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1.
BMC Vet Res ; 17(1): 172, 2021 Apr 23.
Article in English | MEDLINE | ID: mdl-33892731

ABSTRACT

BACKGROUND: Canine distemper virus (CDV) is an enveloped negative-strand RNA virus that exhibits a high mutation rate and continuously expands the range of hosts. Notably, CDV has infected giant panda with spill over from viral reservoirs in canines. Giant pandas (Ailuropoda melanoleuca), especially captive pandas, are known to be susceptible to natural infection with CDV. The high fatality rate of CDV poses a serious threat to the safety of the giant panda population. However, vaccines or drugs for canine distemper in giant pandas have not been developed to date. Therefore, a rapid test that can achieve accurate onsite detection of CDV is important to enable the timely implementation of control measures. In this study, we established a nucleic acid visualization assay for targeting the CDV N gene by using combines reverse transcription recombinase polymerase amplification with a closed vertical flow visualization strip (RT-RPA-VF). RESULTS: The RT-RPA-VF assay does not require sophisticated equipment, and it was determined to provide rapid detection at 35 °C for 30 min, while the limit of detection was 5 × 101 copies/µl RNA transcripts and 100.5 TCID50 ml- 1 viruses. The results showed that the assay was high specific to CDV and had no cross-reactivity with other viruses infecting the giant panda. Compared with RT-qPCR, RT-RPA-VF assay had a sensitivity of 100% and a specificity of 100% in 29 clinical samples. The coincidence rate between RT-RPA-VF and RT-qPCR was 100% (kappa = 1), indicating that the RT-RPA-VF assay possessed good diagnostic performance on clinical samples. CONCLUSIONS: The RT-RPA-VF provides a novel alternative for the simple, sensitive, and specific identification of CDV and showed great potential for point of care diagnostics for captive and wild giant panda.


Subject(s)
Distemper Virus, Canine/genetics , Distemper Virus, Canine/isolation & purification , Distemper/diagnosis , Nucleic Acid Amplification Techniques/veterinary , Ursidae/virology , Animals , Distemper/virology , Nucleic Acid Amplification Techniques/methods , RNA, Viral , Real-Time Polymerase Chain Reaction/veterinary , Reverse Transcription , Sensitivity and Specificity
2.
Arch Virol ; 166(5): 1521-1524, 2021 May.
Article in English | MEDLINE | ID: mdl-33721099

ABSTRACT

Polyomaviruses are ancient DNA viruses that infect several species of animals. While recognition of the family Polyomaviridae has grown rapidly, there are few studies that consider their potential association with disease. Carnivora are a diverse and widespread order affected by polyomaviruses (PyVs) that have co-evolved with their hosts for millions of years. PyVs have been identified in sea lions, raccoons, badgers, Weddell seals, and dogs. We have discovered a polyomavirus, tentatively named "Ursus americanus polyomavirus 1" (UaPyV1) in black bears (Ursus americanus). UaPyV1 was detectable in various tissues of six out of seven bears submitted for necropsy. Based on viral phylogenetic clustering and detection of the virus in multiple individuals, we suggest that black bears are the natural hosts for UaPyV1. In this albeit small group, there is no clear relationship between UaPyV1 infection and any specific disease.


Subject(s)
Polyomavirus Infections/veterinary , Polyomavirus/classification , Tumor Virus Infections/veterinary , Ursidae/virology , Animals , Base Sequence , DNA, Viral/genetics , Genome, Viral/genetics , Phylogeny , Polyomavirus/genetics , Polyomavirus Infections/pathology , Polyomavirus Infections/virology , Tumor Virus Infections/pathology , Tumor Virus Infections/virology , United States , Viral Proteins/genetics
3.
Front Cell Infect Microbiol ; 11: 820144, 2021.
Article in English | MEDLINE | ID: mdl-35198456

ABSTRACT

A feline panleukopenia virus (FPV), Giant panda/CD/2018, was isolated from a captive giant panda with mild diarrhea in 2018 in Chengdu, China, and further identified via indirect immunofluorescence assay (IFA), transmission electron microscopy (TEM) observation, and genetic analysis. Phylogenetic analysis based on the complete VP2 nucleotide sequences showed that it shared high homology with Chinese FPV isolates and grouped within FPV cluster 1. One unique substitution Gly(G)299Glu(E) in the capsid protein VP2 was first identified with Giant panda/CD/2018. The presence of the G299E substitution is notable as it is located on the top region of the interconnecting surface loop 3, which may be involved in controlling the host range and antigenicity of FPV. These findings first demonstrate that FPV with natural point mutation G299E in the VP2 gene is prevalent in giant panda and suggest that etiological surveillance and vaccination among all giant pandas are urgently needed to protect this endangered species against FPV infection.


Subject(s)
Feline Panleukopenia Virus , Parvoviridae Infections , Ursidae , Animals , Animals, Zoo/virology , Capsid Proteins/genetics , China/epidemiology , Diarrhea/veterinary , Diarrhea/virology , Feline Panleukopenia Virus/genetics , Parvoviridae Infections/veterinary , Phylogeny , Ursidae/virology
4.
PLoS One ; 15(12): e0244056, 2020.
Article in English | MEDLINE | ID: mdl-33332429

ABSTRACT

Viral infections were investigated in American black bears (Ursus americanus) from Nevada and northern California with and without idiopathic encephalitis. Metagenomics analyses of tissue pools revealed novel viruses in the genera Circoviridae, Parvoviridae, Anelloviridae, Polyomaviridae, and Papillomaviridae. The circovirus and parvovirus were of particular interest due to their potential importance as pathogens. We characterized the genomes of these viruses and subsequently screened bears by PCR to determine their prevalence. The circovirus (Ursus americanus circovirus, UaCV) was detected at a high prevalence (10/16, 67%), and the chaphamaparvovirus (Ursus americanus parvovirus, UaPV) was found in a single bear. We showed that UaCV is present in liver, spleen/lymph node, and brain tissue of selected cases by in situ hybridization (ISH) and PCR. Infections were detected in cases of idiopathic encephalitis and in cases without inflammatory brain lesions. Infection status was not clearly correlated with disease, and the significance of these infections remains unclear. Given the known pathogenicity of a closely related mammalian circovirus, and the complex manifestations of circovirus-associated diseases, we suggest that UaCV warrants further study as a possible cause or contributor to disease in American black bears.


Subject(s)
Animal Diseases/virology , Circoviridae/pathogenicity , Encephalitis, Viral/virology , Parvoviridae/pathogenicity , Ursidae/virology , Animal Diseases/epidemiology , Animals , Brain/virology , Circoviridae/genetics , Circoviridae/isolation & purification , DNA Barcoding, Taxonomic , Encephalitis, Viral/epidemiology , Liver/virology , Metagenome , Parvoviridae/genetics , Parvoviridae/isolation & purification , Spleen/virology , United States
5.
PLoS One ; 15(12): e0243180, 2020.
Article in English | MEDLINE | ID: mdl-33259561

ABSTRACT

This study investigates the occurrence of erythematous lip lesions in a captive sun bear population in Cambodia, including the progression of cheilitis to squamous cell carcinoma, and the presence of Ursid gammaherpesvirus 1. Visual assessment conducted in 2015 and 2016 recorded the prevalence and severity of lesions. Opportunistic sampling for disease testing was conducted on a subset of 39 sun bears, with histopathological examination of lip and tongue biopsies and PCR testing of oral swabs and tissue biopsies collected during health examinations. Lip lesions were similarly prevalent in 2015 (66.0%) and 2016 (68.3%). Degradation of lip lesion severity was seen between 2015 and 2016, and the odds of having lip lesions, having more severe lip lesions, and having lip lesion degradation over time, all increased with age. Cheilitis was found in all lip lesion biopsies, with histological confirmation of squamous cell carcinoma in 64.5% of cases. Single biopsies frequently showed progression from dysplasia to neoplasia. Eighteen of 31 sun bears (58.1%) had at least one sample positive for Ursid gammaherpesvirus 1. The virus was detected in sun bears with and without lip lesions, however due to case selection being strongly biased towards those showing lip lesions it was not possible to test for association between Ursid gammaherpesvirus 1 and lip squamous cell carcinoma. Given gammaherpesviruses can play a role in cancer development under certain conditions in other species, we believe further investigation into Ursid gammaherpesvirus 1 as one of a number of possible co-factors in the progression of lip lesions to squamous cell carcinoma is warranted. This study highlights the progressively neoplastic nature of this lip lesion syndrome in sun bears which has consequences for captive and re-release management. Similarly, the detection of Ursid gammaherpesvirus 1 should be considered in pre-release risk analyses, at least until data is available on the prevalence of the virus in wild sun bears.


Subject(s)
Lip Diseases/veterinary , Lip/pathology , Ursidae , Animals , Cambodia/epidemiology , Carcinoma, Squamous Cell/epidemiology , Carcinoma, Squamous Cell/pathology , Carcinoma, Squamous Cell/veterinary , Disease Progression , Erythema/epidemiology , Erythema/pathology , Erythema/veterinary , Female , Gammaherpesvirinae/classification , Gammaherpesvirinae/genetics , Gammaherpesvirinae/isolation & purification , Lip Diseases/epidemiology , Lip Diseases/pathology , Lip Neoplasms/epidemiology , Lip Neoplasms/pathology , Lip Neoplasms/veterinary , Male , Phylogeny , Prevalence , Risk Factors , Ursidae/virology
6.
Virus Res ; 277: 197846, 2020 02.
Article in English | MEDLINE | ID: mdl-31870796

ABSTRACT

The presence of a novel adenovirus (AdV) was detected by PCR and sequencing, in the internal organs of a captive polar bear that had died in the Budapest zoo. The virus content of the samples proved to be high enough to allow for conventional Sanger sequencing on PCR-amplified genomic fragments. With this approach, the sequence of the entire genome of the putative polar bear adenovirus 1 (PBAdV-1) was obtained. Although the genome was found to be short, consisting of 27,952 base pairs merely, with a relatively balanced G + C content of 46.3 %, its organisation corresponded largely to that of a typical mastadenovirus. Every genus-common gene could be identified except that of protein IX. The short E3 region of the PBAdV-1 consisted of two novel, supposedly type-specific ORFs only, whereas no homologue of any of the E3 genes, usually conserved in mastadenoviruses, such as for example that of the 12.5 K protein, were present. In the E4 region, only the highly conserved gene of the 34 K protein was found besides two novel ORFs showing no homology to any known E4 ORFs. In silico sequence analysis revealed putative splicing donor and acceptor sites in the genes of the E1A, IVa2, DNA-dependent DNA polymerase, pTP, 33 K proteins, and also of U exon protein, all being characteristic for mastadenoviruses. Phylogenetic calculations, based on various proteins, further supported that the newly-detected PBAdV is the representative of a new species within the genus Mastadenovirus, and may represent the evolutionary lineage of adenoviruses that coevolved with carnivorans.


Subject(s)
Adenoviridae Infections/veterinary , Genome, Viral , Mastadenovirus/classification , Phylogeny , Ursidae/virology , Adenoviridae Infections/virology , Animals , Animals, Zoo/virology , DNA, Viral/genetics , Female , Mastadenovirus/isolation & purification , Sequence Analysis, DNA , Viral Proteins/genetics , Whole Genome Sequencing
7.
Emerg Infect Dis ; 25(12): 2303-2306, 2019 12.
Article in English | MEDLINE | ID: mdl-31742520

ABSTRACT

We report influenza A(H1N1)pdm09 virus infection in a captive giant panda in Hong Kong. The viral load peaked on day 1 and became undetectable on day 5, and an antibody response developed. Genome analysis showed 99.3%-99.9% nucleotide identity between the virus and influenza A(H1N1)pdm09 virus circulating in Hong Kong.


Subject(s)
Influenza A Virus, H1N1 Subtype/classification , Influenza A Virus, H1N1 Subtype/genetics , Orthomyxoviridae Infections/epidemiology , Orthomyxoviridae Infections/virology , Ursidae/virology , Animals , Cell Line , Genome, Viral , Genomics , Hemagglutinin Glycoproteins, Influenza Virus/genetics , Hong Kong/epidemiology , Male , Phylogeny , Viral Load
8.
J Wildl Dis ; 55(2): 499-503, 2019 04.
Article in English | MEDLINE | ID: mdl-30289326

ABSTRACT

We tested sera of 24 free-ranging European brown bears ( Ursus arctos) from six regions of Slovakia for antibodies to 10 viral agents. We tested sera by an indirect fluorescence antibody test for antibodies to canine distemper virus (CDV), canine coronavirus (CCV), canine parvovirus type 2 (CPV-2), canine adenovirus, canine parainfluenza virus type 2 (CPIV-2), and canine herpesvirus type 1 (CHV-1). We used an enzyme-linked immunosorbent assay for detection of antibodies to hepatitis E virus, bluetongue virus, West Nile virus (WNV), and Aujeszky's disease virus (ADV). We detected antibodies to CDV, CHV-1, CPV-2, CPIV-2, CCV, WNV, and ADV in seven (29%), three (12%), two (8%), two (8%), one (4%), one (4%), and one (4%) bear, respectively. Evidence of exposure of free-ranging European brown bears to CCV and ADV has not been reported.


Subject(s)
Antibodies, Viral/blood , Ursidae/virology , Virus Diseases/veterinary , Animals , Seroepidemiologic Studies , Slovakia/epidemiology , Virus Diseases/blood , Virus Diseases/epidemiology , Virus Diseases/virology
9.
Virus Res ; 259: 46-53, 2019 01 02.
Article in English | MEDLINE | ID: mdl-30385363

ABSTRACT

Herpesvirus infection was investigated in black bears (Ursus americanus) with neurological signs and brain lesions of nonsuppurative encephalitis of unknown cause. Visible cytopathic effects (CPE) could only be observed on days 3-5 post-infection in HrT-18G cell line inoculated with bear tissue extracts. The observed CPE in HrT-18G cells included syncytia, intranuclear inclusions, and cell detachments seen in herpesvirus infection in vitro. Herpesvirus-like particles were observed in viral culture supernatant under the electron microscope, however, capsids ranging from 60 nm to 100 nm in size were often observed in viral cultures within the first two passages of propagation. Herpesvirus infection in the bear tissues and tissue cultures were detected by PCR using degenerate primers specific to the DNA polymerase gene (DPOL) and glycoprotein B gene (gB). DNA sequencing of the amplicon revealed that the detected herpesvirus has 94-95% identity to Ursid gammaherpesvirus 1 (UrHV-1) DNA sequences of DPOL. Phylogenetic analysis of DPOL sequences indicates that black bear herpesviruses and UrHV-1 are closely related and have small distances to members of Rhadinovirus. Interestingly, black bear herpesvirus infections were also found in bears without neurological signs. The DPOL DNA sequence of black bear herpesviruses detected in neurological bears were similar to the those detected in the non-neurological bears. However, the gB DNA sequence detected from the neurological bear is different from non-neurological bear and has only 64.5%-70% identity to each other. It is possible that at least two different types of gammaherpesviruses are present in the U. americanus population or several gammaherpesviruses exist in ursine species.


Subject(s)
Animal Diseases/virology , Gammaherpesvirinae/physiology , Herpesviridae Infections/veterinary , Ursidae/virology , Animal Diseases/pathology , Animals , Cell Line , Cytopathogenic Effect, Viral , DNA, Viral , Female , Gammaherpesvirinae/classification , Gammaherpesvirinae/isolation & purification , Gammaherpesvirinae/ultrastructure , Male , Phylogeny , Sequence Analysis, DNA
10.
Arch Virol ; 164(2): 653-656, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30569277

ABSTRACT

The complete genome of a bear picornavirus 1 (BePV-1) in the viscera of an Asian black bear (Ursus thibetanus) from China was characterized using viral metagenomics and RT-PCR/Sanger sequencing. The genome of BePV1 is 6703 nt long, contains a type-IV IRES 5'UTR with the '8-like' motif, encodes a 2053-aa-long polyprotein showing a 3-4-4 organization pattern and two 2A genes. BePV-1 showed the highest overall genome nucleotide sequence identity of 71.7% to a picornavirus genome from an Arctic ringed seal (Phoca hispida) from Canada, classified as a member of the species Aquamavirus A, currently the only one in the genus Aquamavirus. Phylogenetic and genetic distance analyses of P1 and 3D indicated that Asian bear picornavirus (aquamavirus B) represents the second sequenced member of the genus Aquamavirus.


Subject(s)
Picornaviridae Infections/veterinary , Picornaviridae/classification , Picornaviridae/isolation & purification , Seals, Earless/virology , Ursidae/virology , 5' Untranslated Regions , Animals , Base Sequence , China , Genome, Viral , Molecular Sequence Data , Open Reading Frames , Phylogeny , Picornaviridae/genetics , Picornaviridae Infections/virology , RNA, Viral/genetics , Viral Proteins/genetics
11.
J Zoo Wildl Med ; 49(3): 599-608, 2018 Sep.
Article in English | MEDLINE | ID: mdl-30212313

ABSTRACT

Infection by equine herpesvirus (EHV) strains (EHV-1, EHV-9) in ursid species, including polar bears ( Ursus maritimus), has been associated with neurological disease and death. A serosurvey of captive exotic equid and polar bear populations in US Association of Zoos and Aquaria institutions was performed to determine the prevalence of EHV strains using quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA) tests. Equid species surveyed included zebra ( Equus spp.), Przewalski's wild horse ( Equus ferus przewalskii), Persian onager ( Equus hemionus), and Somali wild ass ( Equus africanus somaliensis). A questionnaire regarding husbandry and medical variables was distributed to institutions housing polar bears. No polar bears tested positive for EHVs on qPCR of blood or nasal swabs. No exotic equids tested positive for EHVs on qPCR of blood, but two exotic equids ( n = 2/22; 9%) tested positive for EHVs on qPCR of nasal swabs. On ELISA, polar bears infrequently were positive for EHV-1 ( n = 5/38; 13%). Exotic equids were positive for EHV-4 on ELISA more frequently ( n = 30/43; 70%) than for EHV-1 ( n = 8/43; 19%). Nine institutions submitted samples from both exotic equids and polar bears, two of which had both exotic equids and polar bears positive for EHVs by ELISA. Each of these institutions reported that the polar bear and exotic equid exhibits were within 80 m of each other and that risk factors for fomite transmission between exhibits based on husbandry practices were present. One institution that did not house exotic equids had a polar bear test positive for EHV-1 on ELISA, with no history of exposure to exotic equids. Further testing of captive polar bears and exotic equids is recommended, as is modification of husbandry practices to limit exposure of polar bears to exotic equids.


Subject(s)
Equidae/virology , Herpesviridae/isolation & purification , Ursidae/virology , Animals , Animals, Zoo , Data Collection , Enzyme-Linked Immunosorbent Assay/veterinary , Equidae/blood , Female , Herpesviridae Infections/veterinary , Horse Diseases , Horses , Male , Surveys and Questionnaires , United States , Ursidae/blood
12.
Transbound Emerg Dis ; 65(6): 2049-2056, 2018 Dec.
Article in English | MEDLINE | ID: mdl-30179311

ABSTRACT

Canine adenovirus type 1 (CAdV-1) is responsible for infectious canine hepatitis. The disease has been described in captive American black bear (Ursus americanus) and European brown bear (Ursus arctos arctos), with just one recently reported case in a cub of a free-ranging brown bear (Ursus arctos horribilis) from Alaska. The aim of this work is to summarize findings related to presence and associated mortality of CAdV-1 in 21 free-ranging Cantabrian brown bears (Ursus arctos arctos) submitted to necropsy in Asturias and Castilla y León (northwestern Spain) from 1998 to 2018. On the basis of the anatomopathological findings and laboratory results three free-ranging brown bears died due to infectious canine hepatitis, which is to our knowledge the first description of death due to this disease in free-ranging bears in Europe. Gross lesions consisted of petechial haemorrhages and congestion in different internal organs, haemorrhagic fluid in internal cavities, friable and yellowish liver and thickening of gall bladder. Microscopic lesions were observed mainly in liver, kidney and brain and consisted of multifocal necrosis of cells with presence of basophilic intranuclear inclusions. Immunohistochemical (IHC) and real-time polymerase chain reaction (qPCR) techniques were used to assess the presence of CAdV-1 in paraffin-embedded liver samples. Viral antigens were detected by IHC labelling within hepatocytes and Küppfer cells in the three animals. The presence of viral DNA was confirmed by qPCR in one of them. In order to evaluate the circulation of CAdV-1 in brown bears, a retrospective study was performed using both IHC and qPCR techniques in 11 and 12 additional brown bears, respectively. An extra brown bear was found positive by IHC. This study shows that CAdV-1 surveillance of brown bears and sympatric carnivores should be considered as major concern for the monitoring the population evolution throughout time in this endangered species.


Subject(s)
Adenoviruses, Canine/isolation & purification , Ursidae/virology , Adenoviruses, Canine/genetics , Animals , Autopsy , DNA, Viral/isolation & purification , Dogs , Hepatitis, Infectious Canine/mortality , Liver/virology , Polymerase Chain Reaction , Spain
13.
mSphere ; 3(4)2018 07 25.
Article in English | MEDLINE | ID: mdl-30045965

ABSTRACT

Polar bears in captivity can be exposed to opportunistic pathogens not present in their natural environments. A 4-month-old polar bear (Ursus maritimus) living in an isolated enclosure with his mother in the Tierpark Berlin, Berlin, Germany, was suffering from severe abdominal pain, mild diarrhea, and loss of appetite and died in early 2017. Histopathology revealed severe hepatic degeneration and necrosis without evidence of inflammation or inclusion bodies, although a viral infection had been suspected on the basis of the clinical signs. We searched for nucleic acids of pathogens by shotgun high-throughput sequencing (HTS) from genomic DNA and cDNA extracted from tissue and blood. We identified a novel Mastadenovirus and assembled a nearly complete genome from the shotgun sequences. Quantitative PCR (qPCR) revealed that viral DNA was present in various concentrations in all tissues examined and that the highest concentrations were found in blood. Viral culture did not yield cytopathic effects, but qPCR suggested that virus replication was sustained for up to three passages. Positive immunofluorescence staining confirmed that the virus was able to replicate in the cells during early passage. Phylogenetic analysis demonstrated that the virus is highly divergent compared to other previously identified Mastadenovirus members and basal to most known viral clades. The virus was found only in the 4-month-old bear and not in other captive polar bears tested. We surmised, therefore, that the polar bear was infected from an unknown reservoir, illustrating that adenoviral diversity remains underestimated and that cross-species transmission of viruses can occur even under conditions of relative isolation.IMPORTANCE Cross-species transmission of viral pathogens is becoming an increasing problem for captive-animal facilities. This study highlights how animals in captivity are vulnerable to novel opportunistic pathogens, many of which do not result in straightforward diagnosis from symptoms and histopathology. In this study, a novel pathogen was suspected to have contributed to the death of a juvenile polar bear. HTS techniques were employed, and a novel Mastadenovirus was isolated. The virus was present in both the tissue and blood samples. Phylogenetic analysis of the virus at both the gene and genome levels revealed that it is highly divergent to other known mastadenoviruses. Overall, this study shows that animals in isolated conditions still come into contact with novel pathogens, and for many of these pathogens, the host reservoir and mode of transmission are yet to be determined.


Subject(s)
Adenoviridae Infections/veterinary , Mastadenovirus/classification , Mastadenovirus/isolation & purification , Ursidae/virology , Adenoviridae Infections/virology , Animal Structures/virology , Animals , Animals, Zoo , Berlin , Genome, Viral , Mastadenovirus/genetics , Mastadenovirus/growth & development , Real-Time Polymerase Chain Reaction , Sequence Analysis, DNA , Virus Cultivation , Virus Replication
14.
J Wildl Dis ; 54(3): 642-645, 2018 07.
Article in English | MEDLINE | ID: mdl-29498898

ABSTRACT

We diagnosed infectious canine hepatitis in a free-ranging brown bear ( Ursus arctos horribilis) cub from Alaska, US, found dead in October 2015. Intranuclear inclusion bodies were present in hepatocytes, and immunohistochemistry showed reactivity to adenoviral antigens. Sequencing of the hexon protein of adenovirus showed 100% identity to canine adenovirus 1.


Subject(s)
Adenoviruses, Canine/isolation & purification , Hepatitis, Infectious Canine/pathology , Ursidae/virology , Alaska/epidemiology , Animals , DNA, Viral/genetics , DNA, Viral/isolation & purification , Dogs , Fatal Outcome , Female , Hepatitis, Infectious Canine/epidemiology , Hepatitis, Infectious Canine/virology
15.
Vet Microbiol ; 212: 87-92, 2017 Dec.
Article in English | MEDLINE | ID: mdl-29173594

ABSTRACT

Hepatitis E virus (HEV) is the causative agent of hepatitis E, an emerging infectious disease of humans. HEV infections have also been described in various animal species. Whereas domestic pigs and wild boars are well-known animal reservoirs for HEV, the knowledge on natural HEV infection in zoo animals is scarce so far. Here, we analysed 244 sera from 66 mammal species derived from three zoos in Germany using a commercial double antigen sandwich ELISA. HEV-specific antibodies were detected in 16 animal species, with the highest detection rates in suids (33.3%) and carnivores (27.0%). However, RNA of the human pathogenic HEV genotypes 1-4 was not detected in the serum samples from suids or carnivores. Using a broad spectrum RT-PCR, a ratHEV-related sequence was identified in a sample of a female Syrian brown bear (Ursus arctos syriacus). Subsequent serum samples within a period of five years confirmed a HEV seroconversion in this animal. No symptoms of hepatitis were recorded. In a follow-up investigation at the same location, closely related ratHEV sequences were identified in free-living Norway rats (Rattus norvegicus), whereas feeder rats (Rattus norvegicus forma domestica) were negative for HEV-specific antibodies and RNA. Therefore, a spillover infection of ratHEV from free-living Norway rats is most likely. The results indicate that a wide range of zoo animals can be naturally infected with HEV or HEV-related viruses. Their distinct role as possible reservoir animals for HEV and sources of HEV infection for humans and other animals remains to be investigated.


Subject(s)
Disease Reservoirs/veterinary , Hepatitis Antibodies/blood , Hepatitis E virus/immunology , Hepatitis E/veterinary , Ursidae/virology , Animals , Animals, Zoo , Female , Germany , Hepatitis E/transmission , Hepatitis E/virology , Hepatitis E virus/genetics , Hepatitis E virus/isolation & purification , Humans , Phylogeny , Rats , Seroepidemiologic Studies , Zoonoses
16.
Virol J ; 14(1): 207, 2017 10 27.
Article in English | MEDLINE | ID: mdl-29078783

ABSTRACT

BACKGROUND: Polyomaviruses infect a wide variety of mammalian and avian hosts with a broad spectrum of outcomes including asymptomatic infection, acute systemic disease, and tumor induction. METHODS: Viral metagenomics and general PCR methods were used to detected viral nucleic acid in the samples from a diseased and healthy giant pandas. RESULTS: A novel polyomavirus, the giant panda polyomavirus 1 (GPPyV1) from the nasal cavity of a dead giant panda (Ailuropoda melanoleuca) was characterized. The GPPyV1 genome is 5144 bp in size and reveals five putative open-reading frames coding for the classic small and large T antigens in the early region, and the VP1, VP2 and VP3 capsid proteins in the late region. Phylogenetic analyses of the large T antigen of the GPPyV1 indicated GPPyV1 belonged to a putative new species within genus Deltapolyomavirus, clustering with four human polyomavirus species. The GPPyV1 VP1 and VP2 clustered with genus Alphapolyomavirus. Our epidemiologic study indicated that this novel polyomavirus was also detected in nasal swabs and fecal samples collected from captive healthy giant pandas. CONCLUSION: A novel polyomavirus was detected in giant pandas and its complete genome was characterized, which may cause latency infection in giant pandas.


Subject(s)
Nasal Cavity/virology , Polyomavirus/classification , Ursidae/virology , Animals , Genes, Viral , Genome, Viral , Genomics/methods , Phylogeny , Polyomavirus/genetics , Polyomavirus/isolation & purification , Whole Genome Sequencing
17.
Microbiome ; 5(1): 90, 2017 08 07.
Article in English | MEDLINE | ID: mdl-28780905

ABSTRACT

BACKGROUND: The giant panda (Ailuropoda melanoleuca) is a vulnerable mammal herbivore living wild in central China. Viral infections have become a potential threat to the health of these endangered animals, but limited information related to these infections is available. METHODS: Using a viral metagenomic approach, we surveyed viruses in the feces, nasopharyngeal secretions, blood, and different tissues from a wild giant panda that died from an unknown disease, a healthy wild giant panda, and 46 healthy captive animals. RESULTS: The previously uncharacterized complete or near complete genomes of four viruses from three genera in Papillomaviridae family, six viruses in a proposed new Picornaviridae genus (Aimelvirus), two unclassified viruses related to posaviruses in Picornavirales order, 19 anelloviruses in four different clades of Anelloviridae family, four putative circoviruses, and 15 viruses belonging to the recently described Genomoviridae family were sequenced. Reflecting the diet of giant pandas, numerous insect virus sequences related to the families Iflaviridae, Dicistroviridae, Iridoviridae, Baculoviridae, Polydnaviridae, and subfamily Densovirinae and plant viruses sequences related to the families Tombusviridae, Partitiviridae, Secoviridae, Geminiviridae, Luteoviridae, Virgaviridae, and Rhabdoviridae; genus Umbravirus, Alphaflexiviridae, and Phycodnaviridae were also detected in fecal samples. A small number of insect virus sequences were also detected in the nasopharyngeal secretions of healthy giant pandas and lung tissues from the dead wild giant panda. Although the viral families present in the sick giant panda were also detected in the healthy ones, a higher proportion of papillomaviruses, picornaviruses, and anelloviruses reads were detected in the diseased panda. CONCLUSION: This viral survey increases our understanding of eukaryotic viruses in giant pandas and provides a baseline for comparison to viruses detected in future infectious disease outbreaks. The similar viral families detected in sick and healthy giant pandas indicate that these viruses result in commensal infections in most immuno-competent animals.


Subject(s)
Animals, Wild/virology , Ursidae/virology , Virus Diseases/veterinary , Viruses/genetics , Viruses/isolation & purification , Animals , China/epidemiology , Feces/virology , Metagenomics , Papillomaviridae/classification , Papillomaviridae/genetics , Papillomaviridae/isolation & purification , Phylogeny , Picornaviridae/classification , Picornaviridae/genetics , Picornaviridae/isolation & purification , Virus Diseases/epidemiology , Virus Diseases/virology , Viruses/classification
18.
Zoonoses Public Health ; 64(7): 566-571, 2017 11.
Article in English | MEDLINE | ID: mdl-28646559

ABSTRACT

In 2009, a pandemic influenza A virus (pH1N1) spread globally in humans and infected a broad range of captive animals with close human contact. In February 2014, a pH1N1 virus was isolated from a sloth bear with respiratory signs at a US zoo, demonstrating that recurring epidemics present an ongoing threat to animals, including threatened species. This is the first report of pH1N1 infection in sloth bears. To understand the sloth bear virus within the global context of pH1N1, phylogenetic trees were inferred including full-length sequences from available non-human, non-swine hosts, representing four families in the order Carnivora and one order of birds. A combination of phylogenetic and epidemiological evidence strongly suggests the sloth bear was infected with a human-origin pH1N1 virus, supporting the implementation of biosecurity measures to protect human and animal health.


Subject(s)
Influenza A Virus, H1N1 Subtype/isolation & purification , Orthomyxoviridae Infections/veterinary , Pandemics , Respiratory Tract Infections/veterinary , Ursidae/virology , Animals , Animals, Zoo , Influenza A Virus, H1N1 Subtype/genetics , Orthomyxoviridae Infections/epidemiology , Orthomyxoviridae Infections/virology , Phylogeny , Respiratory Tract Infections/epidemiology , Respiratory Tract Infections/virology , United States/epidemiology
19.
Sci Rep ; 7: 39954, 2017 01 04.
Article in English | MEDLINE | ID: mdl-28051146

ABSTRACT

The recent increase in infectious disease outbreaks has been directly linked to the global loss of biodiversity and the decline of some endangered species populations. Between December 2014 and March 2015, five captive giant pandas died due to canine distemper virus (CDV) infection in China. CDV has taken a heavy toll on tigers and lions in recent years. Here, we describe the first gut microbiome diversity study of CDV-infected pandas. By investigating the influence of CDV infection on gut bacterial communities in infected and uninfected individuals and throughout the course of infection, we found that CDV infection distorted the gut microbiota composition by reducing the prevalence of the dominant genera, Escherichia and Clostridium, and increasing microbial diversity. Our results highlight that increases in intestinal inflammation and changes in the relative abundances of pathogen-containing gut communities occur when individuals become infected with CDV. These results may provide new insights into therapeutics that target the microbiota to attenuate the progression of CDV disease and to reduce the risk of gut-linked disease in individuals with CDV. In addition, our findings underscore the need for better information concerning the dynamics of infection and the damage caused by pathogens in panda populations.


Subject(s)
Distemper Virus, Canine/pathogenicity , Distemper/virology , Ursidae/microbiology , Ursidae/virology , Animals , Gastrointestinal Microbiome , Inflammation/virology , Ursidae/genetics
20.
Sci Rep ; 6: 27518, 2016 06 16.
Article in English | MEDLINE | ID: mdl-27310722

ABSTRACT

We report an outbreak of canine distemper virus (CDV) infection among endangered giant pandas (Ailuropoda melanoleuca). Five of six CDV infected giant pandas died. The surviving giant panda was previously vaccinated against CDV. Genomic sequencing of CDV isolated from one of the infected pandas (giant panda/SX/2014) suggests it belongs to the Asia-1 cluster. The hemagglutinin protein of the isolated virus and virus sequenced from lung samples originating from deceased giant pandas all possessed the substitutions V26M, T213A, K281R, S300N, P340Q, and Y549H. The presence of the Y549H substitution is notable as it is found at the signaling lymphocytic activation molecule (SLAM) receptor-binding site and has been implicated in the emergence of highly pathogenic CDV and host switching. These findings demonstrate that giant pandas are susceptible to CDV and suggest that surveillance and vaccination among all captive giant pandas are warranted to support conservation efforts for this endangered species.


Subject(s)
Distemper Virus, Canine/pathogenicity , Distemper/epidemiology , Distemper/virology , Ursidae/virology , Animals , Binding Sites/genetics , China/epidemiology , Distemper Virus, Canine/genetics , Hemagglutinins/genetics , Signaling Lymphocytic Activation Molecule Family/genetics , Vaccination/methods
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