: Fany Reffuveille, Université de Reims Champagne-Ardenne, France

                Reviewed by: Skander Hathroubi, Humboldt University of Berlin, Germany; Farzad Khademi, Ardabil University of Medical Sciences, Iran

                *Correspondence: Paweł Krzyżek, krojcerpawel@ 123456gmail.com

                This article was submitted to Biofilms, a section of the journal Frontiers in Cellular and Infection Microbiology

                Article
                10.3389/fcimb.2022.868905
                8990135
                35402304
                ca0298ef-51c8-40e6-b400-da65ee529edd
                Copyright © 2022 Krzyżek, Migdał, Grande and Gościniak

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 03 February 2022
                : 07 March 2022
                Page count
                Figures: 7, Tables: 0, Equations: 0, References: 85, Pages: 16, Words: 8874
                Funding
                Funded by: Uniwersytet Medyczny im. Piastów Slaskich we Wroclawiu , doi 10.13039/501100009687;
                Award ID: SUBZ.A130.22.010
                Funded by: Narodowe Centrum Badań i Rozwoju , doi 10.13039/501100005632;
                Award ID: IA/SP/453975/2020
                Categories
                Cellular and Infection Microbiology
                Original Research

                Infectious disease & Microbiology
                helicobacter pylori,antibiotic resistance,multidrug resistance,biofilm formation,biofilm matrix,flow conditions,microfluidic system,bioflux

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