Limited antimicrobial efficacy of oral care antiseptics in microcosm biofilms and phenotypic adaptation of bacteria upon
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ORIGINAL ARTICLE
Limited antimicrobial efficacy of oral care antiseptics in microcosm biofilms and phenotypic adaptation of bacteria upon repeated exposure Sophia R. Schwarz 1 & Stefanie Hirsch 1 & Andreas Hiergeist 2 & Christian Kirschneck 3 & Denise Muehler 1 & Karl-Anton Hiller 1 & Tim Maisch 4 & Ali Al-Ahmad 5 & André Gessner 2 & Wolfgang Buchalla 1 & Fabian Cieplik 1 Received: 3 June 2020 / Accepted: 1 October 2020 # The Author(s) 2020
Abstract Objectives The aims of this study were to investigate the antimicrobial efficacy of antiseptics in saliva-derived microcosm biofilms, and to examine phenotypic adaption of bacteria upon repeated exposure to sub-inhibitory antiseptic concentrations. Methods Saliva-derived biofilms were formed mimicking caries- or gingivitis-associated conditions, respectively. Microbial compositions were analyzed by semiconductor-based 16S rRNA sequencing. Biofilms were treated with CHX, CPC, BAC, ALX, and DQC for 1 or 10 min, and colony forming units (CFU) were evaluated. Phenotypic adaptation of six selected bacterial reference strains toward CHX, CPC, and BAC was assessed by measuring minimum inhibitory concentrations (MICs) over 10 passages of sub-inhibitory exposure. Protein expression profiles were investigated by SDS-PAGE. Results Both biofilms showed outgrowth of streptococci and Veillonella spp., while gingivitis biofilms also showed increased relative abundances of Actinomyces, Granulicatella, and Gemella spp. Antiseptic treatment for 1 min led to no relevant CFUreductions despite for CPC. When treated for 10 min, CPC was most effective followed by BAC, ALX, CHX, and DQC. Stable adaptations with up to fourfold MIC increases were found in E. coli toward all tested antiseptics, in E. faecalis toward CHX and BAC, and in S. aureus toward CPC. Adapted E. coli strains showed different protein expression as compared with the wildtype strain. Conclusion Antiseptics showed limited antimicrobial efficacy toward mature biofilms when applied for clinically relevant treatment periods. Bacteria showed phenotypic adaptation upon repeated sub-inhibitory exposure. Clinical relevance Clinicians should be aware that wide-spread use of antiseptics may pose the risk of inducing resistances in oral bacteria. Keywords Antimicrobial . Biofilm . Microcosm . Antiseptic . Adaptation Sophia R. Schwarz and Stefanie Hirsch contributed equally to this work. * Fabian Cieplik [email protected] 1
Department of Conservative Dentistry and Periodontology, University Hospital Regensburg, Franz-Josef-Strauß-Allee 11, 93053 Regensburg, Germany
2
Institute of Clinical Microbiology and Hygiene, University Hospital Regensburg, Regensburg, Germany
3
Department of Orthodontics, University Hospital Regensburg, Regensburg, Germany
4
Department of Dermatology, University Hospital Regensburg, Regensburg, Germany
5
Department of Operative Dentistry and Periodontology, Center for Dental Medicine, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, Germany
Introduction The predominant mode of bacte
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