Borna disease in horses.

Borna disease is a sporadically occurring, progressive viral polioencephalomyelitis that primarily affects horses and sheep. The etiological agent, Borna disease virus (BDV), is an enveloped, single-stranded RNA virus that has been classified in the new virus family Bornaviridae within the order Mononegavirales. Serological evidence of BDV infection has been found in an increasing number of countries throughout the world. After an incubation period lasting a few weeks to several months, BDV infection can cause locomotor and sensory dysfunction followed by paralysis and death. Borna disease is the result of a virus-induced immunopathological reaction. BDV-specific antibodies and viral RNA have been found in humans with various psychiatric disorders.     

The humoral and cellular immune response of sheep against Borna disease virus in endemic and non-endemic areas

Borna Disease (BD) is a mostly fatal disease of horses and sheep endemic in central Europe. Antibodies to Borna disease virus (BDV) have been described in sheep and other species living in BD non-endemic areas. Meaningful clinical BDV serology is hampered by difficulties in defining serological cut-offs, which require the investigation of populations from endemic areas. Here we studied BD serology in sheep from endemic and non-endemic areas of similar geography in Switzerland. Antibodies to BDV antigens were detected by ELISA and indirect immunofluorescence analysis (IFA) only in sera from 3 of 6 sheep with autopsy confirmed BD. One serum was positive by IFA but not by ELISA, while 2 sera were negative in both assays, indicating that not all diseased animals develop BDV specific antibodies. Six % of clinically healthy animals (6/106) from an endemic area and 2% from a non-endemic area (4/192) had serum antibody to either BDV p40 or p24 as detected by ELISA. None of the animals showed a cellular immune response to BDV p40. In some healthy sheep from the endemic area, serum antibody titers to BDV p24 antigen remained elevated over several months without onset of disease symptoms. Infections with either BDV or related viruses may thus occur at low frequency in sheep from non-endemic areas leading to the production of antibodies to BDV antigens. We further propose viral strain differences or environmental factor(s) may determine the clinical outcome.     

Markers of Borna disease virus infection in cats with staggering disease

Borna disease virus (BDV) is a RNA-virus causing neurological disorders in a wide range of mammals. In cats, BDV infection may cause staggering disease. Presently, staggering disease is a tentative clinical diagnosis, only confirmed at necropsy. In this study, cats with staggering disease were investigated to study markers of BDV infection aiming for improvement of current diagnostics. Nineteen cats fulfilled the inclusion criteria based on neurological signs and pathological findings. In 17/19 cats, BDV infection markers (BDV-specific antibodies and/or BDV-RNA) were found, and antibodies in serum (13/16, 81%) were the most common marker. BDV-RNA was found in 11/19 cats (58%). In a reference population without neurological signs, 4/25 cats were seropositive (16%). The clinical history and neurological signs in combination with presence of BDV infection markers, where serology and rRT-PCR on blood can be helpful tools, improve the diagnostic accuracy in the living cat.     

A serosurvey of Borna disease virus infection in wild rats by a capture ELISA

For a serological diagnostic test for Borna disease (BD), we developed a capture ELISA with specificity and sensitivity based on detection of antibodies against BD virus (BDV) p40 protein. Using our capture ELISA system, the antibody response of rats inoculated intracerebrally with BDV at 4 weeks after birth showed a sharp increase from 1 to 4 weeks postinoculation (p.i.) and a steady level after 5 weeks p.i. To investigate prevalence of BDV infection among wild rats, we examined sera of Rattus norvegicus in Kami-iso town, Oshima district, Hokkaido, suggesting that rats in this area had not been infected by BDV.     

Borna disease in Switzerland and in the principality of Liechtenstein

Borna disease (BD) is a rare immunopathological disorder of the central nervous system (CNS) caused by infection with Borna disease virus (BDV) and histologically characterized by mononuclear encephalomyelitis. BD primarily affects equines and sheep in well defined endemic areas of central Europe, but BDV infections have also been reported in other host species including humans, as well as in non endemic regions. In this paper recent data on the pathogenesis of BD are reviewed and the current situation in Switzerland and the Principality of Liechtenstein is summarized.     

Seroprevalence of Borna disease virus in domestic animals in Xinjiang, China

To investigate the animals infected with Borna disease virus (BDV) in Xinjiang, China, we examined for BDV antibodies in the sera from groups of 20 horses, sheep and cattle, and from 165 wild rodents (18 species) by ELISA and immunoblot. The serological study disclosed the presence of antibodies to both BDV-p24 and -p40 in the horses (20%) and sheep (25%), whereas no apparent positive reaction was detected either in cattle or rodents. The results suggested that BDV is prevalent in horses and sheep in the district investigated.     

Borna disease virus (BDV) infection in cats. A concise review based on current knowledge

Persistent viral infections of the central nervous system have been the subject of intense interest for decades. One of these viral agents has been identified as Borna disease virus (BDV) of the family Bornaviridae. There have been various reports that link BDV to staggering disease in cats, with symptoms that include ataxia and behavioural disorders, and the disease is often referred to as feline Borna disease. Serological and molecular detection of BDV has been reported at a higher prevalence in cats with neurological disorders in comparison to healthy cats. The transmission route(s) of BDV remain largely unknown, and the hypothesis that BDV is a zoonotic agent is yet to be proven. This review summarises the current knowledge on BDV infection in cats and discusses epidemiological aspects of infection.     

Borna disease: current knowledge and virus detection in France

For over two centuries, Borna disease (BD) has been described as a sporadically occurring infectious meningoencephalomyelitis affecting horses and sheep in Central Europe. Over the last decade, the BD epidemiology has been discussed. Firstly, its geographical distribution seems larger than what was previously thought. Secondly, the disease can affect a large number of warm-blooded animal species, including humans. The aetiological agent is the Boma disease virus (BDV), an enveloped, nonsegmented negative-stranded RNA virus classified in the new virus family Bornaviridae (Mononegavirales order). It can induce severe clinical signs of encephalitis with striking behavioural disturbances and may cause death. BDV genome has recently been detected in France in the blood and brain of several animal species (horses, bovines, foxes).     

Demonstration of continuously seropositive population against Borna disease virus in Misaki feral horses,a Japanese strain: a four-year follow-up study from 1998 to 2001

Borna disease virus (BDV)-specific antibodies were monitored in Misaki feral horses annually for 4 years using an electrochemiluminescence immunoassay (ECLIA). Among 130 horses examined, 35 (26.9%) with an ECLIA count above 1000 once or more were judged as BDV seropositive. Throughout the study period, p24 antibodies were more frequent than p40 antibodies in almost all positive animals. Among the 35 seropositive horses, the ECLIA count was consistently high in 12 cases. Eleven horses seroconverted from negative to positive and 7 underwent reversal. The count in the remaining 95 horses (73.1%) remained low for 4 years and these animals were judged as seronegative.     

Prevalence of circulating antibodies to p10, a non-structural protein of the Borna disease virus in cats with ataxia.

Japanese domestic cats were surveyed for circulating antibodies to the plO and p24 proteins of the Borna disease virus (BDV) by Western blotting. Twenty-four of 52 cats (46.2%) with ataxia and other neurologic symptoms of unknown cause were positive for antibodies to BDV p10 and/or p24. In contrast, cats without neurological symptoms gave a significantly lower prevalence of anti-BDV antibodies to p10 and/or p24 (36 of 152 cats, 23.7%). Thirty specific pathogen-free (SPF) cats tested as controls were uniformly negative to BDV pl0 and p24 antigens. These results suggest that BDV may play a role in ataxia in cats. Additionally, our results suggest that it is necessary to use both p10 and p24 as antigens to detect circulating antibodies to BDV in cats.     

Prevalence of Borna disease virus antibodies in healthy Japanese black cattle in Kyushu

Epidemiological studies have demonstrated that asymptomatic infection of Borna disease virus (BDV) is found in various species of animals in Japan. Recent reports have also revealed that neurological diseases caused by this virus could exist in horses, cattle, a dog, and cats in this country. In this study, we investigated seroprevalence of BDV antibodies in Japanese black cows reared in Kyushu, the southernmost main island of Japan, using ELISA and Western-immunoblotting. Of 101 serum samples, 11 (10.9%) and 21(20.7%) sera were identified as having antibodies to the BDV N and P antigens, respectively. Among the positive sera, three cows (2.9%) were seropositive for both of the antigens. Furthermore, interestingly, only female cows showed antibodies to P, whereas N antibodies were detected in male and female cows with a comparative ratio. Together with previous studies, our results indicate that BDV might be widely spread in cattle raised in Japan. Furthermore, this is the first report to show that beef cattle, Japanese black cattle, have antibodies against a possible zoonotic pathogen, BDV.     

Peripheral and intracerebral T cell immune response in cats naturally infected with Borna disease virus.

Borna disease virus (BDV) is a neurotropic agent with capacity to cause encephalomyelitis in a wide range of animal species, including horses and cats. Recent studies also point to a link between BDV and human neuropsychiatric disorders. The pathogenesis of Borna disease (BD) has been proposed to be immune-mediated, mainly through the effects of cytotoxic T cells. We used flow cytometric analysis in order to characterize the peripheral and intracerebral T cell immune response in cats naturally infected with BDV. Our results show the presence of two different CD8+ cell populations (CD8+low and CD8+high) in the blood, spleen and brain of these cats. In the brain, CD8+low cells predominated over CD8+high cells. Since CD8+low cells have been suggested to represent a non-MHC-restricted T cell population, the recruitment of such cells to the brains of BDV-infected cats could possibly be of importance for the clearance of virus from neurones.     

博尔纳病

博尔纳病是最初发生在德国马匹的一种渐进性病毒性脑脊髓炎,可以感染多种动物,甚至可以感染人类,可通过直接或间接接触分泌物或污染物而感染,其发病率较低,病死率较高。临床症状各不相同,但典型的症状表现在精神、感觉、敏感性、活动性以及自主神经系统方面。本病是一种免疫介导性疾病,可以持续感染。临床上难以做出诊断,需要结合实验室检测来确诊。博尔纳病毒作为非反转录病毒,近年发现能"篡改"人类基因组。论文从博尔纳病的病原学、宿主范围、流行病学、临床症状、解剖病变、免疫力、预防控制等方面进行了系统介绍。     

Detection of antibodies to Borna disease virus in Turkish cats by using recombinant p40.

Recombinant p40 produced by baculovirus was used in an ELISA to screen samples of serum taken from 80 cats in Istanbul. The sera were also analysed for feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV). Antibodies to Borna disease virus- (BDV) p40 were detected in 34 (42-5 per cent) of the 80 cats. Seventy-three per cent of the sera which were positive for FIV and 26 per cent of the sera which were negative for FIV had antibodies to BDV. There was no difference in the percentage of sera which were positive for BDV between the cats that were positive or negative for FeLV. Three of the cats had neurological disease and two of these had antibodies to BDV. Six sera with low, medium or high optical densities (ODS) by ELISA were analysed by Western blotting. Only the sera with medium and high ODS reacted specifically with p40 at a dilution of 1 in 1,000.     

Infection of the enteric nervous system by Borna disease virus (BDV) upregulates expression of Calbindin D-28k

Borna disease virus (BDV) is a neurotropic agent infecting distinct neuronal subpopulations in the central nervous system of various mammalian species possibly including humans. Horses, a major natural host for BDV, show gastrointestinal dysfunctions besides characteristic neurological symptoms. Therefore, we hypothesized that enteric neurons may be targets of BDV replication. The presence of BDV-specific antigen in subpopulations of the ENS was investigated. Four-week-old Lewis rats were infected intracerebrally and sacrificed 4-14 weeks post infection (p.i.). BDV-immunoreactive neurons were found in submucous and myenteric neurons of the proximal colon. Fourteen weeks p.i., the proportion of BDV-positive neurons was 44+/-17 and 24+/-7% in the submucous and myenteric plexus, respectively. The majority of BDV-positive myenteric neurons showed immunoreactivity for choline acetyltransferase. Expression of Calbindin D-28k (CALB) was found in 96% of submucous and 67% of myenteric BDV-immunoreactive neurons. Additionally, the number of CALB-immunoreactive neurons was significantly higher in the myenteric plexus of infected rats compared to controls. These data indicate that BDV infects specific subpopulations of enteric neurons. Therefore, the ENS might serve as a site for BDV replication and as an immunoprivileged reservoir for BDV. In addition, upregulation of CALB in neurons of the myenteric plexus is probably induced during BDV-infection.     

Borna disease virus infection in domestic cats: evaluation by RNA and antibody detection

Borna disease virus (BDV) infection has been suggested to cause spontaneous neurological disease in cats referred to as staggering disease. However the evaluation of BDV infection in neurologically asymptomatic cats remained unclear. In the present study, BDV infected, asymptomatic cats in Tokyo were surveyed both by the presence of plasma antibodies against BDV-p24 and -p40 and by RNA detection in peripheral blood mononuclear cells. Seven of 32 domestic cats (21.9%) were serologically or genetically judged to be BDV-infected. Six cats were positive for anti-BDV antibody and two cats were positive for BDV RNA. Within the 2 RNA-positive cats, only one was positive for anti-BDV antibodies. Furthermore, the findings of anti-BDV-p40 and anti-BDV-p24 antibody-positive cats did not completely overlap. These results suggest that there are neurologically asymptomatic domestic cats infected with BDV present in the Tokyo area.     

High prevalence of Borna disease virus in domestic cats with neurological disorders in Japan

A total of 15 (T-1–T-15) domestic cats with neurological disorders in Tokyo area were examined for association with Borna disease virus (BDV). None had detectable antibodies to feline immunodeficiency virus (FIV), feline leukemia virus, feline infectious peritonitis virus and Toxoplasma gondii, and only cat T-8 had detectable antibody to FIV. Serological and molecular epidemiological studies revealed a significantly high prevalence of BDV infection in these cats: antibodies against BDV p24 and/or p40 proteins in 10/15 (66.7%) and p24 and/or p40 RNA in peripheral blood mononuclear cells in 8/15 (53.3%). Further, in situ hybridization and immunohistochemistry analyses of the autopsied brain samples derived from one of the cats (T-15) revealed BDV RNA predominantly in neuronal cells in restricted regions, such as olfactory bulb and medulla of cerebrum. Thus, BDV is present in Japanese domestic cats with neurological disorders at a high prevalence.     

Expression of interferon gamma in the brain of cats with natural Borna disease virus infection

Borna disease virus (BDV) is a neurotropic, negative-stranded RNA virus, which causes a non-suppurative meningoencephalomyelitis in a wide range of animals. In cats, BDV infection leads to staggering disease. In spite of a vigorous immune response the virus persists in the central nervous system (CNS) in both experimentally and naturally infected animals. Since the CNS is vulnerable to cytotoxic effects mediated via NK-cells and cytotoxic T-cells, other non-cytolytic mechanisms such as the interferon (IFN) system is favourable for viral clearance. In this study, IFN-γ expression in the brain of cats with clinical signs of staggering disease (N=12) was compared to the expression in cats with no signs of this disease (N=7) by quantitative RT-PCR. The IFN-γ expression was normalised against the expression of three reference genes (HPRT, RPS7, YWHAZ). Cats with staggering disease had significantly higher expression of IFN-γ compared to the control cats (p-value ≤ 0.001). There was no significant difference of the IFN-γ expression in BDV-positive (N=7) and -negative (N=5) cats having clinical signs of staggering disease. However, as BDV-RNA still could be detected, despite an intense IFN-γ expression, BDV needs to have mechanisms to evade this antiviral immune response of the host, to be able to persist.