First isolation of a rabid bat infected with European bat lyssavirus in Luxembourg.

Rabid bats are regularly reported in Europe, especially in countries that have implemented a bat surveillance network. In May 2013, bat rabies was evidenced for the first time in Luxembourg (southern city of Differdange). The rabies virus, an EBLV-1b strain, was diagnosed in a serotine bat that bit a 29-year-old male person while he was asleep.

Epidemiology and molecular diversity of rabies viruses in Bulgaria.

A 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.

Isolation of Bokeloh bat lyssavirus in Myotis nattereri in France.

Bokeloh bat lyssavirus (BBLV) was found in Myotis nattereri for the first time in northeastern France in July 2012. The complete genome sequence of the virus from the infected Natterer's bat was determined by whole-genome sequencing and compared to that of the first BBLV strain isolated in 2010 in Germany and with those of all currently identified lyssaviruses. The French isolate [KC169985] showed 98.

Comparative assay of fluorescent antibody test results among twelve European National Reference Laboratories using various anti-rabies conjugates.

Twelve National Reference Laboratories (NRLs) for rabies have undertaken a comparative assay to assess the comparison of fluorescent antibody test (FAT) results using five coded commercial anti-rabies conjugates (Biorad, Bioveta, Fujirebio, Millipore, and SIFIN conjugates). Homogenized positive brain tissues infected with various lyssavirus species as well as negative samples were analyzed blindly using a standardized FAT procedure. Conjugates B, C, D, and E were found to be significantly more effective than conjugate A for GS7 (French RABV) diluted samples (1/8 and 1/100) while the frequency of concordant results of conjugates C and D differ significantly from conjugates A, B and E for CVS 27.

Evaluation of an ELISA to detect rabies antibodies in orally vaccinated foxes and raccoon dogs sampled in the field.

The assessment of the efficacy of oral vaccination in wildlife is based on detection in the teeth of a biomarker (tetracycline) which is incorporated in the vaccine bait, and the quantification of rabies antibodies. A blocking ELISA was evaluated and compared with the FAVN test and a validated in-house ELISA, using sera from foxes and raccoon dogs collected following oral vaccination campaigns in France and Estonia. Specificity reached 100% in sera from naïve animals.

Evaluation of a Rapid Immunochromatographic Diagnostic Test for the detection of rabies from brain material of European mammals.

The surveillance of rabies relies on investigations conducted on dead suspected animals or animals showing clinical signs suggestive of rabies. An immunochromatographic method based on lateral flow principle has been evaluated against a collection of brain samples mainly of European mammals including bats. The performance of this new test has been compared to the conventional gold standard methods: the fluorescent Antibody Test (FAT) and the Rapid Tissue Culture Infection Test (RTCIT).

Evaluation of ELISA for detection of rabies antibodies in domestic carnivores.

Serological tests of pets have increased as many rabies-free countries have amended their quarantine measures and adopted a scheme requiring rabies vaccination followed by a serological test. A European directive requires the measurement of neutralising antibodies as proof of protection to allow the free movement of pets within the European Union and between third countries non listed in the list C of regulation 998/2003 and European countries. At present, the recommended neutralisation tests (FAVN test or RFFIT) are time-consuming, expensive and require highly trained technicians as well as special laboratory facilities.

International interlaboratory trials on rabies diagnosis: an overview of results and variation in reference diagnosis techniques (fluorescent antibody test, rabies tissue culture infection test, mouse inoculation test) and molecular biology techniques.

Interlaboratory trials on rabies diagnosis were organised in 2009 and in 2010 by the European Union Reference Laboratory (EURL) for rabies. In 2009, two panels of virus samples were sent to participating laboratories to compare results on reference diagnosis techniques and on RT-PCR. A single panel was sent in 2010 to test FAT (fluorescent antibody test), RTCIT (rabies tissue culture infection test) and RT-PCR techniques.

Experimental infection of foxes with European Bat Lyssaviruses type-1 and 2.

Background: Since 1954, there have been in excess of 800 cases of rabies as a result of European Bat Lyssaviruses types 1 and 2 (EBLV-1, EBLV-2) infection, mainly in Serotine and Myotis bats respectively. These viruses have rarely been reported to infect humans and terrestrial mammals, as the only exceptions are sheep in Denmark, a stone marten in Germany and a cat in France. The purpose of this study was to investigate the susceptibility of foxes to EBLVs using silver foxes (Vulpes vulpes) as a model.

Experimental infection of foxes with European bat lyssaviruses type-1 and -2.

Experimental studies have been undertaken to assess the susceptibility of silver foxes to bat variants of rabies virus, namely European Bat Lyssaviruses (EBLVs). Both EBLV-1 and EBLV-2 have been isolated in European bats since 1954, in Eptesicus serotinus and Myotis species, respectively. Since 2000, the number of reported cases has increased largely due to the improvement of the surveillance of bat rabies virus throughout Europe.

Efficacy of a square presentation of V-RG vaccine baits in red fox, domestic dog and raccoon dog.

Oral vaccination programmes conducted in rabies infected countries from Eastern Europe and Eurasia should not be restricted to foxes but should target other major rabies vectors such as dogs and raccoon dogs as well. The objective of this experimental trial was to assess the protection induced by the vaccine by challenging these different species, which had been previously vaccinated intramuscularly with the square V-RG baits (produced in the US). Different parameters were evaluated such as attractiveness of the baits and induction of neutralising antibodies as an indicator for immunogenicity and protection after rabies challenge.

Efficacy and bait acceptance of vaccinia vectored rabies glycoprotein vaccine in captive foxes (Vulpes vulpes), raccoon dogs (Nyctereutes procyonoides) and dogs (Canis familiaris).

The red fox, dog, and raccoon dog are known to play a major role in the global epidemiology of rabies. These three canid species were used to compare the appetency and efficacy of two commercial bait formats, each containing a single dose of vaccinia-rabies glycoprotein (V-RG) vaccine. Square and rectangular RABORAL V-RG baits were fed to individual caged animal, and results were evaluated using three parameters: bait consumption, induction of rabies virus neutralizing antibodies and protection after a virulent rabies challenge.

The safety and efficacy of the oral rabies vaccine SAG2 in Indian stray dogs.

India is one of the countries with the highest prevalence of human rabies throughout the world. Dogs are primarily responsible for rabies transmission. Among them, stray dogs play a major role in that country.

Means used for terrestrial rabies elimination in France and policy for rabies surveillance in case of re-emergence.

In France, the first case of wildlife rabies was detected in 1968, with the red fox (Vulpes vulpes) as reservoir and vector of terrestrial rabies. The last case was reported at the end of 1998. The maximum infected area amounted to 140,000 km2 in 1989 with a record number of 4,213 infected wild and domestic animals.

Tools for rabies serology to monitor the effectiveness of rabies vaccination in domestic and wild carnivores.

Serology remains the only way to monitor the effectiveness of vaccination of humans and animals against rabies. Many techniques for determining the level of rabies antibodies have been described, including seroneutralisation techniques such as tests for fluorescent antibody virus neutralisation (FAVN) and rapid fluorescent focus inhibition (RFFIT), enzyme-linked immunosorbent assay (ELISA), and in-vivo tests (the mouse neutralisation test, MNT). The need to verify the effectiveness of rabies vaccination has become widespread, particularly in the context of international trading of domestic carnivores from infected to rabies-free territories.

Safety and efficacy of the oral rabies vaccine SAG2 in raccoon dogs.

Oral vaccination programmes in several rabies-infected countries from Northern and Eastern Europe should not be restricted to foxes but should target raccoon dogs as well. The safety, immunogenicity and efficacy of Rabigen SAG2 bait was evaluated in raccoon dogs. Safety of SAG2 was demonstrated after direct instillation (n = 5) or ingestion of a bait (n = 5) using a quantity of virus at least 10 times superior to the field dose.

Elimination of terrestrial rabies in Western European countries.

Since the late 1930s, the red fox (Vulpes vulpes) has been the main vector of rabies in Europe. Practically, decimation of fox population did not prevent the spread of the disease. The only efficient method to control wildlife rabies consisted in using oral vaccination by depositing vaccine baits containing a capsule or a plastic sachet filled with an attenuated anti-rabies liquid vaccine throughout fox habitats.

Development of a qualitative indirect ELISA for the measurement of rabies virus-specific antibodies from vaccinated dogs and cats.

A protocol suitable for the detection of rabies virus-specific antibodies in serum samples from companion animals using an enzyme linked immunosorbent assay (ELISA) is described. This method has been used successfully for the qualitative assessment of rabies virus-specific antibodies in serum samples from a cohort of vaccinated dogs and cats. In two initial field studies, a variable population of field samples from the Veterinary Laboratories Agency (VLA), United Kingdom were tested.