Avian Bornavirus Research Articles

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.

Fluoroscopic evaluation of gastrointestinal transit time in African Grey parrots

Contrast studies are commonly performed to evaluate the gastrointestinal tract. With ever increasing knowledge of Proventricular Dilation Disease (PDD) in psittacines, and its association with avian bornavirus, radiographic assessment of...

The X proteins of bornaviruses interfere with type I interferon signalling

Borna disease virus (BDV) is a neurotropic, negative-stranded RNA virus causing persistent infection and progressive neurological disorders in a wide range of warm-blooded animals. The role of the small non-structural X protein in viral pathogenesis is not completely understood. Here we investigated whether the X protein of BDV and avian bornavirus (ABV) interferes with the type I interferon (IFN) system, similar to other non-structural proteins of negative-stranded RNA viruses. In luciferase reporter assays, we found that the X protein of various bornaviruses interfered with the type I IFN system at all checkpoints investigated, in contrast to previously reported findings, resulting in reduced type I IFN secretion.

The Detection of Avian Bornavirus Within Psittacine Eggs

Avian bornavirus (ABV) is a known cause of proventricular dilatation disease in parrots and encephalitis in waterfowl and is a significant cause of both morbidity and mortality in captive birds. Transmission is thought to occur primarily by the fecal-oral route. In an aviary setting, controlling the disease involves a thorough understanding of the complete transmission cycle, including determining whether vertical transmission occurs. In this study, vertical transmission of ABV was evaluated by using 61 eggs obtained from birds in 2 aviaries where proventricular dilatation disease was prevalent, and the presence of ABV had been confirmed by fecal reverse transcription-polymerase chain reaction by using a primer set designed to detect ABV M protein. The contents of these eggs were then tested for the presence of ABV RNA. Of the eggs tested, 10 were determined to contain ABV RNA. These eggs ranged from apparently nonviable to those that contained developing embryos. ABV was detected in the brain tissue of 2 embryos. It remains to be proven that infected chicks can hatch from these eggs to complete the vertical transmission cycle; however, these findings suggest that vertical transmission of ABV may occur.

Vertical Transmission of Avian Bornavirus in Psittaciformes: Avian Bornavirus RNA and Anti-Avian Bornavirus Antibodies in Eggs, Embryos, and Hatchlings Obtained from Infected Sun Conures (Aratinga solstitialis)

Fertilized eggs were obtained from four pairs of sun conures (Aratinga solstitialis) infected with avian bornavirus (ABV) genotype 2, as determined by the sequence of the P24 gene. ABV RNA could be detected in early embryos of all four pairs. ABV RNA also was detected in brain, liver, and eyes of late-stage embryos of one of the pairs (Pair 4) and in blood of a 2-wk-old hatchling of this pair, demonstrating that vertical transmission can occur. ABV RNA could be detected in the liver but not in the brain or eyes of the late-stage embryos of another pair (Pair 3). Although it could be detected in the undeveloped eggs of the female parent and 8-day-old embryos, bornaviral RNA could not be found in the brain and liver of the late-stage embryos or in feathers and blood of young (5-9-wk-old) hatchlings of a third pair (Pair 2). At 11 wk, ABV RNA could be detected again in feathers and blood of these hatchlings and in the brain of one of the hatchlings of Pair 2 that suddenly died. ABV RNA could however be detected in throat swabs of the 5- and 9-wk-old hatchlings and their parents (Pair 2). Although the continued presence of ABV RNA in feathers and blood below the detection level of the reverse transcription-PCR used cannot be excluded, this result also may be attributable to feeding by the infected parents. Analysis by enzyme-linked immunosorbent assay showed that egg yolks and serum of late-stage embryos contain variable amounts of non-neutralizing anti-ABV-P40, -P10, -P24, and -P16 antibodies, the ratio of which reflected the antibody ratio in the serum of the female parent. Antibodies against the viral glycoprotein, which are considered neutralizing in mammals, and against ABV RNA polymerase were not detected. Whereas 5-wk-old hatchlings of the pair (Pair 2) that produced ABV RNA-free late-stage embryos were free of anti-ABV antibodies, such antibodies could be detected again in the serum of these hatchlings at 9 wk of age, before the age that bornaviral RNA could again be detected in feathers and blood. At 16 wk, these antibodies became abundant. The finding that late-stage embryos, presumably free of ABV RNA, can be obtained from eggs from infected parents suggests that hand- or foster-raising of such birds may be a method to obtain birnavirus-free offspring from some infected birds.

Vertical Transmission of Avian Bornavirus in Psittaciformes: Avian Bornavirus RNA and Anti-Avian Bornavirus Antibodies in Eggs, Embryos, and Hatchlings Obtained from Infected Sun Conures (Aratinga solstitialis)

Vertical Transmission of Avian Bornavirus in Psittaciformes: Avian Bornavirus RNA and Anti-Avian Bornavirus Antibodies in Eggs, Embryos, and Hatchlings Obtained from Infected Sun Conures (Aratinga solstitialis)

Efficient isolation of avian bornaviruses (ABV) from naturally infected psittacine birds and identification of a new ABV genotype from a salmon-crested cockatoo (Cacatua moluccensis)

Avian bornaviruses (ABV) have been discovered in 2008 as the causative agent of proventricular dilatation disease (PDD) in psittacine birds. To date, six ABV genotypes have been described in psittacines. Furthermore, two additional but genetically different ABV genotypes were recognized in non-psittacine birds such as canary birds and wild waterfowl. This remarkable genetic diversity poses a considerable challenge to ABV diagnosis, since polymerase chain reaction (PCR) assays may fail to detect distantly related or as yet unknown genotypes. In this study we investigated the use of virus isolation in cell culture as a strategy for improving ABV diagnosis. We found that the quail fibroblast cell line CEC-32 allows very efficient isolation of ABV from psittacine birds. Isolation of ABV was successful not only from organ samples but also from cloacal and pharyngeal swabs and blood samples collected intra vitam from naturally infected parrots. Importantly, using this experimental approach we managed to isolate a new ABV genotype, termed ABV-7, from a salmon-crested cockatoo (Cacatua moluccensis). Phylogenetic analysis showed that ABV-7 is most closely related to the psittacine genotypes ABV-1, -2, -3, and -4 and clearly distinct from genotypes ABV-5 and -6. Our successful identification of ABV-7 emphasizes the necessity to consider the high genetic diversity when trying to diagnose ABV infections with high reliability and further shows that classical virus isolation may represent a useful diagnostic option, particularly for the detection of new ABV genotypes.

Widespread avian bornavirus infection in mute swans in the Northeast United States.

Avian bornavirus (ABV) matrix (M) genes were detected by RT-PCR on brain tissue obtained from 192 mute swans harvested from several Northeastern states. A RT-PCR product was detected in 45 samples. Sequencing of the PCR products confirmed the presence of ABV belonging to the 'goose' genotype. The prevalence of positive samples ranged from 28% in Michigan to 0% in northern New York State. Two Rhode Island isolates were cultured. Their M, N, and X-P gene sequences closely matched recently published sequences from Canada geese.

Identification of Mixed Infections with Different Genotypes of Avian Bornaviruses in Psittacine Birds with Proventricular Dilatation Disease

Proventricular dilatation disease (PDD) is a fatal, progressive neurological disorder of psittacine birds, which is caused by a single-stranded RNA virus, the avian bornavirus (ABV). The disease pattern includes lymphoplasmacytic inflammation of the central, peripheral and autonomic nervous system. Seven avian bornavirus genotypes have been identified during the last years. So far only monoinfections with a single genotype of ABV have been attributed to PDD cases. However, after a recent survey discovered a case of a double infection with two different ABV genotypes, this seemed to indicate the need for a more systematic search for mixed infections. Brain specimens from 21 psittacine birds affected with PDD were examined. Aim of the investigation was to generate partial ABV sequences of a part of the matrix protein (M) gene and to evaluate whether sequences of more than one ABV genotype were present. RNA was extracted, and subjected to reverse transcriptase PCR with primer pairs generating a partial sequence of the matrix protein (M) gene, followed by a cloning procedure. Ten clones per case were sequenced in order to elucidate whether sequences characteristic for one or more than one genotype were present. In 19 of 21 cases clear M gene sequences could be generated; in two cases nucleic acid amplification failed. Seven birds were infected with ABV 2 and nine with ABV 4, representing the predominant genotypes in Europe. Two cases showed a mixed infection with ABV 2 and ABV 4, and one case a mixed infection with ABV 2 and ABV 6. These results suggest that the molecular cloning method is a useful tool for distinguishing between single and multiple infection events by different ABV genotypes.

Avian bornavirus in the urine of infected birds.

Avian bornavirus (ABV) causes proventricular dilatation disease in multiple avian species. In severe clinical disease, the virus, while primarily neurotropic, can be detected in many organs, including the kidneys. We postulated that ABV could be shed by the kidneys and found in the urine of infected birds. Immunohistochemical staining demonstrated viral N and P proteins of ABV within the renal tubules. We adapted a nonsurgical method of urine collection for use in parrots known to be shedding ABV in their droppings. We obtained urine without feces, and results were compared with swabs of fresh voided feces. Reverse transcription-polymerase chain reaction assay performed on these paired samples from five birds indicated that ABV was shed in quantity in the urine of infected birds, and a single sample was urine-positive and fecal-negative. We suggest that urine sampling may be a superior sample for detection of birds shedding ABV, and advocate that additional birds, known to be shedding or infected with ABV, should be investigated via this method.

Identification of Mixed Infections with Different Genotypes of Avian Bornaviruses in Psittacine Birds with Proventricular Dilatation Disease

Identification of Mixed Infections with Different Genotypes of Avian Bornaviruses in Psittacine Birds with Proventricular Dilatation Disease

Complete Genome Sequence of Avian Bornavirus Genotype 1 from a Macaw with Proventricular Dilatation Disease

Avian bornaviruses (ABV) were first detected and described in 2008. They are the etiologic agents of proventricular dilatation disease (PDD), a frequently fatal neurologic disease of captive parrots. Seven ABV genogroups have been identified worldwide from a variety of sources, and that number may increase as surveillance for novel bornaviruses continues. Here, we report the first complete sequence of a genogroup 1 avian bornavirus (ABV1).

Detection of Avian bornavirus 5 RNA in Eclectus roratus with feather picking disorder

Avian bornavirus (ABV) was discovered recently in parrots with proventricular dilatation disease (PDD), a fatal neurological disease. Although ABV has been shown to be a causative agent of PDD, its virological characteristics are largely unknown. Here we report the detection of ABV genotype 5 RNA in an Eclectus roratus with feather picking disorder (FPD). Interestingly, although the bird was persistently infected with ABV5 for at least 8 months, it had no clinical signs of PDD. Although it remains unclear whether ABV5 is associated with FPD, these findings raise the importance of epidemiological studies of birds with diseases other than PDD.

Follow-Up Investigations on Different Courses of Natural Avian Bornavirus Infections in Psittacines

To study the course of natural avian bornavirus (ABV) infection, 63 psittacines of three bird collections where ABV had been demonstrated were investigated over a period of 1 yr. The psittacines were clinically observed and swabs of crop and cloaca as well as serum samples were collected three separate times at intervals of 2-6 mo. According to the results of detection of ABV RNA by reverse transcriptase polymerase chain reaction (RT-PCR) and of anti-ABV antibodies by indirect immunofluorescence assay (IIFA), 43 of the birds were found to be infected with ABV. Based on variations in virus shedding and antibody production in combination with the occurrence of proventricular dilatation disease (PDD) -related clinical signs, pathological findings, and lethal outcome, four different groups of infected psittacines and a fifth group of noninfected psittacines were identified. Group 1 comprised six birds with various courses of ABV infection and forms of clinical PDD. Groups 2-4 included all birds with subclinical ABV infections: Group 2 contained 13 birds that were consistently (subgroup A, 6 birds) or inconsistently (subgroup B, 7 birds) ABV positive by PCR and serology; group 3 was composed of 13 psittacines exhibiting only anti-ABV antibodies; and 8 birds that had positive ABV RNA detection in crop and cloaca, but did not develop anti-ABV specific antibodies, were classified in group 4. Twenty-three out of the 63 psittacines remained free of detectable ABV RNA or anti-ABV antibodies over the whole observation period (group 5). Based on the results, it seems that birds with high ABV RNA load in crop and cloaca combined with high anti-ABV antibodies have a high risk of the development of PDD, indicating that the humoral antibodies do not protect against the disease. The meaning of the detection of ABV RNA and antibodies at a low and inconsistent level for the single bird as well as for the epidemiology of the ABV infection remained unclear in this field study and needs to be further investigated.

Follow-Up Investigations on Different Courses of Natural Avian Bornavirus Infections in Psittacines

Follow-Up Investigations on Different Courses of Natural Avian Bornavirus Infections in Psittacines

Pathogenesis of Avian Bornavirus in Experimentally Infected Cockatiels

Avian bornavirus (ABV) is the presumed causative agent of proventricular dilatation disease (PDD), a major fatal disease in psittacines. However, the influencing factors and pathogenesis of PDD are not known and natural ABV infection exhibits remarkable variability. We investigated the course of infection in 18 cockatiels that were intracerebrally and intravenously inoculated with ABV. A persistent ABV infection developed in all 18 cockatiels, but, as in natural infection, clinical disease patterns varied. Over 33 weeks, we simultaneously studied seroconversion, presence of viral RNA and antigens, infectious virus, histopathologic alterations, and clinical signs of infection in the ABV-infected birds. Our study results further confirm the etiologic role of ABV in the development of PDD, and they provide basis for further investigations of the pathogenetic mechanisms and disease-inducing factors for the development of PDD.

ボルナウイルス

Bornaviridae is an enveloped animal virus carrying an 8.9 kb non-segmented, negative-strand RNA genome. The genus bornavirus contains two members infecting vertebrates, Borna disease virus (BDV) and avian bornavirus (ABV), which could preferably infect the nervous systems. BDV causes classical Borna disease, a progressive meningoencephalomyelitis, in horses and sheep, and ABV is known to induce proventricular dilatation disease, a fatal disease characterized by a lymphocytic, plasmacytic inflammation of central and peripheral nervous tissues, in multiple avian species. Recent evidences have demonstrated that bornavirus is unique among RNA viruses as they not only establish a long-lasting, persistent infection in the nucleus, but also integrate their own DNA genome copy into the host chromosome. In this review, I outline the recent knowledge about the unique virological characteristics of bornaviruses, as well as the diseases caused by the infection of BDV and ABV.

Vertical Transmission of Avian Bornavirus in Psittacines

Vertical Transmission of Avian Bornavirus in Psittacines

Proventricular Dilatation Disease in Cockatiels (Nymphicus hollandicus) After Infection With a Genotype 2 Avian Bornavirus

An isolate of genotype 2 avian bornavirus (ABV) was recovered from a cockatiel (Nymphicus hollandicus) that was euthanatized for an unrelated lesion and showing no clinical evidence of proventricular dilatation disease (PDD). On histopathologic examination, mild inflammatory lesions were present in the heart and brain, but gastrointestinal lesions characteristic of classic PDD were not observed. To investigate if this ABV2 isolate had reduced virulence, the virus was propagated in duck embryo fibroblasts and inoculated into 2 adult cockatiels by the oral and intramuscular routes. One bird developed clinical signs on day 33 and was euthanatized on day 36. The second challenged bird developed clinical signs on day 41 and was euthanatized on day 45. At necropsy, the proventriculus of both birds was slightly enlarged. Histopathologic examination showed lesions typical of PDD in the brain, spinal cord, heart, adrenal gland, and intestine. A control, uninoculated cockatiel was apparently healthy when euthanatized on day 50. These results show that ABV2 is now the second ABV genotype to be formally shown to cause PDD.

Occurrence of avian bornavirus infection in captive psittacines in various European countries and its association with proventricular dilatation disease

A total of 1442 live birds and 73 dead birds out of 215 bird collections in Spain, Germany, Italy, the UK and Denmark were tested for avian bornavirus (ABV) infection by four different methods. The majority of the birds were psittacines belonging to 54 different genera of the order Psittaciformes. In total, 22.8% of the birds reacted positive for ABV in at least one of the tests. Combined testing of swabs from the crop and cloaca, and serum for the diagnosis of ABV infection in live birds revealed that virus shedding and antibody production coincided in only one-fifth of the positive birds so that the examination of these three samples is recommended for reliable ABV diagnosis. By statistical analysis of this large number of samples, the ABV infection proved to be highly significant (P <0.001) associated with histopathologically confirmed proventricular dilatation disease (PDD) in dead birds as well as with clinically assumed PDD in live birds. However, ABV infection was also detected in psittacines without pathological lesions or clinical signs of PDD. Twelve non-psittacine birds belonging to the genera Aburria, Ciconia, Geopelia, Leucopsar and Pavo were tested negative for ABV infection. Within the order of Psittaciformes, birds belonging to 33 different genera reacted positive for ABV. In 16 of these psittacine genera, the ABV infection was demonstrated for the first time. The present study emphasizes the widespread occurrence of clinically variable ABV infections in Europe by analysing a large number of specimens from a broad range of bird species in several assays.