@misc{FroemmelLehmannRoedigeretal., author = {Fr{\"o}mmel, Ulrike and Lehmann, Werner and R{\"o}diger, Stefan and B{\"o}hm, Alexander and Nitschke, J{\"o}rg and Weinreich, J{\"o}rg and Groß, Julia and Roggenbuck, Dirk and Zinke, Olaf and Ansorge, Hermann and Vogel, Steffen and Klemm, Per and Wex, Thomas and Schr{\"o}der, Christian and Wieler, Lothar H. and Schierack, Peter}, title = {Adhesion of human and animal Escherichia coli strains in association with their virulence-associated genes and phylogenetic origins}, series = {Applied and environmental microbiology}, volume = {79}, journal = {Applied and environmental microbiology}, number = {19}, issn = {1098-5336}, doi = {10.1128/AEM.01384-13}, pages = {5814 -- 5829}, abstract = {Intestinal colonization is influenced by the ability of the bacterium to inhabit a niche, which is based on the expression of colonization factors. Escherichia coli carries a broad range of virulence-associated genes (VAGs) which contribute to intestinal (inVAGs) and extraintestinal (exVAGs) infection. Moreover, initial evidence indicates that inVAGs and exVAGs support intestinal colonization. We developed new screening tools to genotypically and phenotypically characterize E. coli isolates originating in humans, domestic pigs, and 17 wild mammal and avian species. We analyzed 317 isolates for the occurrence of 44 VAGs using a novel multiplex PCR microbead assay (MPMA) and for adhesion to four epithelial cell lines using a new adhesion assay. We correlated data for the definition of new adhesion genes. inVAGs were identified only sporadically, particularly in roe deer (Capreolus capreolus) and the European hedgehog ( Erinaceus europaeus). The prevalence of exVAGs depended on isolation from a specific host. Human uropathogenic E. coli isolates carried exVAGs with the highest prevalence, followed by badger (Meles meles) and roe deer isolates. Adhesion was found to be very diverse. Adhesion was specific to cells, host, and tissue, though it was also unspecific. Occurrence of the following VAGs was associated with a higher rate of adhesion to one or more cell lines: afa-dra, daaD, tsh, vat, ibeA, fyuA, mat, sfa-foc, malX, pic, irp2, and papC. In summary, we established new screening methods which enabled us to characterize large numbers of E. coli isolates. We defined reservoirs for potential pathogenic E. coli. We also identified a very broad range of colonization strategies and defined potential new adhesion genes.}, language = {en} } @misc{SchierackRoedigerKuhletal., author = {Schierack, Peter and R{\"o}diger, Stefan and Kuhl, Christoph and Hiemann, Rico and Roggenbuck, Dirk and Li, Ganwu and Weinreich, J{\"o}rg and Berger, Enrico and Nolan, Lisa K. and Nicholson, Bryon and R{\"o}mer, Antje and Fr{\"o}mmel, Ulrike and Wieler, Lothar H. and Schr{\"o}der, Christian}, title = {Porcine E. coli: Virulence-Associated Genes, Resistance Genes and Adhesion and Probiotic Activity Tested by a New Screening Method}, series = {PLoS One}, volume = {8}, journal = {PLoS One}, number = {4}, doi = {10.1371/journal.pone.0059242}, pages = {13}, abstract = {We established an automated screening method to characterize adhesion of Escherichia coli to intestinal porcine epithelial cells (IPEC-J2) and their probiotic activity against infection by enteropathogenic E. coli (EPEC). 104 intestinal E. coli isolates from domestic pigs were tested by PCR for the occurrence of virulence-associated genes, genes coding for resistances to antimicrobial agents and metals, and for phylogenetic origin by PCR. Adhesion rates and probiotic activity were examined for correlation with the presence of these genes. Finally, data were compared with those from 93 E. coli isolates from wild boars. Isolates from domestic pigs carried a broad variety of all tested genes and showed great diversity in gene patterns. Adhesions varied with a maximum of 18.3 or 24.2 mean bacteria adherence per epithelial cell after 2 or 6 hours respectively. Most isolates from domestic pigs and wild boars showed low adherence, with no correlation between adhesion/probiotic activity and E. coli genes or gene clusters. The gene sfa/foc, encoding for a subunit of F1C fimbriae did show a positive correlative association with adherence and probiotic activity; however E. coli isolates from wild boars with the sfa/foc gene showed less adhesion and probiotic activity than E. coli with the sfa/foc gene isolated from domestic pigs after 6 hour incubation. In conclusion, screening porcine E. coli for virulence associated genes genes, adhesion to intestinal epithelial cells, and probiotic activity revealed a single important adhesion factor, several probiotic candidates, and showed important differences between E. coli of domestic pigs and wild boars.}, language = {en} } @misc{RoedigerLehmannFroemmeletal., author = {R{\"o}diger, Stefan and Lehmann, Werner and Fr{\"o}mmel, Ulrike and B{\"o}hm, Alexander and Nitschke, J{\"o}rg and Burdukiewicz, Michał and Schr{\"o}der, Christian and Dangla, R{\´e}mi and Droniou, Magali and Schierack, Peter}, title = {The Modi Operandi of the VideoScan Platform for the Detection and Analysis of Nucleic Acids. Life Science Day 2013 \& 17. Leibniz Conference of Advanced Science, Freie Universit{\"a}t Berlin, 24.10.2013}, pages = {1}, language = {en} } @incollection{RoedigerSchierackBoehmetal., author = {R{\"o}diger, Stefan and Schierack, Peter and B{\"o}hm, Alexander and Nitschke, J{\"o}rg and Berger, Ingo and Fr{\"o}mmel, Ulrike and Schmidt, Carsten and Ruhland, Mirko and Schimke, Ingolf and Roggenbuck, Dirk and Lehmann, Werner and Schr{\"o}der, Christian}, title = {A highly versatile microscope imaging technology platform for the multiplex real-time detection of biomolecules and autoimmune antibodies}, series = {Molecular diagnostics}, booktitle = {Molecular diagnostics}, editor = {Seitz, Harald and Berger, Ingo}, publisher = {Springer}, address = {Berlin [u.a.]}, isbn = {978-3-642-37690-0}, pages = {35 -- 74}, language = {en} } @misc{FroemmelBoehmNitschkeetal., author = {Fr{\"o}mmel, Ulrike and B{\"o}hm, Alexander and Nitschke, J{\"o}rg and Weinreich, J{\"o}rg and Groß, Julia and R{\"o}diger, Stefan and Wex, Thomas and Ansorge, Hermann and Zinke, Olaf and Schr{\"o}der, Christian and Roggenbuck, Dirk and Schierack, Peter}, title = {Adhesion patterns of commensal and pathogenic Escherichia coli from humans and wild animals on human and porcine epithelial cell lines}, series = {Gut Pathogens}, volume = {5}, journal = {Gut Pathogens}, number = {31}, issn = {1757-4749}, doi = {10.1186/1757-4749-5-31}, pages = {8}, abstract = {Abstract BACKGROUND: Different strategies of colonization or infection by E. coli result in formation of certain adhesion patterns which help also in classifying intestinal E. coli into pathotypes. Little is known about adhesion patterns and host- and tissue adaption of commensal E. coli and about E. coli originating in clinically healthy hosts carrying pathotype-specific virulence-associated genes. FINDINGS: Adhesion pattern of E. coli (n = 282) from humans and from 18 animal species were verified on intestinal human Caco-2 and porcine IPEC-J2 cells and, furthermore, for comparison on human urinary bladder 5637, porcine kidney PK-15 epithelial and HEp-2 cells. The analysis was carried out on 150,000 images of adhesion assays.Adhesion patterns were very diverse; 88 isolates were completely non-adherent, whereas 194 adhered to at least one cell line with the dominant adhesion patterns "diffusely distributed" and "microcolony formation". Adhesion patterns "chains" and "clumps" were also visible. Chain formation was mediated by the presence of epithelial cells. Clump formation was very specific on only the 5637 cell line. All enteropathogenic (eae+) E. coli (EPEC; n = 14) were able to form microcolonies which was cell line specific for each isolate. Most EPEC formed microcolonies on intestinal IPEC-J2 and Caco-2 but several also on urinary tract cells. Shigatoxin-producing (stx+) E. coli (n = 10) showed no specific adhesion patterns. CONCLUSIONS: E. coli isolates were highly diverse. Commensal and pathogenic isolates can adhere in various forms, including diffuse distribution, microcolonies, chains and clumps. Microcolony formation seems to be a global adhesion strategy also for commensal E. coli.}, language = {en} } @misc{RoedigerKramerFroemmeletal., author = {R{\"o}diger, Stefan and Kramer, Toni and Fr{\"o}mmel, Ulrike and Weinreich, J{\"o}rg and Roggenbuck, Dirk and Guenther, Sebastian and Schaufler, Katharina and Schr{\"o}der, Christian and Schierack, Peter}, title = {Intestinal Escherichia coli colonization in a mallard duck population over four consecutive winter season}, series = {Environmental Microbiology}, volume = {17}, journal = {Environmental Microbiology}, number = {9}, issn = {1462-2920}, doi = {10.1111/1462-2920.12807}, pages = {3352 -- 3361}, language = {en} }