TY - CONF A1 - Sobisch, Lydia-Yasmin T1 - Time-kill kinetics reveal heterogeneous tolerance to disinfectants N2 - Background. Disinfection is an important mitigation strategy to control and prevent the spread of infections. Incomplete or incorrect usage of disinfection may promote evolution of resistance against disinfectants and antibiotics. Ideally, disinfection reduces the number of surviving bacteria and the chance for resistance evolution. Resistance describes the ability to grow in previously inhibitory concentrations of an antimicrobial, whereas tolerance is associated with enhanced survival of lethal doses. Individual bacteria from the same population can display considerable heterogeneity in their ability to survive treatment (i.e. tolerance) with antimicrobials, which can result in unexpected treatment failure. Objective. In this study, we investigated six active substances of disinfectants, preservatives, and antiseptics against a population of E. coli to identify the presence of a tolerant subpopulation. Methods. We performed time-kill experiments and analyzed the data with a mathematical model to statistically infer whether the data is best explained by the presence of a tolerant subpopulation. Results. The analysis identified bimodal kill kinetics for benzalkonium chloride, didecyldimethylammonium chloride, and isopropanol. In contrast, kill kinetics by chlorhexidine, glutaraldehyde, and hydrogen peroxide were best explained by unimodal kill kinetics. These findings have implications for the risk of disinfection failure. In addition, we are currently performing adaptive laboratory evolution (ALE) experiments with the different disinfectants to investigate the potential consequences of tolerant sub-populations for the evolution of antimicrobial resistance and tolerance. T2 - FEMS Conference CY - Hamburg, Germany DA - 10.07.2023 KW - Biocide KW - Desinfectant KW - E.coli PY - 2023 AN - OPUS4-58471 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Sobisch, Lydia-Yasmin T1 - Time-kill kinetics reveal heterogeneous tolerance to disinfectants N2 - Background. Disinfection is an important mitigation strategy to control and prevent the spread of infections. Incomplete or incorrect usage of disinfection may promote evolution of resistance against disinfectants and antibiotics. Ideally, disinfection reduces the number of surviving bacteria and the chance for resistance evolution. Resistance describes the ability to grow in previously inhibitory concentrations of an antimicrobial, whereas tolerance is associated with enhanced survival of lethal doses. Individual bacteria from the same population can display considerable heterogeneity in their ability to survive treatment (i.e. tolerance) with antimicrobials, which can result in unexpected treatment failure. Objective. In this study, we investigated six active substances of disinfectants, preservatives, and antiseptics against a population of E. coli to identify the presence of a tolerant subpopulation. Methods. We performed time-kill experiments and analyzed the data with a mathematical model to statistically infer whether the data is best explained by the presence of a tolerant subpopulation. Results. The analysis identified bimodal kill kinetics for benzalkonium chloride, didecyldimethylammonium chloride, and isopropanol. In contrast, kill kinetics by chlorhexidine, glutaraldehyde, and hydrogen peroxide were best explained by unimodal kill kinetics. These findings have implications for the risk of disinfection failure. In addition, we are currently performing adaptive laboratory evolution (ALE) experiments with the different disinfectants to investigate the potential consequences of tolerant sub-populations for the evolution of antimicrobial resistance and tolerance. T2 - Vereinigung Allgemeiner und Angewandter Mikrobiobiologie Jahreskongress 2023 CY - Göttingen, Germany DA - 10.09.2023 KW - Biocide KW - Desinfectant KW - E.coli PY - 2023 AN - OPUS4-58470 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Taghavi Kalajahi, Sara T1 - The effectiveness of cathodic protection (CP) on microbiologically influenced corrosion (MIC) control N2 - Cathodic protection (CP) is an electrochemical method, to manage corrosion in different industries, especially in buried and sub-merged environments. In those environments microorganisms are present and can affect the corrosion process and cause microbiologically influenced corrosion (MIC). Most of the industry standards recommend performing CP using -800 mV (Ag/AgCl), however, if microorganisms capable of initiating MIC are present, for instance sulfate reducing bacteria (SRB), it is recommended to use even more negative potentials. It is assumed that this will provide adequate protection of the metal. Currently there is no information or valid data in the context of CP, on the level of MIC threat and the extent to which more negative potentials can be used to provide adequate protection and not over-protection (due to hydrogen embrittlement threat). Conflicting statements can be found in the literature regarding the effectiveness of CP on MIC, from reducing biofilm attachment to increasing bacterial activity and biofilm attachment. Recently, the development and lower price of molecular microbiological methods (MMM) have opened the door for more effective studies of MIC mechanisms along with other electrochemical methods and surface analysis. In this work, the genetic functionality of biofilms formed in the laboratory under CP conditions is investigated using transcriptomics. Gene expression of SRB biofilms under different CP potentials (-800, -850 and -900 mV) will be studied; comparison with control will allow us to distinguish the specific genes that are differentially expressed, leading to a better understanding of the mechanism of CP to affect bacterial activity and diversity. Keywords - cathodic protection, microbiologically influenced corrosion, molecular microbial methods. T2 - IBBS CY - Berlin, Germany DA - 09.09.2024 KW - Microbiologically influenced corrosion PY - 2024 AN - OPUS4-61981 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Nordholt, Niclas 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 - Koerdt, Andrea T1 - Technical challenges for the investigation of Microbiologically influenced corrosion under laboratory conditions across several sectors N2 - Microbiologically influenced corrosion MIC occurs in different sectors, under various conditions and can significantly contribute to material degradation. The most convincing effect can be observed with sulphate-reducing bacteria (SRB). There are many stakeholders who do not need to be convinced of this as there is ample scientific evidence clearly showing that SRBs corrode metal. However, many believe this is solely due to the produced H2S. Fewer people are aware that SRBs also have other mechanisms that can lead to corrosion, such as electrical MIC on metal (direct electron uptake from the metal used as electron donor). Despite this, many sectors or industries remain skeptical in several areas. When considering other classes or species, the skepticism increases even more. The reason is simple: microbiologist cannot consistently prove MIC on laboratory scale for all corrosive microorganisms. In contrast to materials sciences, where experiments can be narrowed down to a limited number of parameters, microbiology is too complex to simulate simply in the lab. Several factors are mandatory for the growth of microorganisms, consequently increasing the number of parameters. Even when making the system more complex, there are still unknown species in the field, whose growth conditions are not yet understood. We know via 16S rRNA analysis that the species is present, but that is all we know. If these samples are enriched in the lab, many species will not survive and their impact on the whole community cannot be investigated. Therefore, lab tests related to MIC works only for a certain number of microorganisms. This means the key-result for other disciplines are missing, and important numbers for calculating the lifespan of a material can only be provided with limited accuracy. This is neither satisfying nor useful for engineers, material scientist or similar professionals, and it is understandable. However, MIC occurs, and we need to address it in our infrastructure or other important areas. With this presentation, I aim to show the currently available techniques for testing on a laboratory scale. I will use SRBs and methanogenic archaea as examples to highlight the advantages and disadvantages of each method and point out where improvements are needed. The examples will be cross-sectoral and applicable to other sectors as well. This talk will highlight the currently existing challenges of testing MIC in the laboratory and may help generate creative solutions. T2 - MICxUS 2025 CY - Nicosia, Cyprus DA - 21.01.2025 KW - MIC KW - Biocorrosion KW - Laboratory testing KW - Interdisziplinarity KW - Microorganisms PY - 2025 UR - https://www.unic.ac.cy/ehresearch/micxus2025/#tab-98dc74f32bf3b8f95bd AN - OPUS4-63725 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Koerdt, Andrea T1 - Simulating Microbiologically Influenced Corrosion (MIC) at Seabed Environment in Monopile N2 - Ensuring the safety of offshore wind structures (OWS) is critical to guaranteeing their long-term performance and supporting reliable green energy supply. Microbiologically influenced corrosion (MIC) presents a significant challenge, particularly for monopiles in seabed environments. This study investigates the behavior of microorganisms and their impact on the corrosion process of carbon steel within monopiles. To simulate MIC at the sediment/water interface, an in-house experimental column was developed and inoculated with sediment and water samples from the North Sea. The system was operated under varying flow rates to replicate seabed movement conditions. Multiple molecular microbiological methods, surface analysis techniques, and other approaches were employed to assess the effects of different treatments. This study provides insights into MIC mechanisms in offshore environments and supports the development of strategies to monitor MIC in OWS infrastructure T2 - ISMOS10 CY - Nashville, TN, USA DA - 11.08.2025 KW - MIC KW - Environmental Simulation KW - Field testing KW - Marine Environment PY - 2025 AN - OPUS4-64298 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Misra, Archismita T1 - Polyoxometalate Ionic Liquids as Protective Coatings for Industrial Infrastructure and Cultural Heritage against Microbiologically Influenced Corrosion (MIC) N2 - Corrosion of stone and metal due to acid rain and biodeterioration poses significant challenges for industrial and residential infrastructure, as well as cultural heritage, including statues and historical artefacts. A promising mitigation strategy involves thin, transparent films of polyoxometalate-based ionic liquids (POM-ILs) as chemical shields. Stone samples coated with acid-resistant, biocidal POM-ILs exhibited negligible corrosion when exposed to simulated acid rain, in stark contrast to the severe deterioration of unprotected samples. Additionally, their biocidal properties effectively prevent biofilm formation on coated surfaces. Following studies successfully explored the effectiveness of the coating against lampenflora growing in the Pommery Champagne cellar; and the long-term performance of POM-ILs under outdoor environmental conditions. So, POM-ILs have already demonstrated remarkable anticorrosion and antimicrobial properties against aerobic microorganisms and, being water-insoluble, do not leach into aquatic ecosystem. The current research project repurposes the POM-ILs, extending their application to metals, specifically targeting microbiologically influenced corrosion (MIC) in cultural heritage artefacts made of brass, carbon steel, cast iron, and bronze. This involves optimizing nanocoating adhesion to the metal surface and evaluating its protective efficacy against MIC caused by anaerobic microorganisms such as methanogenic archaea and sulfate reducing bacteria (SRB). This presentation will highlight POM-ILs as sustainable, high-performance nanocoatings for biocorrosion mitigation. It will showcase published success stories, discuss ongoing research and proof-of-concept results, and explore future prospects for these advanced materials in safeguarding metal infrastructure and artefacts across different industrial sectors as well as in the context of cultural heritage conservation. T2 - 10th International symposium on applied microbiology and molecular biology in oil systems (ISMOS10) CY - Nashville, Tennessee, USA DA - 11.08.2025 KW - Polyoxometalates KW - Corrosion KW - Microbiologically Influenced Corrosion KW - Ionic Liquid PY - 2025 AN - OPUS4-64548 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Misra, Archismita T1 - Polyoxometalate Ionic Liquids as Protective Coatings for Industrial Infrastructure and Cultural Heritage against Microbiologically Influenced Corrosion (MIC) N2 - Corrosion of stone and metal due to acid rain and biodeterioration poses significant challenges for industrial and residential infrastructure, as well as cultural heritage, including statues and historical artefacts. A promising mitigation strategy involves thin, transparent films of polyoxometalate-based ionic liquids (POM-ILs) as chemical shields. Stone samples coated with acid-resistant, biocidal POM-ILs exhibited negligible corrosion when exposed to simulated acid rain, in stark contrast to the severe deterioration of unprotected samples. Additionally, their biocidal properties effectively prevent biofilm formation on coated surfaces. Following studies successfully explored the effectiveness of the coating against lampenflora growing in the Pommery Champagne cellar; and the long-term performance of POM-ILs under outdoor environmental conditions. So, POM-ILs have already demonstrated remarkable anticorrosion and antimicrobial properties against aerobic microorganisms and, being water-insoluble, do not leach into aquatic ecosystem. The current research project repurposes the POM-ILs, extending their application to metals, specifically targeting microbiologically influenced corrosion (MIC) in cultural heritage artefacts made of brass, carbon steel, cast iron, and bronze. This involves optimizing nanocoating adhesion to the metal surface and evaluating its protective efficacy against MIC caused by anaerobic microorganisms such as methanogenic archaea and sulfate reducing bacteria (SRB). This presentation will highlight POM-ILs as sustainable, high-performance nanocoatings for biocorrosion mitigation. It will showcase published success stories, discuss ongoing research and proof-of-concept results, and explore future prospects for these advanced materials in safeguarding metal infrastructure and artefacts across different industrial sectors as well as in the context of cultural heritage conservation. T2 - Euro-MIC COST Action - Closing Workshop Conference CY - Horsens, Denmark DA - 17.09.2025 KW - Polyoxometalates KW - Corrosion KW - Microbiologically Influenced Corrosion KW - Ionic Liquid PY - 2025 AN - OPUS4-64549 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Keshmiri, Hamid T1 - Plasmon-enhanced diffractive supercells N2 - Multiple diffractive surface modulations can concurrently couple the light to several electromagnetic surface waves. We present a multi-resonant plasmonic supercell structure with a broad range of applicability in harvesting the light over an extensive wavelength range and angles of incidence. T2 - Molecular Plasmonics 2023 CY - Jena, Germany DA - 11.05.2023 KW - Optics PY - 2023 AN - OPUS4-59248 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Physiological and evolutionary consequences of exposing Pseudomonas aeruginosa to biocide-antibiotic combinations N2 - Antimicrobials can exert specific physiological effects when used in combination that are different from those when applied alone. These effects include physiological effects (i.e. synergy, antagonism and suppression) as well as evolutionary effects on the selection of resistant strains (i.e. cross-resistance and collateral sensitivity). While combination effects have been extensively mapped for antibiotic-antibiotic combinations, the combination effects of antibiotics with antimicrobials used as biocides or antiseptics have not been systematically investigated. Here, we investigated the physiological and evolutionary consequences of combinations of antibiotics (meropenem, gentamicin and ciprofloxacin) and substances used as biocides or antiseptics (octenidine, benzalkonium chloride, cetrimonium bromide, chlorhexidine, povidone-iodine, silver) on growth and selection of Pseudomonas aeruginosa. We find prevalent physiological combination effects with synergy occurring 6 times and antagonism occurring 10 times. The effects are specific to the antibiotic-biocide combination with meropenem showing a tendency for antagonism with biocides (6 of 7), while gentamicin has a tendency for synergy (5 of 7). A particular strong antagonism is apparent for the meropenem-chlorhexidine combination, for which we conducted an in-depth study on the underlying molecular mechanism using RNASeq. Moreover, we find widespread effects of the biocide-antibiotic combinations on selection of P. aeruginosa strains resistant to the antibiotics, including cross-resistance and collateral sensitivity. In conclusion, antibiotics and biocides or antiseptics exert physiological and evolutionary combination effects on the pathogen P. aeruginosa. These effects have consequences for the efficacy of both types of substances and for the selection of antimicrobial resistant strains in clinical applications with combined exposure (e.g. wound care, coated biomaterials). T2 - ASM-FEMS World Microbe Forum CY - Online meeting DA - 20.06.2021 KW - Antimicrobial resistance KW - Antagonism KW - Biofilms PY - 2021 AN - OPUS4-53165 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank 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 - Schreiber, Frank 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 - Nordholt, Niclas 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 - Schreiber, Frank A1 - Nordholt, Niclas 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 T1 - NBD-DDA: a tool to study biocide & disinfectant resistance mechanisms N2 - Quaternary ammonium compounds (QACs) are widely used as active agents in disinfectants, antiseptics, and preservatives. Despite being in use since the 1940s, there remain multiple open questions regarding their detailed mode-of-action and the mechanisms that can make bacteria less susceptible to QACs, including phenotypic heterogeneity. Goals To develop a fluorescent quaternary ammonium compound and to use it to investigate the mechanisms underlying QAC susceptibility in bacteria. Material & Methods A fluorescent quaternary ammonium compound, N-dodecyl-N,N-dimethyl-[2-[(4-nitro-2,1,3-benzoxadiazol-7-yl)amino]ethyl] azanium-iodide (NBD-DDA), was synthesized. Chemical structure and purity of NBD-DDA was confirmed by mass spectrometry and NMR spectroscopy. Antimicrobial activity against S. aureus, P. aeruginosa and E. coli was assessed by determination of minimum inhibitory concentrations and time-kill assays. Uptake, heterogeneity and efflux of NBD-DDA in wild-type and efflux-deficient mutants (ΔtolC) were determined, using flow cytometry and fluorometry. Sub-cellular localization of NBD-DDA was determined using confocal laser scanning microscopy. Results NBD-DDA was readily detected by flow cytometry and fluorescence microscopy with standard GFP/FITC-settings. NBD-DDA retained antimicrobial activity comparable to the structurally similar, widely used QAC benzalkonium chloride (BAC). Characteristic time-kill kinetics and increased tolerance of a BAC tolerant E. coli strain against NBD-DDA suggest that the mode of action of NBD-DDA is similar to that of BAC. NBD-DDA was preferentially localized to the cell envelope, which is a primary target of BAC and other QACs. Leveraging NBD-DDA‘s fluorescent properties, it was demonstrated that reduced cellular accumulation underlies BAC tolerance in a BAC tolerant E. coli strain and that NBD-DDA is subject to efflux mediated by TolC. Summary Overall, NBD-DDA’s antimicrobial activity, its fluorescent properties, and its ease of detection render it a powerful tool to study mechanisms of QAC susceptibility and mode-of-action in bacteria, both on the level of populations and single-cells. T2 - VAAM - Jahrestagung 2024 CY - Würzburg, Germany DA - 02.06.2024 KW - Biocides KW - Phenotypic heterogeneity PY - 2024 AN - OPUS4-61177 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Braymer, Joseph T1 - Methanobacterium IM1 as a model organism to study MIC N2 - Microbiologically influenced corrosion (MIC), also known as biocorrosion, is a process where microbes corrode metal surfaces causing detrimental damage to manmade infrastructure. Several studies, mainly under anaerobic conditions, have identified sulfate-reducing bacteria and methanogens as key players in biocorrosion. However, an important issue is that most MIC-related microbes at the moment are non-culturable. In addition, improved technologies (i.e., omics, biochemical, and microscopy methods) are needed to study, detect, simulate, and modulate MIC-related cases. Therefore, the selection of strains that can cause MIC, are culturable, and are genetically accessible, are of high importance for establishing new methods to study MIC on the molecular level. For example, the archaeal strain Methanobacterium IM1 has caught great attention due to its involvement in biocorrosion processes. Genes encoding for a novel enzyme (MIC NiFe-hydrogenase) have been proposed in this methanogen to be directly related to the e T2 - International Biodeterioration and Biodegradation Symposium 19 CY - Berlin, Germany DA - 11.09.2024 KW - Biocorrosion KW - Methanogen KW - Iron KW - Hydrogenase KW - Mechanism PY - 2024 AN - OPUS4-61508 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Mezera, Marek T1 - Influence of the pulse repetition rate on the chemical and morphological properties of laser generated surface structures N2 - Inter-pulse accumulation of heat could affect the chemical and morphological properties of the laser processed material surface. Hence, the laser pulse repetition rate may restrict the processing parameters for specific laser-induced surface structures. In this study, the evolution of various types of laser-induced micro- and nanostructures at various laser fluence levels, effective number of pulses and at different pulse repetition rates (1 – 400 kHz) are studied for common metals/alloys (e.g. steel or titanium alloy) irradiated by near-infrared ultrashort laser pulses (925 fs, 1030 nm) in air environment. The processed surfaces were characterized by optical and scanning electron microscopy (OM, SEM), energy dispersive X-ray spectroscopy (EDX) as well as time of flight secondary ion mass spectrometry (TOF-SIMS). The results show that not only the surface morphology could change at different laser pulse repetition rates and comparable laser fluence levels and effective number of pulses, but also the surface chemistry is altered. Consequences for medical applications are outlined. T2 - European Materials Research Society Spring 2021 Meeting CY - Online meeting DA - 31.05.2021 KW - Laser-induced pariodic surface structures KW - LIPSS PY - 2021 AN - OPUS4-52778 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Bonse, Jörn T1 - Impact of laser-induced periodic surface structures on the bactericidal properties of copper and brass N2 - Surfaces of metallic copper and copper alloys effectively inactivate microorganisms and viruses. However, the exact inactivation mode is still under debate. Main factors are assumed to include direct contact with the metallic surface, influx of Cu(I)/Cu(II) ions and the generation of reactive oxygen species (ROS). Laser-induced periodic surface structures (LIPSS) are frequently reported to act antibacterial, mainly by prevention of bacterial adhesion due to a limited number of possible adhesion points or by increasing the overall surface of intrinsically antibacterial materials. In time-kill experiments with E. coli and S. aureus we analyzed the impact of LIPSS on the toxicity of metallic copper and brass. We also conducted ROS accumulation assays and conclude that the application of LIPSS is not generally straight forward to obtain or improve antibacterial surfaces. Thus, the antibacterial effects of LIPPS. T2 - 2023 Spring Meeting CY - Strasbourg, France DA - 29.05.2023 KW - LIPSS KW - Antimicrobial KW - Reactive oxygen species PY - 2023 AN - OPUS4-58465 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Nordholt, Niclas 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 - Koerdt, Andrea T1 - Exploring Microbial Impacts on Hydrogen Storage: - A Novel System for Corrosion Testing of Gas- and Material Degradation N2 - Underground hydrogen storage (UHS) is a strategic step towards implementing the hydrogen economy. Achieving the required infrastructure by 2050 necessitates advancements in hydrogen-dedicated assets and the evaluation of existing infrastructure. The unique conditions in UHS require an experimental set-up to simulate UHS operating conditions, which allows to assess the readiness of current storage and transmission for hydrogen, and develop new technologies for material-resistance, operational-simulations, and risk-assessments. In addition to the physical/chemical conditions in UHS (e.g., salinity, hydrogen concentration, operating temperature/-pressure, water content), biological threats must also be considered. Therefore, we present here a high-pressure-set-up, developed for research/-industrial testing purposes. Currently, UHS-experiments for microbiologically-influenced-corrosion (MIC) are performed in standard autoclaves with relatively high volumes/pressures; they were primarily designed for material-specific investigations. While these methods provided some useful information for biological questions, they had significant limitations. The novel UHS-simulation-set-up presented here is designed with a controlled independently temperature and pressure. Field samples can be used to mimic geology, water chemistry, construction materials, and microbiological conditions. Most significant advantages of the set-up are: 1. It allows for liquid addition during the test, enabling the study of biocides or the evaluation of operating setups. 2. It permits liquid/-gas sampling during the test, allowing for more efficient monitoring of testing conditions and a better understanding of the process over time. Additionally, a low-release function is added, which is particularly important for studying MIC to avoid negative side effects, on the material (e.g. polymers/corrosion product-layer/cells itself) which might occur due to the fast pressure release T2 - ISMOS10 CY - Nashville, TN, USA DA - 11.08.2025 KW - MIC KW - MISTRAL KW - Corrosion KW - Environmental simulation KW - Metall KW - Polymer PY - 2025 AN - OPUS4-64297 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -