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Intertypic differentiation and detection of intratypic variants among canine and phocid morbillivirus isolates by kinetic neutralization using a novel immunoplaque assay.

Intertypic antigenic differences and the intratypic variability of the closely related canine (CVD) and phocid distemper viruses (PDV) were examined using a molecular (monoclonal antibodies specific for the H- and F-glycoproteins) and a functional (kinetic neutralization, KN) approach. KN studies were carried out using a novel immunoplaque technique which combined conventional plaque assay and antigen-specific enzyme-immunostaining techniques. Morbillivirus isolates of canine and phocid origin clearly formed two separate groups. Minor antigenic differences were also evident within each cluster. A distemper isolate of mustelid origin was distinguishable from both CDV- and PDV-like prototype viruses by kinetic neutralization.

Animals↗

Identification of epitope(s) on the internal virion proteins of rinderpest virus which are absent from peste des petits ruminants virus.

Monoclonal antibodies (MAb) raised against the RBOK vaccine strain of rinderpest virus were characterized by radio-immunoprecipitation (RIPA) and in the indirect ELISA using measles (MV), distemper (CDV), rinderpest (RPV) and peste des petits ruminants viruses (PPRV). Those found to be specific for the matrix (M) protein and the nucleocapsid (N) protein could be classified into different groups on the basis of the anti-morbillivirus MAb classification scheme; a number of these MAb showed a selective recognition of RPV, measles virus and distemper virus, or of different isolates of rinderpest virus, demonstrating that greater inter-isolate variation occurs than was apparent from analyses using polyclonal antisera. One group of anti-F protein MAb (group F1) reacted with all isolates of both RPV and PPRV. A second group of anti-N protein MAb (group N1/A) reacted with all RPV isolates, but not with the PPRV isolates. Furthermore, these group N1/A antibodies reacted strongly with RPV isolates which were upon original isolation of high pathogenicity, but had a weaker reaction against the isolates of this virus which were of low pathogenicity. Thus, MAb against RPV, in particular those against the N protein offered a potential superior to that of molecular analyses for "isolate fingerprinting", the differentiation of RPV from PPRV and the discrimination between rinderpest viruses which had been, upon isolation, of either high or low pathogenicity.

Animals↗

Morbillivirus infections in wildlife (in relation to their population biology and disease control in domestic animals).

The three members of the morbillivirus genus that infect wildlife in ecosystems where domestic animals occur are rinderpest, peste des petits ruminants (PPR) and canine distemper. Data on the relative susceptibility of species of the Order Artiodactyla for rinderpest have been obtained from historical records of outbreaks. Rinderpest in wildlife has only occurred in equatorial and eastern Africa since the great pandemic of 1889-1897. The distributions, densities and population dynamics of susceptible species in this region are described. There has only been one recorded outbreak of PPR in wildlife but the possibility of its occurrence in the future now that it is present in many parts of west and eastern Africa is discussed. Wild carnivora are not likely to be important maintenance hosts for canine distemper but the disease is of significance in free-ranging carnivores and particularly in small populations of endangered susceptible wildlife species. It is also of great significance in zoo populations.

Africa↗

Paget's disease of bone and unvaccinated dogs.

The etiology of Paget's disease is unknown, but several observations suggest a viral cause. Since canine distemper virus is among those proposed to be a causal agent, we investigated the hypothesis that dog ownership may increase the risk of Paget's disease of bone by means of a case control study. One hundred and fifty patients with Paget's disease and 185 controls were interviewed using a structured questionnaire administered by the same interviewer. Participants were asked about their socioeconomic background and their exposure to pets. The risk of Paget's disease was not significantly increased in individuals. However, ownership of a mongrel dog was associated with a significant increase in risk (OR = 1.6; 95% CI = 1.0-2.4) and the risk increased still further with the ownership of an unvaccinated dog (OR = 2.8; 95% CI = 1.8-4.5). Additionally, ownership of cats and birds excluding dog owners, increased the risk of Paget's disease (OR = 2.5; 95% CI = 1.0-5.0; OR = 2.3; 95% CI = 1.0-5.3, respectively). These data suggest that previous exposure to cats, birds, and dogs unvaccinated for canine distemper may increase the risk of an individual developing Paget's disease of bone.

Adult↗

Antibodies to heat shock proteins in dogs with rheumatoid arthritis and systemic lupus erythematosus.

Antibodies to heat shock proteins of the 65 kDa group were demonstrated in canine sera and synovial fluid. This paper reports these antibody measurements in three groups of dogs with joint disease and compares them with those of a control population. Dogs with rheumatoid arthritis (RA) showed higher anti-heat shock proteins (HSP) antibody levels, in both their sera and synovial fluids, compared to the control dogs and these antibodies were predominantly of the IgG and IgM class; there was a significant correlation between IgM anti-HSP65 and IgM-rheumatoid factors. There was also a significant correlation between anti-HSP65 and antibodies to canine distemper virus, but only of the IgM class and the relevance of these antibodies to the overall pathogenesis of canine RA and, in particular, to the presence of canine distemper virus within the joint, are discussed.

Animals↗

Multiple sclerosis and age at infection with common viruses.

Increased risk of multiple sclerosis has been reported among individuals with a history of measles and other common childhood diseases during adolescence, infectious mononucleosis, or exposure to the canine distemper virus. We investigated these associations in a case-control study nested within the Nurses' Health Study (121,700 women traced since 1976) and the Nurses' Health Study II (116,671 women traced since 1989). Age at diagnosis of common viral diseases and birth order were obtained through a questionnaire. Our results include 301 cases with multiple sclerosis and their (up to six) matched controls. Except for infectious mononucleosis, which was a moderate risk factor (odds ratio = 2.1, 95% confidence interval = 1.5-2.9), we found little association between history of common viral diseases or exposure to canine distemper virus and risk of multiple sclerosis. We did find a relation between mumps after 15 years of age (odds ratio = 2.3, 95% confidence interval = 1.2-4.3) or measles after age 15 years of age (odds ratio = 2.8, 95% confidence interval = 0.8-9.1) and multiple sclerosis. Birth order was not materially related to multiple sclerosis. Our findings support the hypothesis that individuals who suffered from infectious mononucleosis, a marker of late infection with the Epstein-Barr virus, have an increased risk of multiple sclerosis. Late infection with other common viruses may also be associated with increased risk.

Adult↗

Multicenter blinded analysis of RT-PCR detection methods for paramyxoviruses in relation to Paget's disease of bone.

UNLABELLED: Conflicting results have been reported on the detection of paramyxovirus transcripts in Paget's disease, and a possible explanation is differences in the sensitivity of RT-PCR methods for detecting virus. In a blinded study, we found no evidence to suggest that laboratories that failed to detect viral transcripts had less sensitive RT-PCR assays, and we did not detect measles or distemper transcripts in Paget's samples using the most sensitive assays evaluated. INTRODUCTION: There is conflicting evidence on the possible role of persistent paramyxovirus infection in Paget's disease of bone (PDB). Some workers have detected measles virus (MV) or canine distemper virus (CDV) transcripts in cells and tissues from patients with PDB, but others have failed to confirm this finding. A possible explanation might be differences in the sensitivity of RT-PCR methods for detecting virus. Here we performed a blinded comparison of the sensitivity of different RT-PCR-based techniques for MV and CDV detection in different laboratories and used the most sensitive assays to screen for evidence of viral transcripts in bone and blood samples derived from patients with PDB. MATERIALS AND METHODS: Participating laboratories analyzed samples spiked with known amounts of MV and CDV transcripts and control samples that did not contain viral nucleic acids. All analyses were performed on a blinded basis. RESULTS: The limit of detection for CDV was 1000 viral transcripts in three laboratories (Aberdeen, Belfast, and Liverpool) and 10,000 transcripts in another laboratory (Manchester). The limit of detection for MV was 16 transcripts in one laboratory (NIBSC), 1000 transcripts in two laboratories (Aberdeen and Belfast), and 10,000 transcripts in two laboratories (Liverpool and Manchester). An assay previously used by a U.S.-based group to detect MV transcripts in PDB had a sensitivity of 1000 transcripts. One laboratory (Manchester) detected CDV transcripts in a negative control and in two samples that had been spiked with MV. None of the other laboratories had false-positive results for MV or CDV, and no evidence of viral transcripts was found on analysis of 12 PDB samples using the most sensitive RT-PCR assays for MV and CDV. CONCLUSIONS: We found that RT-PCR assays used by different laboratories differed in their sensitivity to detect CDV and MV transcripts but found no evidence to suggest that laboratories that previously failed to detect viral transcripts had less sensitive RT-PCR assays than those that detected viral transcripts. False-positive results were observed with one laboratory, and we failed to detect paramyxovirus transcripts in PDB samples using the most sensitive assays evaluated. Our results show that failure of some laboratories to detect viral transcripts is unlikely to be caused by problems with assay sensitivity and highlight the fact that contamination can be an issue when searching for pathogens by sensitive RT-PCR-based techniques.

Base Sequence↗

Morbidity and mortality of red foxes (Vulpes vulpes) and gray foxes (Urocyon cinereoargenteus) admitted to the Wildlife Center of Virginia, 1993-2001.

The medical records of 48 red foxes (Vulpes vulpes) and 35 gray foxes (Urocyon cinereoargenteus) examined at the Wildlife Center of Virginia (Waynesboro, Virginia, USA) from 1993 to 2001 were reviewed. The most common diagnosis in red foxes was orphaned (33%), followed by trauma (27%), undetermined diagnosis (23%), and sarcoptic mange (17%). Trauma (46%) was the most frequent cause of morbidity and mortality in gray foxes followed by orphaned (23%), undetermined (20%), toxoplasmosis (6%), presumptive canine distemper (3%), and rabies (3%). One gray fox had concurrent toxoplasmosis and presumptive canine distemper (3%). Similar diseases were detected in previous studies at a diagnostic laboratory; however in this study, trauma and orphaned animals were more common than infectious diseases. The lack of diagnostic information on some cases limited the usefulness of this study, and more emphasis should be placed on performing postmortem examinations of wildlife presented to wildlife rehabilitation centers.

Animals↗

Antibodies to canine and feline viruses in spotted hyenas (Crocuta crocuta) in the Masai Mara National Reserve.

Spotted hyenas (Crocuta crocuta) are abundant predators in the Serengeti ecosystem and interact with other species of wild carnivores and domestic animals in ways that could encourage disease transmission. Hyenas also have a unique hierarchical social system that might affect the flow of pathogens. Antibodies to canine distemper virus (CDV), feline immunodeficiency virus (FIV), feline panleukopenia virus/canine parvovirus (FPLV/CPV), feline coronavirus/ feline infectious peritonitis virus (FECV/IPV), feline calicivirus (FCV), and feline herpesvirus 1 (FHV1) have been detected in other Serengeti predators, indicating that these viruses are present in the ecosystem. The purpose of this study was to determine whether spotted hyenas also had been infected with these viruses and to assess risk factors for infection. Serum samples were collected between 1993 and 2001 from 119 animals in a single clan for which behavioral data on social structure were available and from 121 hyenas ill several other clans. All animals resided in the Masai Mara National Reserve. Antibodies to CDV, FIV, FPLV/CPV, FECV/FIPV, FCV, and FHV1 were present in 47%, 3.5%, 81%, 36%, 72%, and 0.5% of study hyenas, respectively. Antibody prevalence was greater in adults for FIV and FECV/FIPV, and being a female of high social rank was a risk factor for FIV. Hyenas near human habitation appeared to be at lower risk to have CDV, FIV, and FECV/FIPV antibodies, whereas being near human habitation increased the risk for FPLV/CPV antibodies. Canine (distemper virus and FECV/FIPV antibody prevalence varied considerably over time, whereas FIV, FPLV/CPV, and FCV had a stable, apparently endemic temporal pattern. These results indicate that hyenas might play a role in the ecology of these viruses in the Serengeti ecosystem. The effect of these viruses on hyena health should be further investigated. The lower prevalence of CDV antibody-positive hyenas near human habitation suggests that reservoirs for CDV other than domestic dogs are present in the Serengeti ecosystem.

Africa↗

Immunoglobulin E antibodies to pollens augmented in dogs by virus vaccines.

An inbred "atopic dog colony" was established to study the effect of viruses on immunoregulation of immunoglobulin (Ig) E antibodies. Dogs were selected for high skin reactivity to grass and weed pollens. Their offspring were inoculated with pollen extracts in alum immediately after routine vaccinations (attenuated live-virus vaccines for canine distemper and infectious canine hepatitis, and a killed bacterin for Leptospira). Heat labile antipollen IgE antibodies were measured by passive cutaneous anaphylaxis. Pups vaccinated for canine distemper before being given pollen extracts had many more IgE antibodies than did their control littermates who were not vaccinated until after the last pollen extract injection. This may be a natural example of the "allergic break-through phenomenon."

Adenoviruses, Canine↗

Failure to detect paramyxovirus sequences in Paget's disease of bone using the polymerase chain reaction.

It is widely believed that Paget's disease of bone is due to a "slow virus" infection of osteoclasts with one of the paramyxovirus group. Controversy continues to surround the identity of the virus involved, however, since at different times evidence has been presented implicating measles virus (MV), respiratory syncytial virus (RSV), and canine distemper virus (CDV) as putative infective agents. In this study we used the technique of reverse transcription and polymerase chain reaction (PCR) to screen for paramyxovirus sequences in ribonucleic acid (RNA) extracted from pagetic bone. We were able to detect viral amplification products of the appropriate size in RNA extracted from as few as 50 cells experimentally infected with a wide range of paramyxoviruses, including measles, canine distemper, parainfluenza 3, and respiratory syncytial virus, but we found no evidence of viral products in RNA extracts of affected bone from 10 consecutive patients with Paget's disease. This study fails to support the hypothesis that active infection with one of these or a related paramyxovirus is involved in the pathogenesis of Paget's disease.

Aged↗

Molecular evolution of viral fusion and matrix protein genes and phylogenetic relationships among the Paramyxoviridae.

Phylogenetic relationships among the Paramyxoviridae, a broad family of viruses whose members cause devastating diseases of wildlife, livestock, and humans, were examined with both fusion (F) and matrix (M) protein-coding sequences. Neighbor-joining trees of F and M protein sequences showed that the Paramyxoviridae was divided into the two traditionally recognized subfamilies, the Paramyxovirinae and the Pneumovirinae. Within the Paramyxovirinae, the results also showed groups corresponding to three currently recognized genera: Respirovirus, Morbillivirus, and Rubulavirus. The relationships among the three genera of the Paramyxovirinae were resolved with M protein sequences and there was significant bootstrap support (100%) showing that members of the genus Respirovirus and the genus Morbillivirus were more closely related to each other than to members of the genus Rubulavirus. Both F and M phylogenies showed that Newcastle disease virus (NDV) was more closely related to the genus Rubulavirus than to the other two genera but were consistent with the proposal (B. S. Seal et al., 2000, Virus Res. 66, 1-11) that NDV be classified as a separate genus within the Paramyxovirinae. Both F and M phylogenies were also consistent with the proposal (L. Wang et al., 2000, J. Virol 74, 9972-9979) that Hendra virus be classified as a new genus closely related and basal to the genus Morbillivirus. Rinderpest was most closely related to measles and a more derived virus than to canine distemper virus, phocine distemper virus, or dolphin morbillivirus.

Databases, Nucleic Acid↗

Sporadic bovine meningo-encephalitis-isolation of a paramyxovirus.

Isolation of a viral agent (107) directly from brain explants of a 15-month-old heifer with symptoms of a sporadic encephalomyelitis is described. The virus shares properties with the paramyxovirus family. It grows in a variety of cell cultures from different species, and induces nuclear and cytoplasmic inclusion bodies in infected cells. Nucleocapsids measuring 17 nm in diameter were found in the nucleus and cytoplasm of these cells when studied electron microscopically, thus indicating a close relationship of the agent to the measles-distemper-rinderpest group. No infectious virus was released from infected cells, although alignment of nucleocapsids was observed beneath the cell membrane, and no hemagglutinating activity could be detected with the methods employed. The 107 agent was compared serologically with parainfluenza viruses type 1, 2 and 3, simian virus 5, mumps and Newcastle disease virus (NDV), two bovine respiratory syncytial viruses and measles/subacute sclerosing panencephalitis, distemper and rinderpest viruses, always using 107 virus infected CV1 cells and antiserum of the different viruses in indirect FA tests. Positive FA reactions were observed only with two sera obtained from SSPE patients with high antibody titer to SSPE virus, and with one rabbit-anti-rinderpest serum. The titers of these sera to 107 virus, however, were significantly lower than those against homologous viruses. Five out of 9 sera from randomly selected healthy cattle showed antibody titers between 1:10 and 1:80 to 107 virus in FA tests. The significance of these results is discussed with respect to the epidemiology of SSPE in children and its possible implication with rinderpest in Europe.

Animals↗

Protection of goats against peste des petits ruminants with a vaccinia virus double recombinant expressing the F and H genes of rinderpest virus.

Peste des petits ruminants (PPR) is a viral disease of goats and sheep characterized by necrotizing and erosive stomatitis, enteritis and pneumonia. The causative agent, PPRV, is a member of the family Paramyxoviridae and the genus Morbillivirus. Other members of the genus include rinderpest (RPV), measles, canine distemper and phocid distemper viruses. PPR has a very high rate of morbidity and mortality, and effective control of this disease is of economic importance in Africa, Asia and the Middle East. Currently, there is no safe and effective vaccine available against the disease. The tissue culture rinderpest vaccine (TCRV) protects small ruminants against severe disease; there are, however, clinical problems associated with vaccination. This laboratory has recently developed several effective vaccinia virus recombinant vaccines for rinderpest. These vaccines are easy to administer, inexpensive to produce and heat-stable. Goats were vaccinated with a vaccinia virus double recombinant expressing the haemagglutinin and fusion genes of RPV. Although vaccinated animals developed antibodies (neutralizing and ELISA) to RPV, and not to PPRV, they were completely protected against challenge inoculation with virulent PPRV. This would indicate that protection is most probably due to cell-mediated immunity. Use of the rinderpest double recombinant vaccinia virus in areas of the world where PPRV is endemic would aid in the control and eradication of PPR.

Animals↗

Nucleic acid hybridization analyses confirm the presence of a hitherto unknown morbillivirus in Mediterranean dolphins.

In 1990 an epidemic caused by a morbillivirus was noticed among Mediterranean dolphins. RNA was extracted from the tissues of dolphins and from cell cultures infected with a corresponding dolphin morbillivirus isolate. By nucleic acid hybridization this RNA was compared to RNA extracted from animal tissue or cell cultures infected with canine distemper virus (CDV), phocine distemper virus (PDV) or measles virus (MV). The presence of morbillivirus RNA in the dolphin tissue was demonstrated. Morbillivirus N, P, M and F gene mRNAs were detected in the RNA from dolphin morbillivirus infected cells. These mRNA species seemed to be of approximately the same size as the corresponding mRNA species of CDV, PDV and MV. The results of the comparison demonstrated that the dolphin morbillivirus is genetically different from CDV, PDV and MV. No indication of a close relationship between the dolphin isolate and either CDV, PDV or MV was found.

Animals↗

Morbillivirus infections of aquatic mammals: newly identified members of the genus.

Several disease outbreaks, which have caused the deaths of many thousands of seals and dolphins during the last decade, have now been attributed to infections with newly identified Morbilliviruses. Outbreaks in the late eighties amongst harbour seals (Phoca vitulina) and grey seals (Halichoerus grypus) in northwestern Europe and amongst baikal seals (Phoca sibirica) in Siberia were caused by the newly discovered phocine distemper virus and by a strain of canine distemper virus, respectively. Although closely related these two viruses were not identical. They were more distantly related to the viruses which caused mass mortality amongst striped dolphins (Stenella coeruleoalba) in the Mediterranean sea in the early nineties. This dolphin morbillivirus was shown to be closely related to a virus that was found in harbour porpoises (Phocoena phocoena) which had stranded at the coasts of northwestern Europe in the late eighties: porpoise morbillivirus. The present knowledge of the genetic and antigenic relationships of these apparently new members of the genus Morbillivirus with the established members of the genus is presented. In addition, the origin and epizootiological aspects of these newly discovered viruses are discussed. Finally experimental evidence that environmental pollution may have contributed to the severity and extent of these infections in recent years is presented.

Animals↗

Morbillivirus infection in cetaceans of the western Atlantic.

We report serologic evidence of morbillivirus infection in eleven of fifteen species of odontocete cetaceans from the western Atlantic since 1986. Blood samples were obtained both from free-ranging and stranded animals. Virus neutralizing titers were higher against porpoise and dolphin morbilliviruses than against peste des petits ruminants virus, phocine distemper virus or canine distemper virus (CDV). Serum from five species, tested in a heterologous immunoprecipitation assay using radiolabelled CDV, precipitated the nucleocapsid (N) protein. Clinical morbillivirus infection may potentially impact already threatened species such as the harbour porpoise and precipitate mass strandings of socially cohesive odontocetes.

Animals↗

Editing of morbillivirus P gene transcripts in infected animals.

RNA editing in the Morbillivirus genus in vivo was investigated by applying a polymerase chain reaction-based primer extension technique to measure the edited and non-edited mRNA transcripts. In this genus of the Paramyxoviridae the P gene transcript is altered by the co-transcriptional addition of one extra G residue to produce the mRNA for the V non-structural protein. Using tissues of phocine distemper virus (PDV) infected seals, canine distemper virus (CDV) infected dogs and rinderpest virus (RPV) infected cattle, it was demonstrated that editing occurs in vivo. The P:V mRNA ratios were generally similar to those found in tissue culture infections with the same virus and a minor fraction of transcripts had 2-4 extra G residues. In one seal brain infected with PDV the ratio of P:V mRNA was reversed but no differences were found in the levels of mRNA editing in different tissues from the same animal infected with CDV or RPV. However, variation was seen between animals infected with different isolates of RPV and even between animals infected with the same isolate of RPV.

Animals↗