TY - JOUR A1 - Tavasolyzadeh, Zeynab A1 - Tang, Peng A1 - Hahn, Marc Benjamin A1 - Hweidi, Gada A1 - Nordholt, Niclas A1 - Haag, Rainer A1 - Sturm, Heinz A1 - Topolniak, Ievgeniia T1 - 2D and 3D Micropatterning of Mussel‐Inspired Functional Materials by Direct Laser Writing N2 - AbstractThis work addresses the critical need for multifunctional materials and substrate‐independent high‐precision surface modification techniques that are essential for advancing microdevices and sensing elements. To overcome existing limitations, the versatility of mussel‐inspired materials (MIMs) is combined with state‐of‐the‐art multiphoton direct laser writing (DLW) microfabrication. In this way, 2D and 3D MIM microstructures of complex designs are demonstrated with sub‐micron to micron resolution and extensive post‐functionalization capabilities. This study includes polydopamine (PDA), mussel‐inspired linear, and dendritic polyglycerols (MI‐lPG and MI‐dPG), allowing their direct microstructure on the substrate of choice with the option to tailor the patterned topography and morphology in a controllable manner. The functionality potential of MIMs is demonstrated by successfully immobilizing and detecting single‐stranded DNA on MIM micropattern and nanoarray surfaces. In addition, easy modification of MIM microstructure with silver nanoparticles without the need of any reducing agent is shown. The methodology developed here enables the integration of MIMs in advanced applications where precise surface functionalization is essential. KW - Direct laser writing KW - Mussel-inspired materials KW - Polyglycerol KW - Polydopamine KW - Micropatterning PY - 2023 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-588778 DO - https://doi.org/10.1002/smll.202309394 SN - 1613-6829 SP - 1 EP - 12 PB - Wiley-VCH CY - Weinheim AN - OPUS4-58877 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Bonse, Jörn T1 - A brief survey on open questions about laser-induced periodic surface structures N2 - The processing of laser-induced periodic surface structures (LIPSS) represents a simple and robust way for the nanostructuring of solids that allows creating a wide range of surface functionalities featuring applications in optics, tribology, medicine, energy technologies, etc. While the currently available laser and scanner technology already allows surface processing rates at the m2/min level, industrial applications of LIPSS are sometimes hampered by the complex interplay between the nanoscale surface topography and the specific surface chemistry. This typically manifests in difficulties to control the processing of LIPSS and in limitations to ensure the long-term stability of the created surface functions. This presentation aims to identify some unsolved scientific problems related to LIPSS, discusses the pending technological limitations, and sketches the current state of theoretical modelling. Hereby, it is intended to stimulate further research and developments in the field of LIPSS for overcoming these limitations and for supporting the transfer of the LIPSS technology into industry. T2 - E-MRS Spring Meeting 2022 CY - Online meeting DA - 30.05.2022 KW - Laser-induced periodic surface structures (LIPSS) KW - Surface functionalization KW - Industrial applications KW - Biofilm growth PY - 2022 AN - OPUS4-54929 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schulze-Makuch, D. A1 - Haque, S. A1 - Beckles, D. A1 - Schmitt-Kopplin, P. A1 - Harir, M. A1 - Schneider, Beate A1 - Stumpp, C. A1 - Wagner, D. T1 - A chemical and microbial characterization of selected mud volcanoes in Trinidad reveals pathogens introduced by surface water and rain water N2 - Terrestrial mud volcanoes are unique structures driven by tectonic pressure and fluids from the deep subsurface. These structures are mainly found in active tectonic zones, such as the area near the Los Bajos Fault in Trinidad. Here we report a chemical and microbiological characterization of three mud volcanoes, which included analyses of multiple liquid and solid samples from the mud volcanoes. Our study confirms previous suggestions that at least some of the mud volcano fluids are a mixture of deeper salt-rich water and surficial/precipitation water. No apparent water quality differences were found between sampling sites north and south of a major geological fault line. Microbiological analyses revealed diverse communities, both aerobic and anaerobic, including sulfate reducers, methanogens, carbon dioxide fixing and denitrifying bacteria. Several identified species were halophilic and likely derived from the deeper salt-rich subsurface water, while we also cultivated pathogenic species from the Vibrionaceae, Enterobacteriaceae, Shewanellaceae, and Clostridiaceae. These microorganisms were likely introduced into the mud volcano fluids both from surface water or shallow ground-water, and perhaps to a more minor degree by rain water. The identified pathogens are a major health concern that needs to be addressed. KW - Water stable isotope analysis KW - Mud volcanoe fluids KW - Metabolomics PY - 2020 DO - https://doi.org/10.1016/j.scitotenv.2019.136087 VL - 707 SP - 136087 PB - Elsevier B.V. AN - OPUS4-50499 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Marano, Roberto T1 - A Customizable Procedure for Predicting Antibiotic Resistance Selection on Novel Antimicrobial Coatings N2 - The global burden of antimicrobial resistance (AR) has manifested in an increasing number of deaths attributable to antibiotic-resistant bacteria (ARB) in recent years, with projections indicating a continued rise. Combating AR requires a multifaceted approach, one aspect of which involves preventing the spread of ARB in hospital environments via high-touch surfaces, which are known contributors to nosocomial infections. In conjunction with routine disinfection protocols and infection prevention measures, antimicrobial surfaces or antimicrobial coatings (AC) are increasingly being investigated and implemented to reduce microbial transmission via high-touch surfaces, thereby mitigating their spread in healthcare settings. However, similar to antibiotics, prolonged use of such surfaces may lead to the direct or indirect selection of ARB. To prevent this, targeted tests must be developed to predict potential AR selection before AC implementation. Materials and Methods As part of the Horizon-Europe-funded project "STOP" (Grant Agreement ID: 101057961), a novel antimicrobial coating is being developed and tested to reduce pathogen transfer on surfaces without selecting for ARB. To this end, two bacterial libraries (80 strains each) were assembled from the two species most associated with AR-attributable mortality worldwide, Escherichia coli and Staphylococcus aureus, as identified in the most recent comprehensive study [1]. Each library maintains a 50/50% ratio of strains resistant or sensitive to third-generation cephalosporins (E. coli) and methicillin (S. aureus), respectively—representing two of the most widespread resistance profiles. An adapted ISO 22196 method was developed using a reference benchmark antimicrobial surface (i.e., copper), with stainless steel serving as a control. The goal was to infer potential advantages of copper-unrelated antibiotic resistance phenotypes on the tested surface. Results Metadata, genomic data, and antibiotic susceptibility testing (AST) data were collected for all selected strains in each library, ensuring diversity in sequence types (ST) of clinical relevance, antibiotic resistance gene profiles, and geographical origins. The method was calibrated on copper surfaces using a reference E. coli strain, establishing the initial parameters required for investigators to customize the test for a given AC. Procedural reproducibility was assessed by comparing results from independent operators. Discussion The two libraries are currently being screened against the selected copper surfaces to identify potential associations between the resistance phenotypes of the tested species and their observed survival rates post-exposure. Three mutually exclusive outcomes are anticipated: (i) a statistically significant survival advantage of resistant strains compared to sensitive strains, (ii) an inverse scenario where sensitive strains exhibit higher survival, or (iii) no significant difference between the two groups. Furthermore, post hoc principal component analysis utilizing metadata and AST data may help elucidate genetic traits that confer a survival advantage on the tested AC. Conclusions This test aims to assist developers of antimicrobial coatings and materials in assessing potential selective pressures toward ARB before these products are implemented and evaluated under real-life conditions. T2 - ESB Conference CY - Torino, Italy DA - 07.09.2025 KW - Antimicrobial-resistance KW - Surface antimicrobial testing KW - Materials testing PY - 2025 AN - OPUS4-64569 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Marano, Roberto T1 - A Customizable Procedure for Predicting Antibiotic Resistance Selection on Novel Antimicrobial Coatings N2 - This presentation introduceda new methodological approach developed to test antimocrobials implemented on high touch surfaces, inlcuding preliminary results on its use on reference antimicrobial materials. The seminar was organized by the 'Fast-real' project, funded by the Horizon Europe (project ID: 101159721). T2 - Novel strategies and considerations in fighting pathogens CY - Tartu, Estonia DA - 16.06.2025 KW - Antimicrobial-resistance KW - Surface antimicrobial testing KW - Materials testing PY - 2025 AN - OPUS4-64568 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Nordholt, Niclas A1 - O'Hara, Kate A1 - Resch-Genger, Ute A1 - Blaskovich, M. A1 - Rühle, Bastian A1 - Schreiber, Frank T1 - A fluorescently labelled quaternary ammonium compound (NBD-DDA) to study resistance mechanisms in bacteria 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, including phenotypic heterogeneity, that can make bacteria less susceptible to QACs. To facilitate studies on resistance mechanisms towards QACs, we synthesized a fluorescent quaternary ammonium compound, namely N-dodecyl-N,N-dimethyl-[2-[(4-nitro-2,1,3-benzoxadiazol-7-yl)amino]ethyl]azanium-iodide (NBD-DDA). NBD-DDA is readily detected by flow cytometry and fluorescence microscopy with standard GFP/FITC-settings, making it suitable for molecular and single-cell studies. As a proof-of-concept, NBD-DDA was then used to investigate resistance mechanisms which can be heterogeneous among individual bacterial cells. Our results reveal that the antimicrobial activity of NBD-DDA against Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa is comparable to that of benzalkonium chloride (BAC), a widely used QAC, and benzyl-dimethyl-dodecylammonium chloride (BAC12), a mono-constituent BAC with alkyl-chain length of 12 and high structural similarity to NBD-DDA. 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. As revealed by confocal laser scanning microscopy (CLSM), NBD-DDA is preferentially localized to the cell envelope of E. coli, which is a primary target of BAC and other QACs. Leveraging these findings and NBD-DDA‘s fluorescent properties, we show that reduced cellular accumulation is responsible for the evolved BAC tolerance in the BAC tolerant E. coli strain and that NBD-DDA is subject to efflux mediated by TolC. Overall, NBD-DDA’s antimicrobial activity, its fluorescent properties, and its ease of detection render it a powerful tool to study resistance mechanisms of QACs in bacteria and highlight its potential to gain detailed insights into its mode-of-action. KW - Antimicrobial resistance KW - Bacteria KW - Disinfection KW - Biocides PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-563811 DO - https://doi.org/10.3389/fmicb.2022.1023326 SN - 1664-302X IS - 13 SP - 1 EP - 13 PB - Frontiers Media CY - Lausanne AN - OPUS4-56381 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Wen, Keqing A1 - Gorbushina, Anna A1 - Schwibbert, Karin A1 - Bell, Jérémy T1 - A microfluidic platform for monitoring biofilm formation in flow under defined hydrodynamic conditions N2 - Bacterial adhesion on surfaces of medical, water and food applications may lead to infections, water or food spoilage and human illness. In comparison to traditional static and macro flow chamber assays for biofilm formation studies, microfluidic chips allow in situ monitoring of biofilm formation under various flow regimes, have better environment control and smaller sample requirements. In this work, a novel microfluidic platform is developed to investigate biofilm adhesion under precisely controlled bacteria concentration, temperature, and flow conditions. This platform central unit is a single-inlet microfluidic flow cell with a 5 mm wide chamber designed and tested to achieve ultra-homogenous flow in the central area of chamber. Within this area, defined microstructures are integrated that will disturb the homogeneity of the flow, thus changing bacterial adhesion pattern. Here we present the monitoring of bacterial biofilm formation in a microfluidic chip equipped with a microstructure known as micro-trap. This feature is based on a 3D bacteria trap designed by Di Giacomo et al. and successfully used to sequester motile bacteria. At first, fluorescent particles similar in size to Escherichia coli (E. coli) are used to simulate bacteria flow inside the flow cell and at the micro-trap. The turbulences induced by the trap are analyzed by imaging and particle tracking velocimetry (PTV). Secondly, the model strain E. coli TG1, ideal and well described for biofilm studies, is used to analyze biofilm formation in the micro-trap. Therefore, a stable fluorescent strain E. coli TG1-MRE-Tn7-141 is constructed by using Tn7 transposon mutagenesis according to the method described by Schlechter et al. Sequestering of E. coli cells within the micro-trap was followed using epifluorescence microscopy. The novel microfluidic platform shows great potential for assessment of bacterial adhesion under various flow regimes. The performance of structural feature with respect to the generation of turbulences that promote or reduce bacterial adhesion can be systematically examined. The combination of flow analysis and fluorescent strain injection into the microfluidic chip shows that the micro-trap is useful for capturing bacteria at defined positions and to study how flow conditions, especially micro-turbulences, can affect biofilm formation. It represents a powerful and versatile tool for studying the relation between topography and bacteria adhesion. T2 - International Conference on Miniaturized Systems for Chemistry and Life Sciences CY - Katowice, Poland DA - 15.10.2023 KW - Biofilm KW - E. coli KW - Microfluidics KW - Velocimetry KW - Fluorescence PY - 2023 AN - OPUS4-59593 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Hobmeier, K. A1 - Cantone, M. A1 - Nguyen, Q. A. A1 - Pflüger-Grau, K. A1 - Kremling, A. A1 - Kunte, Hans-Jörg A1 - Pfeiffer, F. A1 - Marin-Sanguino, A. T1 - Adaptation to varying salinity in Halomonas elongata: Much more than ectoine accumulation N2 - The halophilic γ-proteobacterium Halomonas elongata DSM 2581T thrives at salt concentrations well above 10 % NaCl (1.7 M NaCl). A well-known osmoregulatory mechanism is the accumulation of the compatible solute ectoine within the cell in response to osmotic stress. While ectoine accumulation is central to osmoregulation and promotes resistance to high salinity in halophilic bacteria, ectoine has this effect only to a much lesser extent in non-halophiles. We carried out transcriptome analysis of H. elongata grown on two different carbon sources (acetate or glucose), and low (0.17 M NaCl), medium (1 M), and high salinity (2 M) to identify additional mechanisms for adaptation to high saline environments. To avoid a methodological bias, the transcripts were evaluated by applying two methods, DESeq2 and Transcripts Per Million (TPM). The differentially transcribed genes in response to the available carbon sources and salt stress were then compared to the transcriptome profile of Chromohalobacter salexigens, a closely related moderate halophilic bacterium. Transcriptome profiling supports the notion that glucose is degraded via the cytoplasmic Entner-Doudoroff pathway, whereas the Embden-Meyerhoff-Parnas pathway is employed for gluconeogenesis. The machinery of oxidative phosphorylation in H. elongata and C. salexigens differs greatly from that of non-halophilic organisms, and electron flow can occur from quinone to oxygen along four alternative routes. Two of these pathways via cytochrome bo' and cytochrome bd quinol oxidases seem to be upregulated in salt stressed cells. Among the most highly regulated genes in H. elongata and C. salexigens are those encoding chemotaxis and motility proteins, with genes for chemotaxis and flagellar assembly severely downregulated at low salt concentrations. We also compared transcripts at low and high-salt stress (low growth rate) with transcripts at optimal salt concentration and found that the majority of regulated genes were down-regulated in stressed cells, including many genes involved in carbohydrate metabolism, while ribosome synthesis was up-regulated, which is in contrast to what is known from non-halophiles at slow growth. Finally, comparing the acidity of the cytoplasmic proteomes of non-halophiles, extreme halophiles and moderate halophiles suggests adaptation to an increased cytoplasmic ion concentration of H. elongata. Taken together, these results lead us to propose a model for salt tolerance in H. elongata where ion accumulation plays a greater role in salt tolerance than previously assumed. KW - Ectoine KW - Osmoadaptation PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-545608 DO - https://doi.org/10.3389/fmicb.2022.846677 SN - 1664-302X VL - 13 SP - 1 EP - 19 PB - Frontiers Media CY - Lausanne AN - OPUS4-54560 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Koerdt, Andrea A1 - Gerengi, Husnu A1 - Kaya, Ertugrul A1 - M. Solomon, Moses A1 - Snape, Matthew T1 - Advances in the Mitigation of Microbiologically Influenced Concrete Corrosion: A Snapshot N2 - Concrete, a versatile construction material, faces pervasive deterioration due to microbiologically influenced corrosion (MIC) in various applications, including sewer systems, marine engineering, and buildings. MIC is initiated by microbial activities such as involving sulfate-reducing bacteria (SRB), sulfur-oxidizing bacteria (SOB), etc., producing corrosive substances like sulfuric acid. This process significantly impacts structures, causing economic losses and environmental concerns. Despite over a century of research, MIC remains a debated issue, lacking standardized assessment methods. Microorganisms contribute to concrete degradation through physical and chemical means. In the oil and gas industry, SRB and SOB activities may adversely affect concrete in offshore platforms. MIC challenges also arise in cooling water systems and civil infrastructures, impacting concrete surfaces. Sewer systems experience biogenic corrosion, primarily driven by SRB activities, leading to concrete deterioration. Mitigation traditionally involves the use of biocides and surface coatings, but their long-term effectiveness and environmental impact are questionable. Nowadays, it is important to design more eco-friendly mitigation products. The microbial-influenced carbonate precipitation is one of the green techniques and involves incorporating beneficial bacteria with antibacterial activity into cementitious materials to prevent the growth and the formation of a community that contains species that are pathogenic or may be responsible for MIC. These innovative strategies present promising avenues for addressing MIC challenges and preserving the integrity of concrete structures. This review provides a snapshot of the MIC in various areas and mitigation measures, excluding underlying mechanisms and broader influencing factors. KW - MIC KW - Corrosion KW - Concrete KW - Environment PY - 2024 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-619218 DO - https://doi.org/10.3390/ma17235846 VL - 17 IS - 23 SP - 1 EP - 19 PB - MDPI AN - OPUS4-61921 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Nordholt, Niclas T1 - ALEE-AMC: Bacterial resistance evolution towards antimicrobial surfaces and development of a standardized test N2 - Background: Antimicrobial surfaces and coatings (AMCs) are important to protect man-made structures from biodeterioration and biodegradation. Advances in nano-structuring methods hold the promise of a new generation of AMCs. However, the evolution and selection of bacterial resistance to AMCs may threaten their efficacy in the long term. Therefore, according to the EU Biocidal Products Regulation, the risk of resistance development upon exposure to AMCs must be evaluated during product authorization. The same applies to the development of cross-resistances to other substances, for instance biocides and antibiotics. However, no standardized method exists to assess the risk of resistance and cross-resistance development upon exposure to AMCs during the authorization process. Objectives: • To develop a standardizable adaptive laboratory evolution experiment to be performed on AMCs (ALEE-AMC) • To assess performance and robustness of ALEE-AMC in a round robin test, using a copper AMC as reference • To uncover the mechanisms underlying evolution of resistance to copper AMC Materials & Methods: ALEE-AMC was developed based on an approved standard to determine the efficacy of antimicrobial surfaces (ISO 22196). ALEE-MC was performed on an antimicrobial copper surface as reference material and Escherichia coli as model organism. A round robin test was conducted with six participants to evaluate the reproducibility and applicability of ALEE-AMC. Evolved E. coli populations from the round robin partners were collected and subjected to phenotypic (antimicrobial susceptibility testing, ISO 22196) and genotypic (whole genome sequencing) characterization at BAM. Results: The results of the ALEE-AMC round robin test indicate that repeated exposure to copper can select for reduced copper susceptibility. However, failure of individual E. coli lineages to adapt to the copper surfaces was also observed. Evolved E. coli exhibited increased survival upon exposure to copper surfaces. Adaptation to copper did not induce cross-resistance to antibiotics. Whole genome sequencing of the evolved E. coli revealed high diversity of mutations among individual evolved strains, indicating the existence of multiple, underexplored evolutionary pathways towards increased survival of antimicrobial copper surfaces. Conclusion & Significance: ALEE-AMC offers a standardizable platform to assess the risk of resistance development towards novel and existing AMCs. Specifically, using ALEE-AMC in a round robin test, insights into evolvable survival mechanisms to copper AMCs have been gained. These mechanistic insights may be exploited to prevent the evolution against copper AMCs. In future steps, criteria need to be defined to provide guidelines for the authorization of AMCs based on the outcomes of ALEE-AMC T2 - International Biodeterioration and Biodegradation Symposium, Berlin, Germany CY - Berlin, Germany DA - 09.09.2024 KW - Biocides KW - Antimicrobial surfaces KW - Resistance KW - Evolution KW - Standardized test PY - 2024 AN - OPUS4-61176 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Varadarajan, A. A1 - Allan, R. A1 - Valentin, J. A1 - Castañeda Ocampo, O. A1 - Somerville, V. A1 - Buhmann, M. A1 - West, J. A1 - Skipp, Paul A1 - van der Mei, H. A1 - Ren, Q. A1 - Schreiber, Frank A1 - Webb, J. A1 - Pietsch, Franziska A1 - Ahrens, C. T1 - An integrated model system to gain mechanistic insights into biofilm-associated antimicrobial resistance in Pseudomonas aeruginosa MPAO1 N2 - Pseudomonas aeruginosa MPAO1 is the parental strain of the widely utilized transposon mutant collection for this important clinical pathogen. Here, we validate a model system to identify genes involved in biofilm growth and biofilm-associated antibiotic resistance. Our model employs a genomics-driven workflow to assemble the complete MPAO1 genome, identify unique and conserved genes by comparative genomics with the PAO1 reference strain and genes missed within existing assemblies by proteogenomics. Among over 200 unique MPAO1 genes, we identified six general essential genes that were overlooked when mapping public Tn-seq data sets against PAO1, including an antitoxin. Genomic data were integrated with phenotypic data from an experimental workflow using a user-friendly, soft lithography-based microfluidic flow chamber for biofilm growth and a screen with the Tn-mutant library in microtiter plates. The screen identified hitherto unknown genes involved in biofilm growth and antibiotic resistance. Experiments conducted with the flow chamber across three laboratories delivered reproducible data on P. aeruginosa biofilms and validated the function of both known genes and genes identified in the Tn-mutant screens. Differential Protein abundance data from planktonic cells versus biofilm confirmed the upregulation of candidates known to affect biofilm formation, of structural and secreted proteins of type VI secretion systems, and provided proteogenomic evidence for some missed MPAO1 genes. This integrated, broadly applicable model promises to improve the mechanistic understanding of biofilm formation, antimicrobial tolerance, and resistance evolution in biofilms. KW - Biofilms PY - 2020 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-515108 DO - https://doi.org/10.1038/s41522-020-00154-8 VL - 6 IS - 1 SP - Article number: 46 PB - Springer Nature CY - Singapore AN - OPUS4-51510 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schiebel, J. A1 - Noack, J. A1 - Rödiger, S. A1 - Kammel, A. A1 - Menzel, Friederike A1 - Schwibbert, Karin A1 - Weise, Matthias A1 - Weiss, R. A1 - Böhm, A. A1 - Nitschke, J. A1 - Elimport, A. A1 - Roggenbuck, D. A1 - Schierack, P. T1 - Analysis of three-dimensional biofilms on different material surfaces N2 - Biofilms cause complications and high costs in both industry and medicine. Of particular interest are bacterial infections of prosthetic materials, which usually cannot be eliminated due to the high antibiotic resistance known for bacteria forming biofilms. The search for new materials and coatings with lower colonization potential and antibacterial activity is of great importance to reduce biofilm formation. However, there is no standardized procedure to examine the colonization characteristics of bacteria in the Biofilm state in situ. Here, we describe an automated epifluorescence microscopy system for the semi-quantitative analysis of three-dimensional (3D) biofilms on various surfaces. To analyze adherent bacteria, three materials (glass, steel and titanium) were incubated with bacteria in a flow chamber system. After fluorescence staining of the bacteria, automated image capturing, quantification of the bacteria, measurement of the colonized area and determination of the 3D biofilm height were carried out by using novel software. Furthermore, the materials were examined for their surface topography using white light scanning interferometry. Titanium compared to glass showed a significantly higher number of adherent bacteria. We argue that this was due to the higher microroughness of titanium. The colonized area was in accordance with the number of adherent bacteria and was also significantly larger on titanium coupons compared to glass. Maximum 3D biofilm height on glass coupons was significantly lower compared to the ones on steel and titanium. This novel method enables the standardized, automated investigation of the colonization with bacteria on different materials. This approach can considerably support the characterization of new material surfaces and their innovative coatings by analyzing the amount of attached Bacteria and thickness of biofilms in situ and eliminates the need of conventional cultivation. KW - Biofilm KW - Bacterial adhesion KW - Biofilm quantification KW - Automated analysis PY - 2020 DO - https://doi.org/10.1039/D0BM00455C SP - 1 EP - 11 PB - Royal Society of Chemistry AN - OPUS4-50815 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Hobmeier, K. A1 - Goëss, M. C. A1 - Sehr, C. A1 - Schwaminger, S. A1 - Berensmeier, S. A1 - Kremling, A. A1 - Kunte, Hans-Jörg A1 - Pflüger-Grau, K. A1 - Marin-Sanguino, A. T1 - Anaplerotic Pathways in Halomonas elongata: The Role of the Sodium Gradient N2 - Salt tolerance in the γ-proteobacterium Halomonas elongata is linked to its ability to produce the compatible solute ectoine. The metabolism of ectoine production is of great interest since it can shed light on the biochemical basis of halotolerance as well as pave the way for the improvement of the biotechnological production of such compatible solute. Ectoine belongs to the biosynthetic family of aspartate-derived amino-acids. Aspartate is formed from oxaloacetate, thereby connecting ectoine production to the anaplerotic reactions that refill carbon into the tricarboxylic acid cycle (TCA cycle). This places a high demand on these reactions and creates the need to regulate them not only in response to growth but also in response to extracellular salt concentration. In this work, we combine modeling and experiments to analyze how these different needs shape the anaplerotic reactions in H. elongata. First, the stoichiometric and thermodynamic factors that condition the flux distributions are analyzed, then the optimal patterns of operation for oxaloacetate production are calculated. Finally, the phenotype of two deletion mutants lacking potentially relevant anaplerotic enzymes: phosphoenolpyruvate carboxylase (Ppc) and oxaloacetate decarboxylase (Oad) are experimentally characterized. The results show that the anaplerotic reactions in H. elongata are indeed subject to evolutionary pressures that differ from those faced by other gram-negative bacteria. Ectoine producing halophiles must meet a higher metabolic demand for oxaloacetate and the reliance of many marine bacteria on the Entner-Doudoroff pathway compromises the anaplerotic efficiency of Ppc, which is usually one of the main enzymes fulfilling this role. The anaplerotic flux in H. elongata is contributed not only by Ppc but also by Oad, an enzyme that has not yet been shown to play this role in vivo. Ppc is necessary for H. elongata to grow normally at low salt concentrations but it is not required to achieve near maximal growth rates as long as there is a steep sodium gradient. On the other hand, the lack of Oad presents serious difficulties to grow at high salt concentrations. This points to a shared role of these two enzymes in guaranteeing the supply of oxaloacetate for biosynthetic reactions. KW - Metabolic flux analysis KW - Halophilic bacteria KW - Halomonas elongata KW - Metabolic modeling PY - 2020 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-513124 DO - https://doi.org/10.3389/fmicb.2020.561800 VL - 11 SP - 561800 AN - OPUS4-51312 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - RPRT 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 - Annex to: Scientific report 'Impact of the use of azole fungicides, other than as human medicines, on the development of azole-resistant Aspergillus spp.' doi:10.2903/j.efsa.2025.9200 - Annex E - Detailed answer to Term of Reference 5 'Environmental hotspots' and Term of Reference 6 'Prevention and control options' N2 - The widespread use of azole compounds in various sectors has led to the emergence of azole-resistant Aspergillus fumigatus (ARAf), which poses a significant challenge for treating fungal infections, especially in immunocompromised patients. Certain environmental conditions and practices, particularly in agricultural settings and the use of azoles as biocides, have been identified as hotspots for the selection and dispersal of azole-resistant strains of Aspergillus spp. Factors contributing to the selection of resistance include the use of azoles in crop protection, wood preservation and, to a much lesser extent, veterinary medicine. For plant protection products (PPPs), a number of scenarios (green waste of indoor-grown vegetables, uses with the production of wet pomace used as fertiliser, maize or sugar beet silage, and field heaps including flower bulbs) are deemed high risk for hotspot development. Based on EU authorised use patterns, these scenarios are characterised by the hazard characteristics of the azole fungicides in terms of activity against the wild-type Aspergillus spp. compared to resistant strains, substrate characteristics and residue levels, and environmental conditions that promote the growth of the fungus. For biocidal azole applications, products (biocidal product [BP]) for temporary preservation of freshly cut wood have been identified to have the potential for hotspot formation because freshly cut wood allows the growth of Aspergillus spp., and azole concentrations in treated wood are above the predicted no effect concentration (PNEC) for resistance selection (PNECres) and below the minimum inhibitory concentration (MIC) of ARAf for most analysed products on the EU market. Following identification of environmental hotspots, the report recommends measures to prevent the selection of azole-resistant strains in the environment, including controlled storage of organic waste, proper waste management, and responsible use and disposal of azole-treated products. Azole use in veterinary medicinal products (VMPs) represents a very small percentage of total azole use and is unlikely to be a significant source of selection of resistance in the environment. As such, the focus for mitigating resistance should be on other uses of azoles. The report stresses the importance of ongoing surveillance to monitor the presence of ARAf in the environment and to inform risk assessments and management strategies. As industrial chemicals, the azole substances are mostly used as intermediates (precursors) to manufacture yet a different substance, are formulated into a mixture or are reported to be manufactured as active substances in PPP, BP or VMP (therefore already covered above). There are only a few industrial azole substances with widespread use, and as for the moment, there is no evidence from the literature that industrial azoles would be a source of a possible hotspot; thus, the industrial chemicals were not further investigated. There are several areas where further research is needed, including understanding the environmental conditions that support the growth of Aspergillus spp. in different agricultural matrices or on wood, assessing human exposure to resistant strains, regional waste practices and the impact of active substance combinations for azole resistance selection. There is also a need for more comprehensive data on the use and quantities of azole-containing products. Furthermore, industrial substances with widespread use and having antifungal effects, e.g. an antidandruff substance in cosmetics, could be further investigated. Measures were identified that could be implemented with respect to the use of azole fungicides in PPPs as well as in BPs and with respect to the storage, processing and disposal of crop (waste) materials containing azole residues to prevent or minimise the selection of environmental resistance or to minimise the spread of resistant Aspergillus spp. to patients. Any measures that slow down or prevent growth in the presence of azoles, sporulation and dispersal of Aspergillus spp. should be encouraged. A coordinated effort among various stakeholders, including farmers, manufacturers, industrial users, waste managers, regulatory bodies and scientists, is essential to effectively address the challenge of azole resistance in A. fumigatus. 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:b43-652187 UR - https://doi.org/10.5281/zenodo.14223436 DO - https://doi.org/10.5281/zenodo.14223435 SP - 1 EP - 76 PB - Zenodo CY - Geneva AN - OPUS4-65218 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Stephan, Ina A1 - Dimke, Thomas A1 - Guterman, R. A1 - Smith, C. A. A1 - Cataldo, V. A. T1 - Antibacterial and degradable thioimidazolium poly(ionic liquid) N2 - New antibacterial agents are urgently required to fight the emergence of antibiotic-resistant bacteria. We recently synthesized the first thioimidazolium ionene, which has antibacterial properties and can degrade in various media. This dual functionality is crucial in order to limit the environmental impact of these biocides. We have found that our polymer is stronger than benzalkonium chloride (BAC) against Pseudomonas aeruginosa and also readily degrades in the presence of base, while remaining stable in acidic environments. These results highlight a new emerging class of antibacterial degradable polymers. KW - Ionic liquid KW - Antibacterial polymer KW - Amphiphilic polymer KW - Thioimidazolium KW - Degradable polymer KW - Polyioinic liquid PY - 2020 DO - https://doi.org/10.1021/acssuschemeng.0c02666 VL - 8 IS - 22 SP - 8419 EP - 8424 PB - ACS Publications AN - OPUS4-50982 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jaut, Valerie T1 - Antibiotic tolerance of biofilms emerging from multicellular effects of antibiotic efflux N2 - Biofilms are multicellular assemblages of bacteria living in a self-produced extracellular matrix. Different mechanisms, like the development of highly tolerant persister cells or increased expression of efflux pumps make them tolerant. Here we want to investigate the emergency of antimicrobial tolerance of multicellular bacterial populations, through the interplay of efflux-mediated spatial interactions and efflux-linked persistence. To this end, we are combining computational modelling with experimentally observations gained from three types of multicellular assemblages, i.e. colonies on agar, multicellular populations grown in a monolayer microfluidic device, and 3D biofilms grown in flow chambers. We generated fluorescently labeled E. coli strains that differ in their levels of AcrAB-TolC efflux pump activity, an acrB knockout-strain and a strain with inducible expression of acrAB. All strains were characterized in terms of their antimicrobial susceptibility of three antibiotics, tetracycline, kanamycin, ampicillin, and the biocide benzalkonium chloride. The knockout strain shows higher susceptibility than the wild type strain, with highest difference when using benzalkonium chlorid e. To investigate the link between colony structure and spatial patterns of gene expression, the strains were mixed equimolar and grown on agar supplemented with antimicrobials. First results show, antimicrobials affect the structure of sector formation and morphology. Cells grown on tetracycline agar show a more finer sector formation. While kanamycin changes the overall colony structure . We will develop a mathematical model and additional experiments with multicellular assemblages to explain the observed interactions and extrapolate the results to more realistic biofilm models. T2 - SPP Conference CY - Berlin, Germany DA - 06.01.2025 KW - Antimicrobial KW - Resistance KW - Tolerance KW - Efflux KW - E. coli PY - 2025 AN - OPUS4-64563 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jaut, Valerie A1 - Schreiber, Frank T1 - Antibiotic tolerance of biofilms emerging fro multicellular effects of antibiotic efflux N2 - The overarching goal of this project is to develop a predictive model for efflux-mediated antimicrobial tolerance in bacterial multicellular assemblies. Our central hypostasis is that efflux pump activity causes emergent antibiotic tolerance of multicellular bacterial populations, through the interplay of efflux mediated spatial interactions and efflux-linked persistence. To test this hypothesis, we will use a combination of microscopy, microbial killing assays, computational modelling, and data analysis, integrating information from 3 types of multicellular assembly: colonies, cell-to-cell interactions in a monolayer microfluidic device, and 3D flow chamber biofilms. Building on our preliminary observations, we will experimentally characterize the link between colony structure and spatial patterns of efflux gene expression in strains that differ in their levels of efflux. We will develop a mathematical model to test whether local growth inhibition of neighbors due to effluxing cells, coupled with local environment-dependent regulation of efflux, can account qualitatively for these results. By including persister cell formation in our model we will predict, and measure, the emergent function of antimicrobial tolerance in our colonies. To fully understand how tolerance emerges from the interplay between efflux-mediated spatial interactions and efflux-linked persister cell formation, we need quantitative measurements at the single cell level. To this end, we will use a microfluidic setup with cells growing in a monolayer to qualify in detail the dependence of efflux expression and persister cell formation on nutrient conditions, the correlation between efflux and persister formation, and the spatial range of efflux-mediated neighbour growth inhibition. To predict and quantitatively understand the emergent multicellular function of tolerance, we will perform individual-based modelling of biofilm growth, using as input the parameters measured on the single-cell level with our microfluidics experiments. Our simulations will predict biofilm spatial structure development, patterns of efflux and persister formation and, ultimately, tolerance to antimicrobial challenge. These predictions will be directly tested in flow-cell biofilm experiments. We are currently generating acrAB-tolC knockout-strain, without efflux activity, and a strain with an inducible acrAB-tolC efflux pump. To distinguish the different strains under the microscope, they were labeled with genes encoding for different fluorescent proteins. All strains are currently characterized in terms of growth, minimum inhibitory concentration of different antimicrobial substances, colony morphology, and biofilm formation ability. On the theoretical side, we are currently working on modeling the system at various scales and degree of detail, ranging from coarse-grained continuum models to stochastic, individual-based models. Some exploratory work was doe to test existing software for individual-based modelling that may be adapted for our purpose. Furthermore, we are in the process of developing more coarse-grained models. This work involves some physiological modelling and literature search, focusing on working mechanisms of efflux pumps and kinetic models for import and export of antibiotics. T2 - SPP Meeting CY - Jena, Germany DA - 04.10.2023 KW - Antibiotic KW - Bioilm KW - Tolerance KW - Efflux PY - 2023 AN - OPUS4-59245 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jaut, Valerie T1 - Antibiotic tolerance of biofilms emerging from multicellular effects of antibiotic efflux N2 - Biofilms are multicellular assemblies of bacteria living in a self-produced extracellular matrix. One characteristic of biofilms is that they are difficult to kill. Different mechanisms, like the development of persister cells or efflux pumps which pump some antimicrobials out of the cell, make them tolerant. Our central hypothesis is that efflux pump activity causes emergent antimicrobial tolerance of multicellular bacterial populations, through the interplay of efflux-mediated spatial interactions and efflux-linked persistence. To verify the hypothesis, we combine computational modelling with information gained from 3 types of multicellular assemblies. We are currently generating strains that differ in their levels of efflux activity, mixes are then cultivated together in the 3 model systems. In colonies the link between structure and spatial patterns of gene expression will be characterized. Using a microfluidic device, the interactions range of efflux as a response to different antimicrobials will be determined. In a flow chamber a 3D biofilm will be generated, to investigate the biofilm development over time and persister cell formation. All results will be compared with model predictions. T2 - EuroBioFilms2024 CY - Copenhagen, Denmark DA - 25.06.2024 KW - Antibiotic KW - Biofilm KW - Tolerance KW - Efflux PY - 2024 AN - OPUS4-61277 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jaut, Valerie T1 - Antibiotic tolerance of biofilms emerging from multicellular effects of antibiotic efflux N2 - Efflux pumps play an important role in the context of antimicrobial resistance, which is the ability to grow in the presence of antimicrobials. Many of these transporters can be categorized into multidrug efflux pumps, extruding various antimicrobials out of the cells, and thereby leading to antimicrobial resistance. It has been shown that efflux pumps can be linked by global regulators that regulate efflux pump expression affecting cell-to-cell-interactions, membrane integrity and biofilm formation. The aim of this study is to investigate the survival of cells in biofilms upon exposure to antimicrobials through the interplay of efflux-mediated spatial interactions . To this end, we generated fluorescently labeled E. coli strains that differ in their levels of AcrAB-TolC efflux pump activity and an acrB knockout-strain. The strains were characterized in terms of their antimicrobial susceptibility of three antibiotics, tetracycline, kanamycin, ampicillin, and the biocide benzalkonium chloride. The knockout strain shows higher susceptibility than the wildtype strain, with highest difference observed upon exposure to benzalkonium chloride. The results were confirmed with an efflux activity assay, which showed decreased efflux for the knockout strain as compared to the wildtype. Interestingly, adding the efflux inhibitor PAβN at intermediate concentrations induced bimodality in efflux activity in the wildtype. To investigate the link between colony structure and spatial patterns of efflux pump gene expression, strains with different fluorescent labels and efflux activity were mixed in a 1:1 ratio and grown on agar supplemented with antimicrobials at sub-inhibitory concentrations. Analysis of the colonies with fluorescence microcopy shows that the absence of the AcrAB efflux pump affects the structure of sector formation and morphology within the colony. We observed relatively large sectors with similar surface area for high efflux and low efflux cells in the absence of antimicrobials and at low concentrations. In contrast, sectors are disappearing due a strong intermixing of high and low efflux strains with increasing antibiotic concentration, specifically upon exposure to tetracycline and kanamycin. As next steps, we will perform quantitative analysis of colony images, to better interpret the results, develop a mathematical model of interacting cell types and investigate efflux-based interactions at the single-cell level. Taken together, out data suggest that efflux shapes cell-to-cell interactions and these interactions affect the spatial arrangement and the morphology of biofilms. Understanding the dynamics can provide insights into the emergence of resistance of bacterial communities to antimicrobial environments, the complex interplay of resistance, efflux, and biofilm formation, which potentially gives information to combat biofilm resistance. T2 - Multi-Drug Efflux Systems Gordon Research Conference CY - Lucca, Italy DA - 26.04.2025 KW - Antimicrobial KW - Resistance KW - Tolerance KW - Efflux KW - E. coli PY - 2025 AN - OPUS4-64561 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jaut, Valerie T1 - Antibiotic tolerance of biofilms emerging from multicellular effects of antibiotic efflux N2 - Biofilms are multicellular assemblies of bacteria living in a self-produced extracellular matrix. One characteristic of biofilms is that they are difficult to kill. Different mechanisms, like the development of persister cells or efflux pumps which pump some antimicrobials out of the cell, make them tolerant. Our central hypothesis is that efflux pump activity causes emergent antimicrobial tolerance of multicellular bacterial populations, through the interplay of efflux-mediated spatial interactions and efflux-linked persistence. To verify the hypothesis, we combine computational modelling with information gained from 3 types of multicellular assemblies. We are currently generating strains that differ in their levels of efflux activity, mixes are then cultivated together in the 3 model systems. In colonies the link between structure and spatial patterns of gene expression will be characterized. Using a microfluidic device, the interactions range of efflux as a response to different antimicrobials will be determined. In a flow chamber a 3D biofilm will be generated, to investigate the biofilm development over time and persister cell formation. All results will be compared with model predictions. T2 - UNA Workshop CY - Berlin, Germany DA - 29.01.2024 KW - Antibiotic KW - Biofilm KW - Tolerance KW - Efflux PY - 2024 AN - OPUS4-61280 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Marano, Roberto T1 - Antimicrobial coatings: modes of action, microbiological efficacy testing, and resistance evolution N2 - Within the Horizon-Europe funded project “STOP: Surface Transfer Of Pathogens”, Grant agreement 101057961, Working Package 6 is currently working on developing tests aimed at predicting possible selection and/or evolution of antibiotic resistance onto antimicrobial coatings meant for high-touch surfaces. This presentation describes the devised protocols and the preliminary results of their application on copper as a benchmark antimicrobial material. T2 - STOP-project Meeting CY - Sofia, Bulgaria DA - 27.02.2025 KW - Antimicrobial-resistance KW - Surface antimicrobial testing KW - Materials testing PY - 2025 AN - OPUS4-64567 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Marano, Roberto T1 - Antimicrobial coatings: modes of action, microbiological efficacy testing, and resistance evolution N2 - Antimicrobial coatings and materials on high touch surfaces can provide additional level of protection to periodic disinfection cycles by reducing bacterial loads. However, different coatings and materials assert their antimicrobial activity in different ways, and it is not clear whether they could pose detrimental selective pressure towards antibiotic-resistant bacteria. This talk elucidates modes of actions and current knowledge gaps in testing antimicrobial resistance on antimicrobial surfaces.” T2 - STOP Antimicrobial Coating Conference CY - Mons, Belgium DA - 05.12.2024 KW - Antimicrobial-resistance KW - Surface antimicrobial testing KW - Materials testing PY - 2025 AN - OPUS4-64566 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Vareschi, Silvia A1 - Jaut, Valerie A1 - Vijay, Srinivasan A1 - Allen, Rosalind J. A1 - Schreiber, Frank T1 - Antimicrobial efflux and biofilms: an interplay leading to emergent resistance evolution N2 - The biofilm mode of growth and drug efflux are both important factors that impede the treatment of bacterial infections with antimicrobials. Decades of work have uncovered the mechanisms involved in both efflux and biofilm-mediated antimicrobial tolerance, but links between these phenomena have only recently been discovered. Novel findings show how efflux impacts global cellular physiology and antibiotic tolerance, underpinned by phenotypic heterogeneity. In addition efflux can mediate cell-to-cell interactions, relevant in biofilms, via mechanisms including efflux of signaling molecules and metabolites, signaling using pump components and the establishment of local antibiotic gradients via pumping. These recent findings suggest that biofilm antibiotic tolerance and efflux are closely coupled, with synergistic effects leading to the evolution of antimicrobial resistance in the biofilm environment. KW - Evolution KW - Efflux KW - Antibiotics KW - Biofilms KW - Antimicrobial resistance KW - Phenotypic heterogeneity PY - 2025 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-632799 DO - https://doi.org/10.1016/j.tim.2025.04.012 SN - 0966-842X SP - 1 EP - 15 PB - Elsevier Ltd. AN - OPUS4-63279 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schmidt, Selina 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 - Schulz, Anne-Christine T1 - Assessing co-selection of biocide and antibiotic resistance in wastewater microbial communities N2 - Biocides are used for a wide range of purposes, including disinfectants or preservatives. Biocides play a major role in the prevention of microbial infections in healthcare and animal husbandry. The use of biocides often leads to the discharge of active biocidal substances into wastewater streams, causing the exposure of wastewater microbial communities to subinhibitory concentrations. In turn, it is known that wastewater treatment plants (WWTP) are hotspots for antibiotic resistant bacteria. Since similar mechanisms confer resistance to biocides and antibiotics, exposure to biocides can result in co-selection of antibiotic resistant bacteria in WWTP due to biocides. Here, we investigate the magnitude and the drivers of co-selection of antibiotic resistance in natural wastewater microbial communities upon biocide exposure. Microbial communities will be sampled at the WWTP Ruhleben in Berlin and characterized regarding their biocide and antibiotic resistance. Changes in the resistance level after exposure to different biocides will be determined by enumerating resistant and non-resistant E. coli and heterotrophic bacteria on selective plates with and without several biocides and antibiotics. Moreover, we are establishing a synthetic community comprising about 100 environmental E. coli isolates each with different antimicrobial resistance traits. Each isolate will be tagged with a unique DNA-barcode. All isolates will be pooled and exposed to different biocides at various concentrations. The barcode labeling enables us to determine the abundance of each isolate at the beginning and end of the experiment by transposon-tag sequencing. The project results will inform risk assessment of the effects of biocidal residues on antimicrobial resistance selection in WWTP. The project is part of the BIOCIDE consortium funded within the call on Aquatic pollutants by JPI-AMR, JPI-OCEANS and JPI-WATER. T2 - 6th International Symposium onn the Environmental Dimension of Antibiotic Resistance - EDAR 6 CY - Gothenburg, Sweden DA - 22.09.2022 KW - Antibiotic resisitance KW - Biocide KW - Wastewater KW - Risk assesment PY - 2022 AN - OPUS4-56796 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Mezera, Marek T1 - Bacterial adhesion on femtosecond laser-induced periodic surface structures N2 - Biofilm formation in industrial or medical settings is usually unwanted and leads to serious health problems and high costs. Inhibition of initial bacterial adhesion prevents biofilm formation and is, therefore, a major mechanism of antimicrobial action of surfaces. Surface topography largely influences the interaction between bacteria and surfaces which makes topography an ideal base for antifouling strategies and eco-friendly alternatives to chemical surface modifications. Femtosecond laser-processing was used to fabricate sub-micrometric surface structures on silicon and stainless steel for the development of antifouling topographies on technical materials. T2 - Future Tech Week 2020 CY - Online meeting DA - 21.09.2020 KW - Laser-induced periodic surface structures (LIPSS) KW - Bacterial adhesion KW - Biofilm growth KW - Structural color KW - Femtosecond laser processing PY - 2020 UR - http://futuretechweek.fetfx.eu/wp-content/uploads/gravity_forms/2-5432af7ecff9e0243d7383ab3f931ed3/2020/09/BioCombs4Nanofibers_Poster-for-Future_Tech_Week_2020_08-09-2020_with_Reprint-permission_for_upload.pdf AN - OPUS4-51233 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Razkin Bartolomé, Malen T1 - Bacterial adhesion on fs-laser processed laser-induced periodic surface structures N2 - Bacteria are ubiquitous and colonize all types of surfaces, including those in close proximity to humans, such as skin, food, and everyday objects. This raises the question of whether their presence represents a problem to be mitigated or a potential source of benefit to be harnessed, thereby stimulating scientific inquiry into the role of surface-associated bacteria in diverse domains ranging fromhuman health to industrial biotechnology. Aim: The objective of this project is to explore the impact of modifying surface topography on bacterial adhesion behavior. By manipulating the physical characteristics of the substrate, the attachment and detachment dynamics of bacteria can potentially be modified, leading to novel strategies for controlling bacterial colonization in various applications, such as medical devices. Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) were tested on LIPSS-covered Fused Silica samples. T2 - 2023 Spring Meeting · , 2023 · Strasbourg CY - Strasbourg, France DA - 29.05.2023 KW - LIPSS KW - Biofilm KW - fs-laser processing PY - 2023 AN - OPUS4-58456 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Richter, Anja T1 - Bacterial adhesion on ultrashort laser processed surfaces N2 - Bacterial biofilms are multicellular communities adhering to surfaces and embedded in a self-produced extracellular matrix. Due to physiological adaptations and the protective biofilm matrix itself, biofilm cells show enhanced resistance towards antimicrobial treatment. In medical and industrial settings, biofilms on e.g. for implants or for surfaces in food-processing industry can be a fertile source of bacterial pathogens and are repeatedly associated with persisting, nosocomial and foodborne infections. As extensive usage of antibiotics and biocides can lead to the emergence of resistances, various strategies are currently developed, tested and improved to realize anti-bacterial surface properties through surface functionalization steps avoiding antibiotics. In this study, contact-less and aseptic large-area ultrashort laser scan processing is employed to generate different surface structures in the nanometer- to micrometer-scale on technical materials, i.e. titanium-alloy, steel, and polymer. The processed surfaces were characterized by optical and scanning electron microscopy and subjected to bacterial colonization studies with Escherichia coli test strains. For each material, biofilm results of the fs-laser treated surfaces are compared to that obtained on polished (non-irradiated) surfaces as a reference. Depending on the investigated surfaces, different bacterial adhesion patterns were found, suggesting an influence of geometrical size, shape and cell appendages of the bacteria and – above all – the laser-processed nanostructure of the surface itself. T2 - European Materials Research Society Spring Meeting 2021 CY - Online Meeting DA - 31.05.2021 KW - Bacterial adhesion KW - Biofilm formation KW - Ultrashort laser processing KW - Laser-induced periodic surface structures (LIPSS) PY - 2021 UR - https://www.european-mrs.com/laser-material-processing-fundamental-interactions-innovative-applications-emrs AN - OPUS4-52765 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Bonse, Jörn T1 - Bacterial adhesion on ultrashort pulse laser processed surfaces ― more than size matters! N2 - Bacterial biofilms are aggregates of bacterial cells, often attached to a surface and enclosed by a self-produced extracellular matrix which confers increased stress tolerance and resistance to cleaning. Biofilm formation leads to biofouling which gives rise to high costs in numerous technical settings due to biocorrosion and biodegradation. However, biofilms can also be attractive for industrial settings such as wastewater treatment systems or for soil bioremediation processes. Hence, the control of bacterial adhesion to a surface is of major concern. Surface topography strongly influences bacterial adhesion. Therefore, one promising way to achieve bacteria-guiding surfaces lies in the contactless and aseptic large-area laser processing of technical surfaces. We used short and ultrashort pulsed laser systems to generate different surface textures, mainly high-spatial-frequency and low-spatial-frequency laser-induced periodic surface structures, LIPSS (HFSL and LFSL), on Ti, Ti-alloy, steel, and polymers (PET and PE). Pristine (polished) and laser processed samples were subjected to bacterial adhesion experiments with two different Escherichia coli strains and Staphylococcus aureus as test organisms. The bacterial strains differed in their cell wall structure (grampositive vs. gramnegative strains), in size, shape, the occurrence of cell appendages, and in their biofilm forming capabilities. Adhesion patterns were analyzed microscopically and compared regarding the respective test strain and surface topography. Our results revealed that adhesion behavior strongly depends not only on the material’s topography and chemistry, but also on the specific bacterial strain, the presence of cell appendages, and ambient growth conditions. T2 - 13th International Conference on Photoexcited Processes and Applications, ICPEPA-13 CY - Lecce, Italy DA - 14.09.2025 KW - Biofilm KW - Bacterial adhesion KW - Laser-induced periodic surface structures (LIPSS) KW - Ultrashort laser pulses PY - 2025 AN - OPUS4-64166 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Bonse, Jörn T1 - Bacterial adhesion on ultrashort pulse laser processed surfaces ― more than size matters! N2 - Bacterial biofilms are aggregates of bacterial cells, often attached to a surface, and enclosed by a self-produced extracellular matrix which confers increased stress tolerance and resistance to cleaning. Biofilm formation leads to biofouling which gives rise to high costs in numerous technical settings due to biocorrosion and biodegradation. However, biofilms can also be attractive for industrial settings such as wastewater treatment systems or for soil bioremediation processes. Hence, the control of bacterial adhesion to a surface is of major concern. Surface topography strongly influences bacterial adhesion. Therefore, one promising way to achieve bacteria-guiding surfaces lies in the contactless and aseptic large-area laser processing of technical surfaces. We used short and ultrashort pulsed laser systems to generate different surface textures, mainly high-spatial-frequency and low-spatial-frequency laser-induced periodic surface structures, LIPSS (HFSL and LFSL), on Ti, Ti-alloy, steel, and polymers (PET and PE). Pristine (polished) and laser processed samples were subjected to bacterial adhesion experiments with two different Escherichia coli strains and Staphylococcus aureus as test organisms. The bacterial strains differed in their cell wall structure (grampositive vs. gramnegative strains), in size, shape, the occurrence of cell appendages, and in their biofilm forming capabilities. Adhesion patterns were analyzed microscopically and compared regarding the respective test strain and surface topography. Our results revealed that adhesion behavior strongly depends not only on the material’s topography and chemistry, but also on the specific bacterial strain, the presence of cell appendages, and ambient growth conditions. T2 - 13th International LIPSS Workshop CY - Enschede, Netherlands DA - 29.10.2025 KW - Bacterial adhesion KW - Biofilm KW - Laser-induced periodic surface structures (LIPSS) KW - Ultrashort laser pulses PY - 2025 AN - OPUS4-64632 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Nordholt, Niclas T1 - Bacterial resistance evolution on antimicrobial surfaces: Mechanistic insights from a standardizable method N2 - Introduction: Antimicrobial surfaces and coatings (AMCs) are important to prevent the spread of pathogens, especially in hygiene-sensitive areas. However, the evolution and selection of bacterial resistance to AMCs may threaten their efficacy in the long term. In addition, resistance evolution to AMCs may pose the risk for the development of cross-resistance to antibiotics. The assessment of unacceptable resistance risks during the authorization of AMCs is hampered by the lack of standardized test methods that quantify the adaptability of exposed bacteria to AMCs. Objectives: • To develop a standardizable method to determine resistance evolution of bacteria on AMCs (ALEE-AMC) • To assess performance and robustness of ALEE-AMC in a ring trial • To uncover the mechanisms underlying evolution of resistance to a metallic copper AMC • To use ALEE-AMC to assess the evolution of resistance on a novel, nano-particle-based AMC Methods: ALEE-AMC was developed based on an international standard to determine the efficacy of antimicrobial surfaces (ISO 22196). In the ALEE-AMC test, adaptive laboratory evolution is conducted by repeated cycles of AMC exposure and re-growth of surviving cells, selecting for increased survival, followed by isolation of evolved clones. Metallic copper was used as a reference AMC and Escherichia coli as a model microorganism in the ring trial. Evolved E. coli populations from the ring trial partners were subjected to phenotypic (antimicrobial susceptibility testing, ISO 22196) and genotypic (whole genome sequencing) characterization. ALEE-AMC will be used to assess the evolution of resistance on a novel, nano-particle-based AMC currently under development. Findings: The results of the ALEE-AMC ring trial show that repeated exposure to a metallic copper AMC can reproducibly select for reduced copper susceptibility in individual evolutionary lineages across ring trial participants. However, failure to adapt in individual lineages was also observed in all trials. Isolated evolved E. coli clones exhibited increased survival upon exposure to copper surfaces. Adaptation to copper did not induce cross-resistance to antibiotics because the antibiotic susceptibility of copper-adapted clones did not increase above the clinical breakpoint. Whole genome sequencing of the evolved E. coli revealed a high diversity of mutations, including mutations in genes involved in survival to antibiotics. These results indicate the existence of multiple, underexplored evolutionary pathways towards increased survival of antimicrobial copper surfaces. Conclusion: ALEE-AMC offers a standardizable platform to assess the risk of resistance development towards novel and existing AMCs, including nano-particle-based and metallic copper AMCs. Specifically, using ALEE-AMC provided insights into evolvable survival mechanisms to copper AMCs and its consequences for antimicrobial resistance. T2 - FEMS MICRO 2025 CY - Mailand, Italy DA - 14.07.2025 KW - Biocides KW - Antimicrobial surfaces KW - Biocide resistance KW - Standardization KW - ISO 22196 KW - Evolution PY - 2025 AN - OPUS4-63837 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schmidt, Selina T1 - Bacterial resistance evolution towards disinfectants and antimicrobial surfaces and development of a standardized test N2 - Question Disinfectants and antimicrobial surfaces (AMCs) are important tools to prevent the spread of pathogens and antimicrobial resistant bacteria. However, concerns have been raised about the possibility for 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 - a single mechanism conferring resistance to a disinfectant and an antibiotic- and co-resistance - two distinct mechanisms physically linked on e.g. a plasmid. The risk for resistance and cross-resistance during use of biocides (including disinfectants and AMCs) must be evaluated during authorization according to the EU biocidal product regulation. However, to date there is a lack of standardized methods that support risk assessment during the authorization process. Methods We used adaptive laboratory evolution (ALE) experiments which are based on repeated exposure of bacteria to disinfectants or AMCs. The experiments are followed by phenotypic (antimicrobial susceptibility testing) and genotypic (whole genome sequencing) characterization of the evolved strains. The basic idea of these experiments is to expose bacteria to lethal conditions and select for mutants with increased survival. This approach is fundamentally different to other ALE experiments, which commonly select for increased growth at subinhibitory concentrations. However, selection for increased survival represents a selective pressure that more realistically reflects selection under in-use conditions of disinfectants and AMCs. Results First, we studied adaptation of E. coli during repeated disinfection with benzalkonium chloride in a suspension assay. The experiments showed a 2000-fold increase in survival within 5 exposure cycles. The adaptive changes are linked to highly parallel mutations in genes related to lipid A biosynthesis, less negative cell surface charge, reduced growth rate and increased competitive ability in the presence of certain antibiotics. We use the same approach to develop standardizable ALE experiments based upon accepted standards that are used to determine the efficacy of disinfectants (EN 13697) and antimicrobial surfaces (ISO 22196). The results highlight pronounced adaptation of different test strains towards surface disinfection (benzalkonium chloride and isopropanol) and AMCs (copper). Conclusion Bacteria can adapt with increased survival towards lethal stress imposed by disinfectants and AMCs. The adaptive ability of bacteria to disinfectants and AMCs can be determined in a standardized manner. T2 - 74. Jahrestagung der Deutschen Gesellschaft für Hygiene und Mikrobiologie [DGHM] e. V. CY - Berlin, Germany DA - 05.09.2022 KW - Antimicrobial surfaces KW - ISO22196 KW - Antimicrobial resistance KW - Round robin test PY - 2022 AN - OPUS4-56432 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Bakterielle Resistenzen und Toleranzen gegen Desinfektionsmittel und antimikrobielle Oberflächen N2 - Dieser Vortrag beschreibt die Entstehung von bakteriellen Resistenzen und Toleranzen gegen Desinfektionsmittel und antimikrobielle Oberflächen. T2 - Fachtagung für Krankenhaushygiene der Deutschen Gesellschaft für Krankenhaushygiene CY - Essen, Germany DA - 12.05.2023 KW - Antimikrobielle Resistenz KW - Antmikrobielle Oberflächen KW - Standardisierung KW - Biozide PY - 2023 AN - OPUS4-57860 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Beyond Antibiotics – Biocides as Drivers of Environmental AMR N2 - This presentation provides an overview about how biocides drive the evolution of antimicrobial resistance during application and as pollutants in the environment. It also discusses the contribution of biocides for the environmental transmission of AMR. T2 - OneBridge: Making environmental AMR Surveillance Fit for Purpose: Data Integration and the Ecology of Resistance CY - Dresden, Germany DA - 06.10.205 KW - Antimicrobial surfaces KW - Biocides KW - Antimicrobial resistance KW - Standardization PY - 2025 AN - OPUS4-64872 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Olbrich, Adelina-Elisa A1 - An Stepec, Biwen Annie A1 - Wurzler, Nina A1 - Terol, E. C. A1 - Koerdt, Andrea A1 - Meermann, Björn T1 - Beyond corrosion: Development of a single cell-ICP-ToF-MS method to uncover the process of microbiologically influenced corrosion N2 - The development of the microbiologically influenced corrosion ( MIC ) -specific inductively coupled plasma-time of flight-mass spectrometry ( ICP-ToF-MS ) analytical method presented here, in combination with the investigation of steel-MIC interactions, contributes significantly to progress in instrumental MIC analysis. For this, a MIC-specific staining procedure was developed, which ensures the analysis of intact cells. It allows the analysis of archaea at a single cell level, which is extremely scarce compared to other well-characterized organisms. The detection method revealed elemental selectivity for the corrosive methanogenic strain Methanobacterium -affiliated IM1. Hence, the possible uptake of individual elements from different steel samples was investigated and results showed the cells responded at a single-cell level to the different types of supplemented elements and displayed the abilities to uptake chromium, vanadium, titanium, cobalt, and molybdenum from solid metal surfaces. The methods developed and information obtained will be used in the future to elucidate underlying mechanisms, compliment well-developed methods, such as SEM-EDS, and develop novel material protection concepts. KW - Single cell KW - Microbiological influenced corrosion MIC KW - Sc-ICP-ToF-MS KW - Method development KW - Ir DNA staining approach KW - Carbon steel corrosion PY - 2022 DO - https://doi.org/10.1093/mtomcs/mfac083 SN - 1756-591X VL - 14 IS - 11 SP - 1 EP - 15 PB - Oxford University Press CY - Oxford AN - OPUS4-56254 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Keshmiri, Hamid T1 - Bidiffractive leaky-mode biosensor N2 - This study details a thorough analysis of leaky and waveguide modes in biperiodic diffractive nanostructures. By tuning diffraction orders and subsequently confining local density of optical states at two distinct resonance wavelengths, we present a highly sensitive refractive index biosensing platform that can resolve 35.5 to 41.3 nm/RIU of spectral shift for two separate biological analytes. T2 - EMBL Symposium: Seeing is Believing - Imaging the Molecular Processes of Life CY - Heidelberg, Germany DA - 04.10.2023 KW - Optics PY - 2023 AN - OPUS4-59247 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - BIOCIDE N2 - This presentation gives an overview about the BIOCIDE project performed with the Aquatic Pollutants joint call. T2 - Aquatic Pollutants TransNet workshop CY - Online meeting DA - 09.11.2022 KW - Antimicrobial resistance KW - Bacteria KW - Biofilms KW - Biocides KW - Risk assessment KW - Wastewater PY - 2022 AN - OPUS4-56265 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Biocide Resistance - Road to Risk Assessment N2 - This presentation details the current status of biocide resistance risk assessment and provides a roadmap for future activities. T2 - OECD, 6th Meeting of the Working Party on Biocides CY - Paris, France DA - 28.09.2022 KW - Antimicrobial resistance KW - Antimicrobial coating KW - Standardization KW - Biocides KW - Risk assessment PY - 2022 AN - OPUS4-56263 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Sobisch, Lydia-Yasmin T1 - Biocide resistance evolution of corrosion causing sulfate reducing bacteria N2 - SRB are environmentally and industrially important microorganisms. The disadvantage of their metabolic activity (e.g. sulfate reduction) results in the formation of toxic sulfide that leads to microbial influenced corrosion. SRB have been responsible for biocorrosion of ferrous metal. One of mitigation strategy is the use of biocides. However, it has been shown that various bacteria develop antimicrobial resistance due to excessive use of biocides. Thus, a deeper understanding of the evolution of biocide resistance of SRB is necessary. Three commonly used biocides, THPS, BAC, and GLUT were applied to investigate the susceptibility of Desulfovibrio alaskensis G20.The minimum inhibitory and bactericidal concentration and the killing kinetics of the three biocides was determined. These results will be used to conduct evolution experiments to determine the evolution of resistance towards biocides of SRBs. The outcome of this work can be helpful to improve the management of MIC treatments. T2 - Panel, Pitch & Popcorn by EUROMIC CY - Online meeting DA - 21.06.2021 KW - Biocide KW - Evolution KW - Mircobially influcenced corrosion KW - Sulfate reducing bacteria PY - 2021 AN - OPUS4-56940 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 - Nordholt, Niclas T1 - Biocides and phenotypic heterogeneity N2 - An overview of our findings regarding the interplay between phenotypic heterogeneity in bacteria and biocides. T2 - One Health and Antimicrobial Resistance CY - Berlin, Germany DA - 29.01.2024 KW - Biocides KW - Phenotypic heterogeneity KW - Biocide resistance PY - 2024 AN - OPUS4-61174 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Biocides As Drivers For Antimicrobial Resistance Evolution In The Environment N2 - Antimicrobial resistance (AMR) is a global health problem with the environment being an important compartment for the evolution, selection and transmission of AMR. These processes are impacted by pollution with antibiotics. However, biocides used as disinfectants and material preservatives are major pollutants by far excceding the market for antibiotics in terms of mass. Our work shows that biocides have the potential to affect evolutionary processes towards AMR by increasing the rates of de-novo mutation and conjugation. These effects depend on the species and biocidal substance. Importantly, chlorhexidine and quaternary ammonium compounds (QACs) affect rates of mutation and conjugation at environmentally relevant concentrations 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. Furthermore, our work highlights the potential of biocides to contribute to selection and transmission of AMR. We show that the application of biocides, especially QAC disinfectants, leads to the rapid evolution of tolerance (i.e. increased survival) in adaptive laboratory evolution (ALE) experiments. The evolved tolerant strains have a selective advantage in the presence of environmentally-relevant concentrations of antibiotics, which could lead to the stabilization of biocide tolerance in environments where biocides and antibiotics co-occur (e.g. wastewater, animal stables). ALE experiments with biocide tolerant strains indicate a decreased evolvability of resistance to antibiotics. Taken together, our work shows the importance of assessing the contribution of biocides on evolution, selection and transmission of AMR in the environment. T2 - 6th Environmental Dimension of Antibiotic Resistance (EDAR6) CY - Gothenburg, Sweden DA - 22.09.2022 KW - Antimicrobial resistance KW - Antimicrobial coating KW - Biofilms KW - Biocides KW - Risk assessment PY - 2022 AN - OPUS4-56262 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Biocides as drivers for the evolution and selection of antimicrobial resistance N2 - This presentation provides an overview about how biocides drive the evolution of antimicrobial resistance during application and as pollutants in the environment. The presentation shows that biocides can lead to heterogeneous killing, facilitating tolerance evolution. This evolution is related to decreased susceptibility to antibiotics and has potential for co-selection. In contrast, evolved tolerance can limit antibiotic evolvability via epistatic interactions. Moreover, biocides can co-select for antibiotic resistance in wastewater and affect rates of mutation and horizontal gene transfer. Biocides and antibiotics show strong combination effects with consequences for selection of antibiotic resistance. T2 - 12. Dresdner Wasserseminar ' Wasser und Verunreinigung' CY - Dresden, Germany DA - 26.06.2025 KW - Antimicrobial surfaces KW - Biocides KW - Antimicrobial resistance KW - Standardization PY - 2025 AN - OPUS4-64868 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Biocides as drivers for the evolution of antimicrobial resistance N2 - This presentation provides an overview about how biocides drive the evolution of antimicrobial resistance during application and as pollutants in the environment. The presentation shows that biocides can lead to heterogeneous killing, facilitating tolerance evolution. This evolution is related to decreased susceptibility to antibiotics and has potential for co-selection. In contrast, evolved tolerance can limit antibiotic evolvability via epistatic interactions. Moreover, biocides can co-select for antibiotic resistance in wastewater and affect rates of mutation and horizontal gene transfer. Biocides and antibiotics show strong combination effects with consequences for selection of antibiotic resistance. T2 - Novel strategies and considerations in fighting pathogens CY - Tartu, Estonia DA - 16.06.2025 KW - Antimicrobial surfaces KW - Biocides KW - Antimicrobial resistance KW - Standardization PY - 2025 AN - OPUS4-64867 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Biocides as drivers for the evolution of antimicrobial resistance N2 - This presentation provides an overview about how biocides drive the evolution of antimicrobial resistance during application and as pollutants in the environment. The presentation shows that biocides can lead to heterogeneous killing, facilitating tolerance evolution. This evolution is related to decreased susceptibility to antibiotics and has potential for co-selection. In contrast, evolved tolerance can limit antibiotic evolvability via epistatic interactions. Moreover, biocides can co-select for antibiotic resistance in wastewater and affect rates of mutation and horizontal gene transfer. Biocides and antibiotics show strong combination effects with consequences for selection of antibiotic resistance. T2 - Vorstellungsvortrag zur Habilitation FU Berlin CY - Berlin, Germany DA - 15.05.2025 KW - Antimicrobial surfaces KW - Biocides KW - Antimicrobial resistance KW - Standardization PY - 2025 AN - OPUS4-64873 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Biocides as drivers for the Selection and evolution of antimicrobial resistance N2 - Antimicrobial resistance (AMR) is a global health problem with the environment being an important compartment for the evolution, selection and transmission of AMR. These processes are impacted by pollution with antibiotics. However, antimicrobial biocides used as disinfectants and material preservatives are major pollutants exceeding the antibiotic market in terms of chemical diversity and mass. The aim of our work is to understand the mechanisms and risks of biocides for resistance and antibiotic cross-resistance evolution in bacteria to optimize their application and safeguard their efficacy. We use adaptive laboratory evolution experiments, phenotypic characterization, single-cell analysis, whole genome sequencing, and competition experiments to investigate AMR evolution and selection of the model organism E. coli in the presence of biocides. Our work shows that biocides have the potential to affect evolutionary processes towards AMR by increasing the rates of de-novo mutation and conjugation. Importantly, widely used compounds such as chlorhexidine and quaternary ammonium compounds (QACs) affect rates of mutation and conjugation at environmentally relevant concentrations. Furthermore, we show that single-cell phenotypic heterogeneity regarding tolerance (persistence) determines survival against specific biocides including QACs and isopropanol. Mechanistic investigations reveal that known antibiotic persister mechanisms contribute to persister formation to biocides. The evolution of high-level tolerance to different biocides is linked to the initial persister level and the evolution of specific genetically encoded mechanisms related to properties of the cell envelope. Biocide-tolerant strains have a selective advantage in the presence of environmentally-relevant concentrations of antibiotics, which could lead to the stabilization of biocide tolerance in environments where biocides and antibiotics co-occur (e.g. wastewater, animal stables). Taken together, our work shows the importance of assessing the contribution of biocides on evolution and selection of AMR in the environment. T2 - 10th Symposium on Antimicrobial Resistance in Animals and the Environment (ARAE) CY - Berlin, Germany DA - 30.06.2025 KW - Antimicrobial surfaces KW - Biocides KW - Antimicrobial resistance KW - Standardization PY - 2025 AN - OPUS4-64869 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schmidt, Selina 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 - Schmidt, Selina T1 - Biocides Used as Material Preservatives Modify Rates of De Novo Mutation And Horizontal Gene Transfer in Bacteria N2 - Antimicrobial resistance (AMR) is an important global health problem. The environment has been regocnized as an improtant compartment for the occurance, evolution and transmission of AMR. Biocides used as material preservatives are in contact with the environment and natural microbial communities through direct application and passive leaching from protected materials. It has been shown that environmental contaminants, such as antibiotics, metals and pesticides, can affect resistance evolution and spread by modifying the underlying processes of de novo mutagenesis, horizontal gene transfer and selection. However, it is unknown if material preservatives are involved in these processes and thereby can also drive AMR in the environment. Here, we investigate the effect of material preservatives on rates of de novo mutation and horizontal gene transfer (HGT) in Escherichia coli and the model soil microorganisms Acinetobacter baylyi and Bacillus subtilis. To this end, we determined the effects of material preservatives on the mutation rates and HGT with the Luria-Delbrück fluctuation assay and a conjugation assay with the broad host-range plasmid pKJK5 and a transformation assay. Our data shows that the quaternary ammonium compound DDAC, copper, the pyrethroid insecticide permethrin and the azole fungicide propiconazole significantly increase mutation rates in E. coli, whereas A. baylyi and B. subtilis are not significantly affected. Moreover, we show that the carbamate IPBC and the insecticide permethrin affect HGT in a concentration dependent manner. Investigations with reporter strains for bacterial stress response pathways show that induction of the general stress response (rpoS) and components of the SOS response (recA) underlie the effects of most biocides on mutation rates and HGT. Taken together, our data is important for assessing the contribution of biocides on AMR evolution and spread in the environment. T2 - World Microbe Forum (ASM, FEMS) CY - Online meeting DA - 20.06.2021 KW - Biocides KW - Horizontal gene transfer KW - Mutation rate PY - 2021 UR - https://www.abstractsonline.com/pp8/#!/9286/presentation/11692 AN - OPUS4-54211 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 - VIDEO A1 - Schwibbert, Karin A1 - Richter, Anja A1 - Bonse, Jörn T1 - BioCombs4Nanofibers: From nanofibers over spiders to bacteria N2 - This 6 minute long MP4-video presents some key results of the European research project "BioCombs4Nanofibers" to the broader public. Inspired by nature, some concepts of certain types of spiders are transferred to technology in order to develop bacteria-repellent surfaces through laser surface nanostructuring. Funding notice: This study was funded by the European Union's research and innovation program under the FET Open grant agreement No. 862016 (BioCombs4Nanofibers, http://biocombs4nanofibers.eu). KW - Antiadhesive surfaces KW - Laser-induced periodic surface structures (LIPSS) KW - Cribellate spiders KW - Bacterial adhesion tests KW - Bacteria-repellent surfaces PY - 2022 UR - https://download.jku.at/org/7kM/xyU/BioCombs4Nanofibers/D5.6_video%20for%20the%20broader%20public_23.03.2022.mp4 UR - https://www.jku.at/en/biocombs4nanofibers/dissemination/ DO - https://doi.org/10.26272/opus4-54939 PB - Bundesanstalt für Materialforschung und -prüfung (BAM) CY - Berlin AN - OPUS4-54939 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Brischke, C. A1 - Bollmus, S. A1 - Melcher, E. A1 - Stephan, Ina T1 - Biological durability and moisture ynamics of Dawn redwood (Metasequoia glyptostroboides) and Port Orford cedar (Chamaecyparis lawsoniana) N2 - Numerous non-native tree species are given attention with respect to the reforestation of calamity areas in Europe. Among them, several species may form durable wood which can be used for outdoor applications, but differences in wood durability are expected between original and European growth sites. This study aimed at examining the biological durability against wooddestroying fungi and water permeability of German-grown Dawn redwood (Metasequoia glyptostroboides) and Port Orford cedar (Chamaecyparis lawsoniana). The heartwood of both wood species was assigned to durability class 4 (DC 4, less durable) in soil contact and DC 1–4 (very to less durable) against wood-destroying basidiomycetes. However, according to the Meyer-Veltrup model, their material resistance dose was notably higher compared to the reference species Norway spruce (Picea abies), and the resulting service life of above ground structures should be a multiple of the reference. KW - Natural durability KW - Fungal decay KW - Moisture performance KW - Permeability KW - Resistance model KW - Water uptake PY - 2022 DO - https://doi.org/10.1080/17480272.2022.2101941 SN - 1748-0272 SP - 1 EP - 11 PB - Taylor & Francis CY - London AN - OPUS4-55490 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Nordholt, Niclas A1 - van Heerden, J. H. A1 - Bruggeman, F. J. T1 - Biphasic Cell-Size and Growth-Rate Homeostasis by Single Bacillus subtilis Cells N2 - The growth rate of single bacterial cells is continuously disturbed by random fluctuations in biosynthesis rates and by deterministic cell-cycle events, such as division, genome duplication, and septum formation. It is not understood whether, and how, bacteria reject these growth-rate disturbances. Here, we quantified growth and constitutive protein expression dynamics of single Bacillus subtilis cells as a function of cell-cycle progression. We found that, even though growth at the population level is exponential, close inspection of the cell cycle of thousands of single Bacillus subtilis cells reveals systematic deviations from exponential growth. Newborn cells display varying growth rates that depend on their size. When they divide, growth-rate Variation has decreased, and growth rates have become birth size independent. Thus, cells indeed compensate for growth-rate disturbances and achieve growth-rate homeostasis. Protein synthesis and growth of single cells displayed correlated, biphasic dynamics from cell birth to division. During a first phase of variable duration, the absolute rates were approximately constant and cells behaved as sizers. In the second phase, rates increased, and growth behavior exhibited characteristics of a timer strategy. These findings demonstrate that, just like size homeostasis, growth-rate homeostasis is an inherent property of single cells that is achieved by cell-cycle-dependent rate adjustments of biosynthesis and growth. KW - Bacterial cell cycle KW - Single cell microbiology KW - Bacillus subtilis KW - Growth-rate homeostasis KW - Biphasic growth PY - 2020 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-510689 DO - https://doi.org/10.1016/j.cub.2020.04.030 VL - 30 IS - 12 SP - 2238 EP - 2247 PB - Cell Press AN - OPUS4-51068 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Keshmiri, Hamid A1 - Cikes, Domagoj A1 - Samalova, Marketa A1 - Schindler, Lukas A1 - Appel, Lisa-Marie A1 - Urbanek, Michal A1 - Yudushkin, Ivan A1 - Slade, Dea A1 - Weninger, Wolfgang J. A1 - Peaucelle, Alexis A1 - Penninger, Josef A1 - Elsayad, Kareem T1 - Brillouin light scattering anisotropy microscopy for imaging the viscoelastic anisotropy in living cells N2 - Maintaining and modulating mechanical anisotropy is essential for biological processes. However, how this is achieved at the microscopic scale in living soft matter is not always clear. Although Brillouin light scattering (BLS) spectroscopy can probe the mechanical properties of materials, spatiotemporal mapping of mechanical anisotropies in living matter with BLS microscopy has been complicated by the need for sequential measurements with tilted excitation and detection angles. Here we introduce Brillouin light scattering anisotropy microscopy (BLAM) for mapping high-frequency viscoelastic anisotropy inside living cells. BLAM employs a radial virtually imaged phased array that enables the collection of angle-resolved dispersion in a single shot, thus enabling us to probe phonon modes in living matter along different directions simultaneously. We demonstrate a precision of 10 MHz in the determination of the Brillouin frequency shift, at a spatial resolution of 2 µm. Following proof-of-principle experiments on muscle myofibres, we apply BLAM to the study of two fundamental biological processes. In plant cell walls, we observe a switch from anisotropic to isotropic wall properties that may lead to asymmetric growth. In mammalian cell nuclei, we uncover a spatiotemporally oscillating elastic anisotropy correlated to chromatin condensation. Our results highlight the role that high-frequency mechanics can play in the regulation of diverse fundamental processes in biological systems. We expect BLAM to find diverse applications in biomedical imaging and material characterization. KW - Optical and magnetic materials KW - Atomic and molecular physics and optics KW - Electronic PY - 2024 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-595125 DO - https://doi.org/10.1038/s41566-023-01368-w SN - 1749-4885 SN - 1749-4893 VL - 18 IS - 3 SP - 276 EP - 285 PB - Nature Publ. Group CY - London AN - OPUS4-59512 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Barkeshli, Mandana A1 - Stephan, Ina A1 - Shevchuk, Ivan A1 - Soltani, Mojtaba T1 - Characteristics of Sizing Materials Used in Persian Medieval Manuscripts: Physical, Optical, Spectral Imaging, and Fungicidal Properties N2 - In this study, we investigated the diverse range of materials used for sizing in Iranian paper manuscripts during the Timurid (fifteenth century) to Safavid (sixteenth century) and Qajar (nineteenth century) periods. Our approach combined historical analysis with scientific examination of reconstructed sizings. We reconstructed 15 sizing materials based on identified Persian historical recipes and analysed their physical, optical, and spectral characteristics. Additionally, we assessed their behaviour against the mould fungus Aspergillus flavus. The results revealed distinctive properties for each sizing material, shedding light on their potential applications in paper preservation. Furthermore, our investigation demonstrated variations in hygroscopicity, thickness, grammage, and ash content post-sizing. The sizing materials also exhibited different effects on paper reflectance properties. Additionally, our study revealed insights into the impact of sizing on burnished papers, indicating that the mechanical process of burnishing did not significantly alter the chemical composition or spectral properties of the paper, with only minor changes in brightness observed in specific cases. All tested sizing materials supported varying levels of mould growth, indicating potential implications for paper conservation. Our findings provide valuable insights into the historical practices of Iranian paper sizing and offer practical considerations for the preservation of paper manuscripts. KW - Paper sizings KW - Persian historical recipes KW - Physical and optical characteristics KW - Fungicidal property KW - Spectral imaging PY - 2024 DO - https://doi.org/10.1080/00393630.2024.2342647 VL - 69 SP - 519 EP - 536 PB - Informa UK Limited AN - OPUS4-60159 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schulz, Anne-Christine T1 - Co-selection for biocide and antibiotic resistance in microbial wastewater communities N2 - Biocides are used for a wide range of purposes, including disinfectants or preservatives. They play a major role in the prevention of microbial infections in healthcare and animal husbandry. The use of biocides often leads to the discharge of active biocidal substances into wastewater streams, causing the exposure of wastewater microbial communities to subinhibitory concentrations. In turn, it is known that wastewater treatment plants (WWTP) are hotspots for antibiotic resistant bacteria. Since similar mechanisms confer resistance to biocides and antibiotics, exposure to biocides can result in co-selection of antibiotic resistant bacteria in WWTP Here, we want to investigate co-selection processes of antibiotic resistance in natural WWTP microbial communities upon biocide exposure. Microbial communities were sampled at the WWTP Ruhleben in Berlin and characterized regarding their susceptibility against different clinically relevant antibiotics. To investigate the link between biocide exposure and antibiotic resistance, changes in the susceptibility level after exposure to environmentally relevant concentrations of the commonly used biocide didecyldimethylammonium chloride (DDAC) will be determined by enumerating resistant and non-resistant E. coli on selective plates with and without antibiotics and DDAC. In case of antibiotics, clinical breakpoint concentrations according to EUCAST will be used to discriminate between susceptible and resistant strains. In case of DDAC (and biocides in general), clinical breakpoints do not exist. Therefore, we determined a cut-off concentration at which the majority of naturally-occurring E. coli strains cannot grow anymore based on (I) the MIC (minimal inhibitory concentration) distribution, and (II) by plating wastewater communities onto selective indicator agar plates loaded with increasing DDAC concentration. Additionally, antibiotic cross-resistance will be determined by spotting single colonies, isolated from DDAC-selective plates onto antibiotic plates. The results of our experiments will help to determine selective concentrations and to estimate the risk of antibiotic co-selection and cross-resistance in microbial WWTP communities upon biocide exposure. T2 - Annual Conference 2023 of the Association for General and Applied Microbiology CY - Göttingen, Germany DA - 10.09.2023 KW - Antibiotic resisitance KW - Biocide KW - Wastewater KW - Risk assesment PY - 2023 AN - OPUS4-58510 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Kjaervik, Marit A1 - Ramstedt, M. A1 - Schwibbert, Karin A1 - Dietrich, P. M A1 - Unger, Wolfgang T1 - Comparative Study of NAP-XPS and Cryo-XPS for the Investigation of Surface Chemistry of the Bacterial Cell-Envelope N2 - Bacteria generally interact with the environment via processes involving their cell-envelope. Thus, techniques that may shed light on their surface chemistry are attractive tools for providing an understanding of bacterial interactions. One of these tools is Al Kα-excited photoelectron spectroscopy (XPS) with its estimated information depth of <10 nm. XPS-analyses of bacteria have been performed for several decades on freeze-dried specimens in order to be compatible with the vacuum in the analysis chamber of the spectrometer. A limitation of these studies has been that the freeze-drying method may collapse cell structure as well as introduce surface contaminants. However, recent developments in XPS allow for analysis of biological samples at near ambient pressure (NAP-XPS) or as frozen hydrated specimens (cryo-XPS) in vacuum. In this work, we have analyzed bacterial samples from a reference strain of the Gram-negative bacterium Pseudomonas fluorescens using both techniques. We compare the results obtained and, in general, observe good agreement between the two techniques. Furthermore, we discuss advantages and disadvantages with the two analysis approaches and the output data they provide. XPS reference data from the bacterial strain are provided, and we propose that planktonic cells of this strain (DSM 50090) are used as a reference material for surface chemical analysis of bacterial systems. KW - P. Fluorescens KW - Cryo XPS KW - NAP-XPS KW - DSM 5009 PY - 2021 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-525640 DO - https://doi.org/10.3389/fchem.2021.666161 VL - 9 SP - Article 666161 PB - Frontiers CY - Switzerland AN - OPUS4-52564 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Dimitrijević, Stevan P. A1 - Dimitrijević, Silvana B. A1 - Koerdt, Andrea A1 - Ivanović, Aleksandra A1 - Stefanović, Jelena A1 - Stanković, Tanja A1 - Gerengi, Husnu T1 - Comparison of Corrosion Resistance of Cu and Cu72Zn28 Metals in Apricot Fermentation Liquid N2 - The production of fruit brandies is based on distilling fermented fruit juices. Distillation equipment is usually made of copper. In traditional manufacturing, it consists of a boiler (batch) distiller, a boiler (pot), a steam pipe, and a condenser, all of which are made of pure copper. This study determined the corrosion parameters for copper (Cu) and Cu72Zn28 (in wt%) alloy in fermented apricot juice at room temperature. The fermentation process examined in this research utilized natural strains of yeast and bacteria, supplemented by active dry yeast Saccharomyces cerevisiae strains. This research used the following methods: open circuit potential (OCP), linear polarization resistance (LPR), and Tafel extrapolation to identify corrosion parameters. Cu had a 3.8-times-lower value of corrosion current density than brass, and both were within the range of 1–10 μA·cm−2, with an excellent agreement between LRP and Tafel. This study proved that Cu is an adequate material for the distillation of fruit brandies from a corrosion perspective. Despite this, there are occasional reports of corrosion damage from the field. Significant corrosion impacts can arise, as evidenced by laboratory tests discussed in this paper. In the absence of a highly corrosive environment, this study indicates that, to some extent, microbiologically influenced corrosion (MIC) can influence the degradation of the equipment material. KW - MIC KW - Metall KW - Envitonment PY - 2025 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-628911 DO - https://doi.org/10.3390/ma18061253 SN - 1996-1944 VL - 18 IS - 6 SP - 1 EP - 17 PB - MDPI AG AN - OPUS4-62891 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Nordholt, Niclas T1 - Consequences of BAC tolerance for selection and evolution in the presence of antibiotics N2 - The exposure to antimicrobial substances drives the evolution of antimicrobial resistance. 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 at BAM 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. Within our group, I mainly focus on the following two questions: How does phenotypic heterogeneity in bacteria affect the ability to survive treatment with biocides? And what are the consequences of phenotypic heterogeneity for the evolution of resistance to biocides and antibiotics? I will share published and unpublished results which demonstrate that phenotypic heterogeneity can enable the survival of biocide treatment and, through this, facilitate the evolution of AMR. On the other hand, we find that adaptation to a biocide can unexpectedly impair the ability to evolve resistance against an antibiotic. T2 - FEMS summer school for postdocs 2022: Microbial Evolvability Mechanisms: Resistance, Biology, and Strategies to Defeat and Detect CY - Split, Croatia DA - 27.04.2022 KW - Disinfection KW - Biocides KW - Evolution KW - Resistance KW - Biocide tolerance PY - 2022 AN - OPUS4-54846 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 - TY - GEN A1 - Schreiber, Frank A1 - Kanaris, Orestis A1 - Nordholt, Niclas T1 - Consequences of benzalkonium chloride tolerance for selection dynamics and de novode 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−1. 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 - Antimicrobial surfaces KW - Biocides KW - Antimicrobial resistance KW - Antibiotics PY - 2025 UR - https://www.ncbi.nlm.nih.gov/bioproject/?term=PRJNA1282584 PB - National Library of Medicine CY - Bethesda AN - OPUS4-65221 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Kanaris, Orestis T1 - Consequences of benzalkonium chloride tolerance in Escherichia coli: Effects on selection and evolution in the presence of ciprofloxacin N2 - We investigated the selection dynamics between a benzalkonium chloride (BAC)-tolerant Escherichia coli strain (S4) and a sensitive wild type under four conditions: in the absence of antibiotics and in the presence of three different sub-inhibitory concentrations of the antibiotic ciprofloxacin in liquid cultures. The wild type was selected over the BAC-tolerant strain in the absence of antibiotics, while the opposite was observed at all ciprofloxacin concentrations investigated.Furthermore, we assessed the evolvability of resistance of the two strains to inhibitory concentrations of ciprofloxacin by performing a serial dilution evolution experiment with gradually increasing ciprofloxacin concentrations. The wild type had a higher probability to develop resistance to ciprofloxacin than the tolerant strain. By the end of the evolution experiment both strains evolved to grow at the highest ciprofloxacin concentration investigated, which was 2048 ×MIC of the wild type. T2 - 6th international symposium on the environmental dimention of antibiotic resistance-EDAR 6 CY - Gothenburg, Sweden DA - 22.09.2022 KW - Antimicrobial resistance KW - Tolerance KW - Experimental evolution KW - Selection PY - 2022 AN - OPUS4-56808 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Kanaris, Orestis T1 - Consequences of benzalkonium chloride tolerance on the development of antibiotic resistance in E. coli 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-1. 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. T2 - µClub Seminar CY - Berlin, Germany DA - 23.05.2025 KW - Biocides KW - AMR KW - Resistance evolution PY - 2025 AN - OPUS4-64658 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Kanaris, Orestis T1 - Consequences of tolerance to disinfectants on the evolution of antibiotic resistance in E. coli N2 - Biocides are used as disinfectants and preservatives; one important active substance in biocides is benzalkonium chloride (BAC). BAC-tolerant bacterial strains can survive short treatments with high concentrations of BAC. BAC tolerance and resistance have been linked to antibiotic resistance. Here, the selection dynamics between a BAC-tolerant Escherichia coli strain and a sensitive wild type were investigated under four conditions: in the absence of antibiotics and in the presence of three different sub-inhibitory concentrations of the antibiotic ciprofloxacin in liquid cultures. The wild type was selected over the BAC-tolerant strain in the absence of antibiotics, while the BAC-tolerant strain was selected over the wild type at all ciprofloxacin concentrations investigated, with a minimum selection concentration (MSC) of 1/10th of the minimum inhibitory concentration (MIC) of the wild type. Furthermore, the evolvability of resistance of the two strains to inhibitory concentrations of ciprofloxacin was assessed by performing a serial dilution evolution experiment with gradually increasing ciprofloxacin concentrations. The wild type had a higher probability to develop resistance to ciprofloxacin than the tolerant strain. By the end of the evolution experiment both strains evolved to grow at the highest ciprofloxacin concentration investigated, which was 2048 ×MIC of the wild type. The importance of these results is highlighted by the fact that concentrations of ciprofloxacin well above the calculated MSC can be found in environmental samples such as hospital wastewaters and livestock slurry. In turn, BAC is used as a disinfectant in the same settings. Thus, the selection of BAC-tolerant strains at sub-inhibitory concentrations of ciprofloxacin can contribute to the stabilization and spread of BAC-tolerance in natural populations. The prevalence of such strains can impair the effects of BAC disinfections. T2 - µClub seminar series CY - Berlin, Germany DA - 15.12.2023 KW - Antimicrobial resistance PY - 2023 AN - OPUS4-59222 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Vandrich, Jasmina A1 - Pfeiffer, F. A1 - Alfaro Espinoza, Gabriela A1 - Kunte, Hans-Jörg T1 - Contribution of mechanosensitive channels to osmoadaptation and ectoine excretion in Halomonas elongata N2 - For osmoadaptation the halophilic bacterium Halomonas elongata synthesizes as its main compatible solute the aspartate derivative ectoine. H. elongata does not rely entirely on synthesis but can accumulate ectoine by uptake from the surrounding environment with the help of the osmoregulated transporter TeaABC. Disruption of the TeaABC-mediated ectoine uptake creates a strain that is constantly losing ectoine to the medium. However, the efflux mechanism of ectoine in H. elongata is not yet understood. H. elongata possesses four genes encoding mechanosensitive channels all of which belong to the small conductance type (MscS). Analysis by qRT-PCR revealed a reduction in transcription of the mscS genes with increasing salinity. The response of H. elongata to hypo- and hyperosmotic shock never resulted in up-regulation but rather in downregulation of mscS transcription. Deletion of all four mscS genes created a mutant that was unable to cope with hypoosmotic shock. However, the knockout mutant grew significantly faster than the wildtype at high salinity of 2 M NaCl, and most importantly, still exported 80% of the ectoine compared to the wildtype. We thus conclude that a yet unknown system, which is independent of mechanosensitive channels, is the major export route for ectoine in H. elongata. KW - Halomonas elongata KW - Ectoine KW - Osmotic shock KW - Solute excretion KW - Osmoadaptation KW - Mechanosensitive channel KW - MscS PY - 2020 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-507229 DO - https://doi.org/10.1007/s00792-020-01168-y VL - 24 SP - 421 EP - 432 PB - Springer AN - OPUS4-50722 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Koerdt, Andrea T1 - Cost action EURO MIC (CA20130) N2 - Vorstellung des vor kurzen angelaufenen EU-Projekts: COST ACTION EURO MIC ( CA20130). Die BAM hat hierbei die Rolle des Chairs sowie Grant-Holding Institute. COST fördert internationale Netzwerk Aktivitäten. Durch COST Action können neue Kooperationen (Industrie, Akademie oder Politk) entstehen. T2 - Dechema CY - Online meeting DA - 10.11.2021 KW - Netzwerk KW - COST KW - Mikrobiell beeinflusste Korrosion KW - MIC PY - 2021 AN - OPUS4-54023 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Koerdt, Andrea T1 - COST-Action - EURO-MIC (CA20130) European MIC Network - New paths for science, sustainability and standards N2 - Microbiologically Influenced Corrosion (MIC) is a phenomenon that is increasingly becoming a problem for the society. MIC describes the negative effects a material can experience due to the presence of microorganisms. In Europe, several research groups/ other industrial stakeholders are already dealing with MIC. Unfortunately, the discussions are fragmented and the exchange of information is limited. A true transdisciplinary approach is hardly ever experienced, although this would be logical for this material/biology related challenge. USA, Canada and Australia have strong networks, and develop methods, prevention measures and standards, which Europe is forced to use, since nothing similar exists for a network and combined knowledge to design them according to european standards. This makes Europe extremely dependent and, in some cases, the potential measures or standards cannot been used because the suggested solutions are prohibited by European laws (e.g. use of biocides). Therefore, it is important to initiate a new European MIC-network. Europe needs to combine the efforts as experts in different fields and develop prevention measures according to the European rules, in close cooperation with industry and plant operators and owners of critical infrastructure. This COST Action will provide the necessary interaction and communication, knowledge sharing, training of personnel and of researchers of different disciplines. This will bring Europe to a leading role in this process, bringing ideas on an equal level with other nations, considering the values which are important for Europe and attitudes (e.g.environmental protection) and representing greater protection for people, property and the environment. The main aim and objective of the Action is to , in the context of MIC-research/control, encourage a fluent/synergistic collaboration/communication, closing the gap between materials scientists, engineers, microbiologists, chemists and integrity managers to encourage sufficient interaction between academia and industry. This Action will create a common MIC-Network, including the important stakeholders. T2 - EuroCorr CY - Online meeting DA - 19.09.2021 KW - COST Action KW - MIC KW - Interdisciplinary KW - Stakeholder KW - Academia PY - 2021 AN - OPUS4-53382 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Dinter, Adelina-Elisa T1 - Deciphering corrosion processes of MIC organisms - single cell-ICP-ToF-MS analysis of archaea on solid steels N2 - ICP-ToF (time of flight) MS enables the analysis of the multi-element fingerprint of single cells. The single cell ICP-ToF-MS is used in the presented poster for the analysis of archaea involved in microbiologically influenced corrosion (MIC) of steel. By means of sc-ICP-ToF-MS, the possible uptake of individual elements from the respective steel is investigated - the information obtained will be used in the future to elucidate underlying mechanisms and develop possible material protection concepts. The work combines modern methods of analytical sciences with materials. T2 - SALSA - Make & Measure 2021 CY - Online meeting DA - 16.09.2021 KW - Sc-ICP-ToF-MS KW - Single cell analysis KW - Microbiologically influenced corrosion KW - Archaea KW - Poster presentation PY - 2021 AN - OPUS4-53337 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Olbrich, Adelina-Elisa T1 - Deciphering microbiological influenced corrosion processes on steel with single cell-ICP-ToF-MS N2 - Microbiologically influenced corrosion (MIC) is a highly unpredictable process dictated by the environment, microorganisms, and the respective electron source. Interaction pathways between cells and the metal surface remain unclear. The development of this novel single cell-inductively coupled plasma-time of flight-mass spectrometry analytical method and a MIC-specific staining procedure facilitate the investigation of steel-MIC interactions. With this it is possible to analyze the multi-elemental fingerprint of individual cells. The detection method revealed elemental selectivity for the corrosive methanogenic archaeal strain Methanobacterium-affiliated IM1. The interface between material and environmental analysis thus receives special attention, e.g., when considering MIC on solid steel. Hence, the possible uptake of individual elements from different steel samples is investigated. Results showed the cells responded at a single-cell level to the different types of supplemented elements and displayed the abilities to interact with chromium, vanadium, titanium, cobalt, and molybdenum from solid metal surfaces. The information obtained will be used in the future to elucidate underlying mechanisms and develop possible material protection concepts, thus combining modern methods of analytical sciences with materials research. References. T2 - Future WiNS CY - Berlin, Germany DA - 07.12.2022 KW - Single cell KW - Microbiological influenced corrosion MIC KW - Sc-ICP-ToF-MS KW - Method development KW - Ir DNA staining approach KW - Carbon steel corrosion PY - 2022 AN - OPUS4-56567 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Development of a laboratory method to assess resistance development of microorganisms to biocides N2 - Bacteria are exposed to biocides through surface disinfection or by antimicrobial surfaces. These stressful environments provide a strong selective pressure for bacteria to adapt. Here, we describe the development of a laboratory method to assess adaption of bacteria by resistance development in response to surface disinfection and antimicrobial surfaces. T2 - OECD, 5th Meeting of the Working Party on Biocides CY - Online meeting DA - 26.05.2021 KW - Antimicrobial resistance KW - Antimicrobial coating KW - Standardization PY - 2021 AN - OPUS4-53163 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Development of a laboratory method to assess resistance development of microorganisms to biocides N2 - Bacteria are exposed to biocides through surface disinfection or by antimicrobial surfaces. These stressful environments provide a strong selective pressure for bacteria to adapt. Here, we describe the development of a laboratory method to assess adaption of bacteria by resistance development in response to surface disinfection and antimicrobial surfaces. T2 - The International Biodeterioration Research Group (IBRG) Spring Meeting 2021 CY - Online meeting DA - 28.06.2021 KW - Antimicrobial resistance KW - Antimicrobial coating KW - Standardization PY - 2021 AN - OPUS4-53164 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Development of a laboratory method to assess resistance development of microorganisms to biocides – An update N2 - This presentation describes the development of a laboratory method to assess resistance development of microorganisms to biocides and antimicrobial surfaces. T2 - The International Biodeterioration Research Group (IBRG) autumn meeting 2022 CY - Online meeting DA - 11.10.2022 KW - Antimicrobial resistance KW - Antimicrobial coating KW - Standardization KW - Biocides KW - Risk assessment PY - 2022 AN - OPUS4-56264 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank 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 - Dinter, Adelina-Elisa ED - Dinter, Adelina-Elisa T1 - Development of a MIC single archaea-ICP-ToF-MS-method for analysis of various elements in solid steel samples N2 - With the latest ICP-MS technology - ICP-ToF (time of flight)-MS - it is possible to analyze the multi-element fingerprint of individual cells. The interface between material and environmental analysis thus receives special attention, e.g. when considering corrosion processes. Microbiologically influenced corrosion (MIC) is highly unpredictable due to the diversity of microbial communities involved. The development of the MIC-specific ICP-ToF-MS analytical method presented here at the single cell level, in combination with the investigation of steel-MIC interactions, contributes significantly to progress in instrumental MIC analysis and will enable clarification of the processes taking place. The supplied document shows the basis of a four minutes lightning talk. T2 - EuroMIC 2021 CY - Online meeting DA - 22.06.2021 KW - SC-ICP-ToF-MS KW - Single cell analysis KW - Microbiologically influenced corrosion KW - Lightning talk PY - 2021 AN - OPUS4-52905 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Dinter, Adelina-Elisa T1 - Development of a MIC single archaea-ICP-ToF-MS-method for analysis of various elements in solid steel samples N2 - ICP-ToF (Flugzeitanalysator, engl. time of flight)-MS ermöglicht den Multielement Fingerabdruck einzelner Zellen (single cell) zu analysieren. Die single cell-ICP-ToF-MS kommt bei dem vorgestellten Poster bei der Analyse von Archaeen, die an mikrobiell beeinflusster Korrosion (engl. microbiologically influenced corrosion, MIC) von Stahl eine Rolle spielen, zum Einsatz. Mittels sc-ICP-ToF-MS wird die mögliche Aufnahme von einzelnen Elementen aus dem jeweiligen Stahl untersucht – die erhaltenen Informationen fließen zukünftig in die Aufklärung zugrunde liegender Mechanismen sowie Entwicklung möglicher Materialschutzkonzepte ein. Die Arbeiten Verknüpfen moderne Methoden der Analytical Sciences mit Materialien. T2 - Tag der Chemie 2021 CY - Online meeting DA - 06.07.2021 KW - SC-ICP-ToF-MS KW - Single cell analysis KW - Microbiologically influenced corrosion KW - Archaea PY - 2021 AN - OPUS4-52941 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Dinter, Adelina-Elisa T1 - Development of a single cell-ICP-ToF-MS-method for multielement analysis of MIC organisms grown on solid steel samples N2 - The latest ICP-MS technology - ICP-ToF (time of flight)-MS – enables the analysis of the multi-element fingerprint of individual cells. The interface between material and environmental analysis thus receives special attention, e.g., when considering corrosion processes. Microbiologically influenced corrosion (MIC) is a highly unpredictable phenomenon due to the influence of the environment, microbial communities involved and the respective electron source. However, the interaction pathway between cells and the metal surface remains unclear. The development of the MIC-specific ICP-ToF-MS analytical method presented here at the single cell level, in combination with the investigation of steel-MIC interactions, contributes significantly to progress in instrumental MIC analysis and will enable clarification of the processes taking place. For this, a MIC-specific staining procedure was developed. It allows the analysis of archaea at a single cell level and provides information about the interaction of the cells with the staining agent which is extremely scarce compared to other well characterized organisms. Additionally, the single cell ICP-ToF-MS is used for the analysis of archaea involved in MIC of steel. Hence, the possible uptake of individual elements from different steel samples is investigated - the information obtained will be used in the future to elucidate underlying mechanisms and develop possible material protection concepts, thus combining modern methods of analytical sciences with materials. T2 - DAAS Doktorandenseminar 2021 CY - Online meeting DA - 20.09.2021 KW - Sc-ICP-ToF-MS KW - Single cell analysis KW - Microbiologically influenced corrosion KW - Archaea PY - 2021 AN - OPUS4-53340 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - de Araujo Abilio, André A1 - Wolodko, John D. A1 - Eckert, Richard B. A1 - Skovhus, Torben L. T1 - Development of an expert system for assessing failures in oil and gas pipelines due to microbiologically influenced corrosion (MIC) N2 - This study highlights the modeling of an expert system for the classification of internal microbiologically influenced corrosion (MIC) failures related to pipelines in the upstream oil and gas industry. The model is based on artificial neural networks (ANNs) and involves the participation of 15 MIC experts. Each expert evaluated a number of model case studies ranging from MIC- to non-MIC-driven upstream pipeline failures. The model accounts for variations in microbiological testing methods, microbiological sample types, and degradation morphology, among other variables. It also accounts for missing datasets, which is commonly the case in actual failure assessments. The outcome is an expert system model whose outputs are classes (classification ANN) which comprises MIC potential and data confidence. The performances of two approaches are contrasted in this study. One classifies the output in 5 classes, a 5-output classification (5OC) model; the other in 3 classes, a 3-output classification (3OC) model. The 5OC model had an accuracy of 62.0% while the simpler 3OC model had a better accuracy of 74.8%. This modelling exercise has demonstrated that knowledge from experts can be captured in a reasonably effective model to screen for possible MIC failures. It is hoped that this study not only may contribute to a better understanding of the prevalence of MIC in the oil and gas sector, but also may highlight the key areas necessary to improve the diagnosis of MIC failures in the future. KW - Corrosion engineering KW - Oil and gas pipelines KW - Carbon steel KW - Artificial neural networks (ANNs) KW - Failure analysis KW - Microbiologically influenced corrosion (MIC) KW - Expert systems PY - 2024 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-632084 DO - https://doi.org/10.1016/j.engfailanal.2024.108426 SN - 1350-6307 VL - 163 SP - 1 EP - 16 PB - Elsevier Ltd. AN - OPUS4-63208 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Die Entstehung von antimikrobiellen Resistenzen durch die Verwendung von Bioziden N2 - Dieser Vortrag gibt einen Überblick über die BAM und die Aktivitäten im Bereich Biozidresistenz. T2 - Berliner Hochschule für Technik Studiengang Biotechnologie CY - Berlin, Germany DA - 05.05.2023 KW - Antimikrobielle Resistenz KW - Antmikrobielle Oberflächen KW - Standardisierung PY - 2023 AN - OPUS4-57859 LA - deu 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 - GEN 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 E. 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 solely for survival 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 sub-inhibitory 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://www.ncbi.nlm.nih.gov/bioproject/PRJNA1074740 PB - National Library of Biotechnology Information CY - Bethesda AN - OPUS4-61559 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - GEN 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 E. 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 solely for survival 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 sub-inhibitory 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 DO - https://doi.org/10.5061/dryad.2jm63xszx PB - Dryad AN - OPUS4-61558 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Ulbricht, Alexander T1 - Do Microbes like Additively Manufactured Aluminium? N2 - The use of laser-additively manufactured metallic components in the industrial sector is gaining traction, with the process now entering its application phase. Consequently, the materials are exposed to corrosive microbes, which this study examines in order to determine the implications of this exposure. Specifically, the study examines the exposure of laser powder bed fused AlSi10Mg (PBF-LB/AlSi10Mg) components to sulphate-reducing bacteria in an anaerobic environment. Is the specific PBF-LB microstructure a potential factor in determining or aiding biocorrosion? A prismatic specimen was exposed to sulphate-reducing bacteria (SRB) for two weeks. The sample was scanned by X-ray computed tomography (XCT) before and after SRB exposure. The two data sets were registered to each other (using the internal void distribution as markers, due to the corrosion of the specimen’s surface) to enable the evaluation of the corrosion damage. Virtual cuts of the 3D XCT reconstructions show a loss of volume and localised damage. The damage distribution aligns with the hatching pattern of laser exposure from laser powder bed fusion. T2 - Konferenz iCT 2026 CY - Linz, Austria DA - 10.02.2026 KW - Computed tomography KW - Biokorrosion KW - Additive manufacturing KW - PBF-LB KW - Laser powder bed fusion KW - Sulphate-reducing bacteria PY - 2026 AN - OPUS4-65570 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Ulbricht, Alexander T1 - Do Microbes like Additively Manufactured Aluminium? First Details of a Corrosion Test using Sulphate-Reducing Bacteria N2 - Additively manufactured metals become relevant for industrial application. Although many studies on wet corrosion of these metals have been conducted, to the authors knowledge no study seems to contain microbiological corrosion (MIC). In the presented study an experiment was conducted on PBF-LB/AlSi10Mg to test this material's susceptibility for MIC. The tested specimen were analysed using Computed Tomography before and after the MIC experiment to enable a detailed characterisation the damage on the specimens' global and local level. A global reduction of material was observed. In addition, localised damage along process inherent features of the materials microstructure was observed. T2 - Beiratssitzung TF Umwelt CY - Berlin, Germany DA - 17.03.2025 KW - Computed Tomography KW - Additive Manufacturing KW - Biocorrosion KW - Sulphate-reducing Bacteria KW - Microbially influenced corrosion PY - 2025 AN - OPUS4-62772 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schreiber, Frank T1 - Dr. Frank Schreiber and Work on Biocide Resistance at BAM N2 - This talk details the career path of Dr. Frank Schreiber and his work on biocide resistance at BAM. Biocides are antimicrobial products for defined applications (disinfectants, preservatives, pest control). Biocides show heterogeneous killing facilitating resistance/tolerance evolution. Adaptation to biocides has effects on growth and selection. T2 - Vortrag an der Berliner Hochschule für Technik CY - Berlin, Gemany DA - 07.06.2024 KW - Antimicrobial resistance KW - Bacteria KW - Standardization KW - Biocides PY - 2024 AN - OPUS4-61543 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Srivastava, Priyanka A1 - Tavernaro, Isabella A1 - Scholtz, Lena A1 - Genger, C. A1 - Welker, P. A1 - Schreiber, Frank A1 - Meyer, Klas A1 - Resch-Genger, Ute T1 - Dual color pH probes made from silica and polystyrene nanoparticles and their performance in cell studies N2 - Ratiometric green–red fluorescent nanosensors for fluorometrically monitoring pH in the acidic range were designed from 80 nm-sized polystyrene (PS) and silica (SiO2) nanoparticles (NPs), red emissive reference dyes, and a green emissive naphthalimide pH probe, analytically and spectroscopically characterized, and compared regarding their sensing performance in aqueous dispersion and in cellular uptake studies. Preparation of these optical probes, which are excitable by 405 nm laser or LED light sources, involved the encapsulation of the pH-inert red-fuorescent dye Nile Red (NR) in the core of self-made carboxylated PSNPs by a simple swelling procedure and the fabrication of rhodamine B (RhB)-stained SiO2-NPs from a silane derivative of pH-insensitive RhB. Subsequently, the custom-made naphthalimide pH probe, that utilizes a protonation-controlled photoinduced electron transfer process, was covalently attached to the carboxylic acid groups at the surface of both types of NPs. Fluorescence microscopy studies with the molecular and nanoscale optical probes and A549 lung cancer cells confirmed the cellular uptake of all probes and their penetration into acidic cell compartments, i.e., the lysosomes, indicated by the switching ON of the green naphthalimide fluorescence. This underlines their suitability for intracellular pH sensing, with the SiO2-based nanosensor revealing the best performance regarding uptake speed and stability. KW - Sensors KW - Silica and polystyrene nanoparticles KW - pH probe KW - Fluorescence spectroscopy KW - Cell studies KW - Dye KW - Particle synthesis PY - 2023 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-569198 DO - https://doi.org/10.1038/s41598-023-28203-0 SN - 2045-2322 VL - 13 IS - 1 SP - 1321 EP - 1336 PB - Nature CY - London AN - OPUS4-56919 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schmidt, Selina T1 - Effect of biocides on the evolution and spread of resistance 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 - Antimicrobial resistance KW - Horizontal gene transfer KW - Mutation rate PY - 2022 AN - OPUS4-56426 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schmidt, Selina T1 - Effect of biocides on the evolution and spread of resistance N2 - Biocides used as material preservatives are in contact with various environments during direct application or passive leaching from protected materials [1]. Antimicrobial resistance (AMR) is a global health problem and the environment is an important compartment for the evolution and transmission of AMR [2,3]. 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 - RokoCon2022 CY - Berlin, Germany DA - 29.09.2022 KW - Biocide KW - Antimicrobial resistance KW - Tolerance KW - Horizontal gene transfer KW - Mutation rate PY - 2022 AN - OPUS4-56431 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Wittmar, J. A1 - Ohle, Corina A1 - Kunte, Hans-Jörg A1 - Brand, I. T1 - Effect of Ectoine on the Conformation and Hybridization of dsDNA in Monolayer Films: A Spectroelectrochemical Study N2 - Lack of long-time stability of dsDNA-based supramolecular assemblies is an important issue that hinders their applications. In this work, 20 base pairs long dsDNA fragments [(dCdG)20 65%] composed of 65% dCdG and 35% dAdT nucleotides were tethered via a thiol to the surface of a gold electrode. The selfassembled (dCdG)20 -65% monolayer was immersed in solutions containing ectoine, a compatible solute. Electrochemical results showed that these monolayers were stable for one month. In situ IR spectroscopy indicated that ectoine interacts weakly with the phosphate-ribose backbone, dehydrating the phosphate groups and stabilizing the A-DNA conformation. This structural reorganization led to a reorientation of nucleic acid base pairs and a local disruption of the double-helix structure. However, the conformation and orientation of the dsDNA fragment was stable in the KW - Ectoine KW - dsDAN monolayer KW - A-DNA conformation PY - 2021 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-533346 DO - https://doi.org/10.1002/celc.202100816 SN - 2196-0216 VL - 8 IS - 20 SP - 3844 EP - 3854 PB - Wiley-VCH GmbH AN - OPUS4-53334 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Liu, Xiangju A1 - Huang, Yanliang A1 - Li, Jianzheng A1 - Yang, Dan A1 - Xu, Yong A1 - Kunte, Hans-Jörg T1 - Effect of microbial hydrogen consumption on the hydrogen permeation behaviour of AISI 4135 steel under cathodic protection N2 - The feasibility of microbial hydrogen consumption to mitigate the hydrogen embrittlement (HE) under different cathodic potentials was evaluated using the Devanathan-Stachurski electrochemical test and the hydrogen permeation efficiency h. The hydrogen permeation efficiency h in the presence of strain GA-1 was lower than that in sterile medium. The cathodic potential inhibited the adherence of strain GA-1 to AISI 4135 steel surface, thereby reducing the hydrogen consumption of strain GA-1. The adherent GA-1 cells were capable of consuming ‘cathodic hydrogen’ and reducing the proportions of absorbed hydrogen, indicating that it is theoretically possible to control HE by hydrogen-consuming microbes. KW - Energy Engineering and Power Technology KW - Condensed Matter Physics KW - Fuel Technology KW - Renewable Energy, Sustainability and the Environment PY - 2020 DO - https://doi.org/10.1016/j.ijhydene.2019.12.058 SN - 1879-3487 VL - 45 IS - 7 SP - 4054 EP - 4064 PB - Elsevier BV CY - New York, NY AN - OPUS4-59319 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - THES A1 - Schmidt, Selina T1 - Effects of biocides on processes underlying resistance evolution N2 - Antimicrobial resistance (AMR) is a global health problem. It is well known that antibiotics can drive evolutionary processes that underlie antimicrobial resistance (AMR) evolution and spread in clinical and environmental settings. In contrast, less is known about the effects of antimicrobial substances that are used as biocides (i.e. disinfectants and preservatives) on AMR evolution and spread. Biocides are present in various settings, interacting with diverse microbial communities. Therefore, it is crucial to evaluate their role in the evolution and dissemination of antimicrobial resistance. Biocides occur in a wide range of concentrations in various environmental settings. By examining how the various concentrations affect selection mechanisms, we gain insights into potential developments related to antimicrobial resistance. The aim of this PhD thesis is to investigate the effects of biocides on processes underlying resistance evolution. Specifically, the work focused on key mechanisms for resistance spread, resistance evolution, and the effect of selection pressures on evolved resistance mechanisms. The thesis is structured around three major objectives: (i) to determine the effect of biocides on the evolution of resistance by affecting the rate of occurrence of de novo mutations, (ii) to determine the effect of biocides on the spread of resistance genes by modifying the rate of horizontal gene transfer (HGT) processes, and (iii) to investigate the selective drivers of the emergence of antimicrobial resistance in adaptive laboratory evolution (ALE) experiments. De-novo mutations are spontaneous mutations that occur at a certain rate in microorganisms. The effect of biocides at subinhibitory environmentally relevant concentrations on the mutation rate in Acinetobacer baylyi, Bacillus subtilis and Escherichia coli was assessed with the fluctuation assay. The results showed that biocides affected mutation rates in a species and substance dependent matter. 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 E. coli, whereas no increases were identified for B. subtilis and A. baylyi. Horizontal gene transfer refers to diverse mechanisms that mediate the transfer of mobile genetic elements between microorganisms. This work focused on conjugation and transformation. Conjugation is a process whereby a conjugative plasmid is transferred from a donor cell to a recipient cell. Transformation is a process whereby exogenous donor DNA is taken up into a recipient cell and integrated into the recipient’s’ genome. The effects of subinhibitory environmentally relevant biocide concentrations on the conjugation rate of E. coli and the transformation rate of the naturally competent organisms A. baylyi in were assessed. The results showed that benzalkonium chloride (BAC), chlorhexidine and permethrin increased conjugation in E. coli, while none of the biocides increased transformation rates in A. baylyi. To further understand the molecular mechanisms underlying the effects on mutation and conjugation rates, I investigated the induction of the RpoS-mediated general stress and the RecA-linked SOS response upon biocide exposure. The results show a link between the general stress and the SOS response with increased rates of mutation and conjugation, but not for all biocides. One major approach to study the evolutionary response of bacteria to antimicrobials are ALE experiments with growth at subinhibitory concentrations linked to serial subculturing over many generations. Such experiments have been used to study resistance evolution to antibiotics and biocides. However, previous work showed that adaptation to biocide stress may be mediated by different evolutionary drivers. Here, I investigated the contributions of evolution for increased survival as opposed to improved growth in ALE experiments with E. coli exposed to subinhibitory BAC concentrations. Two distinct evolutionary treatments selecting for survival only or survival and growth led to specific evolutionary adaptations apparent in the phenotypes and genotypes of the evolved populations. Populations growing in the presence of BAC evolved increased fitness in the presence of BAC associated with higher resistance to BAC and cross-resistance to antibiotics, while this was not the case for populations evolving for increased survival only. Genotypic characterization by whole genome sequencing of the evolved populations revealed parallelism in mutated genes among replicate populations and distinct differences across treatments. Treatments selecting for survival and growth showed mutations in stress response related genes (hslO and tufA), while selection for survival led to mutations in genes for metabolic regulation (cyaA) and cellular structure (flagella fliJ). In summary, this thesis shows that biocides affect AMR evolution and emphasizes the importance of understanding of how biocides impact the molecular and evolutionary process that underlie AMR evolution. KW - Biocides KW - Antimicrobial resistances KW - Microbial survival mechanisms PY - 2024 UR - https://nbn-resolving.org/urn:nbn:de:kobv:188-refubium-43383-9 SP - 1 EP - 101 PB - Freie Universität CY - Berlin AN - OPUS4-60678 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Deland, Eric T1 - Environmental conditions affect the corrosion product composition of Methanogen induced microbiologically influenced corrosion (Mi-MIC) N2 - Corrosion is a very expensive and serious problem in the different industry sectors, eg. Petroleum-, On- and off-shore, infrastructure. It is estimated that 20% of all corrosion damage is caused by microorganisms or microbiologically influenced corrosion (MIC). Several microorganisms are known to cause corrosion, including sulfate-reducing bacteria, nitrate-reducing bacteria, methanogens etc. For several years, methanogens were regarded as a mild corroder (~0.065 mm/yr), largely due to a lack of detailed investigation on the corrosion mechanism under real-environment simulated conditions. Resulting in the common belief that siderite, a non-conductive compound, is the sole corrosion product (CP) of methanogen-induced MIC (Mi-MIC). To simulate natural environmental conditions, we developed and introduced the multiport flow column system (MFC), a multi-sectional corrosion flow-cell. Using the MFC, we obtained ten times higher corrosion rates than previously reported. With a combination of several analytical techniques, such as ToF-SIMS, SEM-EDS and FIB-SEM, we found strong indication that siderite is not the sole corrosion product of Mi-MIC. The corrosion layers contained phosphorus, oxygen, magnesium, calcium and iron. The differences in the CP between static and dynamic environments demonstrated the impact of testing procedures on the corrosive potential of methanogens. To further verify and deepen our understanding of Mi-MIC, we are currently studying the influence of additional environmental parameters (e.g. pH, salinity, flow rate) on Mi-MIC. Overall, results of this study will expand the current understanding of MIC from both analytical and mechanistic points of view, thus aiding the development of different mitigation strategies for various industry sectors. T2 - ISMOS-8 CY - Online meeting DA - 08.06.2021 KW - Methanogen-induced microbiologically influenced corrosion KW - Carbon steel KW - ToF-SIMS KW - Modelling studies PY - 2021 AN - OPUS4-54041 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Deland, Eric T1 - Environmental conditions impact the corrosion layer composition of Methanogen induced microbiologically influenced corrosion (Mi-MIC) N2 - Microbiologically influenced corrosion (MIC) is a now well-known challenge and affects industry, society and infrastructure. For a long time, the impact of methanogen-induced MIC (Mi-MIC) was underestimated. This was mainly due to the rather low published corrosion rates and the presumed corrosion product siderite, which is not electrically conductive. In our laboratory, we were able to show that this trivialization or underestimation of Mi-MIC was due to the nature of the testing. The static systems used so far do not provide environmentally relevant information about the corrosion rate and the corrosion product, especially for methanogens To further illustrate the importance of the environment on the MIC process, we established a multiport flow column (MFC), to simulate the natural environment. With this method, and in contrast to published results using the static conditions, we obtained ten times higher corrosion rates. We analyzed the corrosion products with a combination of techniques, like ToF-SIMS, SEM-EDS and FIB-SEM, and found strong evidence that siderite is not the sole corrosion product of Mi-MIC. The corrosion layers contain phosphorus, oxygen, magnesium, calcium and iron and lacked on carbon-related species. It is hypothesized that methanogens may have influenced the nucleation process of siderite, converting bicarbonate into carbon dioxide for methanogenesis. This results in increased localized corrosion and reduced siderite formation. To verify and deepen our understanding of Mi-MIC, we are currently studying the influence of additional environmental parameters (e.g. pH, salinity, flow rate etc.) on Mi-MIC and the subsequent impacts on corrosion rates and the corrosion products. Overall, results of this study will expand the current understanding of MIC from both analytical and mechanistic points of view, thus aiding the development of different mitigation strategies for various industry sectors. T2 - Eurocorr 2021 CY - Online meeting DA - 20.09.2021 KW - Methanogen-induced microbiologically influenced corrosion KW - Carbon steel KW - SEM-EDS KW - Modelling studies PY - 2021 AN - OPUS4-54043 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schwibbert, Karin A1 - de Eulate Pérez, Elisa García A1 - Wen, Keqing A1 - Voss, Heike A1 - Thiele, Dorothea A1 - Richter, Anja M. A1 - Bonse, Jörn T1 - Escherichia coli repellence via laser-induced periodic surface structures (LIPSS) on titanium: Critical role of biofilm-associated gene expression N2 - The inconsistency in the observed bacteria-repellent effects of laser-induced surface textures remains a subject of scientific debates. With E. coli TG1 as test strain, we performed biofilm formation assays on ultrashort pulse laser textured titanium. Samples covered with low spatial frequency LIPSS (LSFL) of periods around 750 nm or with high spatial frequency LIPSS (HSFL) featuring smaller periods around 100 nm were processed on polished samples. While LSFL significantly reduce the number of adhering bacteria compared to the polished reference, HSFL do not. Simultaneously, we concentrate our study on bacterial strategies employed for biofilm formation, that is the transcription of the biofilm-associated genes csgB, involved in primary adhesion to a surface, and wza, participating in biofilm maturation. Promoter regions of these genes were cloned into a promotor probe plasmid with the green fluorescent protein GFP+ as reporter and transferred into the test strain. As indicated by the fluorescence intensity of the reporter protein, among the bacteria colonizing LSFL-covered areas, a markedly higher proportion of cells transcribing csgB or wza were detected, compared to the bacterial populations colonizing the polished or HSFL-covered areas. To our knowledge, this is the first study indicating that the antifouling effect of LSFL topographies can be counteracted when bacteria activate biofilm-associated genes. KW - Laser-induced periodic surface structures (LIPSS) KW - Titanium KW - Biofilm KW - Antifouling KW - Gene expression KW - Fluorescence microscopy PY - 2026 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-655572 DO - https://doi.org/10.1016/j.apsusc.2026.166363 SN - 0169-4332 VL - 730 SP - 1 EP - 9 PB - Elsevier B.V. AN - OPUS4-65557 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Koerdt, Andrea T1 - Euro-MIC 2021-2025, our journey, and paths forward N2 - Microbiologically Influenced Corrosion (MIC) refers to the detrimental effects on materials caused by microorganisms, and it is becoming an increasingly significant issue for society. Unlike the USA, Canada, and Australia, Europe has less developed cooperation on MIC. Although several research groups and industrial stakeholders are addressing MIC, discussions remain fragmented, and information exchange is limited. A truly transdisciplinary approach is rarely seen. As a result, Europe often relies on methods, preventive measures, and standards from other regions, as there are no equivalent European standards. This situation makes Europe a) highly dependent, and b) in some cases, unable to use certain measures or standards due to European legal restrictions (e.g., the use of biocides). In 2021, researchers established the “Euro-MIC” network, financially supported by the EU project “COST-Action,” to tackle these issues. Through COST-Action, Euro-MIC aims to facilitate necessary interactions, communication, knowledge sharing, and training for personnel and researchers across various disciplines. COST-Action supports network activities, workshops, training schools, conferences, and more. Euro-MIC aspires to position Europe as a leader in MIC, promoting ideas on par with other nations while upholding European values and ensuring greater protection for people, property, and the environment. In this presentation, I will briefly introduce the principles of COST Action and highlight the significant opportunities provided by this EU-funded project. COST Action fosters interdisciplinarity, networking, training, scientific exchange, and the promotion of young scientists. By showcasing some examples of CA20130 COST ACTION Euro-MIC, I hope to demonstrate that COST Action is not only relevant for addressing MIC but can also be applied to other important topics and sectors. T2 - Unseen Corrosion: Unveiling Hidden Threats and Innovating Monitoring Solutions CY - Bergen, Norway DA - 15.05.2025 KW - COST Action CA20130 KW - MIC KW - Network KW - Corrosion KW - Microorganisms PY - 2025 UR - https://www.norceresearch.no/en/events/unseen-corrosion AN - OPUS4-63726 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Koerdt, Andrea T1 - Euro-MIC COST Action CA20130 Closing Workshop & Conference 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 - Final COST Action Conference - Horsens CY - Horsens, Denmark DA - 17.09.2025 KW - MIC KW - COST Action Euro-MIC KW - Interdisciplinarity PY - 2025 AN - OPUS4-64299 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 - Evolution and mechanistic basis of disinfectant tolerance in E. coli N2 - Disinfectants are important to provide hygiene in sensitive areas, to prevent the spread of infections and to preserve materials from biodeterioration. Bacteria can survive disinfection through phenotypic and genotypic adaptation. Phenotypic heterogeneity may be linked to the ability to evolve disinfectant tolerance. The genetic factors which determine the survival of disinfection remain largely unknown. Here, we investigate the effects of phenotypic heterogeneity on the evolvability of disinfectant tolerance. Furthermore, using a whole-genome CRISPRi-library, we uncover genetic determinants that are important for the survival of disifenction. T2 - µClub Seminar Berlin CY - Berlin, Germany DA - 23.05.2025 KW - Biocides KW - Heterogeneity KW - Biocide resistance KW - Evolution KW - Disinfectants PY - 2025 AN - OPUS4-63833 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Nordholt, Niclas A1 - Kanaris, Orestis A1 - Lewerenz, Dominique A1 - Gödt, Annett A1 - Schreiber, Frank T1 - Evolutionary implications of heterogeneous disinfectant tolerance N2 - Introduction: Effective disinfection is crucial to maintain hygiene and to prevent the spread of infections. Phenotypic heterogeneity in disinfection survival (i.e. tolerance) may result in failure of disinfection, which in turn may foster the evolution of resistance to both disinfectants and antibiotics. However, the consequences of phenotypic heterogeneity for disinfection outcome and resistance evolution are not well understood. Goal: This study investigates the impact of phenotypic heterogeneity on the survival and evolution of Escherichia coli during disinfection with six commonly used substances. Furthermore, the consequences of evolved disinfectant tolerance for antibiotic resistance evolution are studied. Materials & Methods: The extent of population heterogeneity during disinfection is derived by determining time-kill kinetics and analysis with mathematical modelling. The link between population heterogeneity and evolvability of disinfectant tolerance was assessed by laboratory evolution experiments under periodic disinfection. The ability of disinfectant tolerant strains to evolve antibiotic resistance is assessed by serial transfer experiments with increasing concentrations of different antibiotics and by whole genome sequencing. Results: Multi-modal time-kill kinetics in three of the six disinfectants suggest the presence of disinfectant-tolerant subpopulations (i.e. persister cells). Importantly, the ability and extent to evolve population-wide tolerance under periodic disinfection is related with the presence of persister cells and the level of phenotypic heterogeneity during disinfection. Interestingly, the probability of high-level resistance evolution to certain antibiotics is attenuated in disinfectant tolerant strains as compared to the sensitive ancestor. Whole-genome sequencing reveals epistatic interactions between disinfectant tolerance and antibiotic resistance mutations, preventing access to canonical evolutionary paths to resistance. Summary: Our findings suggest that phenotypic heterogeneity can facilitate disinfection survival and the evolution of population wide tolerance, which can impact future antibiotic resistance evolution. T2 - Vereinigung Allgemeiner und Angewandter Mikrobiobiologie Jahreskongress 2023 CY - Würzburg, Germany DA - 02.06.2024 KW - Biocide KW - Resistance KW - Persistence KW - Evolution KW - Herteogeneous phenotypes PY - 2024 AN - OPUS4-60244 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Dinter, Adelina-Elisa T1 - Examination of biological samples by means of single-cell ICP-ToF-MS N2 - Up to now, different analytical methods for single cell analysis exist focusing on key features such as size, shape, morphology and elemental composition. The combination of the latest ICP-MS techniques - ICP-ToF-MS - together with the latest developments in the field of sample delivery - micro droplet generator (MDG) – will allow a Deep insight into the composition and size of cells. Microbiologically influenced corrosion (MIC) is an oxidation of metals affected by the presence or activity (or both) of microorganisms e.g. Shewanella Putrefaciens in biofilms on the surface of the corroding material. As this can happen for example in the soil on iron pipes of water pipes, in oil tanks or on steel sheet piling, there is great interest in MIC research, not only from various industrial sectors, but also from the environmental aspect. T2 - SALSA - Make & Measure CY - Online Meeting DA - 15.10.2020 KW - SC-ICP-ToF-MS KW - Single cell analysis KW - MDG ICP-ToF-MS KW - Microdroplet generator PY - 2020 AN - OPUS4-52441 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Abilio, André T1 - Expert System for Screening Microbiologically Influenced Corrosion (MIC) as Internal Failure Cause in Oil and Gas Upstream Pipelines N2 - The analysis of pipeline failures due to Microbiologically Influenced Corrosion (MIC) is challenging due to the complex interaction of many influencing parameters including pipeline operation conditions, fluid chemistry and microbiology, as well as the analysis of corrosion features and products. To help address this challenge, an expert system was developed to assist non-specialists in screening internal pipeline corrosion failures due to MIC related threats. To accomplish this, 15 MIC subject matter experts (with a total of 355 man-years of accumulated MIC based experience) were recruited to evaluate a total of 65 MIC failure cases based on real-life scenarios. These case study parameters and the expert elicited results were input into an Artificial Neural Network (ANN) model to create a model system which can screen whether a given failure scenario is one of three outcomes: a) failure is likely due to MIC, b) failure is likely not due to MIC, or c) the conclusion is inconclusive (analysis needs more data/information). The model system had an overall accuracy of 74.8% and it showcases that knowledge from subject matter experts can be captured in a reasonably effective way to screen for possible MIC failures. Based on that, this presentation will provide details of the model development process and key results to date. Important considerations regarding the level of confidence of the diagnoses and variation between expert opinion will also be discussed alongside with ideas on how to improve the model for field applicability. T2 - ISMOS 9 CY - Edinburgh, United Kingdom DA - 27.06.2023 KW - Expert System KW - Microbiologically Influenced Corrosion (MIC) KW - Oil and Gas Upstream KW - Failure Investigation KW - Artificial neural networks (ANN) PY - 2023 AN - OPUS4-64603 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Abilio, André T1 - Expert System for Screening Microbiologically Influenced Corrosion asInternal Failure Cause in Oil and Gas Upstream Pipelines N2 - The analysis of pipeline failures due to Microbiologically Influenced Corrosion (MIC) is challenging due to the complex interaction of many influencing parameters including pipeline operation conditions, fluid chemistry and microbiology, as well as the analysis of corrosion features and products. To help address this challenge, an expert system was developed to assist non-specialists in screening internal pipeline corrosion failures due to MIC related threats. To accomplish this, 15 MIC subject matter experts (with a total of 355 man-years of accumulated MIC based experience) were recruited to evaluate a total of 65 MIC failure cases based on real-life scenarios. These case study parameters and the expert elicited results were input into an Artificial Neural Network (ANN) model to create a model system which can screen whether a given failure scenario is one of three outcomes: a) failure is likely due to MIC, b) failure is likely not due to MIC, or c) the conclusion is inconclusive (analysis needs more data/information). The model system had an overall accuracy of 74.8%and it showcases that knowledge from subject matter experts can be captured in a reasonably effective way to screen for possible MIC failures. Based on that, this presentation will provide details of the model development process and key results to date. Important considerations regarding the level of confidence of the diagnoses and variation between expert opinion will also be discussed alongside with ideas on how to improve the model for field applicability. T2 - ISMOS 9 CY - Edinburgh, United Kingdom DA - 27.06.2023 KW - Expert System KW - Microbiologically Influenced Corrosion (MIC) KW - Oil and Gas Upstream KW - Artificial neural networks (ANNs) KW - Failure analysis PY - 2023 AN - OPUS4-64602 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -