Avian Paramyxovirus Serotype Research Articles

Complete sequence of the genome of avian paramyxovirus type 2 (strain Yucaipa) and comparison with other paramyxoviruses

The complete RNA genome sequence of avian paramyxovirus (APMV) serotype 2, strain Yucaipa isolated from chicken has been determined. With genome size of 14,904 nucleotides (nt), strain Yucaipa is consistent with the “rule of six” and is the smallest virus reported to date among the members of subfamily Paramyxovirinae. The genome contains six non-overlapping genes in the order 3′-N-P/V-M-F-HN-L-5′. The genes are flanked on either side by highly conserved transcription start and stop signals and have intergenic sequences varying in length from 3 to 23 nt. The genome contains a 55 nt leader sequence at 3′ end and a 154 nt trailer sequence at 5′ end. Alignment and phylogenetic analysis of the predicted amino acid sequences of strain Yucaipa proteins with the cognate proteins of viruses of all of the five genera of family Paramyxoviridae showed that APMV-2 strain Yucaipa is more closely related to APMV-6 than APMV-1.

Evaluation of Newcastle disease virus immunoassays for waterfowl using a monoclonal antibody specific for the duck immunoglobulin light chain

In the present study a monoclonal antibody (mAb 14A3) was tested for its reactivity against serum immunoglobulin Y (IgY) of several waterfowl species, and subsequently for its applicability as anti-species antibody in common immunoassays. Western blot analyses demonstrated its broad cross-reactivity with the serum IgY light chain of different duck species: Muscovy duck (Cairina moschata), Mallard (Anas platyrhynchos), white-winged wood duck (Asarcornis scutulatus), common pintail (Dafila acuta). Reactivity was also evident with IgY of two swan species—mute swan (Cygnus olor) and black-necked swan (Sthenelides melanocoryphus)—and two goose species—domestic goose (Anser anser var. domestica) and red-breasted goose (Rufibrenta ruficollis). Applying the mAb for Newcastle disease virus (avian paramyxovirus serotype 1 [APMV-1]) test systems, its functionality within indirect immunoassays was evaluated. Using APMV-1-positive sera of domestic geese and Muscovy ducks, mAb 14A3 facilitated specific staining of APMV-1-infected cells in an immunofluorescence test. In addition, it proved to be functional in an indirect enzyme-linked immunosorbent assay (ELISA) and a western blot assay. Thus, the analysed mAb represents an attractive and versatile reagent that offers the opportunity to develop serological tests for waterfowl, allowing a high sample throughput using the ELISA technique or the fine analysis of humoral immune responses using the western blot.

Isolation and pathogenicity identification of avian paramyxovirus serotype 1 (Newcastle disease) virus from a Japanese quail flock in Iran

Newcastle disease virus (NDV) was isolated and identified for the first time from a flock of Japanese quail suspected to ND in Iran. Birds of the flock aged variously, and some of them died from the disease. Loss of appetite, weakness and decrease in egg production, diarrhea and nervous symptoms were the main clinical signs in the birds. Hemorrhagic lesions of the intestinal tracts and proventriculus were found in gross pathology investigations. Samples were taken for bacteriology, parasitology and virology. The results of parasitology and bacteriology examinations were negative. The virus was isolated only from brain samples and was identified as Avian Paramyxovirus type 1 (APMV-1) by HI test using NDV-Specific antibody. The pathogenicity of the virus was identified by mean death time (MDT) in embryonated chicken eggs, intracerebral pathogenicity index (ICPI) and intravenous pathogenicity index (IVPI) in chickens. Values 40, 1.62 and 2.31 were obtained for MDT, ICPI and IVPI respectively. These results indicate that isolated APMV-1 belongs to NDV velogenic strains with severe pathogenicity. According to these findings vaccination against NDV in quail farms of Iran is highly recommended.

Serological Survey on Prevalence of Antibodies to Avian Paramyxovirus Serotype 2 in China

We report the prevalent status of avian paramyxovirus serotype 2 (APMV-2) in China. Between 2003 and 2005, 9156 sera in total were collected and screened for APMV-2 antibodies by using the hemagglutination inhibition assay. The averaged seropositivity ofAPMV-2 for chickens, ducks, peacocks, ostriches, and partridges was 42.9%, 25.1%, 45.8%, 47.6%, and 80.0%, respectively. The results of this survey indicate that the distribution of APMV-2 is very widespread in China and that more attention should be paid to the influence of APMV-2 on poultry production.

Serological Survey on the Prevalence of Antibodies to Avian Paramyxovirus Serotype 2 in China

Serological Survey on the Prevalence of Antibodies to Avian Paramyxovirus Serotype 2 in China

Effects of Polyether Ionophores on the Protective Immune Responses of Broiler Chickens against Angara Disease and Newcastle Disease Viruses

Immunization against Angara disease virus (ADV), a serotype 4 avian adenovirus, and Newcastle disease virus (NDV), an avian paramyxovirus serotype 1, is the mainstay of a broiler vaccination programme, while polyether ionophores usually form an essential component of a broiler medication programme in most parts of India and Pakistan. The role of polyether ionophores in the protective immune responses of broiler chickens vaccinated and challenged with ADV and NDV was investigated. A total of 1600 birds were divided into eight groups of 200 birds each. First four groups were vaccinated against NDV and ADV, while the remaining four served as unvaccinated controls. The first 3 groups of birds were administered salinomycin, monensin and cyclophosphamide (CYP), respectively. The last group served as an untreated control. The same treatment schedule was also followed for the next four unvaccinated groups. The post-vaccination and post-challenge serological responses to NDV and ADV, body and lymphoid organ weight gains, post-challenge survival rate and detection of NDV and ADV in the tissues of infected birds were evaluated. Birds administered salinomycin showed a significant stimulation of protective immune responses against both NDV and ADV as compared to the untreated and CYP-treated birds. Monensin also enhanced the protective immune responses against both viruses but the effect was not statistically significant. Thus, it is concluded that monensin and salinomycin augment the anti-NDV and anti-ADV immune responses in broiler chickens, which supports their use in poultry flocks.

Isolation, Identification, and Comparison of Four Isolates of Avian Paramyxovirus Serotype 2 in China

Four Yucaipa-like viruses of avian paramyxovirus serotype 2 (APMV-2) were isolated in China from the imported Gouldian Finch (Chloebia gouldiae) and broilers in 1998-2002, and were named F4, F6, F8, and NK, respectively. Examined under electron microscope, the isolates were found to be round in shape and varying in size. The results of the hemagglutination inhibition test and indirect enzyme-linked immunosorbent assay (using monoclonal antibodies) showed some differences between the isolates and the reference strain Yucaipa. The isolates derived from chickens had a closer relationship to Yucaipa virus than did those of finches. Sequence comparison of the fusion gene and the haemagglutinin-neuraminidase gene showed similar results, although the variations were lesser among APMV-2 viruses in nucleotide and amino acid sequence. By sequence comparison, it was also revealed that at the molecular level the four virus strains belong to APMV-2, and that two of the strains were isolated from the same group of imported Gouldian Finches.

Third genome size category of avian paramyxovirus serotype 1 (Newcastle disease virus) and evolutionary implications

The goal of the study was to establish if there was a relationship between molecular patterns and virus evolution. Therefore the complete genome sequence of two distinct apathogenic Newcastle disease virus (NDV) strains was determined and a third genome size category, containing 15,198 nucleotides, was recognized. Phylogenetic analysis revealed that two major separations resulting in three genome size categories occurred during the history of NDV. An ancient division in the primordial reservoir (wild waterbird species) led to two basal sister clades, class I and II, with genome sizes 15,198 (due to a 12 nucleotide insert in the phosphoprotein gene) and 15,186 nucleotides, respectively. Ancestors of only class II viruses colonized chicken populations and subsequently converted to virulent forms. These took place more than once and resulted in an early lineage [including genotypes I–IV and H33(W)] with genome size of 15,186 nucleotides. A second division occurred in the 20th century in the secondary (chicken) host. This gave rise to the branching-off of a clade (including recent genotypes V–VIII consisting of only pathogenic viruses) with the concomitant insertion of six nucleotides into the 5′ non-coding region of the nucleoprotein gene thereby increasing the genome size to 15,192 nucleotides.

Detection of Newcastle disease virus using nucleic acid sequence-based amplification

Newcastle disease (ND) is a contagious and widespread avian disease affecting most species of birds. ND virus (NDV) is the only member of the avian paramyxovirus serotype 1 (APMV1) causing ND outbreak in bird flocks. The technique of nucleic acid sequence-based amplification (NASBA) is a potential method to rapidly and reliably detect NDV isolates. Here, we describe an effective and unprecedented method for detecting NDV strains of all pathotypes. A conserved region of the fusion protein gene was used for designing oligonucleotides specific to all NDV pathotypes. The dynamic range of this NDV NASBA detection method is comparable to virus culture and therefore the NDV NASBA method is a potential alternative for NDV screening and surveillance.

Avian paramyxovirus serotype 1 isolates from the spinal cord of parrots display a very low virulence.

The spinal cord of 32 psittacines suffering from proventricular dilatation disease (PDD) was investigated. In six cases, a virus was isolated which upon electron microscopic examination revealed morphological details typical of members of the Paramyxoviridae. All isolates were subsequently characterized as avian paramyxovirus serotype 1 (APMV-1) by type-specific polyclonal antisera. According to their reactivity with APMV-1 specific monoclonal antibodies, the six isolates shared epitopes within the haemagglutinin-neuraminidase spike protein, distinct from pigeon-type paramyxoviruses and the LaSota vaccine strain. This grouping was further corroborated by properties of the haemagglutinin: all isolates showed a very thermosensitive haemagglutination activity and were rapid eluters. Virulence of the APMV-1 isolates in 1-day-old specific pathogen free (spf) chicken was very low, with intracerebral pathogenicity indices between 0 and 0.1. In embryonated spf chicken eggs, psittacine isolates replicated to high titres (10(8.6)-10(10.7) EID50/ml). However, they exhibited a reduced lethality over an observation time of 7 days (10(6.1)-10(8.3) ELD50/ml). In a haemagglutination inhibition test with parrot sera from birds with no history of APMV-1 vaccination, sera reacted preferentially with two isolates compared with APMV-1 vaccine strains LaSota and B1. The other four isolates exhibited a differentiated reaction pattern with the parrot sera, indicating an antigenic inhomogeneity. This is the first report of isolating very low virulent APMV-1 from neuronal tissue of parrots and implications for a possible role in slow progressing disease will be discussed.

Detection and differentiation of pathogenicity of avian paramyxovirus serotype 1 from field cases using one-step reverse transcriptase-polymerase chain reaction

Amplification of avian paramyxovirus serotype 1 (APMV-1)-specific nucleic acid fragments, followed by restriction endonuclease analysis (REA) using Bgl I, was carried out to type strains according to their virulence. Primer sequences were used to amplify a 202 base pair fragment, encompassing the fusion protein cleavage site, in a one-step reverse transcriptase-polymerase chain reaction (RT-PCR) test for detection of a range of field cases and reference strains of APMV-1. Subsequent REA of the amplified fragments enabled differentiation of low virulent lentogenic field and vaccine strains from more virulent mesogenic and velogenic field strains of APMV-1, including pigeon PMV-1. In the present paper, we report the development and application of a one-step RT-PCR test coupled with REA as a fast, specific method for both the detection and typing of APMV-1 from field samples.

Newcastle disease and other avian paramyxovirus

Newcastle disease (ND), caused by avian paramyxovirus serotype 1 (APMV-1) viruses, is included in List A of the Office International des Epizooties. Historically, ND has been a devastating disease of poultry, and in many countries the disease remains one of the major problems affecting existing or developing poultry industries. Even in countries where ND may be considered to be controlled, an economic burden is still associated with vaccination and/or maintaining strict biosecurity measures. The variable nature of Newcastle disease virus strains in terms of virulence for poultry and the different susceptibilities of the different species of birds mean that for control and trade purposes, ND requires careful definition. Confirmatory diagnosis of ND requires the isolation and characterisation of the virus involved. Assessments of virulence conventionally require in vivo testing. However, in vitro genetic characterisation of viruses is being used increasingly now that the molecular basis of pathogenicity is more fully understood. Control of ND is by prevention of introduction and spread, good biosecurity practices and/or vaccination. Newcastle disease viruses may infect humans, usually causing transient conjunctivitis, but human-to-human spread has never been reported. Eight other serotypes of avian paramyxoviruses are recognised, namely: APMV-2 to APMV-9. Most of these serotypes appear to be present in natural reservoirs of specific feral avian species, although other host species are usually susceptible. Only APMV-2 and APMV-3 viruses have made a significant disease and economic impact on poultry production. Both types of viruses cause respiratory disease and egg production losses which may be severe when exacerbated by other infections or environmental stresses. No reports exist of natural infections of chickens with APMV-3 viruses.

Similarity of avian paramyxovirus serotype 1 isolates of low virulence for chickens obtained from contaminated poultry vaccines and from poultry flocks

At present Denmark has the status of a ‘non-vaccinating’ country for Newcastle disease and its poultry population should therefore be free of antibodies to avian paramyxovirus 1 (APMV-1). Three live...

Detection of Newcastle disease virus in organs and faeces of experimentally infected chickens using RT-PCR

Newcastle disease (ND) is a highly contagious avian disease. Rapid diagnosis of ND plays an important role in controlling outbreaks. Until now, time-consuming isolation of ND virus (NDV) in embryonated chicken eggs was used for NDV detection. For rapid diagnosis, reverse transcription-polymerase chain reaction (RT-PCR) with RNA extracted from tissue samples and faeces originating from experimentally and contact-infected chickens was established. Conjunctiva, lung, caecal tonsil and kidney proved to be the most suitable organs. In infected animals, NDV was detected most frequently between day 4 and 6 post-infection. Contact-infected animals gave most positive results between day 6 and 13 after exposure. RT-PCR was also able to reproducibly detect NDV in faecal samples. The RT-PCR did not show any cross-reactivity with other avian paramyxovirus serotypes, and additionally offers the possibility of subsequent sequencing of the amplified DNA allowing pathotyping of the isolate.

Isolation of avian paramyxovirus serotype 3 from domestic fowl in Israel: close antigenic relationship with the psittacine strain of avian paramyxovirus serotype 3.

12. Netherwood T, Chanter N, Mumford JA: 1996, Improved isolation of Clostridium perfringens from foal feces. Res Vet Sci 61:147–151. 13. Rood JI, Cole ST: 1991, Molecular genetics and pathogenesis of Clostridium perfringens. Microbiol Rev 55:621–648. 14. Samuel SC, Hancock P, Leigh DA: 1984, An investigation into Clostridium perfringens enterotoxin-associated diarrhea. J Hosp Infect 18:219–230. 15. Songer JG, Meer RR: 1996, Genotyping of Clostridium perfringens by polymerase chain reaction is a useful adjunct to diagnosis of clostridial enteric disease in animals. Anaerobe 2:197– 203. 16. Stringer MF, Watson GN, Gilbert RJ, et al.: 1985, Faecal carriage of Clostridium perfringens. J Hyg 95:277–288. 17. Twedt DC: 1992, Clostridium perfringens-associated enterotoxicosis in dogs. In: Current veterinary therapy XI: small animal practice, ed. Kirk RW, Bonagura JD, 11th ed., pp. 602–604. WB Saunders, Philadelphia, PA.

An outbreak of Newcastle disease in free-living pheasants (Phasianus colchicus).

The epidemiology of an outbreak of Newcastle disease in a population of approximately 12,000 free-living pheasants (Phasianus colchicus) on the island of Faenø in Denmark in 1996 is described. The mortality during the epizootic was 56%. The spread of the disease between 7 groups of pheasants could be demonstrated over an observation period of 3 weeks. A total of 70 avian paramyxovirus serotype 1 (APMV-1) isolates was made from the flock. The intra cerebral pathogenicity indices of the 4 isolates tested were in the range 1.78-1.88. By means of immunoperoxidase monolayer assay with murine monoclonal antibodies and sequence analysis of an RT-PCR amplified segment of the F0 viral protein it was found, that the virus belonged to the highly virulent C1 antigenic group and that the amino acid sequence at the F0 cleavage site corresponded with the sequences of virulent APMV-1 strains. Based on the epidemiological circumstances it is believed that the virus was transmitted to the pheasants by feral birds.

Isolation and characterization of avian paramyxovirus type 1 (Newcastle disease) viruses from a flock of ostriches (Struthio camelus) and emus (Dromaius novaehollandiae) in Europe with inconsistent serology

During a 95‐day study period in 1995 in Denmark, 18 ostriches in a flock of 77 ostriches and four emus held in quarantine died. Clinical and pathological observations did not indicate the presence of transmissible infectious disease in the flock. Management failures and indoor housing were believed to have contributed significantly to the number of deaths. Samples from 17 of the dead ostriches were examined virologically. Three isolates of avian paramyxovirus serotype 1 (APMV‐1) were obtained from intestines and intestinal contents of dead ostriches submitted for laboratory investigations. In ICPI tests in day‐old chicks values for the three APMV‐1 isolates were in the range 1.63–1.69. Characterization by means of mouse monoclonal antibodies and by restriction site analysis revealed that the three isolates were indistinguishable and similar to APMV‐1 viruses present in a simultaneous epizootic of Newcastle disease in back yard poultry in Denmark. Blood samples were taken from all live birds in the flock after 25 and 95 days of quarantine and all were negative for antibodies to APMV‐1 in haemagglutination inhibition tests. All samples taken after 95 days of quarantine were also negative for antibodies to APMV‐1 in serum neutralization tests performed in chicken embryo cells. Blood samples taken after 95 days of quarantine were tested in a commercial ELISA for antibodies to APMV‐1. In this test 35% of the samples were positive, 35% were border line and 30% were negative.

Detection of Newcastle disease virus-specific antibodies in ostrich sera by three serological methods

Sera from 211 ostriches were tested for the presence of Newcastle disease virus (NDv)-specific antibodies by the virus neutralisation test, the haemagglutination inhibition (HI) test and a recently developed avian...

Mixed paramyxovirus infection of wild and domestic birds in Israel

Five cases of dual isolations of different serotypes of avian paramyxoviruses (APMV) from domestic and wild birds are described: one case of mixed infection by APMV-1 and APMV-4 and four cases of infection by APMV-1 and APMV-2 serotypes. The double infection was proven by consecutive isolations of two viruses from allantoic fluid samples derived from single swabs after their respective treatment by antisera against each suspected virus. The finding of double APMV infection in poultry farms appears to be important for epizootiology and pathogenesis of APMV-caused diseases.

Isolation and characterization of paramyxoviruses from snakes and their relationship to avian paramyxoviruses.

Several viruses were isolated from snakes which died during an outbreak of disease in a zoo. The viruses could be cultivated at 28 degrees C on a snake cell line (VH2, ATCC CCL 140). In cell culture they formed syncytia, leading to destruction of the monolayer. The isolates were sensitive to treatment with chloroform but not to growth in the presence of IUDR and revealed haemagglutinating activity with chicken erythrocytes. These results, combined with electron microscopic examination of the infectious supernatant, confirmed the diagnosis of paramyxoviruses (PMV). Haemagglutination inhibition tests with antisera against avian PMV revealed a serological relationship between the snake viruses and the avian PMV serotypes 1 and 7. Tests for thermostability at 56 degrees C showed a marked decrease of the haemagglutination and infectivity titres in contrast to the avian PMV serotype 7. The multiplication of the snake viruses seemed to be restricted to incubation temperatures below 37 degrees C. At 28 degrees C the snake viruses could be cultivated in several cell cultures. Separation of virus proteins using polyacrylamide gel electrophoresis showed differences between the snake isolates and avian PMVs of serotypes 1 and 7.