Besnoitia besnoiti infection alters both endogenous cholesterol de novo synthesis and exogenous LDL uptake in host endothelial cells

Besnoitia besnoiti, an apicomplexan parasite of cattle being considered as emergent in Europe, replicates fast in host endothelial cells during acute infection and is in considerable need for energy, lipids and other building blocks for offspring formation. Apicomplexa are generally considered as defective in cholesterol synthesis and have to scavenge cholesterol from their host cells for successful replication. Therefore, we here analysed the influence of B. besnoiti on host cellular endogenous cholesterol synthesis and on sterol uptake from exogenous sources. GC-MS-based profiling of cholesterol-related sterols revealed enhanced cholesterol synthesis rates in B. besnoiti-infected cells. Accordingly, lovastatin and zaragozic acid treatments diminished tachyzoite production. Moreover, increased lipid droplet contents and enhanced cholesterol esterification was detected and inhibition of the latter significantly blocked parasite proliferation. Furthermore, artificial increase of host cellular lipid droplet disposability boosted parasite proliferation. Interestingly, lectin-like oxidized low density lipoprotein receptor 1 expression was upregulated in infected endothelial hostcells, whilst low density lipoproteins (LDL) receptor was not affected by parasite infection. However, exogenous supplementations with non-modified and acetylated LDL both boosted B. besnoiti proliferation. Overall, current data show that B. besnoiti simultaneously exploits both, endogenous cholesterol biosynthesis and cholesterol uptake from exogenous sources, during asexual replication

Occurrence of mcr-1 and mcr-2 colistin resistance genes in porcine Escherichia coli isolates (2010–2020) and genomic characterization of mcr-2-positive E. coli

Introduction: The global emergence of plasmid-mediated colistin resistance is threatening the efficacy of colistin as one of the last treatment options against multi-drug resistant Gram-negative bacteria. To date, ten mcr-genes (mcr-1 to mcr-10) were reported. While mcr-1 has disseminated globally, the occurrence of mcr-2 was reported scarcely. Methods and results: We determined the occurrence of mcr-1 and mcr-2 genes among Escherichia coli isolates from swine and performed detailed genomic characterization of mcr-2-positive strains. In the years 2010-2017, 7,614 porcine E. coli isolates were obtained from fecal swine samples in Europe and isolates carrying at least one of the virulence associated genes predicting Shiga toxin producing E. coli (STEC), enterotoxigenic E. coli (ETEC) or enteropathogenic E. coli (EPEC) were stored. 793 (10.4%) of these isolates carried the mcr-1 gene. Of 1,477 additional E. coli isolates obtained from sheep blood agar containing 4 mg/L colistin between 2018 and 2020, 36 (2.4%) isolates were mcr-1-positive. In contrast to mcr-1, the mcr-2 gene occurred at a very low frequency (0.13%) among the overall 9,091 isolates. Most mcr-2-positive isolates originated from Belgium (n = 9), one from Spain and two from Germany. They were obtained from six different farms and revealed multilocus sequence types ST10, ST29, ST93, ST100, ST3057 and ST5786. While the originally described mcr-2.1 was predominant, we also detected a new mcr-2 variant in two isolates from Belgium, which was termed mcr-2.8. MCR-2 isolates were mostly classified as ETEC or ETEC-like, while one isolate from Spain represented an atypical enteropathogenic E. coli (aEPEC; eae+). The ST29-aEPEC isolate carried mcr-2 on the chromosome. Another eight isolates carried their mcr-2 gene on IncX4 plasmids that resembled the pKP37-BE MCR-2 plasmid originally described in Belgium in 2015. Three ST100 E. coli isolates from a single farm in Belgium carried the mcr-2.1 gene on a 47-kb self-transmissible IncP type plasmid of a new IncP-1 clade. Discussion: This is the first report of mcr-2 genes in E. coli isolates from Germany. The detection of a new mcr-2 allele and a novel plasmid backbone suggests the presence of so far undetected mcr-2 variants and mobilizable vehicles.Peer Reviewe

The GadX regulon affects virulence gene expression and adhesion of porcine enteropathogenic Escherichia coli in vitro

The ability of enteropathogenic Escherichia coli (EPEC) to express virulence factor genes and develop attaching and effacing (AE) lesions is inhibited in acidic environmental conditions. This inhibition is due to the activation of transcription factor GadX, which upregulates expression of glutamic acid decarboxylase (Gad). Gad, in turn, produces γ-aminobutyric acid (GABA), which was recently shown to have a beneficial effect on the jejunal epithelium in vitro due to increased mucin-1 levels. In the present study, we sought to test whether forced GadX activation/overexpression abolishes virulence associated features of EPEC and provokes increased GABA production. EPEC strains were isolated from diarrheic pigs and submitted to activation of GadX by acidification as well as gadX overexpression via an inducible expression vector plasmid. GABA concentrations in the growth medium, ability for adhesion to porcine intestinal epithelial cells (IPEC-J2) and virulence gene expression were determined. Growth in acidified media led to increased GABA levels, upregulated gadA/B expression and downregulated mRNA synthesis of the bacterial adhesin intimin. EPEC strains transformed with the gadX gene produced 2.1 to 3.4-fold higher GABA levels than empty-vector controls and completely lost their ability to adhere to IPEC-J2 cells and to induce actin accumulation. We conclude that intensified gadX activation can abolish the ability of EPEC to adhere to the intestinal epithelium by reducing the expression of major virulence genes

Cross-sectional study: prevalence of oedema disease Escherichia coli (EDEC) in weaned piglets in Germany at pen and farm levels

Background Escherichia coli bacteria capable of producing the toxin Stx2e and possessing F18-fimbriae (edema disease E. coli, EDEC) are considered causative agents of porcine oedema disease. This disease, which usually occurs in piglets shortly after weaning, has a high lethality in affected animals and can lead to high economic losses in piglet rearing. The aim of this cross-sectional field study was to determine the prevalence of EDEC in weaned piglets in Germany at pen and farm levels. Results Ninety-nine farms with unknown history of infections with shigatoxin-producing E. coli (STEC) and oedema disease were sampled. On each farm, up to five pens were selected for sampling (n = 481). The piglets in these pens were at an age 1–3 weeks after weaning. Single faecal samples (n = 2405) and boot swabs (n = 479) were collected from the floor. On 50 farms, cotton ropes were additionally used to collect oral fluid samples (n = 185) and rope wash out samples (n = 231) from the selected pens. All samples were analyzed by bacterial culture combined with a duplex PCR for the presence of the corresponding genes stx2e and fedA (major subunit protein of F18 fimbriae). In addition, whole DNA specimens extracted from boot swabs, oral fluid samples, and rope wash out samples were directly examined by duplex PCR for DNA of stx2e and fedA. A pen was classified as positive if at least one of the samples, regardless of the technique, yielded a positive result in the PCR, and farms were considered positive if at least one pen was classified as positive. Overall, genes stx2e and fedA were found simultaneously in 24.9% (95% CI 22.1–29.1%) of sampled pens and in 37.4% (95% CI 27.9–47.7%) of sampled farms. Regardless of the presence of F18-fimbriae, Escherichia coli encoding for Stx2e (STEC-2e) were found in 35.1% (95% CI 31.0–39.1%) of the pens and 53.5% (95% CI 44.4–63.6%) of the farms sampled. Conclusions Escherichia coli strains considered capable to cause oedema disease in swine (EDEC) are highly prevalent in the surveyed pig producing farms in Germany. Due to intermittent shedding of EDEC and a potentially low within-farm prevalence, we recommend a combination of different sampling techniques for EDEC monitoring at pen and farm levels. Further studies are needed to understand which STEC-2e strains really pose the risk of causing severe porcine disease

„Mit anderen Augen“ – Kann ein Film Einstellungen gegenüber Geflüchteten verändern?

Dieser Beitrag beschreibt Ergebnisse einer Studie des Instituts für Demokratie und Zivilgesellschaft im Rahmen des Filmprojekts „Mit anderen Augen“. Im Filmprojekt drehten aus Syrien geflüchtete Frauen gemeinsam mit deutschen Frauen einen Film über ihre ersten Begegnungen. Dieser Film wurde anschließend in der Region vorgestellt und in seiner Wirkung auf das Publikum mittels Fragebogenerhebung untersucht. Die Ergebnisse zeigen: Partizipative (Film-)Projekte können demnach in ländlichen Kontexten, wo nur wenige Kontaktmöglichkeiten zwischen Einheimischen und Geflüchteten bestehen, eine gute Möglichkeit bieten, Integration zu fördern

Interaction of Coxiella burnetii Strains of Different Sources and Genotypes with Bovine and Human Monocyte-Derived Macrophages

Most human Q fever infections originate from small ruminants. By contrast, highly prevalent shedding of Coxiella (C.) burnetii by bovine milk rarely results in human disease. We hypothesized that primary bovine and human monocyte-derived macrophages (MDM) represent a suitable in vitro model for the identification of strain-specific virulence properties at the cellular level. Twelve different C. burnetii strains were selected to represent different host species and multiple loci variable number of tandem repeat analysis (MLVA) genotypes. Infection efficiency and replication of C. burnetii were monitored by cell culture re-titration and qPCR. Expression of immunoregulatory factors after MDM infection was measured by qRT-PCR and flow cytometry. Invasion, replication and MDM response differed between C. burnetii strains but not between MDMs of the two hosts. Strains isolated from ruminants were less well internalized than isolates from humans and rodents. Internalization of MLVA group I strains was lower compared to other genogroups. Replication efficacy of C. burnetii in MDM ranged from low (MLVA group III) to high (MLVA group IV). Infected human and bovine MDM responded with a principal up-regulation of pro-inflammatory cytokines such as IL-1β, IL-12, and TNF-α. However, MLVA group IV strains induced a pronounced host response whereas infection with group I strains resulted in a milder response. C. burnetii infection marginally affected polarization of MDM. Only one C. burnetii strain of MLVA group IV caused a substantial up-regulation of activation markers (CD40, CD80) on the surface of bovine and human MDM. The study showed that replication of C. burnetii in MDM and the subsequent host cell response is genotype-specific rather than being determined by the host species pointing to a clear distinction in C. burnetii virulence between the genetic groups

Recidivism of young prisoners after release from youth prison. An empirical study of the youth imprisonment system of the State of Hesse, years of release 2003 and 2006. Main Report

Zwischen 2006 und 2011 führte ein Forscher/innen-Team der Universitäten Marburg und Tübingen im hessischen Jugendstrafvollzug eine Studie zum Rückfall und zur Akzeptanz sowie der Auswirkung von pädagogischen und sozialtherapeutischen Maßnahmen bei jungen männlichen Gefangenen durch. Dabei wurden die Entlassungsjahrgänge 2003 und 2006 jeweils vollständig und nach dreijähriger Verzögerung mittels Auszügen des Bundeszentralregisters untersucht (quantitative Teilstudie Tübingen). Zudem wurden Befragungen mit Gefangenen durchgeführt, am Anfang, zu Ende und nach der Haft, unter anderem zur Persönlichkeitsentwicklung, sozialen Kompetenz, Struktur nach der Haft, Einschätzung von Maßnahmen, auch durch die Vollzugsbediensteten. Zusätzlich wurden Bewährungshelfer und Bewährungshelferinnen zur Fremdeinschätzung interviewt (qualitative Teilstudie Marburg). In diesem Band werden, im Nachgang zu früheren, und teilweise allgemein zugänglichen, Berichten an das Hessische Ministerium der Justiz, die Befunde der quantitativen Teilstudie in aktualisierter Form vorgestellt, unter Berücksichtigung neuerer Literatur sowie in geändertem Layout. Ergänzende Texte, Tabellen und Schaubilder finden sich in dem Begleitband, d. h. Band 41 dieser Schriftenreihe.From 2006 to 2011 two co-ordinated research teams from the University of Tuebingen and the University of Marburg were intensely studying the Youth Correctional System of the German Federal State of Hesse. The research was concentrated on young male inmates from roughly 14 to 25 years of age, and dealt particularly with the acceptance of educational and treatment measures, their consequences after youth prison release, and the extent and structure of the former inmates´ recidivism, in terms of new convictions resp. penal sentences. The qualitative part of the project (Marburg team) administered semi-structured interviews at the beginning, near the end, and after the release of the young offenders. The researchers asked e. g. for the individual evaluation of the experienced measures, for personality development, social competency, everyday life structure after release in society, and for experiences with the parole service. In addition, prison and probation officers were asked for their impressions. The quantitative part of the project (Tuebingen team) was dealing with two full cohorts of releases, one for the year 2003, the other for the year 2006. Based on administrative documents of the Youth Prisons, and on records of the national criminal registry, the official criminal history of all ex-inmates until their release, and during their individualized terms of exactly three years post-release, was scrutinized and analysed under structural and longitudinal perspectives. Former reports had been delivered to the Hessian Ministry of Justice, and were made afterwards available to interested persons and institutions. With this volume, the Tuebingen team is presenting a thoroughly controlled new edition of the quantitative study results, with some revisions of resp. amendments to the text, and with additions of hitherto published relevant literature. In the companion volume (“Materialienband”= No. 41) to this TüKrim Series additional explanations, tables and figures are being presented

Recidivism of young prisoners after release from youth prison. An empirical study of the youth imprisonment system of the State of Hesse, years of release 2003 and 2006. Supplementary documents (texts, tables, figures)

Dieser Band ist als Ergänzungsband zum Hauptband des Berichts über die Tübinger Teilstudie zum Jugendstrafvollzug Hessen, Entlassungsjahrgänge 2003 und 2006 (TüKrim Band 40) konzipiert. Er enthält, unter anderem, ergänzende Tabellen zu dem im Forschungsbericht verwendeten Schaubildern bzw. Tabellen sowie weitere Berechnungen, exemplarisch einen anonymisierten Bundeszentralregisterauszug, Erläuterungen zu besonderen methodischen oder inhaltlichen Fragen der Auswertung, Details zu den bei der Forschung eingesetzten Codierschemata, und schließlich Grundauszählungen.This volume is conceived as a companion to the Main Report (TüKrim No. 40), regarding the Tübingen part of a coordinated Marburg-Tübingen research project on young inmates in the German State of Hesse youth correctional institutions in the years 2003 and 2006; this part was dealing with two full cohorts of inmates having been released from youth imprisonment, and followed up for an individualised period of exactly three years after regaining liberty. The volume offers additional details of the study, e.g. tables, figures, methodological considerations on excerpts from the national criminal registry, code schemes, and raw results on the study variables

Molecular Features and Antimicrobial Susceptibilities of <i>Streptococcus equi</i> ssp. <i>equi</i> Isolates from Strangles Cases in Indonesia

Strangles, caused by Streptococcus equi ssp. equi (S. equi equi), is a highly infectious and frequent disease of equines worldwide. No data are available regarding the molecular epidemiology of strangles in Indonesia. This study aimed to characterize S. equi equi isolates obtained from suspected strangles cases in Indonesia in 2018. Isolates originated from seven diseased horses on four different farms located in three provinces of Indonesia. Whole genome sequences of these isolates were determined and used for seM typing, multilocus sequence typing (MLST), and core genome MLS typing (cgMLST). Genomes were also screened for known antimicrobial resistance genes and genes encoding for the recombinant antigens used in the commercial Strangvac® subunit vaccine. All seven S. equi equi isolates from Indonesia belonged to ST179 and carried seM allele 166. Isolates differed from each other by only 2 to 14 cgSNPs and built an exclusive sub-cluster within the Bayesian Analysis of Population Structure (BAPS) cluster 2 (BAPS-2) of the S. equi equi cgMLST scheme. All isolates revealed predicted amino acid sequence identity to seven and high similarity to one of the eight antigen fragments contained in Strangvac®. Furthermore, all isolates were susceptible to beta-lactam antibiotics penicillin G, ampicillin, and ceftiofur. Our data suggest that the horses from this study were affected by strains of the same novel sublineage within globally distributed BAPS-2 of S. equi equi. Nevertheless, penicillin G can be used as a first-choice antibiotic against these strains and Strangvac® may also be protective against Indonesian strains

Cross-sectional study: prevalence of oedema disease Escherichia coli (EDEC) in weaned piglets in Germany at pen and farm levels

Abstract Background Escherichia coli bacteria capable of producing the toxin Stx2e and possessing F18-fimbriae (edema disease E. coli, EDEC) are considered causative agents of porcine oedema disease. This disease, which usually occurs in piglets shortly after weaning, has a high lethality in affected animals and can lead to high economic losses in piglet rearing. The aim of this cross-sectional field study was to determine the prevalence of EDEC in weaned piglets in Germany at pen and farm levels. Results Ninety-nine farms with unknown history of infections with shigatoxin-producing E. coli (STEC) and oedema disease were sampled. On each farm, up to five pens were selected for sampling (n = 481). The piglets in these pens were at an age 1–3 weeks after weaning. Single faecal samples (n = 2405) and boot swabs (n = 479) were collected from the floor. On 50 farms, cotton ropes were additionally used to collect oral fluid samples (n = 185) and rope wash out samples (n = 231) from the selected pens. All samples were analyzed by bacterial culture combined with a duplex PCR for the presence of the corresponding genes stx2e and fedA (major subunit protein of F18 fimbriae). In addition, whole DNA specimens extracted from boot swabs, oral fluid samples, and rope wash out samples were directly examined by duplex PCR for DNA of stx2e and fedA. A pen was classified as positive if at least one of the samples, regardless of the technique, yielded a positive result in the PCR, and farms were considered positive if at least one pen was classified as positive. Overall, genes stx2e and fedA were found simultaneously in 24.9% (95% CI 22.1–29.1%) of sampled pens and in 37.4% (95% CI 27.9–47.7%) of sampled farms. Regardless of the presence of F18-fimbriae, Escherichia coli encoding for Stx2e (STEC-2e) were found in 35.1% (95% CI 31.0–39.1%) of the pens and 53.5% (95% CI 44.4–63.6%) of the farms sampled. Conclusions Escherichia coli strains considered capable to cause oedema disease in swine (EDEC) are highly prevalent in the surveyed pig producing farms in Germany. Due to intermittent shedding of EDEC and a potentially low within-farm prevalence, we recommend a combination of different sampling techniques for EDEC monitoring at pen and farm levels. Further studies are needed to understand which STEC-2e strains really pose the risk of causing severe porcine disease