TY - CONF A1 - Schmidt, Selina A1 - Schreiber, Frank T1 - Priming in soil microbial communities mediated by biocide-induced horizontal gene-transfer N2 - Soil microbes are exposed to different environmental stressors originating from various sources. Biocides used as material preservatives can represent environmental stressors since they are in direct contact with the environment including soil. Microorganism in soils can adapt to stress by different mechanisms; for example, by transferring mobile genetic elements via horizontal gene transfer (HGT). Here, we hypothesize that material preservatives can cause increased frequencies of HGT (i.e. altered community permissiveness) facilitating microbial community adaptation to stress. Furthermore, we hypothesize that soil microbial communities are primed by biocide exposure facilitating the response to different types of stresses. We will incubate soil mesocosms with selected biocides to investigate if these compounds promote HGT of plasmids that carry resistance genes in soil microbial communities. Subsequently, we will prime the soil microbial community with sub-inhibitory concentrations of biocides followed by exposure to toxic biocide concentrations or other types of common environmental stresses including metals, antibiotics, and salt. Using suitable control experiments, a shift in the functional response of the primed as compared to the non-primed community would indicate that biocides prime microbial communities via HGT. Together these results will elucidate the effect of biocides on HGT-mediated adaptation to environmental stressors in soil microbial communities. T2 - BAM PhD Day 2018 CY - Berlin, Germany DA - 31.05.2018 KW - Resistance evolution KW - Horizontal gene transfer HGT KW - Biocides KW - Microbiology PY - 2018 AN - OPUS4-46286 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Koerdt, Andrea A1 - An, Biwen Annie A1 - Kleinbub, Sherin A1 - Schreiber, Frank T1 - Investigating the effects of biocides and corrosion inhibitors on corrosive methanogens N2 - Microbiologically influenced corrosion (MIC) is the deterioration of metals due to the metabolic activities of microorganisms. Microorganisms can take electrons directly from the metal surface (EMIC) thereby causing corrosion. Well known culprits of EMIC are: sulfate-reducing bacteria (SRB), acetogens and methanogens. T2 - AMiCI WG2 workshop Berlin CY - Berlin, Germany DA - 7.7.2018 KW - Corrosion KW - Biocides KW - Methanogens PY - 2018 AN - OPUS4-45734 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schmidt, Selina A1 - Schreiber, Frank T1 - Antimicrobial resistance in soil microbes mediated by resistance evolution and horizontal gene transfer (HGT) N2 - Biocides are in contact with soils through direct application and passive leaching from protected materials. Consequently, soil microorganisms are exposed to biocides even though they are not their primary targets. Soil is a large reservoir of microbial diversity and has been hypothesized to be a crucial factor for the evolution and spread of antimicrobial resistance. Currently, there is little knowledge on how biocides used to protect materials affect the evolution and spread of resistance. Thus, our aim is to investigate the risk for the evolution of biocide resistance and cross-resistance to antibiotics. In addition, we aim to elucidate the affect of biocides on the spread of resistance via horizontal gene transfer (HGT). In adaptive laboratory evolution experiments we cultured selected model soil microorganism with representative biocides under selection regimes with increasing and stable biocide concentrations followed by antibiotic and biocide cross-resistance determination. Moreover, we investigate if the selected biocides affect the rates of de novo mutations and HGT of plasmids that carry resistance genes among soil microorganism. Our results show only small increases of biocide resistance during serial transfers under increasing biocide concentrations. One reason for this might be the narrow selective window for biocide resistance due to steep dose-response relationships. Furthermore, our results indicate that a stable low-level biocide regime did not select for high level cross-resistance to antibiotics and other biocides. Moreover, material preservatives affected the rates of HGT via conjugation and the mutation rates at sub-inhibitory concentrations. The results will enable future risk assessment regarding resistance evolution for biocides used as material preservatives. T2 - 4th Evo Eco PhD Meeting CY - Lutherstadt Wittenberg DA - 04.03.2020 KW - Microbiology KW - Biocides KW - Horizontal gene transfer HGT KW - Resistance evolution KW - Antimicrobial resistance PY - 2020 AN - OPUS4-51313 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank A1 - Nordholt, Niclas A1 - Kanaris, Orestis A1 - Schmidt, Selina T1 - Persistence against benzalkonium chloride promotes rapid evolution of tolerance under periodic disinfection N2 - Biocides used as disinfectants are important to prevent the transmission of pathogens, especially during the current antibiotic resistance crisis. This crisis is exacerbated by phenotypically tolerant persister subpopulations which can survive transient antibiotic treatment and facilitate resistance evolution. Despite the transient nature of disinfection, knowledge concerning persistence to disinfectants and its link to resistance evolution is currently lacking. Here, we show that E. coli displays persistence against a widely used disinfectant benzalkonium chloride (BAC). Periodic, persister-mediated failure of disinfection rapidly selects for BAC tolerance. BAC tolerance is associated with reduced cell surface charge and mutations in the novel tolerance locus lpxM. Moreover, the fitness cost incurred by BAC tolerance turned into a fitness benefit in the presence of antibiotics, suggesting a selective advantage of BAC-tolerant mutants in antibiotic environments. Our findings provide a mechanistic underpinning for the faithful application of disinfectants to prevent multi-drug-resistance evolution and to steward the efficacy of biocides and antibiotics. T2 - New Approaches and Concepts in Microbiology CY - Online meeting DA - 07.07.2021 KW - Persistence KW - Biocides KW - Evolution KW - Cross-resistance KW - Biocide tolerance PY - 2021 AN - OPUS4-53168 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Nordholt, Niclas A1 - Schreiber, Frank A1 - Kanaris, Orestis A1 - Schmidt, Selina T1 - Persistence against benzalkonium chloride promotes rapid evolution of tolerance under periodic disinfection N2 - Biocides used as disinfectants are important to prevent the transmission of pathogens, especially during the current antibiotic resistance crisis. This crisis is exacerbated by phenotypically tolerant persister subpopulations which can survive transient antibiotic treatment and facilitate resistance evolution. Despite the transient nature of disinfection, knowledge concerning persistence to disinfectants and its link to resistance evolution is currently lacking. Here, we show that E. coli displays persistence against a widely used disinfectant benzalkonium chloride (BAC). Periodic, persister-mediated failure of disinfection rapidly selects for BAC tolerance. BAC tolerance is associated with reduced cell surface charge and mutations in the novel tolerance locus lpxM. Moreover, the fitness cost incurred by BAC tolerance turned into a fitness benefit in the presence of antibiotics, suggesting a selective advantage of BAC-tolerant mutants in antibiotic environments. Our findings provide a mechanistic underpinning for the faithful application of disinfectants to prevent multi-drug-resistance evolution and to steward the efficacy of biocides and antibiotics. T2 - World Microbe Forum (ASM, FEMS) CY - Online meeting DA - 20.06.2021 KW - Persistence KW - Biocides KW - Evolution KW - Cross-resistance KW - Biocide tolerance PY - 2021 AN - OPUS4-53167 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Risk Assessment of Biocide Resistance N2 - This presentation details approaches for the risk assessment of biocide resistance. Different methods are presented to acquire the necessary data for such risk assessments. T2 - OECD, 7th Meeting of the Working Party on Biocides CY - Leiden, Netherlands DA - 18.09.2023 KW - Antimicrobial resistance KW - Bacteria KW - Standardization KW - Biocides PY - 2023 AN - OPUS4-59062 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Nordholt, Niclas A1 - Gustiherdini, Cahyaruri A1 - Lewerenz, Dominique A1 - Kanaris, Orestis A1 - Schreiber, Frank T1 - The disinfectant glutaraldehyde induces antibiotic tolerance N2 - Glutaraldehyde is widely used as a disinfectant and preservative, but little is known about its effects on bacterial susceptibility to antibiotics and the selection of tolerant phenotypes. We found that short-term exposure to sub-inhibitory levels of glutaraldehyde makes E. coli resistant to high doses of bactericidal antibiotics from different classes. This tolerance is associated with delayed, heterogeneous regrowth dynamics and global transcriptome remodeling. We identified over 1200 differentially expressed genes, including those related to antibiotic efflux, metabolic processes, and the cell envelope. The cells entered a disrupted state likely due to the unspecific mode-of-action of glutaraldehyde. Despite this unregulated response, we identified several differentially expressed genes not previously associated with antibiotic tolerance or persistence that induce antibiotic tolerance when overexpressed alone. These findings highlight how the unspecific mode-of-action of disinfectants can make bacteria temporarily resistant to antibiotics. They have implications for settings where disinfectants and antibiotics are used in close proximity, such as hospitals and animal husbandry, and for the selection dynamics of tolerant pheno- and genotypes in fluctuating environments where microorganisms are exposed to these substances, such as sewage systems. A trade-off arises from overcoming the disrupted state as quickly as possible and maintaining antibiotic tolerance. T2 - Gordon Research Conference - Molecular Mechanisms in Evolution CY - Easton, Massachusetts, United States DA - 25.06.2023 KW - Glutaraldehyde KW - Biocides KW - Tolerance KW - Bacteria KW - Disinfection KW - Heterogeneity KW - Antibiotics PY - 2023 AN - OPUS4-58033 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Nordholt, Niclas T1 - EVOCIDE: Preserving disinfectant efficacy by predicting evolution: exposing the principles of disinfectant survival and adaptation in bacteria N2 - Presentation of the EVOCIDE research proposal work programme.EVOCIDE seeks to gain a systems level understanding of disinfectant survival and evolution in bacteria. T2 - Joint Group Seminar Rolff-McMahon-Armitage-Steiner CY - Berlin, Germany DA - 20.12.2023 KW - Disinfectants KW - Evolution KW - Microbiology KW - Bacteria KW - Biocides PY - 2023 AN - OPUS4-59224 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Nordholt, Niclas T1 - The disinfectant glutaraldehyde induces antibiotic tolerance underpinned by phenotypic heterogeneity and transcriptome remodeling N2 - Glutaraldehyde is widely used as a disinfectant and preservative, but little is known about its effects on bacterial susceptibility to antibiotics and the selection of tolerant phenotypes. We found that short-term exposure to sub-inhibitory levels of glutaraldehyde makes E. coli resistant to high doses of bactericidal antibiotics from different classes. This tolerance is associated with delayed, heterogeneous regrowth dynamics and global transcriptome remodeling. We identified over 1200 differentially expressed genes, including those related to antibiotic efflux, metabolic processes, and the cell envelope. The cells entered a disrupted state likely due to the unspecific mode-of-action of glutaraldehyde. Despite this unregulated response, we identified several differentially expressed genes not previously associated with antibiotic tolerance or persistence that induce antibiotic tolerance when overexpressed alone. These findings highlight how the unspecific mode-of-action of disinfectants can make bacteria temporarily resistant to antibiotics. They have implications for settings where disinfectants and antibiotics are used in close proximity, such as hospitals and animal husbandry, and for the selection dynamics of tolerant pheno- and genotypes in fluctuating environments where microorganisms are exposed to these substances, such as sewage systems. A trade-off arises from overcoming the disrupted state as quickly as possible and maintaining antibiotic tolerance. T2 - µClub Seminar Series CY - Berlin, Germany DA - 26.05.2023 KW - Glutaraldehyde KW - Biocides KW - Tolerance KW - Bacteria KW - Disinfection KW - Heterogeneity PY - 2023 AN - OPUS4-58031 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Nordholt, Niclas T1 - The disinfectant glutaraldehyde induces antibiotic tolerance underpinned by a Disrupted Cellular State and Heterogenous Regrowth Dynamics N2 - Glutaraldehyde is widely used as a disinfectant and preservative, but little is known about its effects on bacterial susceptibility to antibiotics and the selection of tolerant phenotypes. We found that short-term exposure to sub-inhibitory levels of glutaraldehyde makes E. coli resistant to high doses of bactericidal antibiotics from different classes. This tolerance is associated with delayed, heterogeneous regrowth dynamics and global transcriptome remodeling. We identified over 1200 differentially expressed genes, including those related to antibiotic efflux, metabolic processes, and the cell envelope. The cells entered a disrupted state likely due to the unspecific mode-of-action of glutaraldehyde. Despite this unregulated response, we identified several differentially expressed genes not previously associated with antibiotic tolerance or persistence that induce antibiotic tolerance when overexpressed alone. These findings highlight how the unspecific mode-of-action of disinfectants can make bacteria temporarily resistant to antibiotics. They have implications for settings where disinfectants and antibiotics are used in close proximity, such as hospitals and animal husbandry, and for the selection dynamics of tolerant pheno- and genotypes in fluctuating environments where microorganisms are exposed to these substances, such as sewage systems. A trade-off arises from overcoming the disrupted state as quickly as possible and maintaining antibiotic tolerance. T2 - Molecular Mechanisms in Evolution (GRS) Gordon Research Seminar CY - Easton, Massachusetts, USA DA - 24.06.2023 KW - Glutaraldehyde KW - Biocides KW - Tolerance KW - Bacteria KW - Disinfection KW - Heterogeneity KW - Antibiotics KW - AMR PY - 2023 AN - OPUS4-58032 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Resistance evolution towards biocides and antimicrobial surfaces N2 - This presentation describes our work at BAM on resistance evolution towards biocides and antimicrobial surfaces. T2 - Break biofilms workshop CY - Vienna, Austria DA - 16.01.2023 KW - Antimicrobial resistance KW - Bacteria KW - Biofilms KW - Biocides PY - 2023 AN - OPUS4-57857 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank A1 - Schmidt, Selina A1 - Boenke, V. A1 - Stephan, Ina T1 - Development of a laboratory method to assess resistance development of microorganisms to biocides – An update N2 - This presentation describes our efforts at BAM towards the development of a laboratory method to assess resistance development of microorganisms to biocides. T2 - The International Biodeterioration Research Group (IBRG) spring meeting 2023 CY - Online meeting DA - 05.03.2023 KW - Antimicrobial resistance KW - Antimicrobial coating KW - Standardization KW - Biocides PY - 2023 AN - OPUS4-57858 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank A1 - Nordholt, Niclas A1 - Kanaris, Orestis A1 - Schmidt, Selina A1 - Lewerenz, Dominique T1 - Phenotypic and evolutionary responses of bacteria to disinfection N2 - Disinfectants are important to prevent the transmission of pathogens, especially in the face of the current antibiotic resistance crisis. The crisis is further exacerbated by phenotypically tolerant persister subpopulations that can survive transient antibiotic treatment and facilitate resistance evolution. Despite the transient nature of disinfectant application, persistence to disinfectants and its role for the evolution of tolerance and cross-resistance to antibiotics has not been studied. Our work shows that E. coli displays persistence against several widely used disinfectants, including benzalkonium chloride (BAC), didecyldimethylammoniumchlorid (DDAC) and isopropanol. The molecular mechanism of BAC persistence is triggered in stationary phase and affected by several antibiotic persister genes (hipA, tisB, tolC, relA, spoT). Experimental evolution and population dynamic modeling show that repeated failure of disinfection due to persisters rapidly selects for BAC tolerance underpinned by reduced cell surface charge due to mutations in genes related to lipid A acylation (lpxML). Furthermore, evolved BAC tolerance affects the susceptibility to antibiotics, leading to positive selection of disinfectant tolerant strains at environmentally relevant antibiotic concentrations and variations in evolvability of antibiotic resistance due to epistatic effects. These results highlight the need for faithful application of disinfectants to steward their efficacy and the efficacy of antibiotics. A better understanding of the bacterial response to disinfectants is crucial to understand and avert the ongoing antimicrobial resistance crisis. T2 - Gordon Research Conference - Molecular mechanisms of evolution CY - Easton, MA, United States DA - 25.06.2023 KW - Antimicrobial resistance KW - Bacteria KW - Disinfection KW - Biocides PY - 2023 AN - OPUS4-57861 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Nordholt, Niclas A1 - Kanaris, Orestis A1 - Gustiherdini, Cahyaruri A1 - Lewerenz, Dominique A1 - Schreiber, Frank T1 - The disinfectant glutaraldehyde induces antibiotic tolerance underpinned by Phenotypic Heterogeneity and transcriptome remodeling N2 - Glutaraldehyde (GTA) is commonly used to disinfect medical equipment, in animal husbandry and in hydraulic fracturing. Its wide use bears the risk that microorganisms in different environments are exposed to potentially non-lethal doses of glutaraldehyde. To date, little is known about the effects of glutaraldehyde on the susceptibility of bacteria to antibiotics and its role in the selection of tolerant phenotypes. Objectives • To determine the effect of glutaraldehyde exposure on the survival of E. coli, S. aureus and P. aeruginosa to antibiotics • To find the mechanistic basis for antibiotic tolerance upon glutaraldehyde exposure Materials & Methods Four bacterial isolates were exposed to sub-inhibitory glutaraldehyde. Antibiotic tolerance was determined by time-kill assays. Regrowth dynamics (lag times) were determined with ScanLag. E. coli was further investigated, using RNAseq to identify genes and processes involved in antibiotic tolerance. Mutants of candidate genes were screened for their antibiotic tolerance and heterogeneous target gene expression under stressed and unstressed conditions. Results Short-term exposure to sub-inhibitory levels of glutaraldehyde induced tolerance to high doses of bactericidal antibiotics. Tolerance to antibiotics was associated with highly heterogeneous regrowth dynamics and global transcriptome remodeling. Differentially expressed genes represented diverse biological functions and cellular components, including antibiotic efflux, metabolic processes, and the cell envelope. The heterogeneous regrowth dynamics and the diversity of the differentially expressed genes are likely related to the unspecific mode-of-action of glutaraldehyde. Among the many differentially expressed genes, several genes were identified that were not previously associated with antibiotic tolerance or persistence, which, when overexpressed alone, increased antibiotic tolerance. Conclusion Our results highlight how the big advantage of a disinfectant, its unspecific mode-of-action, can induce transient tolerance to antibiotics in bacteria. These findings have implications for 1.) settings where disinfectants and antibiotics are used in proximity, such as hospitals and animal husbandry, and 2.) for the selection dynamics of tolerant bacteria in fluctuating environments because of the trade-off that arises from overcoming the lag phase as fast as possible and maintaining antibiotic tolerance. T2 - VAAM Jahrestagung 2023 CY - Göttingen, Germany DA - 10.09.2023 KW - Glutaraldehyde KW - Disinfectants KW - Biocides KW - Antibiotics KW - Bacteria PY - 2023 AN - OPUS4-58439 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Biocide resistance risk assessment N2 - This presentation details the current status of biocide resistance risk assessment. T2 - Risk assessment of biocide and antibiotic resistance CY - Online meeting DA - 09.03.2022 KW - Antimicrobial resistance KW - Antimicrobial coating KW - Standardization KW - Biocides KW - Risk assessment PY - 2022 UR - https://www.gu.se/en/biocide/risk-assessment-of-biocide-and-antibiotic-resistance AN - OPUS4-56235 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Resistance evolution towards biocides and antimicrobial surfaces N2 - Biocides, including disinfectants and antimicrobial surfaces (AMCs), are important to prevent the spread of pathogens and antimicrobial resistant bacteria via surfaces. However, concerns have been raised about the evolution and selection of resistance against disinfectants and AMCs. In turn, resistance against disinfectants and AMCs can be associated to antibiotic resistance due to cross-resistance and co-resistance. We need to understand the mechanisms and risks of disinfectants and AMCs for resistance and cross-resistance evolution to optimize their application and safeguard their long-term efficacy. We used adaptive laboratory evolution (ALE) experiments based on repeated exposure of bacteria to disinfectants. Our results show that repeated disinfection of E. coli with benzalkonium chloride in suspension results in a 2000-fold increase in survival within 5 exposure cycles. Adaption is linked to the initial presence of persister cells highly tolerant to benzalkonium chloride. We used the same approach to develop standardizable ALE experiments to determine resistance evolution to AMCs. The results highlight rapid adaptation of E. coli and P. aeruginosa towards copper surfaces. Moreover, there are multiple situations in the clinic or in the environment in which biocides and antibiotics co-occur and in which combination effects can shape their antimicrobial activity or their selective effects. Our work with P. aeruginosa shows prevalent combination effects of biocides and antibiotics, ranging from synergy to antagonism and resulting in the selection for or against antibiotic resistant strains. The combination effects are dependent on the biofilm mode-of-growth, manifesting in apparent differences in the structural arrangement of antibiotic sensitive and resistant strains in biofilms exposed to combinations. Furthermore, biocides affect rates of mutation and horizontal gene transfer, thereby having a potential facilitating effect on resistance evolution. Taken together, our work shows that the role of biocides as potential drivers of resistance evolution and selection deserves further study and regulative action. T2 - Eurobiofilms 2022 CY - Palma, Spain DA - 31.08.2022 KW - Antimicrobial resistance KW - Bacteria KW - Biofilms KW - Biocides KW - Antimicrobial surfaces PY - 2022 AN - OPUS4-55608 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Nordholt, Niclas A1 - Schreiber, Frank T1 - Heterogeneous tolerance to biocides and its consequences for the evolution of antimicrobial resistance N2 - The exposure to antimicrobial substances drives the evolution of antimicrobial resistance (AMR). Biocides are antimicrobials used as disinfectants, antiseptics and preservatives. They find application on a large scale in the industrial and medical sector, but also in private households. In terms of mass, the worldwide use of biocides exceeds that of antibiotics. Thus, despite their important role in preventing the spread of pathogens, due to their ubiquity, biocides are suspected to be drivers of the antimicrobial resistance crisis. In our work we try to understand how biocides contribute to the emergence of AMR, what the underlying adaptation principles and mechanisms are and how they compare to those found for antibiotics. Here, I present results which demonstrate that phenotypic heterogeneity in E. coli, namely the formation of tolerant persister cells, can enable the survival of disinfection and consequentially facilitate the evolution of AMR. Using experimental evolution that selects for tolerance, we find that mutations in lipid A biosynthesis arise following periodic treatment with benzalkonium chloride (BAC), a widely used disinfectant. BAC tolerance is associated with additional, diverse mutations as well as changes in the susceptibility to antibiotics from different classes and changes in motility and biofilm formation, suggesting extensive regulatory re-wiring in the evolved clones. Remarkably, we find that fitness costs (growth rate reduction) incurred by BAC tolerance are alleviated in the presence of antibiotics. These findings demonstrate the complexity underlying the adaptation to antimicrobials and highlight the links between persistence to disinfectants and resistance evolution to antimicrobials. T2 - 4th VAAM discussion meeting 'Microbial Cell Biology' CY - Berlin, Germany DA - 09.10.2022 KW - Persistence KW - Biocides KW - Evolution KW - Disinfection KW - Biocide tolerance KW - Heterogeneity PY - 2022 AN - OPUS4-55958 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Nordholt, Niclas A1 - Kanaris, Orestis A1 - Schmidt, Selina A1 - Schreiber, Frank T1 - Persistence against benzalkonium chloride promotes rapid evolution of tolerance under periodic disinfection N2 - The exposure to antimicrobial substances drives the evolution of antimicrobial resistance (AMR). Biocides are antimicrobials used as disinfectants, antiseptics and preservatives. They find application on a large scale in the industrial and medical sector, but also in private households. In terms of mass, the worldwide use of biocides exceeds that of antibiotics. Thus, despite their important role in preventing the spread of pathogens, due to their ubiquity, biocides are suspected to be drivers of the antimicrobial resistance crisis. In our work we try to understand how biocides contribute to the emergence of AMR, what the underlying adaptation principles and mechanisms are and how they compare to those found for antibiotics. Here, I present results which demonstrate that phenotypic heterogeneity in E. coli, namely the formation of tolerant persister cells, can enable the survival of disinfection and consequentially facilitate the evolution of AMR. Using experimental evolution that selects for tolerance, we find that mutations in lipid A biosynthesis arise following periodic treatment with benzalkonium chloride (BAC), a widely used disinfectant. BAC tolerance is associated with additional, diverse mutations as well as changes in the susceptibility to antibiotics from different classes and changes in motility and biofilm formation, suggesting extensive regulatory re-wiring in the evolved clones. Remarkably, we find that fitness costs (growth rate reduction) incurred by BAC tolerance are alleviated in the presence of antibiotics. These findings demonstrate the complexity underlying the adaptation to antimicrobials and highlight the links between persistence to disinfectants and resistance evolution to antimicrobials. T2 - Bacterial Networks (BacNet22) CY - Sant Feliu de Guixols, Spanien DA - 04.09.2022 KW - Persistence KW - Biocides KW - evolution KW - disinfection KW - biocide tolerance PY - 2022 AN - OPUS4-55713 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schmidt, Selina A1 - Schreiber, Frank T1 - Biocides Used as Material Preservatives Modify Rates of de novo Mutation and Horizontal Gene Transfer in Bacteria N2 - Biocides used as material preservatives are in contact with various environments during direct application or passive leaching from protected materials. Antimicrobial resistance (AMR) is a global health problem and the environment is an important compartment for the evolution and transmission of AMR. Soil is an environment with a large reservoir of natural microbial communities and antimicrobial resistance (AMR) genes. Those natural microbial communities are frequently exposed to biocides used as material preservatives. Previous studies have shown that antibiotics, metals and pesticides affect the underlying processes of resistance evolution and spread; namely de novo mutagenesis and horizontal gene transfer by conjugation and transformation in microbial communities. However, it is unknown if active substances used in material preservatives are involved in these processes. We show that biocides used as material preservatives affect rates of mutation and conjugation in microorganism in a species- and substance-dependent manner, while rates of transformation are not directly affected. Our data highlights the importance of assessing the contribution of material preservatives on AMR evolution and spread in the environment. T2 - Microbiome Network Meeting CY - Berlin, Germany DA - 20.07.2022 KW - Biocides KW - Horizontal gene transfer KW - Mutation rate PY - 2022 AN - OPUS4-55263 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schoknecht, Ute T1 - The Impact of Weather Conditions on Biocides in Paints N2 - Aim. The European biocidal products regulation requires environmental risk assessments for biocidal products under service conditions. This is only possible, if processes that lead to biocide emissions into the environment are understood and can be reasonably predicted. Actual emissions due to leaching result from different processes that are affected by material properties and environmental conditions. Transformation of biocides can affect emissions considerably. Leaching and possible transformation of the film preservatives carbendazim, diuron, OIT and terbutryn in paints was observed under laboratory and field conditons in order to investigate the influence of different factors, e.g. water contact and radiation on biocide emissions. The paints contained either white titanium dioxide or a red iron oxide pigment to check whether pigments affect leaching processes. Methods. Test specimens of a water-based styrene acrylic dispersion paint on glass were exposed to UVA-radiation in a UV weathering device and/or exposed to water contact by immersion events on the basis of the European standard EN 16105. Plywood panels were coated and exposed to natural weather conditions in a semi-field experiment. Concentrations of biocides and transformation products were analysed in leachates from laboratory tests, runoff samples and remaining coatings. Results. The impact of weathering conditions on transformation processes differs for the investigated active substances. Water contact, radiation as well as pigments in the paints determine the pattern and amount of transformation products on coatings, leachates and runoff water. Observations from laboratory and field experiments were compared. It proved that results from laboratory tests support understanding of data from field experiments. Detailed knowledge on the fate of active substances under environmental conditions supports manufacturers to develop safe applications of film preservatives and authorities to evaluate its environmental impact. T2 - European Technical Coatings Congress 2022 CY - Kraków, Poland DA - 12.07.2022 KW - Paints KW - Biocides KW - Transformation KW - Emission PY - 2022 AN - OPUS4-55274 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -