Canine Distemper Virus Infection Research Articles

Characterization of a new dog isolate of canine distemper virus from China

Canine distemper virus (CDV) is a highly contagious pathogen of dogs. Vaccination is an effective way to protect dogs from CDV infection, but occasionally fails. In the present study, a wild type (wt) CDV, named XJ2, was isolated from a dead vaccinated dog. The hemagglutinin (H) gene of the XJ2 was amplified and analyzed for the molecular characteristics including N-glycosylation sites, phylogenesis, hydrophobicity and epitopes. The data indicated that XJ2 was a genetic variant strain of CDV. CDV-sero-negative dogs were inoculated intranasally with XJ2, developed severe clinical symptoms and died, suggesting high virulence.

Prevalence of positive antibody test results for canine parvovirus (CPV) and canine distemper virus (CDV) and response to modified live vaccination against CPV and CDV in dogs entering animal shelters

Canine parvovirus (CPV) and canine distemper virus (CDV) infections are relatively common in animal shelters and are important population management issues since the immune status of incoming dogs is usually unknown. This study aimed to determine the prevalence of positive antibody test results for CPV and CDV in incoming dogs aged ≥4 months and to measure antibody response over 2 weeks following vaccination with a modified live vaccine (MLV). Dogs aged 4–24 months entering an adoption-guarantee shelter (Shelter 1, n=51) and aged ≥4 months entering a limited admission shelter (Shelter 2; n=51) were enrolled. Dogs from Shelter 1 had been vaccinated with MLV at a municipal shelter 5 days before enrolment, whereas dogs from Shelter 2 had no known history of vaccination at enrolment. Sera were obtained on day 1, immediately prior to CPV/CDV MLV, and tested using an in-clinic ELISA kit to detect CPV/CDV antibodies. Dogs negative for CPV and/or CDV were retested at day 6–8 and those dogs still negative at day 6–8 were retested at day 13–15. Prior to CPV/CDV MLV on day 1, more dogs tested positive for CPV (Shelter 1 – 68.6%; Shelter 2 – 84.3%) than for CDV (Shelter 1 – 37.3%; Shelter 2 – 41.2%). On day 1, prior to MLV, all spayed/neutered animals tested CPV antibody-positive (n=17/102) and CPV antibody-positive dogs were older than serologically negative dogs (Shelter 1, P=0.0029; Shelter 2, P=0.0042). By day 13–15, almost all dogs were CPV antibody-positive (Shelter 1 – 97.9%; Shelter 2 – 100.0%) and CDV antibody-positive (Shelter 1 – 93.8%; Shelter 2 – 97.8%). MLV induces protective antibody titers against CPV/CDV in almost all dogs after 13–15 days.

Pathogenesis and phylogenetic analyses of canine distemper virus strain ZJ7 isolate from domestic dogs in China

A new isolate of canine distemper virus (CDV), named ZJ7, was isolated from lung tissues of a dog suspected with CDV infection using MDCK cells. The ZJ7 isolate induced cytopathogenic effects of syncytia in MDCK cell after six passages. In order to evaluate pathogenesis of ZJ7 strain, three CDV sero-negative dogs were intranasally inoculated with its virus suspension. All infected dogs developed clinical signs of severe bloody diarrhea, conjunctivitis, ocular discharge, nasal discharge and coughing, fever and weight loss at 21 dpi, whereas the mock group infected with DMEM were normal. The results demonstrated that CDV-ZJ7 strain isolated by MDCK cell was virulent, and the nucleotide and amino acid sequences of strain ZJ7 had no change after isolation by MDCK cell when compared with the original virus from the fresh tissues. Molecular and phylogenetic analyses for the nucleocapsid (N), phosphoprotein (P) and receptor binding haemagglutinin (H) gene of the ZJ7 isolate clearly showed it is joins to the Asia 1 group cluster of CDV strains, the predominant genotype in China.

Increased p75 neurotrophin receptor expression in the canine distemper virus model of multiple sclerosis identifies aldynoglial schwann cells that emerge in response to axonal damage

Gliogenesis under pathophysiological conditions is of particular clinical relevance since it may provide regeneration-promoting cells recruitable for therapeutic purposes. There is accumulating evidence that aldynoglial cells with Schwann cell-like growth-promoting properties emerge in the lesioned CNS. However, the characterization of these cells and the signals triggering their in situ generation have remained enigmatic. In the present study, we used the p75 neurotrophin receptor (p75(NTR) ) as a marker for Schwann cells to study gliogenesis in the well-defined canine distemper virus (CDV)-induced demyelination model. White matter lesions of CDV-infected dogs contained bi- to multipolar, p75(NTR) -expressing cells that neither expressed MBP, GFAP, BS-1, or P0 identifying oligodendroglia, astrocytes, microglia, and myelinating Schwann cells nor CDV antigen. Interestingly, p75(NTR) -expression became apparent prior to the onset of demyelination in parallel to the expression of β-amyloid precursor protein (β-APP), nonphosphorylated neurofilament (n-NF), BS-1, and CD3, and peaked in subacute lesions with inflammation. To study the role of infiltrating immune cells during differentiation of Schwann cell-like glia, organotypic slice cultures from the normal olfactory bulb were established. Despite the absence of infiltrating lymphocytes and macrophages, a massive appearance of p75(NTR) -positive Schwann-like cells and BS-1-positive microglia was noticed at 10 days in vitro. It is concluded that axonal damage as an early signal triggers the differentiation of tissue-resident precursor cells into p75(NTR) -expressing aldynoglial Schwann cells that retain an immature pre-myelin state. Further studies have to address the role of microglia during this process and the regenerative potential of aldynoglial cells in CDV infection and other demyelinating diseases.

Inhibition of viral RNA synthesis in canine distemper virus infection by proanthocyanidin A2

Canine distemper virus (CDV) is a contagious and multisystemic viral disease that affects domestic and wild canines as well as other terrestrial and aquatic carnivores. The disease in dogs is often fatal and no specific antiviral therapy is currently available.In this study, we evaluated the in vitro antiviral activity against CDV of proanthocyanidin A2 (PA2), a phenolic dimer belonging to the class of condensed tannins present in plants. Our results showed that PA2 exerted in vitro antiviral activity against CDV with a higher selectivity index compared to ribavirin, included in our study for the previously tested anti-CDV activity. The time of addition assay led us to observe that PA2 was able to decrease the viral RNA synthesis and to reduce progeny virus liberation, at different times post infection suggesting multiple mechanisms of action including inhibition of viral replicative complex and modulation of the redox milieu. These data suggest that PA2, isolated from the bark of Aesculus hippocastanum, has potential usefulness as an anti-CDV compound inhibiting viral replication.

Epizootic canine distemper virus infection among wild mammals

In the spring of 2007, seven raccoon dogs and a weasel were captured near the city of Tanabe in Wakayama prefecture, Japan. The causative agent of the animals’ death 1–2 days after capture was identified as canine distemper virus (CDV) by virus isolation, immunostaining with an anti-CDV polyclonal antibody, and a commercially available CDV antigen-detection kit. Sequence analysis of hemagglutinin genes indicated the isolated viruses belong to genotype Asia-1 and possess the substitution from tyrosine (Y) to histidine (H) at position 549 that is associated with the spread of CDV to non-canine hosts. A serosurvey for CDV was then conducted among wild animals in the region. The animals assayed consisted of 104 raccoons, 41 wild boars, 19 raccoon dogs, five Sika deer, two badgers, one weasel, one marten, one Siberian weasel and one fox. Virus-neutralization (VN) tests showed that, except for fox and weasel, all of the species assayed had VN antibodies to CDV. Interestingly, 11 of the 41 wild boars (27%) and two of the five Sika deer assayed possessed VN antibodies to CDV. These findings indicate that CDV infection was widespread among wild mammals during this epizootic.

Cell-cell Interaction Appears to Represent a Pivotal Factor for Susceptibility of Adult Canine Schwann Cell-like Brain Glia to Canine Distemper Virus Infection in vitro

Cell-cell Interaction Appears to Represent a Pivotal Factor for Susceptibility of Adult Canine Schwann Cell-like Brain Glia to Canine Distemper Virus Infection in vitro

Canine distemper virus induces apoptosis in cervical tumor derived cell lines

Apoptosis can be induced or inhibited by viral proteins, it can form part of the host defense against virus infection, or it can be a mechanism for viral spread to neighboring cells. Canine distemper virus (CDV) induces apoptotic cells in lymphoid tissues and in the cerebellum of dogs naturally infected. CDV also produces a cytopathologic effect, leading to apoptosis in Vero cells in tissue culture. We tested canine distemper virus, a member of the Paramyxoviridae family, for the ability to trigger apoptosis in HeLa cells, derived from cervical cancer cells resistant to apoptosis. To study the effect of CDV infection in HeLa cells, we examined apoptotic markers 24 h post infection (pi), by flow cytometry assay for DNA fragmentation, real-time PCR assay for caspase-3 and caspase-8 mRNA expression, and by caspase-3 and -8 immunocytochemistry. Flow cytometry showed that DNA fragmentation was induced in HeLa cells infected by CDV, and immunocytochemistry revealed a significant increase in the levels of the cleaved active form of caspase-3 protein, but did not show any difference in expression of caspase-8, indicating an intrinsic apoptotic pathway. Confirming this observation, expression of caspase-3 mRNA was higher in CDV infected HeLa cells than control cells; however, there was no statistically significant change in caspase-8 mRNA expression profile. Our data suggest that canine distemper virus induced apoptosis in HeLa cells, triggering apoptosis by the intrinsic pathway, with no participation of the initiator caspase -8 from the extrinsic pathway. In conclusion, the cellular stress caused by CDV infection of HeLa cells, leading to apoptosis, can be used as a tool in future research for cervical cancer treatment and control.

Urban domestic dog populations as a source of canine distemper virus for wild carnivores in the Coquimbo region of Chile

Urban areas can support dog populations dense enough to maintain canine distemper virus (CDV) and can be a source of infection for rural dogs and free-ranging carnivores. The aim of this study was to investigate the relationships between urban and rural domestic dog and wild carnivore populations and their effects on the epidemiology of CDV to explain retrospectively a CD outbreak in wild foxes in 2003. From 2005 to 2007 a cross-sectional household questionnaire survey was conducted in Coquimbo and Ovalle cities, in three towns and in rural sites along two transects from these cities to the Fray Jorge National Park (FJNP) in the Coquimbo region, Chile. Blood samples were collected from unvaccinated dogs at surveyed households and from free-ranging foxes in rural areas along the transects. The seroprevalence of CDV in domestic dogs was higher in urban than in rural areas and in the later was highest in dogs born before 2001–2002. The seroprevalence of CDV in foxes was higher in areas closer to human settlements. A high seroprevalence in dogs born before 2001–2002 further supports a link between CDV patterns in rural dog and fox populations. In our study area, urban dogs are proposed to be the source of CDV infection to wild carnivores. The large dog population size and density detected in Coquimbo and Ovalle provides optimal conditions for maintaining a large and dense susceptible population of dogs, which can act as a reservoir for highly infectious diseases and could have been the source of infection in the CD outbreak in wild foxes.

Immunohistochemical detection of metalloproteinase-9 (MMP-9), anti-oxidant like 1 protein (AOP-1) and synaptosomal-associated protein (SNAP-25) in the cerebella of dogs naturally infected with spontaneous canine distemper

In most viral infections of the central nervous system (CNS), the integrity of brain extracelluar matrix (ECM), oxidative stress and dysfunction in neuronal transmission may contribute to the observed pathology. The purpose of this study was to investigate the role of these factors in demyelinating canine distemper virus (CDV) infections. Regardless of ECM integrity, the expression of metalloproteinase-9 (MMP-9) was visualized in microglial-like cells, whereas the expression of anti-oxidant like-1 (AOP-1) and synaptosomal associated protein (SNAP-25) was frequently detected in Purkinje cells (r(2) = 0.989; p < 0.05), regardless of whether the lesions were classified as acute or chronic. Increased numbers of immunolabeled microglia-like cells and reactive gliosis were observed in advanced cases of demyelinating CDV, suggesting that the expression of AOP-1 and SNAP-25 is correlated with the ultimate death of affected cells. Our findings bring a new perspective to understanding the role of the AOP-1, MMP-9 and SNAP-25 proteins in mediating chronic leukoencephalitis caused by CDV.

A distinct CDV genotype causing a major epidemic in Alpine wildlife

Canine distemper virus (CDV) infection represents an important conservation threat to many carnivore species and has contributed to the population decline of several wild terrestrial and aquatic mammalian species. Since 2006, the Alpine region of North-Eastern (NE) Italy has been experiencing a severe and widespread outbreak of CDV affecting the wild carnivore population. In this study we performed an extensive phylogenetic and molecular evolutionary analysis of CDV identified during the recent wildlife epidemic in the Alpine region. Our analysis yielded data on the evolutionary dynamics of the Alpine wildlife CDV epidemic and revealed the emergence and spread of a single genetic cluster of CDV. The wide distribution of the novel cluster combined with the identification of a specific amino acid mutation, which is believed to increase the ability of the virus to replicate in a wider host range, raises concerns over the possible implications of the spread of this virus on the conservation of endangered wildlife species.

Atypical necrotizing encephalitis associated with systemic canine distemper virus infection in pups

This report describes the naturally occurring atypical neuropathological manifestation of systemic canine distemper virus (CDV) infection in two 16-day-old Pit Bull pups. CDV-induced changes affected the gray and white matter of the forebrain while sparing the hindbrain. Histologically, there was necrosis with destruction of the nervous parenchyma due to an influx of inflammatory and reactive cells associated with eosinophilic intranuclear inclusion bodies within glial cells. Positive immunoreactivity against CDV antigens was predominantly observed within astrocytes and neurons. RT-PCR was used to amplify CDV-specific amplicons from brain fragments. These findings suggest the participation of CDV in the etiopathogenesis of these lesions.

Canine Distemper Epizootic among Red Foxes, Italy, 2009

Canine Distemper Epizootic among Red Foxes, Italy, 2009

Immunophenotypic Characterization of Canine Histiocytic Sarcoma Cells with and without Canine Distemper Virus Infection

Immunophenotypic Characterization of Canine Histiocytic Sarcoma Cells with and without Canine Distemper Virus Infection

Inhibition of viral RNA synthesis in canine distemper virus infection by proanhocyanidin A2

Inhibition of viral RNA synthesis in canine distemper virus infection by proanhocyanidin A2

Fatal canine distemper infection in a pack of African wild dogs in the Serengeti ecosystem, Tanzania

In 2007, disease related mortality occurred in one African wild dog ( Lycaon pictus) pack close to the north-eastern boundary of the Serengeti National Park, Tanzania. Histopathological examination of tissues from six animals revealed that the main pathologic changes comprised interstitial pneumonia and suppurative to necrotizing bronchopneumonia. Respiratory epithelial cells contained numerous eosinophilic intracytoplasmic inclusion bodies and multiple syncytial cells were found throughout the parenchymal tissue, both reacting clearly positive with antibodies against canine distemper virus (CDV) antigen. Phylogenetic analysis based on a 388 nucleotide (nt) fragment of the CDV phosphoprotein (P) gene revealed that the pack was infected with a CDV variant most closely related to Tanzanian variants, including those obtained in 1994 during a CDV epidemic in the Serengeti National Park and from captive African wild dogs in the Mkomazi Game Reserve in 2000. Phylogenetic analysis of a 335-nt fragment of the fusion (F) gene confirmed that the pack in 2007 was infected with a variant most closely related to one variant from 1994 during the epidemic in the Serengeti National Park from which a comparable fragment is available. Screening of tissue samples for concurrent infections revealed evidence of canine parvovirus, Streptococcus equi subsp. ruminatorum and Hepatozoon sp. No evidence of infection with Babesia sp. or rabies virus was found. Possible implications of concurrent infections are discussed. This is the first molecular characterisation of CDV in free-ranging African wild dogs and only the third confirmed case of fatal CDV infection in a free-ranging pack.

Systemic Sarcocystosis in a Striped Skunk (Mephitis mephitis)

A striped skunk with neurological signs was euthanized and examined via necropsy. Histologically, protozoa were found in multiple tissues. Protozoal schizonts measured 15 to 25 mum in diameter and contained 4 to 6 mum crescent-shaped merozoites. Protozoa were associated with necrosis and inflammation in the lung, brain, liver, and nasal epithelium. Immunohistochemistry labeled protozoa strongly positive for Sarcocystis neurona. Polymerase chain reaction-amplified products from the protozoan were 99.6% identical to the corresponding portion of the nuclear small subunit ribosomal RNA gene of S neurona. S neurona origin was further confirmed by amplifying a 451-base pair DNA fragment from the skunk lung, which differed by just 2 or 3 base pairs from the small subunit ribosomal RNA gene of S neurona. Striped skunks act as intermediate and aberrant hosts for S neurona; however, S neurona has rarely been found in extraneural tissues in any species, and systemic sarcocystosis has not been reported in skunks. Additionally, canine distemper virus infection was confirmed with histopathology and immunohistochemistry. Concurrent canine distemper suggests that immunosuppression may have played a role in S neurona infection in this skunk.

Canine distemper virus persistence in demyelinating encephalitis by swift intracellular cell-to-cell spread in astrocytes is controlled by the viral attachment protein

The mechanism of viral persistence, the driving force behind the chronic progression of inflammatory demyelination in canine distemper virus (CDV) infection, is associated with non-cytolytic viral cell-to-cell spread. Here, we studied the molecular mechanisms of viral spread of a recombinant fluorescent protein-expressing virulent CDV in primary canine astrocyte cultures. Time-lapse video microscopy documented that CDV spread was very efficient using cell processes contacting remote target cells. Strikingly, CDV transmission to remote cells could occur in less than 6 h, suggesting that a complete viral cycle with production of extracellular free particles was not essential in enabling CDV to spread in glial cells. Titration experiments and electron microscopy confirmed a very low CDV particle production despite higher titers of membrane-associated viruses. Interestingly, confocal laser microscopy and lentivirus transduction indicated expression and functionality of the viral fusion machinery, consisting of the viral fusion (F) and attachment (H) glycoproteins, at the cell surface. Importantly, using a single-cycle infectious recombinant H-knockout, H-complemented virus, we demonstrated that H, and thus potentially the viral fusion complex, was necessary to enable CDV spread. Furthermore, since we could not detect CD150/SLAM expression in brain cells, the presence of a yet non-identified glial receptor for CDV was suggested. Altogether, our findings indicate that persistence in CDV infection results from intracellular cell-to-cell transmission requiring the CDV-H protein. Viral transfer, happening selectively at the tip of astrocytic processes, may help the virus to cover long distances in the astroglial network, “outrunning” the host’s immune response in demyelinating plaques, thus continuously eliciting new lesions.

Modulación de la respuesta inmune durante la infección por virus distemper canino: implicancias terapéuticas y en el desarrollo de vacunas

Canine distemper virus (CDV) infection is the main infectious cause of mortality in canines and exotic animals worldwide, and also threatens several endangered species such as the giant panda and big felids. The CDV blocks interferon and cytokines signaling pathways through massive infection of peripheral mononucJear cells and Iymphocytes, which decreases B and T cell proliferation and causes CD4 + Th I depletion. These events explain the severe and long lasting immunosuppression that characterizes CDV infection, which leads to a multisystemic disease with subsequent deleterious opportunistic infections. Complex events such as cell dysfunction caused directly by virus replication or exaggerated immune response triggered by infected cells, contribute to the establishment of diverse and complex neurologic diseases throughout the course of the disease. Considering that several species are affected and that CDV infection has a high morbidity and mortality, the present review highlights the relevance of designing safer vaccines, capable of both inducing strategic immunity and preventing CNS pathology. Additionally, in order to better understand the diverse and dynamic mechanisms involved in the disruption ofthe adaptive immune response, as well as immunity induction during viral infection and vaccination, this review addresses the role of dendritic cells (DC) during CDV infection. Furthermore, we will discuss how DC-based therapies could improve the outcome of these patients in terms of survival and prevention of associated sequels.

From the Clinical to the Molecular Diagnosis of Canine Distemper Disease

Canine distemper virus (CDV) is a member of genus Morbillivirus and family Paramyxoviridae that have a negative single-strand RNA genome (Appel, M.J.G., 1987). Canine distemper is a multi-systemic viral disease of dogs characterized by a wide variety of clinical manifestations and lesions, and several other protozoal, bacterial and viral infections as well as toxin poisoning mimic distemper symptoms (Beineke et al, 2009; Green & Appel, 1998). Therefore, the signs are not always suggestive for canine distemper suspicion, and the etiological diagnosis need to be performed. The most used methods of CDV diagnosis are ELISA, direct and indirect immunofluorescent antibody tests, seroneutralisation, and immunofluorescent techniques to detect viral antigen were commercially available, and the veterinary laboratories usually perform them (Green & Appel, 1998). Unfortunately, the most of these techniques have their diagnostic limitation and sometimes can lead to false positive results. Thirty cases of dogs, different mixed breeds and aged less than one year were investigated for CDV infection by ELISA IgG (ImmunoComb®, Biogal, Israel), ELISA IgM (ImmunoComb®, Biogal, Israel), Immunochromatographic CDV-Ag test (CDV IC, Agrolabo, Italy), and reverse transcription polymerase chain reaction (RT-PCR). The blood serum samples were collected from unvaccinated dogs with nervous manifestations and a history of disease up to one month. Eight (8/30) samples were IgM and IgG positive, fourteen (14/30) samples were only IgG positive, all immunochromatographic antigenic tests and RT-PCR were negative. In our opinion, the presence of CDV-antibodies leads to false-negative results for the antigenic and genomic viral tests. These results suggest that blood serum samples from dogs with nervous manifestations are not useful for antigenic tests and RT-PCR.