Human Borna disease virus 1 (BoDV-1) encephalitis cases in the north and east of Germany.

In 2021, three encephalitis cases due to the Borna disease virus 1 (BoDV-1) were diagnosed in the north and east of Germany. The patients were from the states of Thuringia, Saxony-Anhalt, and Lower Saxony. All were residents of known endemic areas for animal Borna disease but without prior diagnosed human cases. Except for one recently detected case in the state of Brandenburg, all >30 notified cases had occurred in, or were linked to, the southern state of Bavaria. Of the three detected cases described here, two infections were acute, while one infection was diagnosed retrospectively from archived brain autopsy tissue samples. One of the acute cases survived, but is permanently disabled. The cases were diagnosed by various techniques (serology, molecular assays, and immunohistology) following a validated testing scheme and adhering to a proposed case definition. Two cases were classified as confirmed BoDV-1 encephalitis, while one case was a probable infection with positive serology and typical brain magnetic resonance imaging, but without molecular confirmation. Of the three cases, one full virus genome sequence could be recovered. Our report highlights the need for awareness of a BoDV-1 etiology in cryptic encephalitis cases in all areas with known animal Borna disease endemicity in Europe, including virus-endemic regions in Austria, Liechtenstein, and Switzerland. BoDV-1 should be actively tested for in acute encephalitis cases with residence or rural exposure history in known Borna disease-endemic areas.

Infections of horses and shrews with Bornaviruses in Upper Austria: a novel endemic area of Borna disease.

Borna disease, a lethal infection with Borna disease virus-1 (BoDV-1), was diagnosed in four horses from Upper Austria in 2015 and 2016. All cases occurred in winter (two cases in February 2015 and two cases in December 2016), and the maximal distance of the affected stables was 17 km. To demonstrate whether the causative agent was also harbored by its reservoir host, the bicolored white-toothed shrew (Crocidura leucodon), 28 shrews from this geographic area were collected in 2015 and investigated for the presence of BoDV-1. The shrew species were identified according to taxonomic clues and molecular barcodes. Affected horses and all shrews were investigated using histology, immunohistochemistry (IHC) and reverse transcription PCR. The horses exhibited severe nonpurulent encephalitis. Large amounts of BoDV-1 antigen were identified in their CNS. Among the 28 shrews, nine were identified as C. leucodon and 13 as Sorex araneus (Common shrew; Eurasian shrew). Six C. leucodon (66.7%) and one S. araneus (7.7%) had BoDV-1 infections. In accordance with previous findings, the IHC of C. leucodon exhibited a high amount of viral antigen in many neural and extraneural tissues. By contrast, the single positive S. araneus had an exclusively neural staining pattern. Of all positive samples, whole-genome BoDV-1 sequences were generated. The acquired sequences of the affected shrews were not identical to each other and clustered around the sequences of the diseased horses belonging, surprisingly, to the German 'strain V' cluster.

GC-MS-Based Metabonomic Profiling Displayed Differing Effects of Borna Disease Virus Natural Strain Hu-H1 and Laboratory Strain V Infection in Rat Cortical Neurons.

Borna disease virus (BDV) persists in the central nervous systems of a wide variety of vertebrates and causes behavioral disorders. Previous studies have revealed that metabolic perturbations are associated with BDV infection. However, the pathophysiological effects of different viral strains remain largely unknown. Rat cortical neurons infected with human strain BDV Hu-H1, laboratory BDV Strain V, and non-infected control (CON) cells were cultured in vitro. At day 12 post-infection, a gas chromatography coupled with mass spectrometry (GC-MS) metabonomic approach was used to differentiate the metabonomic profiles of 35 independent intracellular samples from Hu-H1-infected cells (n = 12), Strain V-infected cells (n = 12), and CON cells (n = 11). Partial least squares discriminant analysis (PLS-DA) was performed to demonstrate discrimination between the three groups. Further statistical testing determined which individual metabolites displayed significant differences between groups. PLS-DA demonstrated that the whole metabolic pattern enabled statistical discrimination between groups. We identified 31 differential metabolites in the Hu-H1 and CON groups (21 decreased and 10 increased in Hu-H1 relative to CON), 35 differential metabolites in the Strain V and CON groups (30 decreased and 5 increased in Strain V relative to CON), and 21 differential metabolites in the Hu-H1 and Strain V groups (8 decreased and 13 increased in Hu-H1 relative to Strain V). Comparative metabonomic profiling revealed divergent perturbations in key energy and amino acid metabolites between natural strain Hu-H1 and laboratory Strain V of BDV. The two BDV strains differentially alter metabolic pathways of rat cortical neurons in vitro. Their systematic classification provides a valuable template for improved BDV strain definition in future studies.

Borna disease virus infection in cats: ABCD guidelines on prevention and management.

OVERVIEW: Borna disease virus (BDV) has a broad host range, affecting primarily horses and sheep, but also cattle, ostriches, cats and dogs. In cats, BDV may cause a non-suppurative meningoencephalomyelitis ('staggering disease'). INFECTION: The mode of transmission is not completely elucidated. Direct and indirect virus transmission is postulated, but BDV is not readily transmitted between cats. Vectors such as ticks may play a role and shrews have been identified as a potential reservoir host. Access to forested areas has been reported to be an important risk factor for staggering disease. DISEASE SIGNS: It is postulated that BDV may infect nerve endings in the oropharynx and spread via olfactory nerve cells to the central nervous system. A strong T-cell response may contribute to the development of clinical disease. Affected cats develop gait disturbances, ataxia, pain in the lower back and behavioural changes. DIAGNOSIS: For diagnostic purposes, detection of viral RNA by reverse transcription PCR in samples collected from cats with clinical signs of Borna disease can be considered diagnostic. Serology is of little value; cats without signs of Borna disease may be seropositive and yet not every cat with BDV infection has detectable levels of antibodies. HUMAN INFECTION: A hypothesis that BDV infection may be involved in the development of selected neurological disorders in man could not be confirmed. A research group within the German Robert Koch Institute studied the potential health threat of BDV to humans and concluded that BDV was not involved in the aetiology of human psychiatric diseases.

Health care professionals at risk of infection with Borna disease virus - evidence from a large hospital in China (Chongqing).

BACKGROUND: Human Borna disease virus (BDV) infections have recently been reported in China. BDV causes cognitive and behavioural disturbances in animals. The impact on human mental disorders is subject to debate, but previous studies worldwide have found neuropsychiatric patients more frequently infected than healthy controls. A few isolates were recovered from severely depressed patients, but contagiousness of BDV strain remains unknown. METHOD: We addressed the risk of infection in health care settings at the first affiliated hospital of Chongqing Medical University (CQMU), located in downtown Chongqing, a megacity in Southwest China. Between February 2012 and March 2013, we enrolled 1529 participants, of whom 534 were outpatients with major depressive disorder (MDD), 615 were hospital personnel, and 380 were healthy controls who underwent a health check. Infection was determined through BDV-specific circulating immune complexes (CIC), RNA, and selective antibodies (blood). RESULTS: One-fifth of the hospital staff (21.8%) were found to be infected (CIC positive), with the highest prevalence among psychiatry and oncology personnel, which is twice as many as were detected in the healthy control group (11.1%), and exceeds the prevalence detected in MDD patients (18.2%). CONCLUSION: BDV circulates unnoticed in hospital settings in China, putting medical staff at risk and warranting clarification of infection modes and introduction of prevention measures.

Discrepancy in the diagnosis of avian Borna disease virus infection of Psittaciformes by protein analysis of feather calami and enzyme-linked immunosorbent assay of plasma antibodies.

The present study compares diagnosis of avian Borna disease virus (ABV) infection of psittacine birds by Western blot of bornaviral proteins in dried feather stems with the detection of anti-bornaviral protein antibodies to bornaviral proteins in plasma by enzyme-linked immunosorbent assay (ELISA). The detection of ABV proteins P40 and P24 in feather calami by Western blotting was possible even after storage of the dried feathers for several years at ambient temperature. Serological identification of anti-bornaviral antibodies may fail (e.g., in young birds, hatched from infected parents), whereas bornaviral P40 and P24 proteins were detected in feather stems. This failure can last at least 10 months after the birds are hatched. In some older birds (>5 years), ABV protein was only detectable in the brain, but not in some peripheral tissues, suggesting that the immune system had succeeded in removing the infecting ABV from tissues outside the brain. These results show that a combination of feather stem analysis for the presence of bornaviral proteins by Western blot combined with serological detection of anti-bornaviral antibodies by ELISA is the most reliable procedure for the detection of a bornaviral infection.

Borna disease virus (BDV) infection in psychiatric patients and healthy controls in Iran.

BACKGROUND: Borna disease virus (BDV) is an evolutionary old RNA virus, which infects brain and blood cells of humans, their primate ancestors, and other mammals. Human infection has been correlated to mood disorders and schizophrenia, but the impact of BDV on mental-health still remains controversial due to poor methodological and cross-national comparability. METHOD: This first report from the Middle East aimed to determine BDV infection prevalence in Iranian acute psychiatric disorder patients and healthy controls through circulating immune complexes (CIC), antibodies (Ab) and antigen (pAg) in blood plasma using a standardized triple enzyme immune assay (EIA). Samples of 314 subjects (114 psychiatric cases, 69 blood donors, and 131 healthy controls) were assayed and data analyzed quantitatively and qualitatively. RESULTS: CICs revealed a BDV prevalence of one third (29.5%) in healthy Iranian controls (27.5% controls; 33.3% blood donors). In psychiatric patients CIC prevalence was higher than in controls (40.4%) and significantly correlating with bipolar patients exhibiting overt clinical symptoms (p = 0.005, OR = 1.65). CIC values were significantly elevated in bipolar (p = 0.001) and major depressive disorder (p = 0.029) patients as compared to controls, and in females compared to males (p = 0.031). CONCLUSION: This study supports a similarly high prevalence of subclinical human BDV infections in Iran as reported for central Europe, and provides again an indication for the correlation of BDV infection and mood disorders. Further studies should address the morbidity risk for healthy carriers and those with elevated CIC levels, along with gender disparities.

Heat stress is a potent stimulus for enhancing rescue efficiency of recombinant Borna disease virus.

Recently developed vector systems based on Borna disease virus (BDV) hold promise as platforms for efficient and stable gene delivery to the central nervous system (CNS). However, because it currently takes several weeks to rescue recombinant BDV (rBDV), an improved rescue procedure would enhance the utility of this system. Heat stress reportedly enhances the rescue efficiency of other recombinant viruses. Here, heat stress was demonstrated to increase the amount of BDV genome in persistently BDV-infected cells without obvious cytotoxicity. Further analyses suggested that the effect of heat stress on BDV infection is not caused by an increase in the activity of BDV polymerase. More cells in which BDV replication occurs were obtained in the initial phase of rBDV rescue by using heat stress than when it was not used. Thus, heat stress is a useful improvement on the published rescue procedure for rBDV. The present findings may accelerate the practical use of BDV vector systems in basic science and the clinic and thus enable broader adoption of this viral vector, which is uniquely suited for gene delivery to the CNS.

The bicolored white-toothed shrew Crocidura leucodon (HERMANN 1780) is an indigenous host of mammalian Borna disease virus.

Borna disease (BD) is a sporadic neurologic disease of horses and sheep caused by mammalian Borna disease virus (BDV). Its unique epidemiological features include: limited occurrence in certain endemic regions of central Europe, yearly varying disease peaks, and a seasonal pattern with higher disease frequencies in spring and a disease nadir in autumn. It is most probably not directly transmitted between horses and sheep. All these features led to the assumption that an indigenous virus reservoir of BDV other than horses and sheep may exist. The search for such a reservoir had been unsuccessful until a few years ago five BDV-infected shrews were found in a BD-endemic area in Switzerland. So far, these data lacked further confirmation. We therefore initiated a study in shrews in endemic areas of Germany. Within five years 107 shrews of five different species were collected. BDV infections were identified in 14 individuals of the species bicolored white-toothed shrew (Crocidura leucodon, HERMANN 1780), all originating from BD-endemic territories. Immunohistological analysis showed widespread distribution of BDV antigen both in the nervous system and in epithelial and mesenchymal tissues without pathological alterations. Large amounts of virus, demonstrated by presence of viral antigen in epithelial cells of the oral cavity and in keratinocytes of the skin, may be a source of infection for natural and spill-over hosts. Genetic analyses reflected a close relationship of the BDV sequences obtained from the shrews with the regional BDV cluster. At one location a high percentage of BDV-positive shrews was identified in four consecutive years, which points towards a self-sustaining infection cycle in bicolored white-toothed shrews. Analyses of behavioral and population features of this shrew species revealed that the bicolored white-toothed shrew may indeed play an important role as an indigenous host of BDV.

Evidence for natural Borna disease virus infection in healthy domestic animals in three areas of western China.

Borna disease virus (BDV) is a non-cytolytic, neurotropic RNA virus that can infect many vertebrate species, including humans. To date, BDV infection has been reported in a range of animal species across a broad global geographic distribution. However, a systematic epidemiological survey of BDV infection in domesticated animals in China has yet to be performed. In current study, BDV RNA and antibodies in 2353 blood samples from apparently healthy animals of eight species (horse, donkey, dog, pig, rabbit, cattle, goat, sheep) from three areas in western China (Xinjiang province, Chongqing municipality, and Ningxia province) were assayed using reverse transcription qPCR (RT-qPCR) and ELISA assay. Brain tissue samples from a portion of the BDV RNA- and/or antibody-positive animals were subjected to RT-qPCR and western blotting. As a result, varying prevalence of BDV antibodies and/or RNA was demonstrated in various animal species from three areas, ranging from 4.4 % to 20.0 %. Detection of BDV RNA and/or antibodies in Chongqing pigs (9.2 %) provided the first known evidence of BDV infection in this species. Not all brain tissue samples from animals whose blood was BDV RNA and/or antibody positive contained BDV RNA and protein. This study provides evidence that BDV infection among healthy domestic animal species is more widespread in western China than previously believed.

Evidence for Borna disease virus infection in neuropsychiatric patients in three western China provinces.

Borna disease virus (BDV) is a non-cytolytic, neurotropic RNA virus that can infect a wide variety of vertebrate species from birds and primates to humans. Several studies have been carried out to investigate whether BDV is associated with neuropsychiatric diseases. However, this association is still inconclusive. Two panels of subjects consisting of 1,679 various neuropsychiatric patients and healthy people from three western China provinces were enrolled in this study. BDV p24 or p40 RNA in peripheral blood mononuclear cells (PBMCs) were detected in the first panel of 1,481 subjects using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and cerebrospinal fluid (CSF) samples from the BDV RNA-positive individuals were subjected to BDV p24 antibodies testing by enzyme-linked immunosorbent assay (ELISA). BDV p24 or p40 RNA in PBMCs and p24 antibodies in plasma were detected in the second panel of 198 subjects by RT-qPCR and Western blot. A higher prevalence for BDV RNA was demonstrated in patients with viral encephalitis (6.70%), Guillain-Barré syndrome (6.70%), schizophrenia (9.90%) and chronic fatigue syndrome (CFS) (12.70%) compared to healthy controls in the first panel. CSF p24 antibodies were demonstrated in three viral encephalitis patients, two schizophrenia patients and two major depressive disorder (MDD) patients. The prevalences of p24 antibodies in plasma from patients with viral encephalitis (13.24%), multiple sclerosis (25.00%) and Parkinson's disease (22.73%) were significantly higher than healthy controls. This study demonstrates that BDV infection also exists in humans from three western China provinces, and suggests the involvement of the contribution of BDV in the aetiology of Chinese patients with some neuropsychiatric disorders, including viral encephalitis, schizophrenia, CFS, multiple sclerosis and Parkinson's disease.

Epidemiology and host spectrum of Borna disease virus infections.

Borna disease virus (BDV) has gained lot of interest because of its zoonotic potential, ability to introduce cDNA of its RNA transcripts into host genomes, and ability to cause severe neurobehavioural diseases. Classical Borna disease is a progressive meningoencephalomyelitis in horses and sheep, known in central Europe for centuries. According to current knowledge, BDV or a close relative also infects several other species, including humans at least occasionally, in central Europe and elsewhere, but the existence of potential 'human Borna disease' with its suspected neuropsychiatric symptoms is highly controversial. The recent detection of endogenized BDV-like genes in primate and various other vertebrate genomes confirms that at least ancient bornaviruses did infect our ancestors. The epidemiology of BDV is largely unknown, but accumulating evidence indicates vectors and reservoirs among small wild mammals. The aim of this review is to bring together the current knowledge on epidemiology of BDV infections. Specifically, geographical and host distribution are addressed and assessed in the critical light of the detection methods used. We also review some salient clinical aspects.

Landscape features and reservoir occurrence affecting the risk for equine infection with Borna disease virus.

Borna disease (BD) is a severe endemic and fatal disorder caused by the neurotropic Borna disease virus (BDV) which mainly occurs in horses and sheep. Borna disease virus belongs to the order Mononegavirales, which includes many reservoir-bound viruses with high zoonotic and pathogenic properties including the filoviruses and lyssaviruses. Clinically manifest BD occurs in endemic areas of Germany, Switzerland, Liechtenstein, and Austria. A seasonal accumulation of cases in spring and summer, incidences that vary from year to year, and the recent detection of BDV in bicolored shrews (Crocidura leucodon) in Swiss endemic areas argue for a natural reservoir. We established a geographic information system analysis of the distribution of 485 equine BD cases in Bavarian (Germany) endemic areas and of the occurrence of 285 records of C. leucodon captured in Bavaria. Boosted regression trees were used to identify driving factors of habitat choice and virus prevalence. The distribution model of C. leucodon and the prevalence model for BDV had very good accuracy. Mean annual precipitation <900 mm, mean annual temperatures of 8 C, elevation <350 m, low forest cover, and a high percentage of urban fabric and arable land describe the optimal habitat for C. leucodon. Occurrence probability of C. leucodon was significantly higher in Bavarian BDV-endemic areas than in random areas in Bavaria. The prevalence of BD was higher in urban areas with annual mean precipitation of 800-900 mm, annual mean temperature of 8 C, and elevation >500 m. Our results indicate that the distribution model can accurately predict BD occurrence. Based on these results, practical safety precautions could be derived. The BDV model represents a suitable system for reservoir-bound, neurotropic Mononegavirales because it allows analyzing ecologic and biologic aspects that determine virus abundance, maintenance in reservoir species, and transmission to end host species.

Serological evidence for infections with Borna disease virus in Turkey.

Distribution of Borna disease virus (BDV) infection outside endemic areas has been studied in several countries. We examined serum samples for anti-BDV antibodies in purebred racing horses and other domestic animals in Turkey. In total serum samples of 437 animals including 282 horses, 50 sheep, 25 goats, 50 cattle, and 30 cats were tested by indirect immunofluorescence assay (IFA). Anti-BDV antibodies were detected in 4.9% of horses, 12% of sheep, 4% of goats, 14% of cattle and 6.6% of cats. No statistical difference was observed between seroprevalence in Arabic and English purebred horses from four different racing centers (p > 0.05). Antibody titers ranged between 1:10 and 1:320. The highest antibody titers were found in sheep and horses and the lowest titer in cattle. Clinical symptoms of Borna disease were not observed in any animal of any species examined. This study confirms the presence of anti-BDV antibodies in racing horses as well as cat population in Turkey. Moreover anti-BDV antibodies are demonstrated for the first time in sheep, goats and cattle in Turkey.

Birds and bornaviruses.

In 2008, avian bornaviruses (ABV) were identified as the cause of proventricular dilatation disease (PDD). PDD is a significant condition of captive parrots first identified in the late 1970s. ABV infection has subsequently been shown to be widespread in wild waterfowl across the United States and Canada where the virus infects 10-20% of some populations of ducks, geese and swans. In most cases birds appear to be healthy and unaffected by the presence of the virus; however, infection can also result in severe non-suppurative encephalitis and lesions similar to those seen in parrots with PDD. ABVs are genetically diverse with seven identified genotypes in parrots and one in canaries. A unique goose genotype (ABV-CG) predominates in waterfowl in Canada and the northern United States. ABV appears to be endemic in North American waterfowl, in comparison to what appears to be an emerging disease in parrots. It is not known whether ABV can spread between waterfowl and parrots. The discovery of ABV infection in North American waterfowl suggests that European waterfowl should be evaluated for the presence of ABV, and also as a possible reservoir species for Borna disease virus (BDV), a related neurotropic virus affecting horses and sheep in central Europe. Although investigations have suggested that BDV is likely derived from a wildlife reservoir, for which the shrew and water vole are currently prime candidates, we suggest that the existence of other mammalian and avian reservoirs should not be discounted.

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.

The influence of Borna disease viral infection on dairy cow reproduction.

We investigated the influence of Borna disease virus (BDV) infection on the clinical state of dairy cows. Sera from 149 cows were examined using enzyme-linked immunosorbent assay and western blotting detect antibodies to the BDV-nucleoprotein antigen. Among 149 investigated cows, 25 (16.8%) showed a positive reaction to BDV antigen. No significant difference existed in milk production or medical history between seropositive and seronegative cows. Although the estrus cycle appeared normal even in the seropositive cows, the frequency of artificial insemination and calving-to-conception intervals significantly increased in seropositive cows. Therefore, fertilization failure was recognized in the BDV-antibody positive cows.