Assessing the occurrence of the novel zoonotic variegated squirrel bornavirus 1 in captive squirrels in Germany —A prevalence study

The newly described zoonotic variegated squirrel bornavirus 1 (VSBV‐1) in German squirrel holdings has been associated with the death of three private owners and one zoo animal caretaker (confirmed cases). Epidemiological investigations were severely impeded by the general lack of data on holdings of the putative reservoir hosts, the family Sciuridae. To fill this lack of data for detailed epidemiological investigations of the captive squirrel population, a register of private and zoological squirrel holdings was established. The findings show a broad variety of kept species and their frequency distribution. By contacting the different stakeholders via Web‐based social groups and societies, information passed in both directions so that disease awareness could be raised and participants could be recruited for further studies. Cross‐sectional studies revealed a prevalence of VSBV‐1‐positive subpopulations of 0% (95% CI 0%–6.2%) among private squirrel collections and 1.9% (95% CI: 0%–9.9%) among zoos in Germany. The approach presented here can be transferred to other populations of non‐traditional pets, which may be equally difficult to monitor, in the case of an emerging zoonotic infectious disease.Peer Reviewe

Variegated squirrel bornavirus 1 in squirrels, Germany and the Netherlands

We screened squirrels in Germany and the Netherlands for the novel zoonotic variegated squirrel bornavirus 1 (VSBV-1). The detection of VSBV-1 in 11 squirrels indicates a considerable risk for transmission to humans handling those animals. Therefore, squirrels in contact with humans should routinely be tested for VSBV-1

The GimA locus of extraintestinal pathogenic E. coli (ExPEC): reductive evolution correlates with habitat and pathovar

Untersuchungen an humanen Gehirnendothelzellen (HBMEC - brain microvascular endothelial cells) sowie an Tierversuchsmodellen mit neugeborenen Ratten und Hühnern konnten zeigen, dass das ibeA-Gen an der Pathogenese von systemischen E. coli-Infektionen beteiligt ist. IbeA ist in einer 20,3 kb großen chromosomalen Region lokalisiert, die von den Genen yjiD und yjiE flankiert wird. Diese Region ist als genetische Insel beschrieben und wird GimA (genomic island of meningitic E. coli containing ibeA) genannt. GimA besteht aus vier Operons (GimA1-4). Die E. coli, die die systemischen Infektionen auslösen, gehören den Pathovaren APEC (aviär pathogene) und NMEC (Neugeborenenmeningits- assoziierte E. coli) aus der Gruppe der extraintestinal-pathogenen E. coli (ExPEC) an. Beispielsweise ist ibeA in die Überwindung der Blut-Hirn-Schranke, einem wesentlichen Schritt in der Pathogenese der Meningitis beim Neugeborenen, involviert. Der putative Insertionslokus von GimA wurde mittels Long-Range-PCR und DNS-DNS-Hybridisierung mit dem Ziel untersucht, die Phylogenie von GimA aufzudecken. Weiterhin sollte der Inselcharakter von GimA eingehender analysiert werden. Die Untersuchungen wurden an insgesamt 410 E. coli-Isolaten durchgeführt. Im Probenumfang waren die Pathovare APEC, NMEC, UPEC (uropathogene) und SepEC (Septikämie-assoziierte E. coli) sowie Isolate aus Stuhlproben bzw. Kotproben von klinisch gesunden Menschen und Tieren enthalten. Darüber hinaus waren die 72 E. coli-Stämme der EcoR-Sammlung (E. coli reference collection) in den 410 Isolaten enthalten. Es konnte neben einer Variante des Insertionslokus, die ein vollständiges GimA (~20.3 kb) (16.1%) (GimA+) und einer, die keinerlei GimA-assoziierte Strukturen (64.4%) (GimA-) enthält, eine dritte Variante des Lokus identifiziert werden, die ein 342 bp großes Restfragment von GimA aufweist (19.5%) (GimA-remnant). Das vollständige GimA und das GimA-remnant waren fast ausschließlich mit der Phylogruppe B2 assoziiert. Zusätzlich trat das GimA-remnant bei Stämmen auf, die zum Multi-Lokus-Sequenztyp ST117 gehören und der rekombinanten Phylogruppe ABD zugeordnet sind. Die weiteren Phylogruppen (A, B1, AxB1, ABD (mit Ausnahme des ST117) und D) waren GimA-. Weiterhin konnten signifikante Assoziationen zwischen dem Vorkommen des vollständigen GimA und den Pathovaren APEC und NMEC gefunden werden. Das GimA-remnant war dagegen signifikant assoziiert mit UPEC sowohl menschlichen als auch tierischen Ursprungs. Ähnlich der Rolle, die ibeA in der Neugeborenenmeningitis-Pathogenese spielt, könnte dies einen möglichen Beitrag dieser Struktur zur Pathogenese von durch UPEC verursachten Harnwegsinfektionen andeuten. Eine Sequenzanalyse des ibeA-Gens ergab eine strikte Assoziation des Gens mit dem phylogenetischen Hintergrund. Dies könnte ein Hinweis darauf sein, dass GimA ein originärer Bestandteil der Phylogruppe B2 ist. Ferner zeigten die Untersuchungen, dass ibeA innerhalb eines definierten phylogenetischen Hintergrundes (ST95-Komplex) keinem positiven Selektionsdruck ausgesetzt ist. Dementsprechend scheint die Evolution von ibeA aufgrund von neutraler Selektion zu erfolgen. Obwohl einige Kriterien für genetische Inseln erfüllt werden, sprechen diese Befunde gegen eine Mobilität von GimA und folglich auch gegen die Annahme, GimA sei eine genetische Insel. GimA scheint eher ein ursprünglicher Teil der Phylogruppe B2 zu sein. Um die Zugehörigkeit von GimA zum „core genome“ zu verdeutlichen sollte diese genetische Region besser als GimA-Lokus angesprochen werden Die Existenz von zwei weiteren Varianten des GimA-Lokus (GimA-remnant und GimA-) deutet auf eine Größenreduktion des Genoms in Form einer reduktiven Evolution hin. Dieser Prozess wird möglicherweise durch Veränderungen der pathogenetischen Bedeutung von einzelnen GimA-Komponenten in verschiedenen Habitaten verursacht. Bei von APEC und NMEC ausgelösten systemischen Infektionen konnten verschiedene Untersuchungen einen Beitrag zur Pathogenese zeigen. Einige GimA-Bestandteile sind für die Translokation der Blut-Hirn-Schranke von Bedeutung (ibeRAT – GimA4), während die übrigen in die Stressantwort des Erregers im Habitat „Blut“ involviert zu sein scheinen (GimA1-3). Der Harnapparat ist im Gegensatz zum Blut ein mit weniger Stressoren für E. coli behaftetes Habitat. Ensprechend könnten Gene, die die Überwindung der Blut-Hirn-Schranke vermitteln, an Bedeutung für E. coli verloren haben und in einem Prozess der Genomoptimierung deletiert worden sein. Lediglich das GimA-remnant blieb erhalten. Dies deutet an, dass diese Struktur bzw. ihre Erhaltung einen Selektionsvorteil bietet. Das GimA-remnant konnte als ORF identifiziert werden und könnte folglich an der Pathogenese von Harnwegsinfektionen beteiligt sein. Weitere Untersuchungen besonders zur Funktionalität des GimA-remnant sind notwendig. Die dritte Variante des GimA-Lokus kann auf zweierlei Wegen entstanden sein: (i) GimA ist in einem einzigen Schritt vollständig verloren gegangen oder (ii) im Anschluss an seine Entstehung das GimA-remnant ebenfalls deletiert worden. Im Falle von UPEC könnte letztere Möglichkeit das Resultat eines fortgesetzten Genomoptimierungsprozesses sein. Die resultierenden Stämme vermeiden eine Immunantwort des Wirtes und können somit als asymptomatische Besiedler im Harnapparat verbleiben.Studies on human brain microvascular endothelial cells (HBMEC) and on in vivo models (newborn rats and chickens) demonstrated that in particular ibeA is involved in the pathogenesis of several extraintestinal pathogenic E. coli (ExPEC) pathovars, including newborn meningitic (NMEC) and avian pathogenic E. coli (APEC). E. g. it is required for translocation of the brain blood barrier, a crucial step in the pathogenesis of newborn meningitis. IbeA is located on a 20.3 kbp chromosomal region flanked by yjiD and yjiE, termed GimA. GimA has also been reported to be a genetic island, consisting of four operons (GimA1-4). To unravel the phylogeny of GimA and to investigate its island character, the putative insertion locus of GimA was determined via Long Range PCR and DNA-DNA hybridization in 410 E. coli isolates, including APEC, NMEC, UPEC (uropathogenic), SepEC (septicemia associated E. coli) and human and animal fecal isolates as well as in 72 strains of the E. coli reference collection (EcoR). In addition to a complete GimA (~20.3 kb) (16.1%) (GimA+) and a locus lacking GimA (64.4%) (GimA-) a third pattern containing a 342 bp remnant of GimA could be found (19.5%) (GimA-remnant). The complete GimA and the GimA-remnant were almost exclusively associated with phylogroup B2. Additionally, the GimA remnant occurs among strains belonging to multilocus sequence type ST117 which is assigned to the recombinant phylogroup ABD. The remaining phylogroups (A, B1, AxB1, ABD (with the exception of ST117), and D) lack any GimA related structure. Moreover, significant associations between the complete GimA and the pathovars APEC and NMEC on the one hand and between the GimA-remnant and UPEC strains on the other hand could be identified. Alike the known contribution of ibeA to the pathogenicity of newborn meningitis, these findings may suggest a possible involvement of this structure in uropathogenicity. A detailed analysis of the ibeA sequences revealed a rigorous association to the phylogenetic background of the strains suggesting GimA originating from phylogroup B2. Moreover, this gene was not under positive selection in strains belonging to the ST95 complex. Thus, the evolution of ibeA seems to be driven by neutral selection, arguing against GimA being mobile. Although common criteria for genetic islands are partially fulfilled, GimA rather seems to be an ancestral part of phylogroup B2. In view of this affiliation to the core genome instead of being an island, it would be appropriate to assign this genomic region the name GimA locus. The existence of two other patterns reflects a genome reduction in a reductive evolution- like manner. The process of reduction in this genomic region may be driven by a shift in pathogenic importance of the GimA components in certain habitats. For systemic infections caused by APEC and NMEC a complete GimA is required. Some components mediate the entrance to the central nervous system (ibeRAT – GimA4), while the others are involved in stress response within the blood stream (GimA1-3). In contrast, the urinary tract is a completely different habitat exhibiting considerably fewer stressors for E. coli. Thus, genes allowing the translocation of the brain blood barrier as well as stress response genes may have lost its importance. Consequently, these genes might have been deleted in a genome optimization process and merely a remnant of GimA maintains. This structure may provide selective advantage and therefore maintains in the E. coli-chromosome. This GimA-remant could be identified as an ORF and is highly associated with UPEC. Hence, it may be involved in uropathogenicity. Further investigations are needed to determine the function of the GimA-remnant. The third variant (GimA-) could have emerged from both (i) directly by the loss of the complete GimA in a one step deletion event and (ii) subsequently following the reduction process leading to the GimA-remnant, respectively. The latter one could in the case of UPEC be the result of an ongoing genome optimization process leading to attenuated strains. Such strains avoid to provoke host response and may therefore maintain in the urinary tract as asymptomatically colonizers

Distribution of ESBL/AmpC-Escherichia coli on a Dairy Farm

The aim of the study was to determine the prevalence of ESBL/AmpC-producing Escherichia (E.) coli and to investigate their on-farm distribution on an exemplary dairy farm. For this purpose, sample sizes were calculated, and fecal samples were collected from cattle of all ages and analyzed for the presence of ESBL/AmpC-E. coli using selective media supplemented with cefotaxime. These antibiotic-resistant bacteria were detected in 22.5% of the samples tested. The prevalence was highest in the calf age group, in which 100% of the collected fecal samples were positive. With increasing age, the prevalence decreased in the other sample groups. While ESBL/AmpC E. coli could still be detected in young stock (15%) and breeding heifers (5%), no resistant pathogens could be detected in adult animals. Whole-genome sequencing of the ESBL/AmpC-E. coli isolates revealed, first, that all isolates were ESBL producers (CTX-M-1 and CTX-M-15) and, second, that ST362, which is known as a biofilm producer, was dominant in the calves (85%, n = 17). Based on these results and the evaluation of a questionnaire, possible causes for the occurrence of ESBL/AmpC-E. coli were discussed and recommendations for the reduction in transmission were formulated. Unlike most German dairy farms, no waste milk feeding was apparent; therefore, factors reducing ESBL/AmpC-E. coli are primarily related to an improvement in hygiene management to prevent biofilms, e.g., in nipple buckets, but also to question the use of antibiotics, e.g., in the treatment of diarrheic calves

Prevalence and Risk Factors for ESBL/AmpC-E. coli in Pre-Weaned Dairy Calves on Dairy Farms in Germany

The objectives of this study were to ascertain the fecal ESBL/AmpC-E. coli prevalence and to detect risk factors for their occurrence in young pre-weaned calves and their dams on large dairy farms in Germany. From 2018–2019 we investigated 2816 individual fecal samples from pre-weaned dairy calves and their dams, representing seventy-two farms (mean = 667 milking cows) from eight German federal states. To assess possible risk factors associated with ESBL/AmpC-E. coli prevalence in calves and dams, a questionnaire was performed, collecting management data. We observed an ESBL/AmpC-E. coli prevalence of 63.5% (95% CI: 57.4–69.5) among the sampled calves and 18.0% (95% CI: 12.5–23.5) among the dams. On all farms, at least one positive sample was obtained. To date, this is the highest ESBL/AmpC-E. coli prevalence observed in dairy herds in Europe. Feeding with waste milk was identified as a significant risk factor for a high prevalence of ESBL/AmpC-E. coli in calves. Many calves at large dairies in Germany are fed with waste milk due to the large amounts generated as a result of antibiotic dry-off routines and mastitis treatment with antibiotics. Other notable risk factors for high ESBL/AmpC-E. coli in calves were the general fitness/health of dams and calves, and the quality of farm hygiene. Taken together, these findings suggest that new or improved approaches to animal health management, for example, antibiotic dry cow management (selective dry cow therapy) and mastitis treatment (high self-recovery), as well as farm hygiene, should be researched and implemented

Longitudinal Study on Extended-Spectrum Beta-Lactamase-E. coli in Sentinel Mallard Ducks in an Important Baltic Stop-Over Site for Migratory Ducks in Germany

Antimicrobial resistance (AMR) is a serious global health threat with extended-spectrum beta-lactamase (ESBL)-producing Enterobacterales as the most critical ones. Studies on AMR in wild birds imply a possible dissemination function and indicate their potential role as sentinel animals. This study aimed to gain a deeper insight into the AMR burden of wild waterfowl by sampling semi-wild mallard ducks used as sentinels and to identify if AMR bacteria could be recommended to be added to the pathogens of public health risks to be screened for. In total, 376 cloacal and pooled fecal samples were collected from the sentinel plant over a period of two years. Samples were screened for ESBL-carrying E. coli and isolates found further analyzed using antimicrobial susceptibility testing and whole-genome sequencing. Over the sampling period, 4.26% (16/376) of the samples were positive for ESBL-producing E. coli. BlaCTX-M-1 and blaCTX-M-32 were the most abundant CTX-M types. Although none of the top global sequence types (ST) could be detected, poultry-derived ST115 and non-poultry-related STs were found and could be followed over time. The current study revealed low cases of ESBL-producing E. coli in semi-wild mallard ducks, which proves the suitability of sentinel surveillance for AMR detection in water-associated wildlife

Legislative Documents

Also, variously referred to as: House bills; House documents; House legislative documents; legislative documents; General Court documents

Wirksamkeit von Impfstrategien gegen Salmonelleninfektionen

In Legehennenbeständen wurde die Schutzwirkung von vier Impfprogrammen, die der Zentralverband der Deutschen Geflügelwirtschaft e.V. empfiehlt und einer Impfung nach der Hühner-Salmonellen-Verordnung untersucht. Unterschiede in den Impfschemata konnten unter den gewählten Versuchsbedingungen nicht festgestellt werden. Eine Impfung gegen Salmonellen garantiert keine vollständige Elimination des Erregers, sondern eine Reduktion der Besiedelung der Organe sowie der Ausscheidung und der Eikontamination. Auch eine zusätzliche Impfung mit Inaktivatimpfstoffen erbrachte keinen effektiveren Schutz vor Salmonella Enteritidis und Salmonella Thyphimurium. Die Schutzwirkung einer Impfung ist am besten, wenn die Salmonellen-Exposition gering ist