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PubMedDate not supplied

Characterization of Newcastle disease viruses isolated from field cases in Japan.

Seven Newcastle disease viruses isolated in Japan from 1930 to 1984 were cloned on chicken embryo fibroblasts (CEFs) and characterized biologically. All seven produced two or more types of plaques on CEFs. The plaques were classified into four types. Plaque cloning was carried out five times, and 22 cloned viruses were established. The biological characters of the cloned viruses suggested that the strains contain different clones and that their clones are different even among close cases, such as G strain and H strain.

Animals↗
PubMed1993

Local antibody forming cell responses to the Hitchner B1 and Ulster strains of Newcastle disease virus.

The Hitchner B1 and Ulster strains of Newcastle disease virus (NDV) replicated to high titre in the Harderian gland (HG) after eye-drop infection. The Harderian gland then became the major site of antiviral IgA-antibody-forming cells (AFC) in the body and their number correlated to the level of antiviral IgA antibody in the tears. The spleen, HG and femoral bone marrow all contained comparable levels of antiviral IgG-AFC and IgM-AFC after two intra-ocular inoculations of virus, whereas the caecal tonsil and bursa contained few AFC despite the local replication of the Ulster strain of NDV leading to high titres of virus in the faeces. Vaccines of the Hitchner B1 strain of NDV were much less effective at inducing antibody by the intranasal compared with intra-ocular route and no virus was re-isolated after intranasal vaccination. The intravenous inoculation of inactivated Iscoms of NDV could stimulate the spleen, but not the Harderian gland to the same extent as a live virus.

Animals↗
PubMedDate not supplied

Detection and differentiation of strains of Newcastle Disease Virus by complement fixation.

A complement-fixation test to detect Newcastle disease virus with antiserum produced in guinea pigs is described. Methodology is given for serum production and for standardization of the test. The test was used to differentiate 13 strains of Newcastle disease virus. Velogenic strains, including isolants form 1970-71 disease outbreaks in California, Florida, and Texas, were poor complement-fixing antigens, whereas lentogenic strains, including LaSota, Hitchner, and England F, were strong complement-fixing antigens. Mesogenic strains ranged from weak to strong in complement-fixing capabilities. This test can be used to differentiate velogenic field isolants from vaccine strains such as LaSota, Hitchner, and Roakin.

Animals↗
PubMedDate not supplied

Familial differences in antibody response of broiler chickens to vaccination with attenuated and inactivated Newcastle disease virus vaccine.

Genetic differences in immune response to Newcastle disease virus (NDV) were studied in 4-week-old broilers, vaccinated with attenuated (live) or inactivated NDV. The experiment included 370 chicks from two farms distributed among 22 sire families and 60 dam families. Results in chicks from both farms were similar. Survival after challenge was closely related to titer level. The genetic correlation between day-7 and day-12 titers (attenuated virus) was 1.0. Significant differences were found between sire families in both sorts of vaccinations. Heritabilities based on the sire variance components for attenuated and inactivated virus vaccinations were respectively 0.31 and 0.60. The genetic correlation between them was 0.49. Nevertheless, it is concluded that selection for response to NDV based on inactivated virus may be most effective in improving response to attenuated NDV vaccinations.

Animals↗
PubMed1975

Phospholipids in Newcastle Disease Virus infected cells.

Infection of chicken cells with Newcastle Disease Virus modifies phosphatidylserine and phosphatidylcholine synthesis in the host cell. The virion contains cellular phospholipids synthesized both before and after infection. Relative concentration of various labeled phospholipids in the virus differ from those in the corresponding cells and their surface membranes. Late in infection, fragments of membranes with a distribution of labeled phospholipids similar but not identical to that of the virus can be found in the supernatant of infected cells. The significance of these findings is discussed in relation to the origin of viral phospholipids and the intervention of the host cell membrane in the assembly of the viral envelope.

Animals↗
PubMedDate not supplied

Progression of tracheal lesions in turkeys exposed by aerosol to LaSota strain of Newcastle disease virus.

Five-week-old turkeys were exposed by aerosol to the LaSota strain of Newcastle disease virus. Poults were killed on days 2, 4, 6, 8, 10, 12, and 14 postexposure, and tracheas were processed for virus quantitation and histologic examination. Newcastle disease virus was recovered at a high titer from all tracheas collected 2, 4, and 6 days postexposure. The initial tracheal lesion observed on day 2 was swelling of ciliated columnar and mucous gland cells. Some of the affected cells contained intracytoplasmic inclusions. Cell swelling and degeneration were followed by epithelial cell proliferation, fibrinopurulent exudation, and lymphocytic infiltration. Epithelial cell proliferation was most severe on days 4 and 6, when tracheas were lined with several layers of immature cells. Lymphoid nodules appeared on day 6 and persisted up to day 14. From day 8 on, there was regression of the proliferative lesion accompanied by differentiation of the immature epithelium. By day 14, the tracheal mucosa regained its normal histologic appearance.

Aerosols↗
PubMed1980

Vaccination of chickens against Newcastle disease with live and inactivated Newcastle disease virus.

Chickens were vaccinated and revaccinated with inactivated Newcastle disease (ND) vaccines from 2 different sources and also with LaSota strain, live Newcastle disease virus (NDV). Both inactivated vaccines induced higher virus neutralizing (VN) and hemagglutination-inhibition (HI) titers than the LaSota virus. One of the inactivated preparations was found superior to the other by both the VN and HI tests. However, poor protection from apparent virus replication, virus shed, and transmission occurred after challenge with a velogenic NDV strain in those vaccinated with each of the inactivated vaccines. In contrast, LaSota virus, given by the eye drop route, produced excellent protection by the same criteria. In one of the groups of chickens, gross lesions of airsacculitis were seen after vaccination with an inactivated vaccine and subsequent challenge. Revaccination with inactivated vaccines did not enhance the protection of the respiratory tract but did result in an anamnestic serological response (VN and HI). In the post-challenge period, the use of tracheal swabs proved more sensitive as an indicator of virus shed than did cloacal swabs with the velogenic NDV strain used. The practical implications of observations made from the trials are discussed.

Animals↗
PubMedDate not supplied

Development of a virosome vaccine for Newcastle disease virus.

In an effort to protect chickens against Newcastle disease (ND), a nonreplicating virosome vaccine was produced by solubilization of Newcastle disease virus (NDV) with Triton X-100 followed by detergent removal with SM2 Bio-Beads. Biochemical analysis indicated that the NDV virosomes had similar characteristics as the parent virus and contained both the fusion and hemagglutinin-neuraminidase proteins. To target the respiratory tract, specific-pathogen-free chickens were immunized intranasally and intratracheally with the NDV virosome vaccine. This vaccine was compared with a standard NDV (LaSota) live-virus vaccine for commercial poultry. Seroconversion (> or = four fold increase in hemagglutination inhibition [HI] antibody titers) was achieved in all birds vaccinated with the virosome vaccine. Upon lethal challenge with a velogenic NDV strain (Texas GB), all birds receiving either vaccination method were protected against death. Antibody levels against NDV, as determined by enzyme-linked immunosorbent assay and HI titer, were comparable with either vaccine and increased after virus challenge. These results demonstrate the potential of virosomes as an effective tool for ND vaccination.

Animals↗
PubMed1993

RNA editing in Newcastle disease virus.

The co-transcriptional editing of the Newcastle disease virus (NDV) P gene has been studied by sequence analysis of cloned viral genomic RNA and mRNA. Evidence has been obtained for the specific insertion of non-templated G nucleotides, the consequence of which is the generation of three populations of P gene-derived mRNAs. The three populations encode proteins (P, V and W) which have a common N-terminal region, but which utilize three different reading frames at their C termini. Paradoxically, NDV edits its P gene mRNA by the insertion of non-templated G residues in a manner similar to Sendai and measles viruses (P-->V editing) despite its apparent closer evolutionary relationship to the simian virus type 5, mumps and related group of viruses which edit a V genomic sequence to generate an mRNA to encode a functional P protein (V-->P editing).

Amino Acid Sequence↗
PubMed1974

Molecular weight determination of Sendai and Newcastle disease virus RNA.

The molecular weights of Sendai and Newcastle disease virus RNA were estimated by sedimentation in sucrose gradients and by length measurements in the electron microscope under both denaturing and nondenaturing conditions. Sedimentation analyses under denaturing conditions yielded molecular weight estimates of 2.3 x 10(6) to 2.6 x 10(6), whereas length measurements yielded estimates of 5.2 x 10(6) to 5.6 x 10(6) for both denatured and nondenatured viral RNA. It would appear that the conditions of denaturation used (99% dimethyl sulfoxide at 26 C, and reaction with 1.1 M formaldehyde for 10 min at 60 C) do not equally denature parainfluenza virus RNA and other RNAs, such as cellular rRNA, 45S rRNA precursor, and R17 RNA.

Animals↗
PubMed2004

Loss of N-linked glycosylation from the hemagglutinin-neuraminidase protein alters virulence of Newcastle disease virus.

The hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) is an important determinant of its virulence. We investigated the role of each of the four functional N-linked glycosylation sites (G1 to G4) of the HN glycoprotein of NDV on its pathogenicity. The N-linked glycosylation sites G1 to G4 at residues 119, 341, 433, and 481, respectively, of a moderately pathogenic NDV strain Beaudette C (BC) were eliminated individually by site-directed mutagenesis on a full-length cDNA clone of BC. A double mutant (G12) was also created by eliminating the first and second glycosylation sites at residues 119 and 341, respectively. Infectious virus was recovered from each of the cDNA clones of the HN glycoprotein mutants, employing a reverse genetics technique. There was a greater delay in the replication of G4 and G12 mutant viruses than in the parental virus. Loss of glycosylation does not affect the receptor recognition by HN glycoprotein of NDV. The neuraminidase activity of G4 and G12 mutant viruses and the fusogenicity of the G4 mutant virus were significantly lower than those of the parental virus. The fusogenicity of the double mutant virus (G12) was significantly higher than that of the parental virus. Cell surface expression of the G4 virus HN was significantly lower than that of the parental virus. The antigenic reactivities of the mutants to a panel of monoclonal antibodies against the HN protein indicated that removal of glycosylation from the HN protein increased (G1, G3, and G12) or decreased (G2 and G4) the formation of antigenic sites, depending on their location. In standard tests to assess virulence in chickens, all of the glycosylation mutants were less virulent than the parental BC virus, but the G4 and G12 mutants were the least virulent.

Antibodies, Monoclonal↗
PubMed2004

Molecular characterization and phylogenetic study of newcastle disease virus isolates from recent outbreaks in eastern Uganda.

Newcastle disease virus isolates from chickens in eastern Uganda in 2001 were found to be velogenic by fusion protein cleavage site sequence analysis and biological characterization; the intracerebral pathogenicity index was 1.8. Analysis of their hemagglutinin-neuraminidase protein gene sequences revealed a novel genotype unrelated to those that caused previous outbreaks.

Amino Acid Sequence↗
PubMed1985

Specificity of antibodies to Newcastle disease virus.

Specificity of antibodies to Victoria strain of Newcastle disease virus (NDV) found in infectious mononucleosis (IM) and other pathologic sera was investigated by agglutination of NDV-modified human O red blood cells, as well as by immunodiffusion and enzyme immunoassay with various preparations of the virus. These studies clearly demonstrated that the NDV antibodies are distinct from P-B or H-D antibodies. The unexpected observation that guinea pig kidney (GPK) tissues absorbed NDV antibodies allowed their classification into a group of 'GPK-positive' heterophile antibodies. The simultaneous occurrence of the NDV antibodies and H-D antibodies in IM and other diseases suggests the possibility that multiple new antigenic determinants, especially those of carbohydrate nature, may appear due to the alteration of self-antigens as a result of various pathologic processes.

Antibodies, Viral↗
PubMed2003

Characterization of a recombinant Newcastle disease virus expressing the green fluorescent protein.

A recombinant Newcastle disease virus (NDV) expressing the green fluorescent protein (GFP) was generated by applying reverse genetics techniques. The GFP open reading frame flanked by NDV transcription start and stop sequences was inserted between the fusion (F)- and hemagglutinin-neuraminidase genes in a full-length cDNA clone of NDV. This plasmid transcribing antigenome RNA was cotransfected with helper plasmids expressing viral nucleoprotein, phosphoprotein and large protein into cells stably expressing T7 RNA polymerase. The rescued virus was first propagated in embryonated eggs and the allantoic fluid was used to infect cells. Northern blot analysis of RNA isolated from infected cells demonstrated the proper transcription of the introduced GFP-mRNA. The appearance of GFP in live infected cells confirmed further the recovery of a recombinant NDV (rNDVGFP1) expressing the reporter gene. The expression of the heterologous gene was maintained stably for at least five passages in embryonated eggs. The replication kinetics in embryonated eggs and pathogenicity in chickens of rNDVGFP1 did not differ significantly from that of the parent virus. Using GFP autofluorescence, virus infected cells could be tracked easily in native preparations, organ explants and primary tracheal cell cultures. Taken together, these data demonstrate the use of GFP-expressing recombinant NDV for analysis of NDV dissemination and pathogenesis and indicate the potential usefulness of NDV as a vaccine vector.

Animals↗
PubMed2004

Heat inactivation of avian influenza and Newcastle disease viruses in egg products.

Avian influenza (AI) and Newcastle disease (ND) viruses are heat labile viruses, but exact parameters for heat inactivation at egg pasteurization temperatures have not been established. In this study we artificially infected four egg products with two AI (one low [LP] and one high pathogenicity [HP]) and three ND (two low and one highly virulent) viruses, and determined inactivation curves at 55, 57, 59, 61 and 63 degrees C. Based on D(t) values, the time to inactivation of the viruses was dependent on virus strain and egg product, and was directly related to virus titre, but inversely related to temperature. For all temperatures, the five viruses had the most rapid and complete inactivation in 10% salt yolk, while the most resistant to inactivation was HPAI virus in dried egg white. This study demonstrated that the LPAI and all ND viruses were inactivated in all egg products when treated using industry standard pasteurization protocols. By contrast, the HPAI virus was inactivated in liquid egg products but not in dried egg whites when using the low-temperature industry pasteurization protocol.

Animals↗
PubMed2006

[Rescue of a recombinant Newcastle disease virus expressing the green fluorescent protein].

A recombinant Newcastle disease virus (NDV) expressing the green fluorescent protein (GFP) was generated by applying reverse genetics techniques. The GFP open reading frame flanked by NDV transcription start and stop sequences was inserted between the phosphoprotein (P) and matrix protein (M) in a full-length cDNA clone of NDV Lasota vaccine strain. This plasmid transcribing antigenome RNA was cotransfected with helper plasmids expressing viral nucleoprotein, phosphoprotein and large protein into cells stably expressing T7 RNA polymerase. The rescued virus was first propagated in 10-day-old embryonated eggs and the allantoic fluid was used to infect primary chicken embryo fibroblasts (CEF) cells. The appearance of GFP in live infected cells confirmed further the recovery of a recombinant NDV (rNDV-GFP) expressing this reporter gene. Nine successive passages in embryonated chicken eggs were performed. Allantoic fluid samples were then titrated by a microtiter plate HA test. HA positive ailantoic fluid were used for further egg passages. All the allantoic fluid samples were titrated by end point dilutions and infected cells were examined for the presence of GFP expression. To analyze virus growth, 10-day-old embryonated SPF chicken eggs were inoculated with 1 x 10(4) EID50 rNDV or rNDV-GFP. At 24,48,72 and 96 h p.i. the allantoic fluid of inoculated eggs containing live embryos was harvested and clarified by centrifugation. Supernatants were used for titration of EID50 in 10-day-old embryonated SPF chicken eggs. rNDV and rNDV-GFP grew to similar titers (10(9) EID50/mL). In order to test the virulence of rNDV-GFP, infectious allantoic fluid of rNDV-GFP were inoculated into embryonated SPF chicken eggs at 1 x 10(6) EID50. No dead embryonated egg was found within 96 hours. The replication kinetics and pathogenicity in SPF embryonated eggs of rNDV-GFP did not differ significantly from that of the parent virus. LaSota is a widely used NDV live vaccine strain. The reverse genetic system established for this LaSota vaccine strain provided a useful platform for development of novel live viral vector vaccines in future.

Animals↗
PubMed1987

Pathogenicity and immunogenicity in chickens of Newcastle disease viruses isolated from wild ducks. Brief report.

Three Newcastle disease viruses (NDV) isolated from wild ducks in Japan were evaluated for their biological activities, pathogenicity and immunogenicity against one-day-old chickens. One isolate was of the mesogenic type and the other two were of the lentogenic type for chicken. The lentogenic isolates could induce enough immunity in chickens to protect them from challenge with a virulent strain of NDV.

Animals↗
PubMed1990

[Immunologic response in bursectomized and non-bursectomized chickens inoculated with Newcastle disease virus (NDV) after administration of sheep erythrocytes (SRBC) or bovine serum albumin (BSA)].

Astra S hybrid chickens, bursectomized within 48 hours of life, were administered the following antigens: virus of Newcastle disease (NDV) in the fourth week of life and sheep red blood cells (SRBC) or bovine serum albumin (BSA) 10-12 days later. Level of antibodies to the antigens was determined in the sixth week of the birds' life. Administration of SRBC to non-bursectomized chickens inoculated with NDV brought about, in comparison with the chickens not administered SRBC, several-time increase in the titre of anti-NDV antibodies, the anti-SRBC antibodies in chickens inoculated and not inoculated with NDV stayed at the same level. Now, administration of BSA to non-bursectomized chickens inoculated with NDV, in comparison with those not stimulated with BSA, brought about a slight decrease in the titre of anti-NDV antibodies. Bursectomized chickens appeared to exhibit much weaker reaction to the three antigens than the non-bursectomized ones. Administration of SRBC to bursectomized chickens inoculated with NDV exerted no influence on the level of anti-NDV antibodies but, in comparison with those not inoculated with NDV, the titre of anti-SRBC antibodies in them increased twice. Inoculation of bursectomized chickens with NDV followed by administration of BSA brought about, in comparison with those not administered BSA, an increase in the titre of anti-NDV antibodies. At the same time bursectomized chickens inoculated with NDV exhibited a lower level of anti-BSA agglutinins in comparison with the not inoculated ones.

Animals↗