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[Newcastle disease virus heredity mutation and correlation of HN and F gene].

Prevailed Newcastle disease virus isolates were collected during 1999-2005 in China. These isolates were purified by CEF plaque assay and replicated in SPF embryos. The fusion protein (F) gene and hemagglutinin-neuraminidase (HN) gene of these isolated viruses were cloned and sequenced. Some of the F gene and HN gene sequences from GenBank were also used in this study. The homologies of nucleotide and amino acids were compared and correlations were analyzed by SPSS8.0 software among different length sequences of the F gene or HN gene. The nucleotide homologies and correlation among the F gene and HN gene were also analyzed. The results indicated there are good correlation among different length sequences of the F gene or HN gene and the F genome or HN genome (r > or = 0.973). There was also good correlations among different length amino acids of NDV F protein or HN protein (0.911< or = r < or = 0.968). But, there was only a less correlation between the whole F gene and HN gene (r = 0.312). The heredity mutation of HN genes had the character of geographical areas. The sequences of HN gene in Chinese isolates had an identity of more than 97%. But there was only 79.2% - 80.7% in HN nucleotide homology among the Chinese isolates and La Sota (vaccine).

Animals↗

The effect of a mesogenic and a lentogenic Newcastle disease virus strain on Burkitt lymphoma Daudi cells.

The destructive effect of Newcastle disease virus (NDV) strains on Burkitt lymphoma Daudi cells was investigated. Interaction of an active and UV-inactivated mesogenic strain (Roakin), as well as an active attenuated lentogenic strain (B1), grown in the allantoic sac of embryonated eggs, at high multiplicity, caused inhibition in cellular DNA synthesis and arrest in cell multiplication, eventually killing of the cells. The lentogenic strain cultivated in chicken fibroblasts exhibited only a moderate activity. The mechanism of the cytolytic effect is presumably linked to the increase in cell membrane permeability indicated by the elevation in 51Cr release. Thus it appears that the massive adsorption and/or penetration of viral particles, active or UV-inactivated (or possibly a toxic component that resides in the virion), damages the plasma membrane and may be responsible for the killing of the cells.

Adsorption↗

Induction of NO synthesis in macrophages by Newcastle disease virus is associated with activation of nuclear factor-kappa B.

Newcastle disease virus (NDV) has received much attention recently because of its non-specific immune stimulating potential and its various anti-tumor activities. Here we describe that NDV induces synthesis of NO and causes an activation of nuclear factor-kappa B (NF-kappa B) in murine macrophages. These reactions were part of an activation process which included also stimulation of adenosine deaminase and inhibition of 5'-nucleotidase. NDV-mediated NO synthesis and NF-kappa B activation were blocked by an antioxidant (butylated hydroxyanisole), by an inhibitor of protein tyrosine kinase (genistein) and of protein kinase A (H-89), but not by an inhibitor of protein kinase C (staurosporin). These data suggest that signalling requirements of NF-kappa B activation and NO production in NDV-treated macrophages are similar.

Adenosine Deaminase↗

Analysis of matrix protein gene nucleotide sequence diversity among Newcastle disease virus isolates demonstrates that recent disease outbreaks are caused by viruses of psittacine origin.

Nucleotide sequence analysis was completed for isolates of Newcastle disease virus (NDV; avian paramyxovirus 1) from 1992 outbreaks in cormorants and turkeys. These isolates were of the neurotropic velogenic type. The cormorant and turkey NDV isolates had the fusion protein cleavage sequence 109SRGRRQKR/FVG119, as opposed to the consensus sequence 109SGGRRQKR/FIG119 of most known velogenic NDV isolates. The R for G substitution at position 110 may be unique for the cormorant and turkey isolates. For comparative purposes, nucleotide sequencing and analysis of the conserved matrix protein gene coding region were completed for isolates representing all pathotypes. Phylogenetic relationships demonstrated that there are two major groups of NDV isolates. One group includes viruses found in North America and worldwide, such as B1, LaSota, Texas/GB, and Beaudette/C. The second group contains isolates, such as ulster/2C, Australia/Victoria, and Herts/33, considered exotic to North America. Within this second group are viruses of psittacine origin. The viruses from 1992 outbreaks of Newcastle disease in North America, and an isolate thought to have caused the major outbreak in southern California during the 1970s, are most closely related to an NDV isolate of psittacine origin.

Amino Acid Sequence↗

Effect of probe-site mismatches on detection of virulent Newcastle disease viruses using a fusion-gene real-time reverse transcription polymerase chain reaction test.

Virulent forms of Newcastle disease virus (NDV) are a major concern for poultry producers around the world and the rapid diagnosis of an outbreak is crucial to any control program. A validated real-time reverse transcription-PCR test (fusion test) directed at the fusion-cleavage site of NDV was developed to differentiate virulent Newcastle disease virus strains from those of low virulence, however one virulent isolate, Dove/Italy/2736/2000, escaped detection during the initial evaluation of the test. The objectives of this study were to determine how this isolate differed from other detectable isolates, to identify other isolates that may fail detection, and to characterize the effect of specific probe-site mutations on the fusion test at a range of annealing temperatures. Using a virulent NDV isolate (Game fowl/US(CA)/2002) as a backbone that has 100% identity to the fusion-test probe, specific changes were made to the fusion-test probe-site to reflect the unique mismatches found in Dove/Italy/2736/2000 and other selected regions of the probe. Mutated clones with mismatches unique to Dove/Italy/2736/2000 at positions 6, 13, and 14 were not detected until annealing temperatures were lowered to 50 degrees C. Those detected at 58 degrees C contained 1-2 mismatches (position 1 and 6, 13 and 14, or 14 only) although increased cycle threshold values compared to the parent clone indicated decreased sensitivity. Data from this study predicts that the fusion test may fail to detect some viruses among lineage 4b and potential solutions to identify this subset of viruses include lowering the annealing temperature or modifying the probe.

Animals↗

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.

Animals↗

Construction of a pigeonpox virus recombinant: expression of the Newcastle disease virus (NDV) fusion glycoprotein and protection of chickens against NDV challenge.

A pigeonpox transfer plasmid was constructed by cloning a 2.5 kb DNA fragment containing the viral thymidine kinase (TK) gene in the psp65 plasmid. The vaccinia virus P11K promoter followed by the NDV fusion (F) gene was inserted in the TK gene. The F gene was transferred to the viral genome by homologous recombination in pigeonpox virus infected CEF cells, transfected with the recombinant plasmid. Recombinant viruses were selected with BUdR and screened for their ability to induce fusion between adjacent cells. Because of the unexpected growth advantage of the TK+ WT over the TK- recombinants, viral purification was needed to obtain stable recombinants expressing a glycosylated and cleaved F protein. Vaccination of chickens by the follicular method induced high anti-F antibody titers and good protection against challenge with the virulent Italian NDV strain. Half of the oculonasal vaccinated chickens showed anti F antibodies and also half of them were protected. Although protection seems to be correlated with antibody titers, no neutralizing antibodies were found.

Animals↗

[Properties of local velogenic strains of Newcastle disease virus in poultry].

Studies were carried out on four strains of the Newcastle disease virus, isolated in Bulgaria and referred, as typed by the routinely used methods, to the velogenic group. The strains proved to be hemagglutinin - thermostable. Two of the local virus strains (Rosa and Toutrakan) belonged to the velogenic-neurotropic pathogenic type, and two (Bregovo and Haskovo) - to the velogenic - viscerotropic pathogenic one. All four strains were slowly eluated from chicken erythrocytes. Full immunologic similarity was established between these strains by means of the live vaccines used in this country for the specific prevention of Newcastle disease, produced with the lentogenic strains Hitchner B1 and La Sota and the mesogenic Komarov.

Animals↗

Rapid in vitro assessment of the virulence of Newcastle disease virus isolates using the ligase chain reaction.

The ligase chain reaction was used to assess the virulence of isolates of Newcastle disease virus. In the main study, 18/18 virulent isolates whose nucleotide sequences that code for the cleavage site and fusor peptide regions were known, successfully ligated oligonucleotides in a primer mix for virulent viruses termed VPM. Five of these isolates yielded a more intense ligated product with a second primer mix for virulent viruses called VPM1. No ligation was evident with eight avirulent isolates in tests with VPM or VPM1, however, each of these viruses did yield a strong ligated product with the primer mix for avirulent viruses (AVPM) as did one virulent isolate considered to be a mixture. Two virulent Australian isolates, 1238/1998 and 1248/1998, showed low but seemingly specific ligation with AVPM. In a blind study, 8/9 virulent isolates whose sequences were unknown ligated primers in VPM. Three avirulent and one virulent isolate, the latter again probably a mixture, ligated primers in AVPM. Ligation of oligonucleotides in VPM and AVPM was detectable in mixtures where virulent and avirulent isolates represented 0.1% and 0.01% by volume respectively of the viral population. The results indicate that LCR offers a potential in vitro alternative to current in vivo tests for virulence determination of Newcastle disease virus isolates.

Ligase Chain Reaction↗

[Study of the immunogenic properties of 3 lentogenic strains of the Newcastle disease virus isolated in Bulgaria].

Three local strains of lentogenic type of the Newcastle disease were used to produce live liquid vaccines. These were given per os to broiler chickens in order to test their immunogenicity (first vaccination at the age of 25 days and vaccination when 70 days old). The immunity thus acquired was followed serologically by the titer of the antihemagglutinins and the resistance manifested by the birds at the control infection with a proven lethal dose of a highly virulent velogenic strain of the Newcastle disease virus. The results obtained were compared with those observed with the use of liquid vaccines of the strains Hitchner-B1 and La Soto. It was concluded that the vaccines of the local Newcastle disease strains are completely harmless and immunogenic. The vaccinal strains are excreted by the immunized birds and serve to immunize the contact control chickens.

Age Factors↗

Newcastle disease virus in Taiwan: II, Relationship between polykaryocytosis and virus virulence.

Fifteen selcted local isolates and five known Newcastle disease virus strains were examined for their cytopathic effects in chick embryo kidney (CEK) cells, egg-infectious units in chick embryos (CE), virulence by mean death time, intracerebral and intravenous pathogenicity indexes for CE and chicks, and ability to cause polykaryocytosis of fusion from within (FFWI) or fusion from without (FFWO) in CEK and BHK-21 monolayer cells. The capacity of the different virus strains to induce cell FFWI at 15 hr post-infection was related to their virulence for CE and chicks, but cell FFWO did not seem to be any relationship with the virulence of the strains.

Animals↗

A mutant CHO-K1 strain with resistance to Pseudomonas exotoxin A is unable to process the precursor fusion glycoprotein of Newcastle disease virus.

RPE.40, a mutant strain of CHO-K1 cells isolated for resistance to Pseudomonas exotoxin A and cross-resistant to alphaviruses, is also highly resistant to virulent strains of Newcastle disease virus. The resistance of RPE.40 cells to Newcastle disease virus results from the failure to cleave the viral envelope precursor glycoprotein Fo to fusion glycoprotein F1 at the consensus sequence (Lys/Arg)-Arg-Gln-(Lys/Arg)-Arg.

ADP Ribose Transferases↗