A Step Forward in Molecular Diagnostics of Lyssaviruses – Results of a Ring Trial among European Laboratories

Rabies is a lethal and notifiable zoonotic disease for which diagnostics have to meet the highest standards. In recent years, an evolution was especially seen in molecular diagnostics with a wide variety of different detection methods published. Therefore, a first international ring trial specifically designed on the use of reverse transcription polymerase chain reaction (RT-PCR) for detection of lyssavirus genomic RNA was organized. The trial focussed on assessment and comparison of the performance of conventional and real-time assays. In total, 16 European laboratories participated. All participants were asked to investigate a panel of defined lyssavirus RNAs, consisting of Rabies virus (RABV) and European bat lyssavirus 1 and 2 (EBLV-1 and -2) RNA samples, with systems available in their laboratory. The ring trial allowed the important conclusion that conventional RT-PCR assays were really robust assays tested with a high concordance between different laboratories and assays. The real-time RT-PCR system by Wakeley et al. (2005) in combination with an intercalating dye, and the combined version by Hoffmann and co-workers (2010) showed good sensitivity for the detection of all RABV samples included in this test panel. Furthermore, all used EBLV-specific assays, real-time RT-PCRs as well as conventional RT-PCR systems, were shown to be suitable for a reliable detection of EBLVs. It has to be mentioned that differences were seen in the performance between both the individual RT-PCR systems and the laboratories. Laboratories which used more than one molecular assay for testing the sample panel always concluded a correct sample result. Due to the markedly high genetic diversity of lyssaviruses, the application of different assays in diagnostics is needed to achieve a maximum of diagnostic accuracy. To improve the knowledge about the diagnostic performance proficiency testing at an international level is recommended before using lyssavirus molecular diagnostics e.g. for confirmatory testing

Palaeomagnetism of the Ordovician dolerites of the Crozon Peninsula (France)

In order to obtain a Lower Palaeozoic pole for the Armorican Massif and to test the origin of the Ibero-Armorican arc, the Ordovician dolerites of the Crozon peninsula have been palaeomagnetically studied. The samples show a multicomponent magnetization which has been revealed by AF and thermal demagnetization and thoroughly investigated with rock magnetic experiments, polished section examinations and K/Ar dating. Four groups of directions have been recognized, often superimposed on each other in an individual sample. One component (D) has always the lowest blocking temperatures and coercivities and is considered to be of viscous origin, acquired recently in situ or in the laboratory during storage. Two components (A and B) are interpreted to be of secondary origin and to correspond to the observed K/Ar age distribution between 300 and 190 Myr. These ages represent the time interval between two regional thermo-tectonic events, associated with the Hercynian orogeny and the intrusion of dykes related to the early opening of the Central Atlantic Ocean and the Bay of Biscay. A fourth component (C) could be of Ordovician or younger Palaeozoic age; it is not clear whether the age of the magnetization is pre- or post-folding, but a pre-folding age would yield a direction of magnetization similar to Ordovician results from the Iberian peninsula. The latter interpretation suggests a fairly high palaeolatitude, which is in agreement with a glacio-marine postulated for sediments overlying the dolerite sills.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/73214/1/j.1365-246X.1983.tb03785.x.pd

Active surveillance of bat rabies in France: A 5-year study (2004–2009)

Active surveillance of bats in France started in 2004 with an analysis of 18 of the 45 bat species reported in Europe. Rabies antibodies were detected in six indigenous species, mainly in Eptesicus serotinus and Myotis myotis, suggesting previous contact with the EBLV-1 rabies virus. Nineteen of the 177 tested bats were shown serologically positive in seven sites, particularly in central and south-western France. Neither infectious viral particles nor viral genomes were detected in 173 and 308 tested oral swabs, respectively. The presence of neutralising antibodies in female bats (18.6%) was significantly higher than in males (5.6%)

In Vitro and In Vivo Isolation and Characterization of Duvenhage Virus

A fatal human case of Duvenhage virus (DUVV) infection in a Dutch traveller who had returned from Kenya was reported in 2007. She exhibited classical symptoms of rabies encephalitis with distinct pathological findings. In the present study we describe the isolation and characterization of DUVV in vitro and its passage in BALB/c mice. The virus proved to be neuroinvasive in both juvenile and adult mice, resulting in about 50% lethality upon peripheral infection. Clinical signs in infected mice were those of classical rabies. However, the distribution of viral antigen expression in the brain differed from that of classical rabies virus infection and neither inclusion bodies nor neuronal necrosis were observed. This is the first study to describe the in vitro and in vivo isolation and characterization of DUVV

Eliminating Rabies in Estonia

The compulsory vaccination of pets, the recommended vaccination of farm animals in grazing areas and the extermination of stray animals did not succeed in eliminating rabies in Estonia because the virus was maintained in two main wildlife reservoirs, foxes and raccoon dogs. These two species became a priority target therefore in order to control rabies. Supported by the European Community, successive oral vaccination (OV) campaigns were conducted twice a year using Rabigen® SAG2 baits, beginning in autumn 2005 in North Estonia. They were then extended to the whole territory from spring 2006. Following the vaccination campaigns, the incidence of rabies cases dramatically decreased, with 266 cases in 2005, 114 in 2006, four in 2007 and three in 2008. Since March 2008, no rabies cases have been detected in Estonia other than three cases reported in summer 2009 and one case in January 2011, all in areas close to the South-Eastern border with Russia. The bait uptake was satisfactory, with tetracycline positivity rates ranging from 85% to 93% in foxes and from 82% to 88% in raccoon dogs. Immunisation rates evaluated by ELISA ranged from 34% to 55% in foxes and from 38% to 55% in raccoon dogs. The rabies situation in Estonia was compared to that of the other two Baltic States, Latvia and Lithuania. Despite regular OV campaigns conducted throughout their territory since 2006, and an improvement in the epidemiological situation, rabies has still not been eradicated in these countries. An analysis of the number of baits distributed and the funding allocated by the European Commission showed that the strategy for rabies control is more cost-effective in Estonia than in Latvia and Lithuania

A rootless suture and the loss of the roots of a mountain chain: The Variscan belt of NW Iberia

Ophiolites of different Paleozoic ages occur in North-West (NW) Iberia in a rootless suture representing the remnants of the Rheic Ocean. Associated allochthonous terranes in the hanging- and foot-walls of the suture derive from the former margins, whereas the relative autochthon corresponds to the Paleozoic passive margin of northern Gondwana. The Paleozoic tectonic evolution of this part of the circum-Atlantic region is deduced from the stratigraphical, petrological, structural and metamorphic evolution of the different units and their ages. The tectonic reconstruction covers from Cambro-Ordovician continental rifting and the opening of the Rheic Ocean to its Middle to Upper Devonian closure. Then, the Variscan Laurussia–Gondwana convergence and collision is briefly described, from its onset to the late stages of collapse associated with the demise of the orogenic roots. [RÉSUMÉ] Une suture sans racines et la perte des racines d’une chaîne montagneuse : la chaîne varisque du Nord-Ouest de l’Ibérie. Des ophiolites d’âges différents affleurent dans le Nord-Ouest de l’Ibérie dans une suture sans racines, témoin de l’océan Rhéïque. Les terrains allochtones sur et sous la suture dérivent de ses deux marges, tandis que l’autochtone relatif appartient à la marge passive du Nord de Gondwana. On peut déduire l’évolution des plaques dans cette partie de la région circum-Atlantique à partir des données stratigraphiques, pétrologiques, structurales, métamorphiques et géochronologiques. Cette évolution inclut le développement d’un rift continental et l’ouverture de l’océan Rhéïque pendant le Cambro-Ordovicien ainsi que sa fermeture au Dévonien moyen à supérieur. On décrit aussi l’évolution de la convergence et collision varisque entre Laurussia et Gondwana, du début jusqu’aux derniers stades d’un effondrement associé à la perte des racines orogéniques

Epidemiology and molecular diversity of rabies viruses in Bulgaria

SUMMARYA health emergency situation occurred in Bulgaria in 2007 when positive rabies cases were notified in Sofia district in the central-western part of the country, suggesting a southward spread of the disease for the first time in the last 10 years. Phylogenetic analysis on 49 isolates sampled between 2009 and 2011 showed, for the first time, evidence of the existence of NEE and D clustered lineages in Bulgaria. Their geographical distribution clearly reveals the permeability of natural barriers, as already suggested by the disease spread that occurred across the Balkan mountain range in 2007. The monitoring and passive surveillance programmes conducted since the first 2009 oral vaccination campaign, the spatio-temporal evolution of the disease in the country since 2007, and the need for further investigation of the role of jackals in virus dispersion are discussed.</jats:p