The excavation at Domerschulstrasse in Würzburg - 1300 years of history of the town

Ausgelöst durch einen Archiv- und Bibliotheksneubau der Diözese Würzburg fanden zwischen Mitte Januar und Anfang Juli 1999 umfangreiche archäologische Ausgrabungen auf dem betroffenen Grundstück statt. Bei dieser Stadtkerngrabung handelte es sich um die erste flächige archäologische Untersuchung in Würzburg, die zudem noch in unmittelbarer Nähe zum Dom, also im Kernbereich der mittelalterlichen Stadt, stattfand. Während der Kampagne konnten zahlreiche Bebauungsstrukturen dokumentiert werden und ein umfangreiches Fundspektrum aus Keramik, Glas, Metall, Bein/ Knochen und Stein geborgen werden. Das vorrangige Ziel der Dissertation bestand in der Klärung der Baugeschichte auf dem ausgegrabenen Areal samt sämtlicher Vorgängerbebauung. Es sollten Fragen zur Entwicklungsgeschichte Würzburgs seit dem 8./ 9. Jahrhundert sowie zu Bebauungsstrukturen und Entwicklung der mittelalterlichen Stadt beantwortet werden. Ein Schwerpunkt bestand in der Klärung von offenen Fragen hinsichtlich der Verteidigungsanlagen und Befestigungssystemen Würzburgs im Früh- und Hochmittelalter. Die Bearbeitung des reichhaltigen Fundgutes versprach wichtige Ergebnisse zu mittelalterlicher Sachkultur, Wirtschaft, Handel und Handwerk zu liefern. Nach Abschluss der Auswertung existieren nun von Seiten der Mittelalterarchäologie her teilweise ganz neue und erweiterte Erkenntnisse zu einzelnen Aspekten der Würzburger Stadtgeschichte, die bisher überwiegend von historischer Seite beleuchtet worden ist. Zu den herausragendsten Befunden zählen die Reste zweier bisher unbekannter Verteidigungsanlagen aus spätmerowingischer bzw. karolingisch-ottonischer Zeit. Besonders hervorzuheben ist dabei, dass erstmals das Vorhandensein einer karolingisch-ottonischen Domimmunitätsbefestigung, also einer Umwehrung und Abgrenzung des engeren Dombereiches nachgewiesen werden konnte. Die historische Stadtgeschichtsforschung hatte eine solche Domburg in Würzburg bisher verneint. Diese beiden frühen Befestigungen komplettieren den Ablauf der städtischen Verteidigungsanlagen vom Frühmittelalter bis in die Neuzeit. Zu den weiteren wichtigen Befunden auf dem Grabungsareal zählt eine romanische, in die Mitte des 12. Jh. zu datierende Hofanlage, bestehend aus einem steinernen, großen, rechteckigen und unterkellertem Gebäude samt Nebengebäuden. Die Baugeschichte dieser Hofanlage mit diversen Um- und Anbaumaßnahmen ( vor allem im 13. und 14. Jh.) konnte bis in die jüngere Neuzeit lückenlos nachvollzogen werden. Die Verbindung mit schriftlichen Quellen erlaubte Aussagen über Nutzung und Bewohner der Gebäude zu den unterschiedlichsten Zeiten. Neben diesen besonderen Befunden wurden beispielsweise auch noch Latrinen, Brunnen und diverse Handwerksöfen untersucht. Die Bearbeitung und Auswertung des Fundmaterials erbrachte unter anderem wichtige Ergebnisse zur Formentwicklung der Keramik vom 8. Jh. bis in die Renaissance. Es konnten Aussagen zu Handel und Handwerk, aber auch zu bestimmten sozialen Strukturen innerhalb der Stadt getroffen werden. Besonders bedeutend sind Kleinfunde des 9./ 10. Jh., die die hohe Bedeutung Würzburgs zu dieser Zeit belegen und auf weite Handelsbeziehungen bis nach Norddeutschland und Dänemark hinweisen. Nach Auswertung aller Befunde und Funde ergibt sich für den ausgegrabenen Bereich eine Bau- und Siedlungsgeschichte mit bedeutenden Befunden, die über einen Zeitraum von fast 1300 Jahren zurückverfolgt werden konnte. Selbst in diesem "kleinen" Bereich zeigt sich ausschnitthaft die Entwicklung der im Früh- und Hochmittelalter bedeutenden Stadt Würzburg. Zuzutreffen scheint auch eine Bemerkung aus dem 11. Jh., nämlich,'dass es den Würzburgern zur Natur geworden sein, abzureißen und zu bauen, Quadratisches durch Rundes zu ersetzen'. Und das von der Merowinger- bis in die Neuzeit.From January to July 1999 the first big excavation in Würzburg was made at Domerschulstrasse No. 17. The excavation area is located very close to the cathedral in the oldest part of the medieval town. During the works numerous housing structures were documented and a lot of finds were saved. Most of the finds are pieces of pottery but also glas, metal artefacts and bone items. The prior intention of the excavation was the review of the building history at this location. There were a few questions concerning the development of Würzburg since the 8th and 9th centuries and its medieval town. One of the main aims was the review of the history and chronology of the defensive fortifications in medieval times. The work on the finds was expected to give important results concerning the social structures, the economy, the trade, and the crafts at the time. After the conclusion of the interpretation and the analysis medieval archaeology has shown some new and advanced results on some points of the history of Würzburg, that had mostly been written by historians up until now. One of the most spectacular findings were the remains of two walls which had been unknown until then. Both of them are parts of old defensive fortifications. The first is to be dated before the 9th century. The second one was built in the middle of the 9th century. This wall and the moat protected and surrounded the clerical area around the cathedral until the end of the 10th century. These two fortifications complete the sequence of fortifications from early medieval times until modern times. Another finding at the site is a romanesque building, made of stone, from the middle of the 12th century with a huge cellar and some annexes. The history of the building with some reconstructions and attachments in the 13th and 14th century was traced till modern times, too. In conjunction with historical documents this gave important results concerning the use of the houses and the history of their residents during the different times. The work on the finds gave fundamental results concerning the development of objects that were used. Especially the work on the pottery generated a chronology of styles, forms, manufacturing, and trading from the 8th century until modern times. There are finds from the 9th and 10th centuries that are very significant and show the great importance of Würzburg at this time. They also indicate contact and trade relations to northern Germany and Scandinavia. After the interpretation history at the excavated site with fortifications and buildings can be traced back for nearly 1300 years. Even this small archaeological excavation gave new and important insights into the history of Würzburg

Evaluation of antibiotic resistance dissemination by wastewater treatment plant effluents with different catchment areas in Germany

The study quantified the abundances of antibiotic resistance genes (ARGs) and facultative pathogenic bacteria (FPB) as well as one mobile genetic element in genomic DNA via qPCR from 23 different wastewater treatment plant (WWTP) effluents in Germany. 12 clinically relevant ARGs were categorized into frequently, intermediately, and rarely occurring genetic parameters of communal wastewaters. Taxonomic PCR quantifications of five FPB targeting Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, and enterococci were performed. The WWTPs differed in their catchment areas being impacted by hospitals, food processing companies, or housing areas only. The total discharges of the analyzed ARGs and FPB were found to cluster independently of the sizes of the WWTPs with a maximum difference of two log units within one cluster. Initially, quantitative data evaluations revealed no significant difference between ARG categories and WWTP catchment areas. More distinct correlations became obvious with a Pearson correlation approach, where each single taxonomic marker is compared to each ARG target. Here, increased correlation of FPB (i.e. E. coli, K. pneumoniae, P. aeruginosa, and enterococci) with clinically relevant ARGs of the category of rarely occurring resistance genes (blaNDM-1, vanA) was found in WWTP effluents being influenced by hospital wastewaters

Engrailed 1 deficiency induces changes in ciliogenesis during human neuronal differentiation

A key pathological feature of Parkinson's Disease (PD) is the progressive degeneration of dopaminergic neurons (DAns) in the substantia nigra pars compacta. Considering the major role of EN1 in the development and maintenance of these DAns and the implications from En1 mouse models, it is highly interesting to study the molecular and protective effect of EN1 also in a human cellular model. Therefore, we generated EN1 knock-out (ko) human induced pluripotent stem cell (hiPSCs) lines and analyzed these during neuronal differentiation. Although the EN1 ko didn't interfere with neuronal differentiation and generation of tyrosine hydroxylase positive (TH+) neurons per se, the neurons exhibited shorter neurites. Furthermore, mitochondrial respiration, as well as mitochondrial complex I abundance was significantly reduced in fully differentiated neurons. To understand the implications of an EN1 ko during differentiation, we performed a transcriptome analysis of human neuronal precursor cells (hNPCs) which unveiled alterations in cilia-associated pathways. Further analysis of ciliary morphology revealed an elongation of primary cilia in EN1-deficient hNPCs. Besides, also Wnt signaling pathways were severely affected. Upon stimulating hNPCs with Wnt which drastically increased EN1 expression in WT lines, the phenotypes concerning mitochondrial function and cilia were exacerbated in EN1 ko hNPCs. They failed to enhance the expression of the complex I subunits NDUFS1 and 3, and now displayed a reduced mitochondrial respiration. Furthermore, Wnt stimulation decreased ciliogenesis in EN1 ko hNPCs but increased ciliary length even further. This further highlights the relevance of primary cilia next to mitochondria for the functionality and correct maintenance of human DAns and provides new possibilities to establish neuroprotective therapies for PD

Antibiotic-resistant bacteria, antibiotic resistance genes, and antibiotic residues in wastewater from a poultry slaughterhouse after conventional and advanced treatments

Slaughterhouse wastewater is considered a reservoir for antibiotic-resistant bacteria and antibiotic residues, which are not sufficiently removed by conventional treatment processes. This study focuses on the occurrence of ESKAPE bacteria (Enterococcus spp., S. aureus, K. pneumoniae, A. baumannii, P. aeruginosa, Enterobacter spp.), ESBL (extended-spectrum β-lactamase)-producing E. coli, antibiotic resistance genes (ARGs) and antibiotic residues in wastewater from a poultry slaughterhouse. The efficacy of conventional and advanced treatments (i.e., ozonation) of the in-house wastewater treatment plant regarding their removal was also evaluated. Target culturable bacteria were detected only in the influent and effluent after conventional treatment. High abundances of genes (e.g., bla$_{TEM}$, bla$_{CTX-M-15}$, bla$_{CTX-M-32}$, bla$_{OXA-48}$, bla$_{CMY}$ and mcr-1) of up to 1.48 × 10$^{6}$ copies/100 mL were detected in raw influent. All of them were already significantly reduced by 1–4.2 log units after conventional treatment. Following ozonation, mcr-1 and bla$_{CTX-M-32}$ were further reduced below the limit of detection. Antibiotic residues were detected in 55.6% (n = 10/18) of the wastewater samples. Despite the significant reduction through conventional and advanced treatments, effluents still exhibited high concentrations of some ARGs (e.g., sul1, ermB and bla$_{OXA-48}$), ranging from 1.75 × 10$^{2}$ to 3.44 × 10$^{3}$ copies/100 mL. Thus, a combination of oxidative, adsorptive and membrane-based technologies should be considered

In Vivo Ligands of MDA5 and RIG-I in Measles Virus-Infected Cells

RIG-I-like receptors (RLRs: RIG-I, MDA5 and LGP2) play a major role in the innate immune response against viral infections and detect patterns on viral RNA molecules that are typically absent from host RNA. Upon RNA binding, RLRs trigger a complex downstream signaling cascade resulting in the expression of type I interferons and proinflammatory cytokines. In the past decade extensive efforts were made to elucidate the nature of putative RLR ligands. In vitro and transfection studies identified 5'-triphosphate containing blunt-ended double-strand RNAs as potent RIG-I inducers and these findings were confirmed by next-generation sequencing of RIG-I associated RNAs from virus-infected cells. The nature of RNA ligands of MDA5 is less clear. Several studies suggest that double-stranded RNAs are the preferred agonists for the protein. However, the exact nature of physiological MDA5 ligands from virus-infected cells needs to be elucidated. In this work, we combine a crosslinking technique with next-generation sequencing in order to shed light on MDA5-associated RNAs from human cells infected with measles virus. Our findings suggest that RIG-I and MDA5 associate with AU-rich RNA species originating from the mRNA of the measles virus L gene. Corresponding sequences are poorer activators of ATP-hydrolysis by MDA5 in vitro, suggesting that they result in more stable MDA5 filaments. These data provide a possible model of how AU-rich sequences could activate type I interferon signaling

Effect of glyphosate, its metabolite AMPA, and the glyphosate formulation Roundup® on brown trout (Salmo trutta f. fario) gut microbiome diversity

Glyphosate is used worldwide as a compound of pesticides and is detectable in many environmental compartments. It enters water bodies primarily through drift from agricultural areas so that aquatic organisms are exposed to this chemical, especially after rain events. Glyphosate is advertised and sold as a highly specific herbicide, which interacts with the EPSP synthase, an enzyme of the shikimate metabolism, resulting in inhibition of the synthesis of vital aromatic amino acids. However, not only plants but also bacteria can possess this enzyme so that influences of glyphosate on the microbiomes of exposed organisms cannot be excluded. Those influences may result in subtle and long-term effects, e.g., disturbance of the symbiotic interactions of bionts with microorganisms of their microbiomes. Mechanisms how the transformation product aminomethylphosphonic acid (AMPA) of glyphosate might interfere in this context have not understood so far. In the present study, molecular biological fingerprinting methods showed concentration-dependent effects of glyphosate and AMPA on fish microbiomes. In addition, age-dependent differences in the composition of the microbiomes regarding abundance and diversity were detected. Furthermore, the effect of exposure to glyphosate and AMPA was investigated for several fish pathogens of gut microbiomes in terms of their gene expression of virulence factors associated with pathogenicity. In vitro transcriptome analysis with the fish pathogen Yersinia ruckeri revealed that it is questionable whether the observed effect on the microbiome is caused by the intended mode of action of glyphosate, such as the inhibition of EPSP synthase activity

Reduction of Antibiotic Resistant Bacteria During Conventional and Advanced Wastewater Treatment, and the Disseminated Loads Released to the Environment

The occurrence of new chemical and microbiological contaminants in the aquatic environment has become an issue of increasing environmental concern. Thus, wastewater treatment plants (WWTPs) play an important part in the distribution of so-called new emerging pathogens and antibiotic resistances. Therefore, the daily loads released by the WWTP were calculated including a model system for the distribution of these loads within the receiving water body. UV-, as well as ozone-treatment in separate or in combination for wastewater treatment were under investigation aiming at the reduction of these loads. Here, the impact of these treatments on the DNA integrity via antibody staining and PCR efficiencies experiments were included. All three facultative pathogenic bacteria [enterococci (23S rRNA), Pseudomonas aeruginosa (ecfX), and Escherichia coli (yccT)] and seven clinically relevant antibiotic resistance genes (ARGs) (mecA (methicillin resistance gene), ctx-M32 (β- lactame resistance gene), ermB (erythromycine resistance gene), blaTEM (β- lactame resistance gene), sul1 (sulfonamide resistance gene), vanA (vancomycin resistance gene), and intI1 (Integrase1 gene) associated with mobile genetic elements were detected in wastewaters. Different reduction efficiencies were analyzed during advanced wastewater treatments. ARGs were still found to be present in the effluents under the parameters of 1.0 g ozone per g dissolved organic carbon (DOC) and 400 J/m2, like ctx-M32, ermB, blaTEM, sul1, and intI1. Especially UV radiation induced thymidine dimerization which was analyzed via antibody mediated detection in the metagenome of the natural wastewater population. These specific DNA alterations were not observed during ozone treatment and combinations of UV/ozone treatment. The dimerization or potential other DNA alterations during UV treatment might be responsible for a decreased PCR efficiency of the 16S rRNA amplicons (176, 490, and 880 bp fragments) from natural metagenomes compared to the untreated sample. This impact on PCR efficiencies was also observed for the combination of ozone and UV treatment