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.     

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.     

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).     

Bat Rabies in Canada 1963-1967

Six hundred and twenty-eight insectivorous bats originating from seven provinces were submitted to this Institute for rabies diagnosis between August 1, 1963 and December 31, 1967. Brain tissue was examined by the fluorescent antibody technique and the mouse infectivity test was carried out with brain, salivary gland, interscapular adipose tissue and kidney samples. Rabies virus was detected in 44 bats, 29 of which were from Ontario, 12 from British Columbia and three from Manitoba. Most of the positive cases were diagnosed in summer months. Seven species were represented among the specimens found to be rabid; there were 32 big brown bats, three hoary bats, three silver-haired bats, two little brown bats, one eastern pipistrelle, one Keen myotis and one red bat. Another bat which was not identified also proved to be infected with rabies.     

Identification of focus areas for Australian Bat Lyssavirus potential exposure prevention in the Metro North Hospital and Health Service region

Australian Bat Lyssavirus (ABLV) is a fatal rabies‐like disease spread to humans from bats. All people who report bat bites or scratches in Queensland are considered potentially exposed to ABLV and are followed up and treated to prevent ABLV. Preventing members of the public intentionally interacting with bats will reduce the number of potential exposures to ABLV. In order to target public health messaging, this study examines the epidemiology of potential ABLV exposures in Metro North Hospital and Health Service (HHS), a region of Queensland that encompasses metro and rural areas in the south‐east of the state. People who intentionally handled bats during the study period were more likely to be adult (93%), male (60%), scratched (51%) by a megabat (72%) and been potentially exposed while rescuing the bat when it was trapped or injured (72%). The number of potential exposures reported in Queensland has increased since 2013; the same year, a Queensland child died of ABLV. Seasonally, exposures are more common during bat breeding and nursing periods when bats are more active (summer to autumn). Although there were more notifications in a band stretching north from the inner city to northern metro suburbs, notification rates were higher in large rural statistical areas in the north of the HHS. These data will be used to develop geographically targeted ABLV prevention messaging for the general public.     

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.     

Bat Rabies in Ontario

Rabies has been diagnosed for the first time in the bat population of Ontario. In the course of a study involving 72 bats from 24 counties of the province, five big brown bats (E. fuscus) were found to be infected with rabies through the mouse inoculation test. At the present time, it does not look as if bats have been connected with the epizootic of sylvatic rabies in Ontario.

La rage est apparue pour la première fois chez les chauves-souris en Ontario. Au cours d'une étude qui a porté sur 72 de ces animaux provenant de 24 comtés de la province, l'inoculation d'animaux de laboratoire a permis confirmer la présence de la maladie chez cinq grosses chauves-souris brunes (E. fuscus). A date, il ne semble toutefois pas que les chauves-souris soient impliquées dans l'épizootie de rage sylvatique qui sévit en Orntario.

     

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.     

Recurrent Temporary Paralysis Reported After Human Rabies Post‐Exposure Prophylaxis

Adverse events can occur after rabies post‐exposure prophylaxis (PEP), and linkage to causality is often difficult to determine. We report a case of recurrent temporary paralysis that began immediately after the initiation of rabies PEP in a man exposed to a bat. The recurrent temporary paralysis first occurred in the patient after his initial dose and then again after day 3 of his rabies PEP. The PEP was terminated prior to a serologic response. The patient continued to experience numerous discrete episodes of temporary paralysis for over two years.     

Exposure of hooded capuchin monkeys (Cebus apella cay) to a rabid bat at a zoological park.

On 27 May 1999, a big brown bat (Eptesicus fuscus) was discovered on an island exhibit at the Denver Zoo that contained a troop of 15 hooded capuchin monkeys (Cebus apella cay). The monkeys were attacking the bat when it was discovered. The bat was collected and humanely euthanatized without direct handling and submitted to the Colorado Department of Public Health and Environment Virology Laboratory for rabies evaluation. The monkeys had not been vaccinated against rabies virus. The next day, the laboratory confirmed that the bat was positive for rabies. The recommendations from the Colorado Department of Public Health and Environment and the Centers for Disease Control and Prevention were to euthanatize the monkeys or quarantine them and comply with the human nonvaccinated postexposure protocol. A 1-ml dose of a killed rabies vaccine was administered i.m. in the hip on each of days 2, 7, 12, 19, and 33 postexposure, and a single dose of human rabies immune globulin was administered i.m. 5 days postexposure. Blood was collected under anesthesia in order to evaluate the immune response after rabies vaccination from six monkeys 5 days postexposure, six monkeys 19 days postexposure (five of the six monkeys were the same monkeys bled 5 days postexposure), 15 monkeys 67 days postexposure, and 13 monkeys approximately 1 yr postexposure. All of the monkeys developed and maintained levels of rabies virus neutralizing antibody above 0.05 IU/ml by 67 days postexposure. Although a serologic titer of 0.05 IU/ml indicates an adequate human response after rabies vaccination, no similar information is available for nonhuman primates. To date, none of the monkeys has succumbed to rabies.     

Faeces as a novel material to estimate lyssavirus prevalence in bat populations

Rabies is caused by infection with a lyssavirus. Bat rabies is of concern for both public health and bat conservation. The current method for lyssavirus prevalence studies in bat populations is by oral swabbing, which is invasive for the bats, dangerous for handlers, time‐consuming and expensive. In many situations, such sampling is not feasible, and hence, our understanding of epidemiology of bat rabies is limited. Faeces are usually easy to collect from bat colonies without disturbing the bats and thus could be a practical and feasible material for lyssavirus prevalence studies. To further explore this idea, we performed virological analysis on faecal pellets and oral swabs of seven serotine bats (Eptesicus serotinus) that were positive for European bat 1 lyssavirus in the brain. We also performed immunohistochemical and virological analyses on digestive tract samples of these bats to determine potential sources of lyssavirus in the faeces. We found that lyssavirus detection by RT‐qPCR was nearly as sensitive in faecal pellets (6/7 bats positive, 86%) as in oral swabs (7/7 bats positive, 100%). The likely source of lyssavirus in the faeces was virus excreted into the oral cavity from the salivary glands (5/6 bats positive by immunohistochemistry and RT‐qPCR) or tongue (3/4 bats positive by immunohistochemistry) and swallowed with saliva. Virus could not be isolated from any of the seven faecal pellets, suggesting the lyssavirus detected in faeces is not infectious. Lyssavirus detection in the majority of faecal pellets of infected bats shows that this novel material should be further explored for lyssavirus prevalence studies in bats.     

Experimental infection of Artibeus intermedius with a vampire bat rabies virus

Experimental infection of Artibeus intermedius, the great fruit-eating bat, was performed with vampire bat rabies isolates. Bats (n = 35) were captured in the wild and quarantined prior to experimental infection. No rabies antibodies were detected by rapid fluorescent focus inhibition test (RFFIT) prior to infection. Three doses of rabies virus (RV) and three different routes of infection were used. One out of 35 bats died without showing any clinical signs at day 14 and was positive for rabies. None of the 34 other bats showed clinical signs for rabies, but high antibody titers were detected post-inoculation, suggesting either innate immune response to the vampire bat rabies virus or possible pre-exposure to RV and inoculation leading to a booster effect. Rabies virus was detected by hemi-nested RT-PCR (hnRT-PCR) in the brain (n = 3), stomach (n = 1) of bats that were negative by immunofluorescence and that survived rabies infection. The bat that died on day 14 was positive by hnRT-PCR on the brain, heart and liver. These results suggest that either previous non-lethal exposure to RV or natural low susceptibility to vampire bat viruses somehow protected Artibeus intermedius from clinical rabies infection leading to a marginal lethality effect on this bats species population in the wild.     

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.     

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.     

First isolation of EBLV-2 in Germany

In Europe, rabies in bats is caused by European Bat Lyssavirus (EBLV) type 1 (EBLV-1) or type 2 (EBLV-2) which form two distinct genotypes (gt 5 and 6) within the genus Lyssavirus of the family of Rhadoviridae. Spill-over infections of EBLV in humans have caused fatal rabies encephalitis and highlighted the relevance of this wildlife disease for public health. The vast majority of the 831 European bat rabies cases reported between 1977 and 2006 were identified as EBLV-1. Only few virus isolates originating from Switzerland, The Netherlands and the United Kingdom were characterized as EBLV-2. Here we report the first EBLV-2 case detected in Germany in a Daubenton's bat (Myotis daubentonii) in August 2007. The bat showed clinical signs of disorders of the central nervous system and subsequently tested positive for rabies. The virus was isolated and characterized as EBLV-2 based on its antigen pattern and by nucleotide sequencing. Phylogenetic analysis indicated an association to EBLV-2 isolates from Switzerland which correlates with the origin of the bat close to the Swiss border.     

Experimentally induced rabies in four cats inoculated with a rabies virus isolated from a bat

Four cats were inoculated IM with rabies virus isolated from the salivary gland of a naturally infected big brown bat (Eptesicus fuscus). The 4 cats developed clinical signs of rabies after a median incubation period of 42 days. The median duration of clinical illness was 5 days. Results of fluorescent antibody evaluation, mouse inoculation, and tissue culture isolation indicated large differences in virus concentrations in various areas of the CNS of individual cats. These differences also were observed between cats. Rabies virus was isolated from the salivary glands and saliva of 2 cats; urinary bladder was the only other nonneural tissue found infected. Our observations indicated that cat rabies can be caused by bat rabies virus; that cats thus infected have infectious saliva during aggressive behavior and can therefore transmit the disease; and that adequate specimens of hippocampus, cerebellum, and brain stem are essential for reliable determination of rabies infection. The findings support recommendations for regular rabies vaccination of cats, even in areas of rabies-free terrestrial mammals.     

Detection of the Severe Acute Respiratory Syndrome‐Related Coronavirus and Alphacoronavirus in the Bat Population of Taiwan

Bats have been demonstrated to be natural reservoirs of severe acute respiratory syndrome coronavirus (SARS CoV) and Middle East respiratory syndrome (MERS) CoV. Faecal samples from 248 individuals of 20 bat species were tested for partial RNA‐dependent RNA polymerase gene of CoV and 57 faecal samples from eight bat species were tested positive. The highest detection rate of 44% for Scotophilus kuhlii, followed by 30% for Rhinolophus monoceros. Significantly higher detection rates of coronaviral RNA were found in female bats and Scotophilus kuhlii roosting in palm trees. Phylogenetic analysis classified the positive samples into SARS‐related (SARSr) CoV, Scotophilus bat CoV 512 close to those from China and Philippines, and Miniopterus bat CoV 1A‐related lineages. Coronaviral RNA was also detected in bat guano from Scotophilus kuhlii and Myotis formosus flavus on the ground and had potential risk for human exposure. Diverse bat CoV with zoonotic potential could be introduced by migratory bats and maintained in the endemic bat population in Taiwan.