TY - CONF A1 - Kanaris, Orestis T1 - Identification of metabolic niches and their association to the persistence of antibiotic resistant bacteria in wastewater N2 - Wastewater treatment plants (WWTP) are hotspots for the development and spread of antimicrobial resistance. Antimicrobial resistant bacteria (ARB) can persist in the environment for long periods of time, despite metabolic fitness costs that often arise with resistance. Recent research efforts are striving to uncover the role of bacterial metabolism for the ecology and evolution of antibiotic resistance. The aim of this study is to understand the ecological mechanisms, which allow ARB to persist in the environment. More specifically, we aim to identify metabolic niches that can select for and against resistant bacteria. 62 E. coli strains isolated from different WWTPs with different levels of resistance to 14 antibiotics and 3 disinfectants were assembled, sequenced, and phenotypically characterized. Next, genome scale metabolic models (GEMs) were constructed, and the growth of the strains was simulated in the presence of 298 different carbon sources. Furthermore, the growth rates of the isolates were measured in the presence of 3 carbon sources to verify the model predictions. Competition experiments with synthetic microbial communities consisting of a selection of 10 WWTP isolates, 5 of which were antibiotic resistant and 5 sensitive, were carried out in minimal medium with different carbon sources. Population dynamics modelling was used to simulate the competition of isolates under different conditions. The isolates have a wide range of susceptibility to the antibiotics, while disinfectants result in a narrower range of susceptibility. GEMs identified 40 carbon sources that can be utilized for growth only by a portion of all the isolates. The prediction accuracy of the GEMs was 93% in the case of D-malate. A range of WWTP isolates were identified which use D-malate as carbon source and are susceptible to specific antibiotics. In contrast, antibiotic-resistant WWTP isolates were identified that did use sucrose as carbon source but not D-malate. Competition experiments demonstrated that changing the carbon source of the medium from sucrose to D-malate resulted in selection against the resistant isolates. Modelling the competition between isolates under different conditions suggests that adding a carbon source to a bacterial community under specific conditions could exclude resistant bacteria from a microbial community. Our data suggest that changing the available carbon source could shift the selection advantage between resistant and susceptible bacterial strains. If this strategy is confirmed experimentally in complex microbial communities, it could be applied to reduce the number of ARB in environments such as wastewater. T2 - FEMS micro Konferenz CY - Milan, Italy DA - 14.07.2025 KW - AMR KW - Wastewater KW - Genome-scale metabolic model PY - 2025 AN - OPUS4-64659 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Kanaris, Orestis T1 - Metabolic niches and persistence of antibiotic resistant bacteria in the environment N2 - Background and Aim: Wastewater treatment plants are considered as hotspots for the development and spread of antimicrobial resistance. Antimicrobial resistant bacteria (ARB) can persist in the environment for long periods of time, despite metabolic fitness costs that can arise with resistance. We are interested in understanding the mechanisms, which allow ARB to persist in the environment. More specifically, we want to identify metabolic niches that can select for resistant bacteria. Procedure/Method: 62 E. coli strains isolated from different WWTPs in Norway were used. The isolates have different levels of resistance to 14 antibiotics. The susceptibility of the isolates to 3 disinfectants was determined. In addition, genome scale metabolic models (GEMs) were constructed and the growth of the strains was simulated in the presence of 198 different carbon sources. Furthermore, the growth rates of the isolates were measured in the presence of 3 carbon sources. Findings/Results: Many of the isolates have high resistance to multiple antibiotics but only few of the isolates have higher minimum inhibitory concentrations to the disinfectants, compared to an E. coli laboratory strain. With the GEMs, we identified 40 carbon sources that can be utilized for growth only by a portion of all the isolates. The prediction accuracy of the GEMs was 93% in the case of D-Malate. A group of 10 isolates was identified of which 5 isolates are resistant to ciprofloxacin, gentamicin and tetracycline and can grow on Sucrose but not on D-Malate and the other 5 isolates are susceptible to the same 3 antibiotics but can grow on D-Malate and not on Sucrose. Implications/Applications: Our data suggest that changing the available carbon source could shift the selection advantage between resistant and susceptible bacterial strains. If this strategy is confirmed experimentally, it could be applied to reduce the number of ARB in environments such as wastewater. T2 - International Biodeterioration and Biodegradation Symposium 19 CY - Berlin, Germany DA - 09.09.2024 KW - Antimicrobial resistance KW - Wastewater PY - 2024 AN - OPUS4-61412 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Kanaris, Orestis T1 - Metabolic niches and persistence of antibiotic resistant bacteria in the environment N2 - Background and Aim: Wastewater treatment plants are considered as hotspots for the development and spread of antimicrobial resistance. Antimicrobial resistant bacteria (ARB) can persist in the environment for long periods of time, despite metabolic fitness costs that can arise with resistance. We are interested in understanding the mechanisms, which allow ARB to persist in the environment. More specifically, we want to identify metabolic niches that can select for resistant bacteria. Procedure/Method: 62 E. coli strains isolated from different WWTPs in Norway were used. The isolates have different levels of resistance to 14 antibiotics. The susceptibility of the isolates to 3 disinfectants was determined. In addition, genome scale metabolic models (GEMs) were constructed and the growth of the strains was simulated in the presence of 198 different carbon sources. Furthermore, the growth rates of the isolates were measured in the presence of 3 carbon sources. Findings/Results: Many of the isolates have high resistance to multiple antibiotics but only few of the isolates have higher minimum inhibitory concentrations to the disinfectants, compared to an E. coli laboratory strain. With the GEMs, we identified 40 carbon sources that can be utilized for growth only by a portion of all the isolates. The prediction accuracy of the GEMs was 93% in the case of D-Malate. A group of 10 isolates was identified of which 5 isolates are resistant to ciprofloxacin, gentamicin and tetracycline and can grow on Sucrose but not on D-Malate and the other 5 isolates are susceptible to the same 3 antibiotics but can grow on D-Malate and not on Sucrose. Implications/Applications: Our data suggest that changing the available carbon source could shift the selection advantage between resistant and susceptible bacterial strains. If this strategy is confirmed experimentally, it could be applied to reduce the number of ARB in environments like wastewater. T2 - Conference on the Environmental dimentions of antimicrobial resistance 7 CY - Montreal, Canada DA - 26.05.2024 KW - Antimicrobial resistance KW - Wastewater PY - 2024 AN - OPUS4-61410 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 - CONF A1 - Frenzel, Olivia T1 - Identification of Metabolites and Transformation Products of Bisphenols N2 - Bisphenol A (BPA) is used in Polycarbonate, Polyacrylic resins, Polysulfones, Epoxy resins, and Polyetherimides. It is also used in recycled Polyvinyl chloride [1–3]. BPA has been classified as a substance of very high concern (SVHC) under REACH [4] due to its endocrine disrupting properties. The German competent authorities want to reduce the content of BPA in the environment. Possible substitutes for BPA are Bisphenol B (BPB), Bisphenol E (BPE), Bisphenol F (BPF), or Bisphenol S (BPS), as they are similar in structure. BPA-based materials have a wide area of application especially outside. These outdoor applications are exposed to different external influences, including physical, biological, mechanical and chemical influences. This results in damage and aging of the material with leaching or migration into the environment. There, the substance is transformed by various transformation processes. The emerging metabolites and transformation products (TPs) can have different properties than the parent substance. Understanding the fate and behavior of the emerging pollutants is very important. Therefore, different transformation products of selected bisphenols will be generated and analyzed: To investigate the fate of different bisphenols in water treatment plants technical transformation products [5] will be generated by chlorination, ozonization, the Fenton reaction, and UV-radiation. For the investigation of the fate of Bisphenols in surface water global radiation will be simulated with UVA-lamps Simulation of phase-I-metabolism with an electrochemical cell coupled to mass spectrometry (EC-MS). T2 - Anakon 2023 CY - Vienna, Austria DA - 11.04.2023 KW - Transformation products KW - Bisphenols KW - Metabolites KW - Wastewater KW - Environment PY - 2022 AN - OPUS4-57374 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 - 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 - Frenzel, Olivia T1 - Identification of transformation products and metabolites of selected Bisphenols N2 - Bisphenol A (BPA) is widely used in many Polymers. Many outdoor applications are made from BPA-based materials which are exposed to a variety of environmental impacts, like temperature, solar radiation, rain, and others. This could lead to damaging and aging of the material which might cause leaching or migration of pollutants into the environment. To understand the fate and behavior different transformation products of selected Bisphenols will be generated and analyzed. T2 - Umwelt2022 CY - Emden, Germany DA - 05.09.2022 KW - Environment KW - Bisphenols KW - Transformation products KW - Metabolites KW - Wastewater PY - 2022 AN - OPUS4-56203 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Frenzel, Olivia T1 - Identification of Metabolites and Transformation Products of Bisphenols N2 - Bisphenol A (BPA) is widely used in Polycarbonate (PC), Polyacrylic resins (PAR), Polysulfones (PSU), Epoxy resins (ER), and Polyetherimides (PEI). It is also used in recycled Polyvinyl chloride (PVC) [1–3]. These BPA-containing materials have a wide area of application, especially outside. All outdoor applications are exposed to a variety of environmental impacts, like temperature, solar radiation (physical influences), chemical influences (rain or ozone), biological influences (microorganisms), and mechanical influences (hail, sand). These impacts are damaging and aging the material which can be followed by leaching or migration of pollutants like Bisphenol A into the environment. Understanding the fate and behavior of the released pollutants is very important. Therefore, different transformation products of selected Bisphenols will be generated and analyzed. T2 - 33rd International Symposium on Chromatography – ISC 2022 CY - Budapest, Hungary DA - 18.09.2022 KW - Environment KW - Transformation products KW - Bisphenols KW - Metabolites KW - Wastewater PY - 2022 AN - OPUS4-56202 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Sichler, Theresa Constanze A1 - Adam, Christian A1 - Barjenbruch, M. A1 - Montag, D. A1 - Mauch, Tatjana A1 - Sommerfeld, Thomas A1 - Ehm, J.H. T1 - Variation of the element composition of municipal sewage sludges in the context of new regulations on phosphorus recovery in Germany N2 - Phosphorus (P) recovery is obligatory for all sewage sludges with more than 20 g P/kg dry matter (DM) from 2029 in Germany. Nine wastewater treatment plants (WWTPs) were chosen to investigate variations of phosphorus contents and other parameters in sewage sludge over the year. Monthly sewage sludge samples from each WWTP were analyzed for phosphorus and other matrix elements (C, N, H, Ca, Fe, Al, etc.), for several trace elements (As, Cr, Mo, Ni, Pb, Sn) and loss of ignition. Among the nine WWTPs, there are four which have phosphorus contents both above and below the recovery limit of 20 g/kg DM along the year. Considering the average phosphorus content over the year, only one of them is below the limit. Compared to other matrix elements and parameters, phosphorus fuctuations are low with an average of 7% over all nine WWTPs. In total, only hydrogen and carbon are more constant in the sludge. In several WWTPs with chemical phosphorus elimination, phosphorus fuctuations showed similar courses like iron and/or aluminum. WWTPs with chamber flter presses rather showed dilution efects of calcium dosage. As result of this study, monthly phosphorus measurement is highly recommended to determine whether a WWTP is below the 20 g/kg DM limit. KW - Sewage sludge KW - Phosphorus recovery KW - Wastewater KW - Phosphorus elimination PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-557499 DO - https://doi.org/10.1186/s12302-022-00658-4 SN - 2190-4707 VL - 34 IS - 1 SP - 1 EP - 12 PB - Springer Nature CY - Berlin AN - OPUS4-55749 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -