Lleida Bat Lyssavirus isolation in Miniopterus schreibersii in France

Bat rabies cases are attributed in Europe to five different Lyssavirus species of 16 recognized Lyssavirus species causing rabies. One of the most genetically divergent Lyssavirus spp. has been detected in a dead Miniopterus schreibersii bat in France. Brain samples were found positive for the presence of antigen, infectious virus and viral RNA by classical virological methods and molecular methods respectively. The complete genome sequence was determined by next‐generation sequencing. The analysis of the complete genome sequence confirmed the presence of Lleida bat lyssavirus (LLEBV) in bats in France with 99.7% of nucleotide identity with the Spanish LLEBV strain (KY006983).     

Evidence of Australian bat lyssavirus infection in diverse Australian bat taxa

Historically, Australia was considered free of rabies and rabieslike viruses. Thus, the identification of Australian bat lyssavirus (ABLV) in 1996 in a debilitated bat found by a member of the public precipitated both public health consternation and a revision of lyssavirus taxonomy. Subsequent observational studies sought to elaborate the occurrence and frequency of ABLV infection in Australian bats. This paper describes the taxonomic diversity of bat species showing evidence of ABLV infection to better inform public health considerations. Blood and/or brain samples were collected from two cohorts of bats (wild‐caught and diagnostic submissions) from four Australian states or territories between April 1996 and October 2002. Fresh brain impression smears were tested for ABLV antigen using fluorescein‐labelled anti‐rabies monoclonal globulin (CENTOCOR) in a direct fluorescent antibody test; sera were tested for the presence of neutralising antibodies using a rapid fluorescent focus inhibition test. A total of 3,217 samples from 2,633 bats were collected and screened: brain samples from 1,461 wild‐caught bats and 1,086 submitted bats from at least 16 genera and seven families, and blood samples from 656 wild‐caught bats and 14 submitted bats from 14 genera and seven families. Evidence of ABLV infection was found in five of the six families of bats occurring in Australia, and in three of the four Australian states/territories surveyed, supporting the historic presence of the virus in Australia. While the infection prevalence in the wild‐caught cohort is evidently low, the significantly higher infection prevalence in rescued bats in urban settings represents a clear and present public health significance because of the higher risk of human exposure.     

Veterinary surgeon's guide to Australian bat lyssavirus

Veterinary surgeons in Australia must be aware of the emerging viral diseases and their potential effects on public health generally and, more specifically, on the veterinary profession. Australian bat lyssavirus was identified in 1996 and causes rabies-like disease in bats and humans. Two humans from Queensland have died of Australian bat lyssavirus encephalitis. Surveillance has shown that all Australian bats must be considered carriers of this new virus, therefore protective apparel should be used when handling bats. The pre-exposure regimen of inactivated rabies vaccine (Pasteur Mérieux) provides protection against infection. As part of the preventive regimen, at risk groups, such as veterinary surgeons, should seriously consider pre-exposure rabies vaccination. The post-exposure protocol involves administration of human rabies immunoglobulins and five intramuscular injections of the inactivated rabies vaccine.     

Detection of European bat lyssavirus 2 (EBLV-2) in a Daubenton's bat (Myotis daubentonii) from Magdeburg, Germany

In Europe bat rabies in Daubenton's bats (Myotisdaubentonii) and in Pond bats (Myotis dasycneme) caused by the European bat lyssavirus 2 (EBLV-2) has been confirmed in less than 20 cases to date. Here we report the second encounter of this virus species in Germany. A Daubenton's bat found grounded in the zoological garden in Magdeburg died shortly after. In the frame of a retrospective study the bat carcass was eventually transferred to the national reference laboratory for rabies at the Friedrich-Loeffler-Institute for rabies diagnosis. Lyssavirus was isolated and characterized as EBLV-2.     

Spill‐over of European Bat Lyssavirus Type 1 into a Stone Marten (Martes foina) in Germany

European bat lyssavirus type 1 (EBLV‐1, genotype 5) is known to endemically circulate in insectivorous bat populations in Germany. In August 2001, a rabies suspect stone marten (Martes foina) was found in the city of Burg (Saxony‐Anhalt, Germany) and was sent to the regional veterinary laboratory for routine rabies diagnosis. Whereas brain samples repeatedly tested negative in the fluorescent antibody test for classical rabies virus (genotype 1), the mouse inoculation test and the rabies tissue culture inoculation test yielded positive results. Rabies viral RNA was also detected in the stone marten brain sample both by nested and heminested RT‐PCR specific for the nucleoprotein gene and for the nucleoprotein phosphoprotein junction of rabies virus. The amplification products were sequenced to genotype the isolate. Sequence data obtained from the first‐round RT‐PCR products were analysed and the suspect stone marten isolate was confirmed as a rabies related virus (EBLV‐1a). Phylogenetic comparison with sequences from recent genotype five isolates from Germany and Denmark showed that it was closely related to a previous isolate of EBLV‐1 from a serotine bat in Saxony‐Anhalt obtained in the same year in an area adjacent to the place where the EBLV‐1 infected stone marten was found. Both EBLV‐1 isolates share a 99.5% identity. This is the first report of an EBLV‐1a spill‐over from an insectivorous bat into wildlife in Europe.     

Isolation of a European bat lyssavirus type 2 from a Daubenton's bat in the United Kingdom

European bat lyssavirus type 2 (EBLV-2) has been isolated once previously from a bat in the UK in June 1996. In September 2002, a Daubenton's bat (Myotis daubentonii) found in Lancashire developed abnormal behaviour, including unprovoked aggression, while it was in captivity. Brain samples from the bat were tested for virus of the Lyssavirus genus, which includes EBLV-2 (genotype 6), and classical rabies virus (genotype 1). A positive fluorescent antibody test confirmed that it was infected with a lyssavirus, and PCR and genomic sequencing identified the virus as an EBLV-2a. Phylogenetic comparisons with all the published sequences from genotype 6 showed that it was closely related to the previous isolate of EBLV-2 in the UK and suggested links to isolates from bats in The Netherlands. The isolation of EBLV-2 from a bat found on the west coast of England provides evidence that this virus may be present within the UK Daubenton's bat population at a low prevalence level.     

Salmonella,Campylobacter, Clostridium difficile,and anti‐microbial resistant Escherichia coli in the faeces of sympatric meso‐mammals in southern Ontario,Canada

The role of free‐ranging wildlife in the epidemiology of enteropathogens causing clinical illness in humans and domestic animals is unclear. Salmonella enterica and anti‐microbial resistant bacteria have been detected in the faeces of raccoons (Procyon lotor), but little is known about the carriage of these bacteria in other sympatric meso‐mammals. Our objectives were to: (a) report the prevalence of Salmonella and associated anti‐microbial resistance, Campylobacter spp, Clostridium difficile, and anti‐microbial resistant Escherichia coli in the faeces of striped skunks (Mephitis mephitis) and Virginia opossums (Didelphis virginiana) in southern Ontario; and (b) compare the prevalence of these bacteria in the faeces of these meso‐mammal hosts with raccoons from a previously reported study. Faecal swabs were collected from striped skunks and Virginia opossums on five swine farms and five conservation areas from 2011 to 2013. Salmonella was detected in 41% (9/22) and 5% (5/95) of faecal swabs from Virginia opossums and striped skunks, respectively. None of the Salmonella serovars carried resistance to anti‐microbials. The prevalence of Campylobacter spp., C. difficile, and anti‐microbial resistant E. coli ranged from 6% to 22% in striped skunk and Virginia opossums. Using exact logistic regression, Salmonella was significantly more likely to be detected in faecal swabs of Virginia opossums than skunks and significantly less likely in faecal swabs from skunks than raccoons from a previously reported study. In addition, Campylobacter spp. was significantly more likely to be detected in raccoons than opossums. Salmonella Give was detected in 8/9 (89%) of Salmonella‐positive Virginia opossum faecal swabs. Our results suggest that striped skunks and Virginia opossums have the potential to carry pathogenic enteric bacteria in their faeces. The high prevalence of Salmonella Give in Virginia opossum faecal swabs in this study as well as its common occurrence in other Virginia opossum studies throughout North America suggests Virginia opossums may be reservoirs of this serovar.     

Prevention educational program of human rabies transmitted by bats in rain forest preserved area of southern Brazilian coast

Guaraqueçaba city is a rain forest environmental protected area located on the southern coast of Brazil. Recently, the local Animal Health Service has noticed haematophagous bats feeding from humans and domestic animals, as well as bat colonies located in houses and public schools. In 2007, two non‐haematophagous bats were tested positive by direct immunofluorescence for rabies in a nearby city. Native fauna and environmental laws protect non‐haematophagous bats in Brazilian preserved areas such as Guaraqueçaba, making non‐haematophagous bat population control almost impossible. Accordingly, the aim of this study was to evaluate a simple and feasible educational protocol applied by a multi‐institutional task force in local elementary schools to prevent rabies transmitted by bats. Information was transmitted to children by video, lectures and oral question–answer section; evaluation was made by written questionnaires to teachers and students. Interinstitutional task force included public and animal health public services, a federal university and the city secretary of environment, of education, of agriculture and of animal health, and also participation of local community. Information was effectively absorbed by children when evaluated just after being given. As important, questionnaires showed that handling and playing with bats at day time was common in several elementary school students, exposing themselves to what may represent higher risk of rabies transmission than haematophagous bat feeding directly from humans. Training of teachers and students may effectively prevent rabies by bats in such communities. Insertion of this subject into science content of local elementary school educational programme was proposed in order to establish a continuing education programme on rabies transmitted by bats in environmental preserved areas.     

Histopathology and immunohistochemistry of bats infected by Australian bat lyssavirus

OBJECTIVE: To describe the lesions and distribution of viral antigens in bats infected by Australian bat lyssavirus. DESIGN: A retrospective histopathological and immunohistochemical study of bats naturally infected with the virus. PROCEDURE: Tissues from 37 infected bats were examined. Nineteen flying foxes (fruit bats) and two insectivorous bats were examined in detail. Brains of another 16 flying foxes were poorly fixed and were examined less fully. RESULT: Lesions varied considerably between individuals and, where present, were mostly those of nonsuppurative meningoencephalomyelitis and ganglioneuritis similar to lesions seen in rabies and rabies-like diseases. The number of cells with intracytoplasmic inclusion bodies (Negri bodies) was variable; none were seen in some bats. Intracytoplasmic vacuolation of neurons was a common finding. Lesions occurred throughout the central nervous system but were most frequent and severe in the hippocampus, thalamus and midbrain, and medulla oblongata and pons. Indirect immunoperoxidase tests for lyssavirus antigen reactions varied in intensity and distribution, but also occurred mostly in the hippocampus, thalamus and midbrain, and medulla oblongata and pons. In peripheral tissues, reactions were seen in autonomic ganglia, in nerve plexuses of the gastrointestinal tract, in nervous tissues within muscles and immediately adjacent to individual muscle fibres, in an adrenal medulla, and in epithelial tissues in one of eight salivary glands examined. CONCLUSION: The main lesion in Australian bat lyssavirus infection is nonsuppurative inflammation similar to that seen in rabies and other rabies-like diseases, except that the number of Negri bodies is more variable. Reactions to immunoperoxidase tests for lyssavirus vary in intensity and distribution and may occur in both central and peripheral nervous systems. These reactions do not always occur in the salivary glands, even if brain infection is present.     

Genetic characterization of rabies viruses isolated from frugivorous bat (Artibeus spp.) in Brazil

In Latin America, rabies cases related to frugivorous bats have been reported since 1930's. Recently, two viruses isolated from Artibeus lituratus were proved to be vampire bat variants by monoclonal antibodies panels [2], but their genetic information is not well known. In this report, four rabies viruses were isolated from frugivorous bats (Artibeus spp.) in Brazil and their nucleoprotein gene sequences were determined. These isolates were found to be genotype 1 of lyssavirus and showed the maximum nucleotide sequence homology of 97.6-99.4% with vampire bat-related viruses in Brazil [6]. These results indicate that the Brazilian frugivorous bat rabies viruses in this study are closely related to vampire bat-related viruses that play a main role in rabies virus transmission to livestock in Brazil.     

No Virological Evidence for an Influenza A ‐ like Virus in European Bats

New members of the influenza A virus genus have been detected recently in bats from South America. By molecular investigations, using a generic real‐time RT‐PCR (RT‐qPCR) that detects all previously known influenza A virus subtypes (H1‐H16) and a newly developed RT‐qPCR specific for the South American bat influenza‐like virus of subtype H17, a total of 1571 samples obtained from 1369 individual bats of 26 species from Central Europe were examined. No evidence for the occurrence of such influenza viruses was found. Further attempts towards a more comprehensive evaluation of the role of bats in the ecology and epidemiology of influenza viruses should be based on more intense monitoring efforts. However, given the protected status of bats, not only in Europe, such activities need to be embedded into existing pathogen‐monitoring programs.     

Increased detection of rabies virus in bats in Ceará State (Northeast Brazil) after implementation of a passive surveillance programme

The intensification of dog, cat and livestock vaccination campaigns significantly reduced rabies cases in humans and domestic animals in Ceará State, Brazil. However, sylvatic animals—bats (order Chiroptera), wild canids, raccoons and non‐human primates— remain as reservoirs for the virus. Our hypothesis is that surveillance and monitoring of rabies virus in bats, especially passive surveillance, is of fundamental importance, besides the implementation of health education and strengthening of surveillance actions in humans exposed to aggressions. Thus, we assessed the occurrence of rabies virus in animals focusing on bats, before and after launching of the Sylvatic Rabies Surveillance Program in 2010. Surveillance data from the 184 municipalities of Ceará State were analysed, collected during the periods 2003–2010 (active surveillance) and 2011–2016 (passive surveillance), respectively. A total of 13,543 mammalian samples were received for rabies diagnosis from 2003 to 2016. Of these, 10,960 were from dogs or cats (80.9%), 1,180 from bats (8.7%), 806 from other sylvatic animals (foxes, marmosets, raccoons; 6.0%) and 597 from herbivores (cattle, goats, sheep, equines, pigs; 4.4%). A total of 588 (4.3%) samples were positive for rabies. About 8.4% (99/1,180) of the bat samples were infected with rabies virus, 92 (92.9%) of these were from non‐haematophagous bat species and 7 (7.1%) from haematophagous species. The number of bat samples received and infection rates increased considerably, after a shift from active surveillance (9/355 [2.5%] samples positive), to passive surveillance (90/825 [10.9%] samples positive). Surveillance of rabies virus in bats is fundamental for human and domestic animal health in Ceará State. Bats have to be considered as targets in surveillance and control programmes. Virus lineages should be characterized to increase knowledge on transmission dynamics of sylvatic rabies virus to domestic animals and the human population, and to provide additional evidence for planning and implementation of improved control measures.     

Bat rabies in alberta 1979-1982

The infection rate among eight species of bats submitted for rabies diagnosis in Alberta during 1979-82 was 4.6%. Prevalence of rabies was greatest (24%) for hoary bats Lasiurus cinereus, while the big brown bat Eptesicus fuscus was the species in which rabies was most commonly diagnosed, and the species submitted most frequently for rabies diagnosis was the little brown bat Myotis lucifugus. The rabies infection rate among male hoary bats was significantly greater than in either sex of all other submitted species. The frequency of rabies diagnosis in hoary bats submitted during 1979-82 was also significantly higher than in those submitted between 1971 and 1978. There has been a significant decrease in the rabies prevalence or infection rate of little brown bats since 1971-78.     

Trends in national surveillance for rabies among bats in the United States (1993-2000)

OBJECTIVE: To describe surveillance trends and epidemiologic features of rabies in bats in the United States, focusing on 3 bat species primarily associated with variants of the rabies virus that affect humans. DESIGN: Retrospective study. ANIMALS: 31,380 bats. PROCEDURE: Data on rabies for bats identified to species and reported by state laboratories from 1993 to 2000 were analyzed, focusing on silver-haired, eastern pipistrelle, and Brazilian free-tailed bats. Categoric variables were derived from other provided information. RESULTS: Data were reported from 37 states during the study interval; complete species-specific data were not reported by any state for the entire interval. Bats primarily associated with rabies virus variants affecting humans were more likely to yield positive test results for rabies (22.7%), compared with all other bats (5.5%) in most seasons and from most regions of the United States. However, certain other bat species had higher percentages of positive results. Risk of positive results was highest in the fall and highest among bats originating in the southwestern United States. CONCLUSIONS AND CLINICAL RELEVANCE: Increased risk of rabies among certain groups of bat species was consistently found across seasons and most geographic regions of the United States. Results were in general agreement with those of previous studies conducted within smaller geographic regions. There are ongoing efforts to improve surveillance of rabies in bats, although surveillance is incomplete in some regions.     

Neotropical Bats from Costa Rica harbour Diverse Coronaviruses

Bats are hosts of diverse coronaviruses (CoVs) known to potentially cross the host–species barrier. For analysing coronavirus diversity in a bat species‐rich country, a total of 421 anal swabs/faecal samples from Costa Rican bats were screened for CoV RNA‐dependent RNA polymerase (RdRp) gene sequences by a pancoronavirus PCR. Six families, 24 genera and 41 species of bats were analysed. The detection rate for CoV was 1%. Individuals (n = 4) from four different species of frugivorous (Artibeus jamaicensis, Carollia perspicillata and Carollia castanea) and nectivorous (Glossophaga soricina) bats were positive for coronavirus‐derived nucleic acids. Analysis of 440 nt. RdRp sequences allocated all Costa Rican bat CoVs to the α‐CoV group. Several CoVs sequences clustered near previously described CoVs from the same species of bat, but were phylogenetically distant from the human CoV sequences identified to date, suggesting no recent spillover events. The Glossophaga soricina CoV sequence is sufficiently dissimilar (26% homology to the closest known bat CoVs) to represent a unique coronavirus not clustering near other CoVs found in the same bat species so far, implying an even higher CoV diversity than previously suspected.     

Pathogenesis studies with Australian bat lyssavirus in grey-headed flying foxes (Pteropus poliocephalus)

OBJECTIVE: To examine the susceptibility of the grey-headed flying fox (Pteropus poliocephalus) to Australian bat lyssavirus (ABL), and to provide preliminary observations on the pathogenesis of the disease in flying foxes. PROCEDURE: Ten flying foxes were inoculated intramuscularly with ABL, and four with a bat-associated rabies virus. Inoculated animals were observed daily, and clinical samples collected every 9 to 14 days. Animals with abnormal clinical signs were euthanased, and samples collected for histological, serological, virological and immunohistological examinations. At 3 months post inoculation (PI), all survivors were euthanased, and each submitted to a similar examination. RESULTS: Three ABL-inoculated flying foxes, and two rabies-inoculated animals developed abnormal clinical signs between 15 and 24 days PI. All three ABL-inoculated animals had histological lesions consistent with a lyssavirus infection, and lyssaviral antigen was identified in the central nervous system (CNS) of each. Virus was isolated from the brain of two affected animals. Of the rabies-inoculated flying foxes, both had histological lesions and viral antigen in the CNS. Virus was recovered from the brain of only one. None of the five affected flying foxes developed anti-lyssavirus antibodies, but, by 3 months PI, five of the seven ABL-inoculated survivors, and one of the two rabies virus-inoculated survivors, had seroconverted. The dynamics of the immune responses were quite variable. CONCLUSIONS: The response of flying foxes to ABL, administered by a peripheral route of inoculation, was similar to that of bats inoculated peripherally with bat-derived rabies viruses.     

Genetic analysis of European bat lyssavirus type 1 isolates from France

European bat lyssavirus type 1a (EBLV-1a) was first identified in central France from a serotine bat (Eptesicus serotinus) collected at the end of 2002. Rabies was diagnosed by reference rabies diagnosis methods and molecular tools. Phylogenetic analysis of 14 viral isolates obtained from French bats infected with EBLV-1 between 1989 and the end of 2002 against 47 nucleoprotein sequences showed a north-west to east distribution of EBLV-1a virus and a south to north distribution of EBLV-1b virus, isolates of which could be divided into two groups: group 1 in north-eastern France and group 2 in central and north-western France.     

Characteristics of bat rabies in Alberta.

Rabies in bats was monitored in Alberta from 1971 to 1978 Big brown bats replaced silver-haired bats as the species most frequently reported rabid during these years. Rabies infection was comparatively high among little brown bats in central Alberta in 1973 and has subsequently declined. Only one rabid little brown bat was discovered in southern Alberta which is populated by a different subspecies. Outbreaks of rabies in little brown and big brown bat colonies tended to be brief events. Observations of free-ranging bats with probable furious rabies suggested that bats do not generally identify humans as targets for attack. Independent trends in infection rates suggested that spread of rabies is primarily intraspecific but there is evidence that migratory bats play a role in introduction and maintenance of rabies in northern temperate bat communities. The dynamics of bat rabies in Alberta are discussed.     

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. The man received rapidly a post‐exposure RABV treatment and was put under strict medical supervision.