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Dog lymphocyte cultures facilitate the isolation and growth of virulent canine distemper virus.

Optimal conditions for the isolation and growth of virulent canine distemper virus (CDV) in canine thymic and peripheral blood lymphocyte cultures were determined. Peak virus titers were seen from 3 to 6 days postinoculation of lymphocytes and depended on the multiplicity of infection. Dog lymphocytes were at least as susceptible as canine macrophages to infection with virulent CDV. Virus replication in lymphocytes resulted in higher virus titers than in dog lung macrophages. Peripheral blood lymphocytes (PBL) from CDV-immune dogs were as susceptible to CDV as were PBL from susceptible dogs.

Animals↗

Immunohistochemical detection of canine distemper virus in haired skin, nasal mucosa, and footpad epithelium: a method for antemortem diagnosis of infection.

A reliable antemortem diagnostic method is needed for determining infection with canine distemper virus (CDV). The utility of immunohistochemical detection of CDV antigen was examined was examined for samples of nasal and footpad epithelium and haired skin in dogs with and without detectable CDV antigen in the lung and/or brain. Tissues from 57 dogs at risk of CDV infection were tested. Viral antigen was found in the lung and/or brain of 28 dogs. Among these dogs, viral antigen was demonstrated in the epithelial cells of the nasal mucosa in 24 of 27 dogs, in the footpad epithelium in 24 of 26 dogs, and in the haired skin of the dorsal neck in 26 of 27 dogs. Among the 29 dogs without CDV antigen in either the lung or brain, 1 dog had positive staining for viral antigen in the skin and nasal mucosa. Biopsies of haired skin of the dorsal neck, which is relatively simple to sample, can be used for immunohistochemical testing for acute and subacute infection with CDV.

Animals↗

Induction of chronic neurologic disease in mice with canine distemper virus.

The capacity of a mouse-adapted strain of canine distemper virus (CDV) to induce central nervous system (CNS) disease in weanling mice was investigated. Lethality of infection was found to be mouse-strain-dependent. In sensitive strains, an acute meningoencephalomyelitis developed. Brain tissue from acutely ill animals demonstrated numerous foci of viral antigen, and extracts yielded infectious virus. Mice of resistant strains, notably the SJL strain, survived the effects of acute infection, appeared well for several weeks, and then began to develop signs of subacute CNS disease. Preliminary histopathologic examination of brain and cord from acutely ill animals revealed prominent perivascular mononuclear cell infiltrates, mononuclear cell meningitis, and gliosis. These features were also found in the subacute disease, where, however, the lesions were less severe. Also, in the latter, virus antigen could not be demonstrated. The results indicate that CDV infection of mice may provide a promising model system for the study of virus-induced chronic CNS disease.

Animals↗

Use of B95a cells for isolation of canine distemper virus from clinical cases.

Canine distemper virus (CDV) was readily isolated at high rate with marked cytopathic effect (CPE) in B95a cells, a marmoset lymphoid cell line, from the peripheral blood leukocytes, cerebrospinal fluid cells and brain of dogs. Difference in type of CPE, i.e. syncytium type and round-cell one, among the virus isolates indicate the presence of heterogeneity of virus populations in prevalent CDV. Thus, this cell system is expected to be useful for ecological studies on CDV in the field.

Animals↗

Tyzzer's disease complicated with distemper in a puppy.

A Pomeranian puppy which died from diarrhea and nasal discharge showed catarrhal pneumonia, acute enteritis and focal liver necrosis. Slender bacilli were detected within ileal enterocytes and hepatocytes. A double infection with a distemper virus and Tyzzer's organism at a cellular level was seen within the ileal enterocytes.

Animals↗

Detection of canine distemper virus nucleocapsid protein gene in canine peripheral blood mononuclear cells by RT-PCR.

For a rapid diagnosis of canine distemper virus (CDV) infection, the reverse transcription-PCR (RT-PCR) was carried out to detect CDV nucleoprotein (NP) gene from canine peripheral blood mononuclear cells (PBMCs). Two sets of primers were targeted to two regions of NP gene of CDV Onderstepoort strain. The NP gene fragments were well amplified by the RT-PCR in each of the RNA extracts from Vero cells infected with 6 laboratory strains of CDV including Onderstepoort strain, and from PBMCs of a dog experimentally infected with CDV. The amplified NP gene was detected in 17 of 32 samples from dogs which were clinically suspected for CDV infection at veterinary hospitals. No RT-PCR product was found in 52 samples from healthy dogs including 40 specific pathogen free beagles vaccinated with an attenuated live virus-vaccine for CDV and 12 stray dogs. The RT-PCR provides a fast, sensitive, and supplementary method for the diagnosis of CDV infection in dogs.

Animals↗

Epidemiological observations on recent outbreaks of canine distemper in Tokyo area.

Recent outbreaks of canine distemper virus (CDV) infection in Tokyo area were investigated on the basis of clinical features and serological test. The affected dogs were clinically classified into two groups; dog with respiratory and gastrointestinal signs associated with central nervous system (CNS) signs, and those with CNS signs alone. Of 62 dogs examined, 34 belonged to the former and 28 to the latter. In immunoperoxidase assay, anti-Onderstepoort strain of CDV serum reacted at a low level against 2 field isolates of CDV. These results suggested the presence of different types of CDV population in the field.

Animals↗

Serological analysis of canine distemper virus using an immunocapture ELISA.

As a rapid and sensitive method to detect canine distemper virus (CDV), an immunocapture enzyme-linked immunosorbent assay (ELISA) was performed. The sensitivity and specificity of the immunocapture ELISA were considered to be high enough. Virus neutralizing (VN) test was also established using the immunocapture ELISA. By using this test, the different cross VN titers between sera of dogs experimentally infected with the Onderstepoort strain and those with a field isolate of CDV were observed.

Animals↗

Molecular identification of a recent type of canine distemper virus in Japan by restriction fragment length polymorphism.

Restriction fragment length polymorphism analysis was used to differentiate recent field viruses of canine distemper virus (CDU) from vaccine strains. Virus genomes were amplified by using reverse transcriptase-polymerase chain reaction in part of the haemagglutinin gene. After digestion with EcoRV, the PCR products of recent field isolates were cut into two fragments that differ from the uncut form of old strains including all of vaccine strains. This method could be applied to fresh or stored brains, spleens and peripheral blood mononuclear cells of infected dogs. This molecular approach is useful for determining the causative agent of postvaccinated CDV infection.

Animals↗

Listeriosis in a raccoon dog (Nyctereutes procyonoides) associated with canine distemper.

A wild raccoon dog (Nyctereutes procyonoides) that manifested severe illness and died was examined. Necropsy revealed severe emaciation, systemic icterus and petechial hemorrhages on the mucous membranes. Histopathologically, necroses were seen in the liver and brain stem associated with meningitis. Eosinophilic intranuclear inclusion bodies were observed in the spleen and intestinal mucosa, and eosinophilic intracytoplasmic inclusion bodies were seen in transitional epithelium in the bladder. Listeria monocytogenes 4b was isolated from the liver, spleen, kidneys and lungs, and the pathogen was also detected in the liver and brain stem immunohistopathologically. The disease was diagnosed as listeriosis associated with canine distemper virus infection in a raccoon dog.

Animals↗

Sequence analysis of the genes encoding the phosphoprotein of recent isolates of canine distemper virus in Japan.

The nucleotide sequences of the phosphoprotein (P) of canine distemper virus (CDV) strains isolated between 1992 and 1996 in Japan were determined. This is the first report of the complete sequences of the P genes of recently prevalent CDV strains. The deduced amino acid sequences of the P, C and V proteins showed that in the new Japanese isolates, these proteins have approximately 93%, 90-91% and 92% identities with those of the Onderstepoort vaccine strain, respectively. The predicted functional regions were conserved. RNA editing resulting in a shift to the open reading frame (ORF) of the V protein was shown to occur with the same efficiency in both the field isolates and vaccine strain.

Amino Acid Sequence↗

Cytotoxic T-lymphocyte activity specific for hemagglutinin (H) protein of canine distemper virus in dogs.

Cytotoxic T-lymphocyte (CTL) responses to hemagglutinin (H) protein of canine distemper virus (CDV) were evaluated in dogs using the replication-deficient adenovirus protein expression system. Skin fibroblasts were isolated from two dogs and were infected with recombinant adenovirus bearing the CDV-H gene (Ade-CDVH). CTL assay was performed using fibroblasts expressing CDV-H protein as target cells and peripheral blood lymphocytes (PBL) collected from the same dogs one week after immunization of CDV as effector cells. Specific cytotoxic activity was observed against autologous but not heterologous fibroblasts expressing CDV-H protein. These results indicate that the CTL epitope(s) were localized in the H protein.

Adenoviridae↗

Prevalence of canine distemper virus, feline immunodeficiency virus and feline leukemia virus in captive African lions (Panthera leo) in Japan.

Sero-prevalences of canine distemper virus (CDV), feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) were evaluated in 20 captive lions in two Japanese zoos. Anti-CDV antibody was detected in 13 of 20 lions. We could pursue antibody responses against CDV in three lions back to 1996. Sera collected in 1996 were negative for anti-CDV antibody, therefore, all of them showed sero-conversion in 2000. This result suggested that the epidemic of CDV infection in this zoo might have happened between 1996 and 2000. The lions were also examined for FIV and FeLV infections. We had no evidence for FeLV infection but eight lions were sero-positive for anti-FIV antibody.

Animals↗

Immunohistochemical analysis of two strains of lion (Panthera leo)-adapted canine distemper virus in ferrets (Mustela putorius furo).

Canine distemper virus (CDV) caused epizootics in lions (Panthera leo) in Tanzania's Serengeti National Park in 1994 and in captive lions and other Panthera spp. in the USA in 1991-1992. In this study, immunohistochemistry was used to compare viral distribution in tissues collected from ferrets (Mustela putorius furo) inoculated with one of the two lion-derived CDV isolates, either from Serengeti (A94-11/13) or from California (A92-27/20). The California isolate resulted in severe morbidity in all nine ferrets, whereas the Serengeti isolate resulted in severe morbidity in five of the nine ferrets. A slightly higher proportion of infected cells was found in many tissues in the Serengeti isolate-inoculated ferrets. These findings indicate that the pathogenicity of the California isolate is not directly related to the number of infected cells.

Animals↗

Immunohistochemical investigation of cerebellum in dogs infected with canine distemper virus.

The cerebella of 21 dogs with canine distemper virus (CDV) infection and four normal dogs were examined histopathologically and immunohistochemically. Cerebella of CDV-infected dogs showed nonsuppurative demyelinating encephalomyelitis, classified as acute, subacute or chronic. Immunolocalisation of CDV antigen also confirmed the infection. Tissues were examined for co-localisation of the CDV antigen with either an astrocyte-specific marker, glial fibrillary acidic protein (GFAP), or an oligodendrocyte-specific marker, galactocerebroside (GalC). Immunoreactive cells were counted in demyelinating areas of the white matter. The number of astrocytes (GFAP positive) was significantly (p < 0.05) higher in CDV-infected dogs compared to controls. In contrast, the number of oligodendrocytes (GalC positive) was significantly (p < 0.001) lower in CDV-infected dogs and was much lower in chronic cases (p < 0.05). Approximately 41% of astrocytes and 17% of oligodendrocytes were immunoreactive for CDV. The ratio of CDV-infected oligodendrocytes and astrocytes remained almost constant during the progression of the disease (P > 0.05). In conclusion, CDV infects both astrocytes and oligodendrocytes. The gradual loss of oligodendrocytes is most likely responsible for the progressive demyelination in CDV infection. Astrocytosis in CDV infection should be further investigated if it occurs to stimulate oligodendrocytes for myelin production to compensate for the loss or to induce oligodendrocyte degeneration.

Animals↗

Neospora caninum infection in a free-ranging raccoon (Procyon lotor) with concurrent canine distemper virus infection.

During a canine distemper virus (CDV) outbreak in raccoons (Procyon lotor) from Cook County, Illinois, a juvenile female suffering from seizures was killed and necropsied. Gross and histologic findings of necrotizing encephalitis and proliferative bronchopneumonia were attributed to CDV infection and considered the cause of clinical signs. A section of cerebellum stained immunohistochemically for Neospora caninum revealed an approximately 40 microm diameter, round to oval cyst with a 2- to 3-microm-thick wall and filled with 1-2 microm diameter, round to oval bradyzoites. Polymerase chain reaction (PCR) results were positive for N. caninum using DNA extracted from the brain. Specific PCR for the closely related organisms Toxoplasma gondii and Hammondia heydorni yielded negative results. This case report provides histologic, immunohistochemical, and molecular evidence that raccoons are a naturally occurring intermediate host of N. caninum.

Animals↗

Serologic evaluation, efficacy, and safety of a commerical modified-live canine distemper vaccine in domestic ferrets.

OBJECTIVE: To determine efficacy and safety of a commercial modified-live canine distemper virus (CDV) vaccine used for prophylaxis in domestic ferrets. ANIMALS: Sixteen 16-week-old neutered male ferrets. PROCEDURES: Equal groups of ferrets were inoculated subcutaneously at 16 and 20 weeks of age with saline (0.9% NaCl) solution or a vaccine derived from the Onderstepoort CDV strain and attenuated in a primate cell line. Live virulent CDV was administered to all ferrets intranasally and orally 3 weeks after the second inoculation. Clinical signs and body weights were monitored regularly during the study. Blood samples for serologic examination were drawn prior to each inoculation, before challenge exposure, and 10, 15, and 21 days after exposure. Blood samples for reverse transcriptase polymerase chain reaction (RT-PCR) were obtained 5 days after the first vaccination, and 5, 10, 15, and 21 days after challenge exposure. RESULTS: After challenge exposure, control ferrets had significantly more clinical signs and weight loss, compared with vaccinates. All vaccinated ferrets survived, whereas all control ferrets died. The RT-PCR assay was successful in detecting CDV in blood and fresh or formalin-fixed tissues from infected ferrets. CONCLUSIONS AND CLINICAL RELEVANCE: Findings suggest that the vaccine when given SC to domestic ferrets as directed is safe and protective against challenge exposure with virulent CDV. The RT-PCR assay may simplify detection of CDV in fresh and fixed tissues.

Animals↗

Clinical use of serum parvovirus and distemper virus antibody titers for determining revaccination strategies in healthy dogs.

OBJECTIVE: To assess whether serum canine parvovirus (CPV) and canine distemper virus (CDV) antibody titers can be used to determine revaccination protocols in healthy dogs. DESIGN: Case series. ANIMALS: 1,441 dogs between 6 weeks and 17 years old. PROCEDURE: CPV and CDV antibody titers in serum samples submitted to a commercial diagnostic laboratory were measured by use of indirect fluorescent antibody (IFA) tests. On the basis of parallel measurements of CPV and CDV serum antibody titers in 61 paired serum samples determined by use of hemagglutination inhibition and serum neutralization methods, respectively, we considered titers > or = 1:5 (IFA test) indicative of an adequate antibody response. RESULTS: Age, breed, and sex were not significantly associated with adequate CPV- or CDV-specific antibody responses. Of 1,441 dogs, 1,370 (95.1%) had adequate and 71 (4.9%) had inadequate antibody responses to CPV, whereas 1,346 of 1,379 (97.6%) dogs had adequate and 33 (2.4%) had inadequate responses to CDV. Vaccination histories were available for 468 dogs (468 for CPV, 457 for CDV). Interval between last vaccination and antibody measurement was 1 to 2 years for the majority (281/468; 60.0%) of dogs and 2 to 7 years for 142 of 468 (30.3%) dogs. Interval was < 1 year in only 45 of 468 (9.6%) dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The high prevalence of adequate antibody responses (CPV, 95.1%; CDV, 97.6%) in this large population of dogs suggests that annual revaccination against CPV and CDV may not be necessary.

Age Factors↗