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    Item type:Publication, Access status: with files ,
    Implementation of SARS-CoV-2 wastewater surveillance systems in Germany- Pilot study in the federal state of Thuringia
    (Multidisciplinary Digital Publishing Institute (MDPI), 2026-01-24)
    Kaller, Felix
    ;
    ; ; ;
    Reichel-Kühl, Katarina
    Since the COVID-19 pandemic, wastewater monitoring has become an additional tool in the surveillance of infectious diseases. Many EU countries put wastewater surveillance systems (WSS) in place to track SARS-CoV-2 and its variants and other pathogens, such as the influenza virus or Respiratory syncytial virus (RSV). In Germany, several research and pilot projects funded by the EU, the Federal Ministry of Education and Research, the Federal Ministry of Health, and projects at Federal State level have been launched in the last four years. In Germany, wastewater monitoring was not implemented as a public health tool before the COVID-19 pandemic, but in September 2022, it has been legally determined in the German infection protection act (Infektionsschutzgesetz, IfSG). As Germany is a federal state, competencies in epidemic management partly belong to the 16 federal states (“Länder”). In the federal states, the local health authorities at the county (“Kreise”) level also have specific risk management and communication competencies. Furthermore, WSS has been incorporated into the revised Urban Wastewater Treatment Directive (EU) 2024/3019. For this reason, the federal states and local health authorities play a pivotal role in successfully implementing wastewater monitoring as a supplementary component of disease surveillance in Germany. Between November 2021 and August 2022, the federal state of Thuringia, Germany, supported a pilot study to implement a surveillance system for SARS-CoV-2-RNA in wastewater of 23 wastewater treatment plants in 17 counties in Thuringia. Here, we describe the study design and the system behind the logistics and the planning, and we provide an overview of the options for involving the public health service. Furthermore, the possibilities for IT concepts and approaches to innovative AI solutions are shown. We also aim to explore the feasibility and potential barriers to further implementing wastewater surveillance as a supplementary public health tool in Thuringia.
    Publicationtype: Journal Article
      19  21
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    Influence of varying pre-culture conditions on the level of population heterogeneity in batch cultures with an escherichia coli triple reporter strain
    (Multidisciplinary Digital Publishing Institute (MDPI), 2023-07) ; ; ; ;
    When targeting robust, high-yielding bioprocesses, phenomena such as population heterogeneity have to be considered. Therefore, the influence of the conditions which the cells experience prior to the main culture should also be evaluated. Here, the influence of a pre-culture medium (complex vs. minimal medium), optical density for inoculation of the main culture (0.005, 0.02 and 0.0125) and harvest time points of the pre-culture in exponential growth phase (early, mid and late) on the level of population heterogeneity in batch cultures of the Escherichia coli triple reporter strain G7BL21(DE3) in stirred-tank bioreactors was studied. This strain allows monitoring the growth (rrnB-EmGFP), general stress response (rpoS-mStrawberry) and oxygen limitation (nar-TagRFP657) of single cells through the expression of fluorescent proteins. Data from batch cultivations with varying pre-culture conditions were analysed with principal component analysis. According to fluorescence data, the pre-culture medium had the largest impact on population heterogeneities during the bioprocess. While a minimal medium as a pre-culture medium elevated the differences in cellular growth behaviour in the subsequent batch process, a complex medium increased the general stress response and led to a higher population heterogeneity. The latter was promoted by an early harvest of the cells with low inoculation density. Seemingly, nar-operon expression acted independently of the pre-culture conditions.
    Publicationtype: Journal Article
    Scopus© Citations 1  24  112
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    Biochemical characterization of recombinant isocitrate dehydrogenase and its putative role in the physiology of an acidophilic micrarchaeon
    (Multidisciplinary Digital Publishing Institute (MDPI), 2021-11-09) ; ;
    Despite several discoveries in recent years, the physiology of acidophilic Micrarchaeota, such as “Candidatus Micrarchaeum harzensis A_DKE”, remains largely enigmatic, as they highly express numerous genes encoding hypothetical proteins. Due to a lacking genetic system, it is difficult to elucidate the biological function of the corresponding proteins and heterologous expression is required. In order to prove the viability of this approach, A_DKE’s isocitrate dehydrogenase (MhIDH) was recombinantly produced in Escherichia coli and purified to electrophoretic homogeneity for biochemical characterization. MhIDH showed optimal activity around pH 8 and appeared to be specific for NADP+ yet promiscuous regarding divalent cations as cofactors. Kinetic studies showed KM-values of 53.03 _ 5.63 _Mand 1.94 _ 0.12mMand kcat-values of 38.48 _ 1.62 and 43.99 _ 1.46 s1 resulting in kcat/KM-values of 725 _ 107.62 and 22.69 _ 2.15 mM1 s1 for DL-isocitrate and NADP+, respectively. MhIDH’s exceptionally low affinity for NADP+, potentially limiting its reaction rate, can likely be attributed to the presence of a proline residue in the NADP+ binding pocket, which might cause a decrease in hydrogen bonding of the cofactor and a distortion of local secondary structure.
    Publicationtype: Journal Article
    Scopus© Citations 1  95  234
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    Improving the cathodic biofilm growth capabilities of kyrpidia spormannii EA-1 by undirected mutagenesis
    (Multidisciplinary Digital Publishing Institute (MDPI), 2020-12-30) ; ; ;
    The biotechnological usage of carbon dioxide has become a relevant aim for future processes. Microbial electrosynthesis is a rather new technique to energize biological CO2 fixation with the advantage to establish a continuous process based on a cathodic biofilm that is supplied with renewable electrical energy as electron and energy source. In this study, the recently characterized cathodic biofilm forming microorganism Kyrpidia spormannii strain EA-1 was used in an adaptive laboratory evolution experiment to enhance its cathodic biofilm growth capabilities. At the end of the experiment, the adapted cathodic population exhibited an up to fourfold higher biofilm accumulation rate, as well as faster substratum coverage and a more uniform biofilm morphology compared to the progenitor strain. Genomic variant analysis revealed a genomically heterogeneous population with genetic variations occurring to various extends throughout the community. Via the conducted analysis we identified possible targets for future genetic engineering with the aim to further optimize cathodic growth. Moreover, the results assist in elucidating the underlying processes that enable cathodic biofilm formation.
    Publicationtype: Journal Article
    Scopus© Citations 10  88  167
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    Accelerated electro-fermentation of acetoin in escherichia coli by identifying physiological limitations of the electron transfer kinetics and the central metabolism
    (Molecular Diversity Preservation International, 2020-11-23) ; ;
    Anode-assisted fermentations offer the benefit of an anoxic fermentation routine that can be applied to produce end-products with an oxidation state independent from the substrate. The whole cell biocatalyst transfers the surplus of electrons to an electrode that can be used as a non-depletable electron acceptor. So far, anode-assisted fermentations were shown to provide high carbon efficiencies but low space-time yields. This study aimed at increasing space-time yields of an Escherichia coli-based anode-assisted fermentation of glucose to acetoin. The experiments build on an obligate respiratory strain, that was advanced using selective adaptation and targeted strain development. Several transfers under respiratory conditions led to point mutations in the pfl, aceF and rpoC gene. These mutations increased anoxic growth by three-fold. Furthermore, overexpression of genes encoding a synthetic electron transport chain to methylene blue increased the electron transfer rate by 2.45-fold. Overall, these measures and a medium optimization increased the space-time yield in an electrode-assisted fermentation by 3.6-fold.
    Publicationtype: Journal Article
    Scopus© Citations 3  68  170