TY - JOUR A1 - Franziska, Pietsch A1 - Nordholt, Niclas A1 - Heidrich, Gabriele A1 - Schreiber, Frank T1 - Prevalent Synergy and Antagonism Among Antibiotics and Biocides in Pseudomonas aeruginosa N2 - Antimicrobials can exert specific physiological effects when used in combination that are different from those when applied alone. 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 effects of combinations of antibiotics (meropenem, gentamicin, and ciprofloxacin) and substances used as biocides or antiseptics [octenidine, benzalkonium chloride, cetrimonium bromide, chlorhexidine, Povidone-iodine, silver nitrate (AgNO3), and Ag-nanoparticles] on the planktonic growth rate of Pseudomonas aeruginosa. Combination effects were investigated in growth experiments in microtiter plates at different concentrations and the Bliss interaction scores were calculated. Among the 21 screened combinations, we find prevalent combination effects with synergy occurring six 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). In conclusion, antibiotics and biocides or antiseptics exert physiological combination effects on the pathogen P. aeruginosa. These effects have consequences for the efficacy of both types of substances and potentially for the selection of antimicrobial resistant strains in clinical applications with combined exposure (e.g., wound care and coated biomaterials). KW - Synergy KW - Antagonism KW - Suppression KW - Biocides KW - Antibiotics KW - Pseudomonas aeruginosa PY - 2021 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-520845 DO - https://doi.org/10.3389/fmicb.2020.615618 VL - 11 SP - Article 615618 PB - Frontiers CY - Lausanne AN - OPUS4-52084 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Valentin, Jules D. P. A1 - Altenried, Stefanie A1 - Varadarajan, Adithi R. A1 - Ahrens, Christian H. A1 - Schreiber, Frank A1 - Webb, Jeremy S. A1 - van der Mei, Henny C. A1 - Ren, Qun T1 - Identification of Potential Antimicrobial Targets of Pseudomonas aeruginosa Biofilms through a Novel Screening Approach N2 - Pseudomonas aeruginosa is an opportunistic pathogen of considerable medical importance, owing to its pronounced antibiotic tolerance and association with cystic fibrosis and other life-threatening diseases. The aim of this study was to highlight the genes responsible for P. aeruginosa biofilm tolerance to antibiotics and thereby identify potential new targets for the development of drugs against biofilm-related infections. By developing a novel screening approach and utilizing a public P. aeruginosa transposon insertion library, several biofilm-relevant genes were identified. The Pf phage gene (PA0720) and flagellin gene (fliC) conferred biofilm-specific tolerance to gentamicin. Compared with the reference biofilms, the biofilms formed by PA0720 and fliC mutants were completely eliminated with a 4-fold-lower gentamicin concentration. Furthermore, the mreC, pprB, coxC, and PA3785 genes were demonstrated to play major roles in enhancing biofilm tolerance to gentamicin. The analysis of biofilm-relevant genes performed in this study provides important novel insights into the understanding of P. aeruginosa antibiotic tolerance, which will facilitate the detection of antibiotic resistance and the development of antibiofilm strategies against P. aeruginosa. KW - Antimicrobial resistance KW - Bacteria KW - Biofilms KW - Pseudomonas aeruginosa PY - 2023 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-570205 DO - https://doi.org/10.1128/spectrum.03099-22 SP - 1 EP - 5 PB - ASM Journals AN - OPUS4-57020 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Chacón, Luz A1 - Kuropka, B. A1 - González-Tortuero, E. A1 - Schreiber, Frank A1 - Rojas-Jiménez, K. A1 - Rodríguez-Rojas, A. T1 - Mechanisms of low susceptibility to the disinfectant benzalkonium chloride in a multidrug-resistant environmental isolate of Aeromonas hydrophila N2 - Excessive discharge of quaternary ammoniumdisinfectants such as benzalkonium chloride (BAC) into aquatic systems can trigger several physiological responses in environmental microorganisms. In this study, we isolated a less-susceptible strain of Aeromonas hydrophila to BAC, designated as INISA09, froma wastewater treatment plant in Costa Rica. We characterized its phenotypic response upon exposure to three di􀀀erent concentrations of BAC and characterizedmechanisms related to its resistance using genomic and proteomic approaches. The genome of the strain, mapped against 52 di􀀀erent sequenced A. hydrophila strains, consists of approximately 4.6Mb with 4,273 genes. We found a massive genome rearrangement and thousands of missense mutations compared to the reference strain A. hydrophila ATCC 7966. We identified 15,762 missense mutations mainly associated with transport, antimicrobial resistance, and outer membrane proteins. In addition, a quantitative proteomic analysis revealed a significant upregulation of several efflux pumps and the downregulation of porins when the strain was exposed to three BAC concentrations.Other genes related tomembrane fatty acid metabolism and redox metabolic reactions also showed an altered expression. Our findings indicate that the response of A. hydrophila INISA09 to BAC primarily occurs at the envelop level, which is the primary target of BAC. Our study elucidates the mechanisms of antimicrobial susceptibility in aquatic environments against a widely used disinfectant and will help better understand howbacteria can adapt to biocide pollution. To our knowledge, this is the first study addressing the resistance to BAC in an environmental A. hydrophila isolate. We propose that this bacterial species could also serve as a new model to study antimicrobial pollution in aquatic environments. KW - Antimicrobial resistance KW - Bacteria KW - Disinfection KW - Biocides PY - 2023 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-578555 DO - https://doi.org/10.3389/fmicb.2023.1180128 SN - 1664-302X VL - 14 SP - 1 EP - 18 PB - Frontiers SA CY - Lausanne AN - OPUS4-57855 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schmidt, Selina B. I. A1 - Täschner, Tom A1 - Nordholt, Niclas A1 - Schreiber, Frank T1 - Differential Selection for Survival and for Growth in Adaptive Laboratory Evolution Experiments With Benzalkonium Chloride N2 - Biocides are used to control microorganisms across different applications, but emerging resistance may pose risks for those applications. Resistance to biocides has commonly been studied using adaptive laboratory evolution (ALE) experiments with growth at subinhibitory concentrations linked to serial subculturing. It has been shown recently that Escherichia coli adapts to repeated lethal stress imposed by the biocide benzalkonium chloride (BAC) by increased survival (i.e., tolerance) and not by evolving the ability to grow at increased concentrations (i.e., resistance). Here, we investigate the contributions of evolution for tolerance as opposed to resistance for the outcome of ALE experiments with E. coli exposed to BAC. We find that BAC concentrations close to the half maximal effective concentration (EC50, 4.36 μg mL−1) show initial killing (~40%) before the population resumes growth. This indicates that cells face a two‐fold selection pressure: for increased survival and for increased growth. To disentangle the effects of both selection pressures, we conducted two ALE experiments: (i) one with initial killing and continued stress close to the EC50 during growth and (ii) another with initial killing and no stress during growth. Phenotypic characterization of adapted populations showed that growth at higher BAC concentrations was only selected for when BAC was present during growth. Whole genome sequencing revealed distinct differences in mutated genes across treatments. Treatments selecting for survival‐only led to mutations in genes for metabolic regulation (cyaA) and cellular structure (flagella fliJ), while treatments selecting for growth and survival led to mutations in genes related to stress response (hslO and tufA). Our results demonstrate that serial subculture ALE experiments with an antimicrobial at subinhibitory concentrations can select for increased growth and survival. This finding has implications for the design of ALE experiments to assess resistance risks of antimicrobials in different scenarios such as disinfection, preservation, and environmental pollution. KW - Antimicrobial resistance KW - Bacteria KW - Standardization KW - Biocides PY - 2024 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-615496 DO - https://doi.org/10.1111/eva.70017 VL - 17 IS - 10 SP - 1 EP - 11 PB - Wiley AN - OPUS4-61549 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - He, Zhiming A1 - Dechesne, Arnaud A1 - Schreiber, Frank A1 - Zhu, Yong-Guan A1 - Larsson, Joakim A1 - Smets, Barth T1 - Understanding Stimulation of Conjugal Gene Transfer by Nonantibiotic Compounds: How Far Are We? N2 - A myriad of nonantibiotic compounds is released into the environment, some of which may contribute to the dissemination of antimicrobial resistance by stimulating conjugation. Here, we analyzed a collection of studies to (i) identify patterns of transfer stimulation across groups and concentrations of chemicals, (ii) evaluate the strength of evidence for the proposed mechanisms behind conjugal stimulation, and (iii) examine the plausibility of alternative mechanisms. We show that stimulatory nonantibiotic compounds act at concentrations from 1/1000 to 1/10 of the minimal inhibitory concentration for the donor strain but that stimulation is always modest (less than 8-fold). The main proposed mechanisms for stimulation via the reactive oxygen species/SOS cascade and/or an increase in cell membrane permeability are not unequivocally supported by the literature. However, we identify the reactive oxygen species/SOS cascade as the most likely mechanism. This remains to be confirmed by firm molecular evidence. Such evidence and more standardized and high-throughput conjugation assays are needed to create technologies and solutions to limit the stimulation of conjugal gene transfer and contribute to mitigating global antibiotic resistance. KW - Antibiotic resistance KW - Horizontal gene transfer KW - Conjugation KW - Chemicals PY - 2024 DO - https://doi.org/10.1021/acs.est.3c06060 SP - 1 EP - 14 PB - ACS Publications AN - OPUS4-60105 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - European Food Safety Authority (EFSA), A1 - European Centre for Disease Prevention and Control (ECDC), A1 - European Chemicals Agency (ECHA), A1 - European Environment Agency (EEA), A1 - European Medicines Agency (EMA), A1 - European Commission's Joint Research Centre (JRC), T1 - Scientific report - Impact of the use of azole fungicides, other than as human medicines, on the development of azole‐resistant Aspergillus spp. N2 - The use of azoles in the European Union and European Economic Area (EU/EEA) other than as human medicines has raised concerns about emergence and spread of azole‐resistant Aspergillus species. EU agencies, with the support of JRC, reviewed the evidence and provided conclusions and recommendations on this topic. Although incomplete, data from 2010 to 2021 showed that around 120,000 tonnes of azoles were sold in EU/EEA for uses other than as human medicines. The majority are used as plant protection products (119,000 tonnes), with a stable temporal trend. Evidence supported a link between environmental azole exposure and cross‐resistance selection to medical azoles in Aspergillus species (primarily shown for A. fumigatus). Prevalence of azole‐resistant A. fumigatus in human A. fumigatus infections ranges from 0.7% to 63.6% among different disease presentations and geographic regions; mortality rates range from 36% to 100% for invasive aspergillosis (IA). It was concluded that azole usage outside the human domain is likely or very likely to contribute to selection of azole‐resistant A. fumigatus isolates that could cause severe disease like IA. Environmental hotspots for resistance selection were identified, including stockpiling of agricultural waste and their possible use as soil amendment/fertiliser for certain agricultural crops (for plant protection products) and freshly cut wood (for biocides). Recommendations were formulated on measures to prevent and control selection of azole resistance in A. fumigatus, including implementation of good agricultural/horticultural practices, proper agricultural and wood waste storage and management, and on approval of new azole fungicides or renewal of existing fungicides. Recommendations on topics to be covered by studies provided when submitting applications for the approval of azole fungicides were listed. For the evaluation of such studies within the approval procedure, a preliminary framework for risk assessment was developed and should be further refined. Data gaps and uncertainties were identified, alongside with respective recommendations to address them. KW - Antimicrobial surfaces KW - Biocides KW - Antimicrobial resistance KW - Azoles KW - Fungi KW - Wood preservatives PY - 2025 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-652175 DO - https://doi.org/10.2903/j.efsa.2025.9200 SN - 1831-4732 VL - 23 IS - 1 SP - 1 EP - 35 PB - Wiley CY - Hoboken, NJ AN - OPUS4-65217 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schmidt, Selina A1 - Rodríguez-Rojas, A. A1 - Rolff, J. A1 - Schreiber, Frank T1 - Biocides used as material preservatives modify rates of de novo mutation and horizontal gene transfer in bacteria N2 - Antimicrobial resistance (AMR) is a global health problem with the environment being an important compartment for the evolution and transmission of AMR. Previous studies showed that de-novo mutagenesis and horizontal gene transfer (HGT) by conjugation or transformation – important processes underlying resistance evolution and spread - are affected by antibiotics, metals and pesticides. However, natural microbial communities are also frequently exposed to biocides used as material preservatives, but it is unknown if these substances induce mutagenesis and HGT. Here, we show that active substances used in material preservatives can increase rates of mutation and conjugation in a species- and substance-dependent manner, while rates of transformation are not increased. The bisbiguanide chlorhexidine digluconate, the quaternary ammonium compound didecyldimethylammonium chloride, the metal copper, the pyrethroid-insecticide permethrin, and the azole-fungicide propiconazole increase mutation rates in Escherichia coli, whereas no increases were identified for Bacillus subtilis and Acinetobacter baylyi. Benzalkonium chloride, chlorhexidine and permethrin increased conjugation in E. coli. Moreover, our results show a connection between the RpoS-mediated general stress and the RecA-linked SOS response with increased rates of mutation and conjugation, but not for all biocides. Taken together, our data show the importance of assessing the contribution of material preservatives on AMR evolution and spread. KW - Mutation rate KW - Horizontal gene transfer KW - Biocides PY - 2022 DO - https://doi.org/10.1016/j.jhazmat.2022.129280 SN - 0304-3894 VL - 437 SP - 1 EP - 13 PB - Elsevier CY - Amsterdam AN - OPUS4-55261 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - An, Biwen Annie A1 - Deland, Eric A1 - Sobol, Oded A1 - Yao, Jizheng A1 - Skovhus, T. L. A1 - Koerdt, Andrea T1 - The differences in the corrosion product compositions of Methanogen-induced microbiologically influenced corrosion (Mi-MIC) between static and dynamic growth conditions N2 - Currently, corrosion rates (CR) and/or corrosion products (CP) obtained for methanogen-induced microbiologically influenced corrosion (Mi-MIC) on carbon steel are mainly analyzed from static-incubations. By using a multiport-flow-column, much higher CRs (0.72 mm/yr) were observed, indicating static-incubations are not suitable for determining the corrosive potential of Mi-MIC. With the combination of various analytical methods (ToF-SIMS/SEM-EDS/SEM-FIB) and contrary to previously published data, we observed that CPs contained phosphorus, oxygen, magnesium, calcium and iron but lacked carbon-related species (e.g. siderite). Overall, siderite nucleation is disrupted by methanogens, as they convert aqueous bicarbonate into carbon dioxide for methanogenesis resulting in increased localized corrosion. KW - Carbon steel KW - Modelling studies KW - SIMS KW - SEM KW - Reactor conditions KW - Microbiologically influenced corrosion PY - 2020 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-517632 DO - https://doi.org/10.1016/j.corsci.2020.109179 SN - 0010-938X VL - 180 SP - 9179 PB - Elsevier AN - OPUS4-51763 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Li, M. A1 - Schlaich, C. A1 - Zhang, J. A1 - Donskyi, Ievgen A1 - Schwibbert, Karin A1 - Schreiber, Frank A1 - Xia, Y. A1 - Radnik, Jörg A1 - Schwerdtle, T. A1 - Haag, R. T1 - Mussel-inspired multifunctional coating for bacterial infection prevention and osteogenic induction N2 - Bacterial infection and osteogenic integration are the two main problems that cause severe complications after surgeries. In this study, the antibacterial and osteogenic properties were simultaneously introduced in biomaterials, where copper nanoparticles (CuNPs) were generated by in situ reductions of Cu ions into a mussel-inspired hyperbranched polyglycerol (MI-hPG) coating via a simple dip-coating method. This hyperbranched polyglycerol with 10 % catechol groups’ modification presents excellent antifouling property, which could effectively reduce bacteria adhesion on the surface. In this work, polycaprolactone (PCL) electrospun fiber membrane was selected as the substrate, which is commonly used in biomedical implants in bone regeneration and cardiovascular stents because of its good biocompatibility and easy post-modification. The as-fabricated CuNPs-incorporated PCL membrane [PCL-(MI-hPG)-CuNPs] was confirmed with effective antibacterial performance via in vitro antibacterial tests against Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), and multi-resistant E. coli. In addition, the in vitro results demonstrated that osteogenic property of PCL-(MI-hPG)-CuNPs was realized by upregulating the osteoblast-related gene expressions and protein activity. This study shows that antibacterial and osteogenic properties can be balanced in a surface coating by introducing CuNPs. KW - Mussel-inspired coating KW - CuNPs KW - Multi-resistant bacteria KW - Antibacterial KW - Antifouling KW - Osteogenesis PY - 2021 DO - https://doi.org/10.1016/j.jmst.2020.08.011 SN - 1005-0302 VL - 68 SP - 160 EP - 171 PB - Elsevier Ltd. AN - OPUS4-51519 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Kanaris, Orestis A1 - Sobisch, Lydia-Yasmin A1 - Gödt, Annett A1 - Schreiber, Frank A1 - Nordholt, Niclas T1 - Consequences of benzalkonium chloride tolerance for selection dynamics and de novo resistance evolution driven by antibiotics N2 - Biocides are used in large amounts in industrial, medical, and domestic settings. Benzalkonium chloride (BAC) is a commonly used biocide, for which previous research revealed that Escherichia coli can rapidly adapt to tolerate BAC-disinfection, with consequences for antibiotic susceptibility. However, the consequences of BAC tolerance for selection dynamics and resistance evolution to antibiotics remain unknown. Here, we investigated the effect of BAC tolerance in E. coli on its response upon challenge with different antibiotics. Competition assays showed that subinhibitory concentrations of ciprofloxacin—but not ampicillin, colistin and gentamicin—select for the BAC-tolerant strain over the BAC-sensitive ancestor at a minimal selective concentration of 0.0013–0.0022 µg/mL. In contrast, the BAC-sensitive ancestor was more likely to evolve resistance to ciprofloxacin, colistin and gentamicin than the BAC-tolerant strain when adapted to higher concentrations of antibiotics in a serial transfer laboratory evolution experiment. The observed difference in the evolvability of resistance to ciprofloxacin was partly explained by an epistatic interaction between the mutations conferring BAC tolerance and a knockout mutation in ompF encoding for the outer membrane porin F. Taken together, these findings suggest that BAC tolerance can be stabilized in environments containing low concentrations of ciprofloxacin, while it also constrains evolutionary pathways towards antibiotic resistance. KW - AMR KW - Resistance evolution KW - Resistance selection PY - 2026 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-653842 DO - https://doi.org/10.1038/s44259-025-00170-8 SN - 2731-8745 VL - 4 IS - 1 SP - 1 EP - 13 PB - Springer Science and Business Media LLC AN - OPUS4-65384 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -