TY - CONF A1 - Gorbushina, Anna T1 - A genetic toolbox for exploring the life style of the rock-inhabiting black fungus Knufia petricola N2 - After their discovery on rock surfaces in cold and hot deserts, a polyphyletic group of ascomycetous black fungi was found to dominate a range of hostile environments – natural and man-made, from salterns to dishwashers, roofs and solar panels. Together with bacteria and algae they may establish subaerial biofilms and cause weathering of the surfaces they grow on. Their impressive survival abilities as well as their constitutive protective pigmentation and cluster-like microcolony organisation are similar in environmental isolates as well as in heat-tolerant opportunistic pathogens of animals and humans. The exact genetic properties that ensure their survival in extreme environments can be studied if some black fungi were amenable to genetic manipulations. We selected the rock-inhabiting fungus Knufia petricola (class Eurotiomycetes, order Chaetothyriales) that grows moderately in axenic culture and exhibits all the characteristics of microcolonial black fungi such as yeast-like cell growth, absence of reproductive structures and constitutive dihydroxynaphthalene (DHN) melanogenesis (Nai et al. 2013, Fungal Genet Biol). We developed protocols to efficiently generate and transform protoplasts resulting in stable homokaryotic transformants by targeting genes involved in pigment synthesis and expressing fluorescent reporter genes. Hence, endogenous and foreign genes can be expressed from episomal AMA1-containing plasmids and genome-integrated DNA constructs. Moderate rates of homologous recombination allow for both ectopic and targeted integrations. CRISPR-Cas9 was further validated as a strategy for obtaining selection marker-free mutants and silencing via RNA interference as an approach to study essential genes. Availability of this genetic toolbox and an annotated genome sequence of the strain A95 is paving the way for studying interactions of K. petricola with environmental stressors, material surfaces, soil matrices and phototrophic symbionts. T2 - European Conference on Fungal Genetics (ECFG15) CY - Rome, Italy DA - 17.02.2020 KW - black fungi KW - genomics KW - genetics PY - 2020 AN - OPUS4-50593 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 - CONF A1 - Erdmann, Eileen T1 - A random mutagenesis approach to elucidate the biology of extremotolerant black fungi N2 - Microcolonial black fungi ubiquitously inhabit sun-exposed natural and man-made surfaces of our planet. To promote genetic studies, which are hindered by slow growth, lack of sexual cycles and transformation difficulties, CRISPR/Cas9-based genetic tools were implemented (Erdmann et al. 2022, Front Fungal Biol). Now efficient targeted mutagenesis of the rock inhabitant Knufia petricola (Eurotiomycetes/Chaetothyriales) - as a representative of the polyphyletic group of black fungi - enables the elucidation of extremotolerance, oligotrophism, unusual types of cell division, mineral weathering and symbiotic interactions. Still more progress on assigning functions to yet unknown genes can be expected if a forward genetics approach is available. We chose the two-component Activator/ Dissociation (Ac/Ds) transposon system from maize for generating a collection of insertional mutants by in-vivo mutagenesis of K. petricola. For the optimal use of this genetic tool, an inducible promoter for the expression of the Ac transposase (AcTPase) and by this the regulatable transposition of the resistance cassette-containing Ds transposon is desired. However, endogenous promoters for nitrate assimilation and galactose catabolism - often used in fungi for regulatable gene expression - are not inducible by their substrates in K. petricola suggesting that the regulatory networks for nutrient acquisition differ significantly in oligotrophic fungi. Therefore, the metabolism-independent Tet-on system was combined with the AcTPase coding sequence and subsequently transformed into Ds-carrying K. petricola strains. In total, four auxotrophic Ac|Ds starter strains containing the Ds transposon at different position of ade2 or ura3 were generated. The cultivation of these strains with doxycycline for induction of TET::Ac and subsequent selection of cells on ADE/URA-lacking media resulted in prototrophic colonies (revertants) for some but not all Ac|Ds strains. Currently, the transposition events in the obtained revertants are studied to validate the procedure. First amplicon sequencing of excision sites revealed footprint patterns, proving the transposon jumped. T2 - 16th European Conference on Fungal Genetics CY - Innsbruck, Austria DA - 05.03.2023 KW - Rock-inhabiting fungi KW - Foward genetics KW - In-vivo mutagenesis PY - 2023 AN - OPUS4-57144 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 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 - Bonnerot, Olivier T1 - Are X-rays safe for manuscripts’ materials? N2 - In the last decade, applications of X-rays to the study of manuscripts significantly spread in both diversity and extent. They range from writing material analysis, mostly with X-ray fluorescence (XRF), permitting non-invasive characterization of inks and pigments used, to the investigation of the origin of writing supports. In addition, XRF mapping has proved to be an invaluable tool for recovering erased text. Finally, computed-tomography (CT) has shown potential in virtually unrolling rolls, making text readable without using-damaging mechanical methods. Despite their growing use, little attention has been paid to the side effects of such analytical tools. We observed irreversible parchment colour changes during some experiments on dead-sea scrolls with synchrotron radiation sources. Furthermore, partial photo-reduction of iron under high intensity beam during X-ray absorption near edge structure spectroscopy (XANES) measurements of iron-gall ink on paper has been reported several times [5,6]. Such phenomena have mostly been overlooked so far, although there is an increasing awareness of the necessity to study them. We conducted experiments at the Deutsches Elektronen-Synchrotron (DESY) facilities to investigate X-ray induced structural alteration of paper and parchment to see whether the presence of absorption centres (ink and pigments) has an impact. In addition to better understanding degradation processes, we are aiming to define an appropriate methodology of analysis of manuscripts with a tolerable risk of damage. The first results concerning X-ray induced damage of cellulose materials have already been presented at the Synchrotron Radiation and Neutrons in Art and Archaeology (SR2A). We are focusing here on the results on parchment materials. T2 - Cultural and Natural Heritage Workshop CY - Grenoble, France DA - 22.01.2020 KW - X-rays KW - Manuscripts KW - Parchment KW - Synchrotron PY - 2020 AN - OPUS4-50305 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 - Cook, Jake Thomas T1 - Assessment of concrete bioreceptivity in algal biofilm green façade systems N2 - Algal biofilm façades are an alternative to traditional green façades which can help to improve biodiversity and air quality within cities. They present a low maintenance approach in which subaerial algae are grown directly on concrete substrates. The intrinsic bioreceptivity of the substrate is a critical factor in successful facade colonisation. Existing research has identified several environmental and material properties which influence concrete bioreceptivity, however a consensus has yet to be made on which properties are most influential and how the interaction between properties may promote algal biofilm growth under specific conditions. T2 - International Conference on Bio-Based Building Materials CY - Vienna, Austria DA - 21.06.2023 KW - Concrete KW - Façade KW - Bio-receptive KW - Extracellular polymeric substances KW - Fractional factorial PY - 2023 UR - https://www.rilem.net/agenda/5th-international-conference-on-bio-based-building-materials-1501 AN - OPUS4-58976 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Kalbe, Ute T1 - Assessment of Microplastic and Contaminant Emissions from Artificial Turf Sports Pitches N2 - Due to their advantages over natural gras, artificial turf sports pitches are well established in popular sports and their number is increasing every year. This raises questions about potential pollutant emissions and possible risks to the environment due to its transfer to soil and groundwater. The main sources of emissions are the rubber infill granules and the abrasion of grass fibers. While estimates on the transfer of pollutants to the soil are already available, there is still insufficient data on the discharge of microplastics from artificial turf systems and thus on the assessment of its relevance for the soil. To close this gap, the pollutant emissions of three artificial turf scenarios - past (fossil-based, rubber infill), present (most frequently installed in Germany, rubber infill) and future (turf with recycled grass fibers, no rubber infill) - were compared in different ageing states (unaged, accelerated, and real time aged). Accelerated ageing consisting of UV weathering and mechanical stress was applied to simulate the outdoor weathering of turf systems during their service life span of approx. 15 years. Emissions of microplastics and environmentally relevant pollutants were sampled simultaneously using newly developed, innovative lysimeters. The microplastics contents were determined using Thermal Extraction-Desorption Gas Chromatography/Mass Spectrometry (TED-GC/MS). In addition, the concentrations of PAH and heavy metals were determined. T2 - Jahrestag der Deutschen Bodenkundlichen Gesellschaft CY - Tübingen, Germany DA - 14.09.2025 KW - Artificial Turf KW - Microplastics KW - Contaminants PY - 2025 AN - OPUS4-65022 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Kittner, Maria T1 - Assessment of Microplastic Emissions from Artificial Turf Sports Pitches N2 - Following the recommendation of the European Chemicals Agency, on 25 September 2023 the European Commission passed a comprehensive new regulation to reduce emissions of microplastics (MP) into the environment, which includes the sale and use of intentionally added MP.1,2 This also applies to the application of synthetic rubber granulate infill in artificial turf systems and will ultimately have an impact on recreational sports. In Germany, rubber granulate made of ethylene-propylene-diene-monomer rubber (EPDM) is currently predominantly used. So far, there is no sufficient database for estimating MP emissions from artificial turf pitches into the environment and thus their relevance as a source of MP pollution.3 This topic is controversially discussed due to the complexity of sampling and analytics. To close this research gap, this project has the goal to determine mass balances for the emissions of MP from artificial turf pitches to allow an estimation of the amount of MP released per artificial turf pitch and year. Within this study, MP emissions of three artificial turf scenarios at different time states (unaged, artificially aged and real-time aged) are compared: the past (old turf: fossil based, synthetic infill), present (most commonly installed in Europe: fossil based, EPDM infill), and the future (turf with recycled gras fibres, no synthetic infill). To simulate the outdoor weathering during the lifespan of an artificial turf of approx. 15 years, brand-new artificial turf and EPDM rubber granulate were accelerated aged by means of UV weathering and mechanical stress. Potential MP emissions into surface and groundwater are simulated by lysimeter and shake experiments. MP mass contents are subsequently determined by Thermal Extraction Desorption Gas Chromatography/Mass Spectrometry. Using special microfilter crucibles allows the estimation of the particle sizes of the emitted MP, which is a fundamental requirement for an assessment of potential health hazards for humans. T2 - SETAC 2024 CY - Seville, Spain DA - 05.05.2024 KW - Mikroplastik KW - TED-GC/MS KW - Lysimeter KW - PAK KW - Schwermetalle PY - 2024 AN - OPUS4-60014 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 - Prajapati, Harshit Kumar T1 - Bees got a virus? N2 - The poster was presented at the long night of sciences 2023, to spread awareness about the Deformed Wing Virus - a leading cause of honeybee colony losses worldwide. We also describe our research contributions to understanding the evolutionary dynamics of this virus. T2 - Lange Nacht der Wissenschaften 2023 CY - Berlin, Germany DA - 17.06.2023 KW - Evolution KW - Varroa destructor KW - Honeybees KW - Virus PY - 2023 AN - OPUS4-61717 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -