Identification of factors influencing the Puumala virus seroprevalence within its reservoir in aMontane Forest Environment.

Puumala virus (PUUV) is a major cause of mild to moderate haemorrhagic fever with renal syndrome and is transmitted by the bank vole (Myodes glareolus). There has been a high cumulative incidence of recorded human cases in South-eastern Germany since 2004 when the region was first recognized as being endemic for PUUV. As the area is well known for outdoor recreation and the Bavarian Forest National Park (BFNP) is located in the region, the increasing numbers of recorded cases are of concern. To understand the population and environmental effects on the seroprevalence of PUUV in bank voles we trapped small mammals at 23 sites along an elevation gradient from 317 to 1420m above sea level. Generalized linear mixed effects models(GLMEM) were used to explore associations between the seroprevalence of PUUV in bank voles and climate and biotic factors. We found that the seroprevalence of PUUV was low (6%-7%) in 2008 and 2009, and reached 29% in 2010. PUUV seroprevalence was positively associated with the local species diversity and deadwood layer, and negatively associated with mean annual temperature, mean annual solar radiation, and herb layer. Based on these findings, an illustrative risk map for PUUV seroprevalence prediction in bank voles was created for an area of the national park. The map will help when planning infrastructure in the national park (e.g., huts, shelters, and trails)

Re-emergence of tularemia in Germany: Presence of <it>Francisella tularensis </it>in different rodent species in endemic areas

<p>Abstract</p> <p>Background</p> <p>Tularemia re-emerged in Germany starting in 2004 (with 39 human cases from 2004 to 2007) after over 40 years of only sporadic human infections. The reasons for this rise in case numbers are unknown as is the possible reservoir of the etiologic agent <it>Francisella (F.) tularensis</it>. No systematic study on the reservoir situation of <it>F. tularensis </it>has been published for Germany so far.</p> <p>Methods</p> <p>We investigated three areas six to ten months after the initial tularemia outbreaks for the presence of <it>F. tularensis </it>among small mammals, ticks/fleas and water. The investigations consisted of animal live-trapping, serologic testing, screening by real-time-PCR and cultivation.</p> <p>Results</p> <p>A total of 386 small mammals were trapped. <it>F. tularensis </it>was detected in five different rodent species with carrier rates of 2.04, 6.94 and 10.87% per trapping area. None of the ticks or fleas (n = 432) tested positive for <it>F. tularensis</it>. We were able to demonstrate <it>F. tularensis-</it>specific DNA in one of 28 water samples taken in one of the outbreak areas.</p> <p>Conclusion</p> <p>The findings of our study stress the need for long-term surveillance of natural foci in order to get a better understanding of the reasons for the temporal and spatial patterns of tularemia in Germany.</p

A new permanent cell line derived from the bank vole (Myodes glareolus) as cell culture model for zoonotic viruses

<p>Abstract</p> <p>Background</p> <p>Approximately 60% of emerging viruses are of zoonotic origin, with three-fourths derived from wild animals. Many of these zoonotic diseases are transmitted by rodents with important information about their reservoir dynamics and pathogenesis missing. One main reason for the gap in our knowledge is the lack of adequate cell culture systems as models for the investigation of rodent-borne (robo) viruses <it>in vitro</it>. Therefore we established and characterized a new cell line, BVK168, using the kidney of a bank vole, <it>Myodes glareolus, </it>the most abundant member of the <it>Arvicolinae </it>trapped in Germany.</p> <p>Results</p> <p>BVK168 proved to be of epithelial morphology expressing tight junctions as well as adherence junction proteins. The BVK168 cells were analyzed for their infectability by several arbo- and robo-viruses: Vesicular stomatitis virus, vaccinia virus, cowpox virus, Sindbis virus, Pixuna virus, Usutu virus, Inkoo virus, Puumalavirus, and Borna disease virus (BDV). The cell line was susceptible for all tested viruses, and most interestingly also for the difficult to propagate BDV.</p> <p>Conclusion</p> <p>In conclusion, the newly established cell line from wildlife rodents seems to be an excellent tool for the isolation and characterization of new rodent-associated viruses and may be used as <it>in vitro-</it>model to study properties and pathogenesis of these agents.</p

Nagetier-übertragene Zoonosen: Beispiele aus Untersuchungen in Süd- und Westdeutschland

Zusammenfassung Nagetiere und andere Kleinsäuger können eine Vielzahl von Krankheitserregern, RNA- und DNA-Viren, Bakterien und Parasiten, auf den Menschen übertragen, die teilweise lebensbedrohliche Erkrankungen hervorrufen. In der folgenden Übersicht soll erstmals ein Überblick über Ergebnisse aus drei Untersuchungen in Deutschland gegeben werden: eine Studie in drei Landkreisen Bayerns von 2001-2004, Untersuchungen in einem Freilandgehege im Rahmen eines Tularämieausbruchs in Niedersachsen im Jahr 2005, und schließlich im Jahr 2007 eine Untersuchung an einem Truppenübungsplatz in Baden-Württemberg. Es wurde dabei exemplarisch die Verbreitung von Zoonoseerregern in Nagetieren und anderen Kleinsäugern näher untersucht, von drei Viren (Hantaviren, Kuhpockenvirus, Frühsommer- Meningo-Enzephalitis-Virus) und vier Bakterien (Leptospiren, Francisellen, Borrelien und Rickettsien). Die hier zusammengefassten Erkenntnisse sind ein erster wichtiger Schritt auf dem Weg zur Erstellung von Verbreitungskarten für die genannten humanpathogenen Zoonoseerreger in ihren Reservoirwirten und der Definition von entsprechenden Risikogebieten. Diese Arbeit soll zudem einen Beitrag leisten, einen Anstoß zu verstärkter Zusammenarbeit von Zoologen, Ökologen, Virologen, Human- und Veterinärmedizinern, Mikrobiologen, Parasitologen, Genetikern, Epidemiologen, Forstwissenschaftlern und Klimaforschern zu geben

Double-stranded RNA-activated protein kinase PKR of fishes and amphibians: Varying the number of double-stranded RNA binding domains and lineage-specific duplications

BackgroundDouble-stranded (ds) RNA, generated during viral infection, binds and activates the mammalian anti-viral protein kinase PKR, which phosphorylates the translation initiation factor eIF2alpha leading to the general inhibition of protein synthesis. Although PKR-like activity has been described in fish cells, the responsible enzymes eluded molecular characterization until the recent discovery of goldfish and zebrafish PKZ, which contain Z-DNA-binding domains instead of dsRNA-binding domains (dsRBDs). Fish and amphibian PKR genes have not been described so far.ResultsHere we report the cloning and identification of 13 PKR genes from 8 teleost fish and amphibian species, including zebrafish, demonstrating the coexistence of PKR and PKZ in this latter species. Analyses of their genomic organization revealed up to three tandemly arrayed PKR genes, which are arranged in head-to-tail orientation. At least five duplications occurred independently in fish and amphibian lineages. Phylogenetic analyses reveal that the kinase domains of fish PKR genes are more closely related to those of fish PKZ than to the PKR kinase domains of other vertebrate species. The duplication leading to fish PKR and PKZ genes occurred early during teleost fish evolution after the divergence of the tetrapod lineage. While two dsRBDs are found in mammalian and amphibian PKR, one, two or three dsRBDs are present in fish PKR. In zebrafish, both PKR and PKZ were strongly upregulated after immunostimulation with some tissue-specific expression differences. Using genetic and biochemical assays we demonstrate that both zebrafish PKR and PKZ can phosphorylate eIF2alpha in yeast.ConclusionConsidering the important role for PKR in host defense against viruses, the independent duplication and fixation of PKR genes in different lineages probably provided selective advantages by leading to the recognition of an extended spectrum of viral nucleic acid structures, including both dsRNA and Z-DNA/RNA, and perhaps by altering sensitivity to viral PKR inhibitors. Further implications of our findings for the evolution of the PKR family and for studying PKR/PKZ interactions with viral gene products and their roles in viral infections are discussed