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PubMed1999

[Strain differentiation of the Newcastle disease virus by reverse transcriptase-polymerase chain reaction and sequencing population exchange].

A system for detection and strain differentiation of Newcastle disease virus (NDV) by reverse transcription of polymerase chain reaction (RT-PCR) (isolation of RNA, choice of primers for nested PCR, and purification of PCR products) and sequencing is developed and optimized. A nucleotide sequence of gene F site, coding for the F2/F1 cleavage site of F0 fusion protein and including several hypervariable regions, is determined for 10 Russian strains and vaccine strains. The data indicate a replacement of NDV populations in Russia and a rapid evolution of the virus. The origin of pathogenic NDV strains which have been circulating up to the present time is still unknown.

Base Sequence↗
PubMedDate not supplied

Scanning electron microscopy of tracheal epithelium of chickens infected with velogenic viscerotropic Newcastle disease virus.

Ultrastructural changes in the tracheal epithelium of chickens infected intranasally with velogenic viscerotropic Newcastle disease virus were examined by scanning electron microscopy. Hypertrophy of the mucus-secreting, or goblet, cells was the first sign of change, followed by disoriented and deformed cilia, hemorrhage, and hyperplasia of goblet cells accompanied by an increase in mucus. By day 7 postinfection, there was a marked decrease in the number of ciliated cells. Submucosal glands and some collagen fibers were exposed to the surface, an indication of loss of the epithelial cells. Macrophages and cell debris were abundant, and hyperplasia of the basal cells was evident in the later stages of infection, probably in an attempt to regenerate the lost epithelium. However, all chickens died 10 days postinfection, before any further work could be done.

Animals↗
PubMed1974

Polyadenylate sequences on Newcastle disease virus mRNA synthesized in vivo and in vitro.

Polyadenylate [poly(A)] sequences are associated with the 35 and 50S Newcastle disease virus (NDV)-specific RNAs as well as all six to seven of the 18-22S NDV-specific messenger RNAs extracted from infected chicken embryo cells. The poly(A) associated with the 18-22S RNA has an average size of 120 to 130 nucleotides. The 18-22S RNA synthesized in vitro by NDV's virion-bound polymerase contains six to seven species of the same size and relative proportions as its intracellular counterpart. This in vitro synthesized 18-22S RNA also contains covalently linked poly(A) sequences which, although variable in size, are usually larger and more heterogeneous than those from the infected cell. In vitro RNA synthesis is supported not only by magnesium (at an optimal concentration of mM) but by manganese (at an optimal concentration of 0.5 to 1.0 mM) as well. However, the major product made in the presence of manganese, although sedimenting at 18 to 22S, differs somewhat from the product made in the presence of magnesium.

Adenosine↗
PubMed1997

Disulfide bond formation is a determinant of glycosylation site usage in the hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus.

Determinants of glycosylation site usage were explored by using the hemagglutinin-neuraminidase (HN) glycoprotein of the paramyxovirus Newcastle disease virus. The amino acid sequence of the HN protein, a type II glycoprotein, has six N-linked glycosylation addition sites, G1 to G6, two of which, G5 and G6, are not used for the addition of carbohydrate (L. McGinnes and T. Morrison, Virology 212:398-410, 1995). The sequence of this protein also has 13 cysteine residues in the ectodomain (C2 to C14). Mutation of either cysteine 13 or cysteine 14 resulted in the addition of another oligosaccharide chain to the protein. These cysteine residues flank the normally unused G6 glycosylation addition site, and mutation of the G6 site eliminated the extra glycosylation found in the cysteine mutants. These results suggested that failure to form an intramolecular disulfide bond resulted in the usage of a normally unused glycosylation site. This conclusion was confirmed by preventing cotranslational disulfide bond formation in cells by using dithiothreitol. Under these conditions, the wild-type protein acquired extra glycosylation, which was eliminated by mutation of the G6 site. These results suggest that localized folding events on the nascent chain, such as disulfide bond formation, which block access to the oligosaccharyl transferase are a determinant of glycosylation site usage.

Amino Acid Sequence↗
PubMed1979

Resistance of chickens to challenge with the virulent Herts 33 strain of Newcastle disease virus induced by prior infection with serologically distinct avian paramyxoviruses.

Results indicate that some degree of protection from challenge by Newcastle disease virus (NDV)/Herts 33 was conferred on chickens by prior infection with PMV/turkey/Wisconsin/68, PMV/turkey/Ontario/6661/68, PMV/Netherlands/449/75 and PMV/parakeet/England/39/78 viruses, all of which are serologically related but distinguishable from NDV. Except for one bird which survived challenge three weeks after infection with Robin/Hiddensee/19/75, no protection was seen in chickens infected with other unrelated avian paramyxoviruses. In contrast to infection with NDV-B1, birds protected by infection with avian paramyxoviruses showed large increases in NDV haemagglutination inhibition (HI) titres after challenge. In these birds considerable increases in the homologous HI titres were also seen after challenge.

Animals↗
PubMed1982

[Precipitinogen activity of Newcastle disease virus and humoral immunity in birds].

Studies was the precipitinogenic activity of the strains La Sota, 'H', and 'II' of the Newcastle disease virus, using chick embryos. It was found that all three of the strains produce precipitinogens. An antigen was obtained for the precipitation reaction in agar gel and the demonstration of precipitins that were specific for the ND virus in the blood serum of birds. Contrary to the hemagglutinins and precipitinogens proved heatresistant. There was a correlation between the antihemagglutinins and the precipitinogens in the blood serum of the immune birds. The low percent of birds (up to 12%) having precipitins in their serum spoke of the development of solid immunity against Newcastle disease within the flock. On the other hand, the high percent of birds having precipitins (over 50%) indicated that the infection persisted, and because of the immunity present it fairly often assumed atypical course or was manifested with low mortality rate.

Animals↗
PubMedDate not supplied

Antibody responses to La Sota strain vaccines of Newcastle disease virus in ostriches (Struthio camelus) as detected by enzyme-link ed immunosorbent assay.

Because of the fact that South Africa is a Newcastle disease virus (NDV)-endemic country, major concerns exist that the export of ostrich meat could transmit velogenic strains of this disease. The ability to transmit the virus could be reduced by effective vaccination of South African ostriches. In this study, two vaccination trials were conducted to assess serum antibody production in response to vaccination with La Sota strain NDV vaccines. To this end, a commercially available chicken anti-NDV enzyme-linked immunosorbent assay (ELISA) was modified for the detection of anti-NDV antibodies in ostrich serum. The results obtained with this ELISA were verified by comparison with an indirect ELISA. In the first trial, ostriches were immunized subcutaneously four times with different volumes of an inactivated vaccine and their immune response was determined from 2.5 mo up to the ideal slaughter age of 14 mo. Results indicated that ostriches responded in a dose-dependent manner and gave support for the vaccination schedule currently recommended to South African farmers. In a second trial, immunization by eyedrop with a live La Sota vaccine of 5-wk-old ostriches did not elicit a humoral immune response. The results indicate that it is highly unlikely that ostriches that have been vaccinated according to the recommended vaccination schedule can transmit the virus.

Animals↗
PubMedDate not supplied

Cellular and humoral response of in ovo-bursectomized chickens to experimental challenge with velogenic Newcastle disease virus.

Humorally deficient, in ovo-bursectomized (Bx) and sham-Bx chickens were vaccinated twice, 1 month apart, with Newcastle disease virus (NDV) Roakin strain and challenged with a velogenic viscerotropic NDV strain via the oronasal route. Hemagglutination-inhibition and seroneutralization tests showed that Bx chickens had reduced antibody-mediated immunity to virus infection. In contrast, they had significantly higher cell-mediated immunity (CMI) before challenge, as estimated simultaneously by determination of blastogenic capacity of peripheral blood lymphocytes induced by phytohemagglutinin and by specific antigen stimulation. After virus challenge, there was transitory inhibition of CMI based on marked reductions in levels of stimulation indices, and this impairment in CMI was supported by persistence of virus in Bx chickens for longer periods. Bx chickens resisted challenge, even though antibody titers were well below those considered predictive of resistance to challenge, suggesting that CMI provides a degree of resistance to velogenic NDV.

Animals↗
PubMedDate not supplied

Pathology of the trachea in turkeys exposed by aerosol to lentogenic strains of Newcastle disease virus.

Five groups of 4-week-old turkey poults were each infected by aerosol with a different lentogenic strain of Newcastle disease virus. Four days postinfection, sections of tracheas were collected for histopathologic characterization and virus titration. The most prominent lesions were fibrinopurulent exudate in tracheal lumens, hyperplasia of epithelial cells, and infiltration by lymphocytes. All strains multiplied to high titers and produced similar microscopic lesions, but the number of poults with severe microscopic lesions varied among groups.

Aerosols↗
PubMed1969

Cells persistently infected with newcastle disease virus: I. Properties of mutants isolated from persistently infected L cells.

The strain of Newcastle disease virus (NDV(pi)) present in persistently infected L cells differed markedly from the Herts strain (NDV(0)) used to initiate the infection. NDV(pi) produced small plaques (less than 1 mm) in chick embryo cell cultures, whereas the wild type (NDV(0)) produced large plaques (2 to 3 mm). The two viruses differed in a number of additional properties. Whereas 80% of adsorbed NDV(0) eluted from chicken red blood cells at 37 C, only about 20% of NDV(pi) was recovered under similar conditions. There was no significant difference in the neuraminidase content of the two viruses. The infectivity of NDV(0) was stable for 1 hr at 48 C, whereas 99.9% of the infectivity of NDV(pi) was destroyed. The two viruses also differed in lethality for chick embryos; NDV(pi) had significantly reduced lethality for 9-day-old chick embryos when compared to NDV(0). In contrast to NDV(0), which produced an abortive infection in L cells, NDV(pi) not only replicated effectively and destroyed these cells, but also induced significantly higher quantities of interferon than did NDV(0). These data furnished additional evidence for the lack of relationship of interferon production to abortive infection of L cells with NDV(0). In contrast, interferon was found to play a significant role in the maintenance of persistent infection.

Journal Article↗
PubMed1980

[Comparative pathomorphological studies of turkeys and hens experimentally infected with Newcastle disease virus].

Comparative pathomorphologic studies were carried out with 40 birds (29 turkeys and 11 hens) infected per os with 0.5 cc embryonal fluid 10(5) ELD 50 each with two Newcastle disease virus strains (Texas -- standard and 7/5 isolated from dead hens and typed as viscerotropic). It was found that the pathoanatomical picture is negative for all birds experimentally infected with the Texas strain. Haemorrhagic necrotic modification of the glandular stomach and intestine were found in birds infected with strain 7/5. The pathoanatomical finding is better expressed in hens compared with that for turkeys. The histologic modification in birds infected with strain Texas were manifested mostly by nonsuppurative encephalitis. Haemorrhagic-diphtheroid inflammation of the alimentary tract, haemorrhages in the viscera and unreactive necrotic foci in the spleen as well as poorly manifested inflammatory changes in the central nervous system were found in birds infected with strain 7/5. No pathoanatomical modifications for Newcastle disease were found in turkeys, analogous to hens, infected with velogenic neurotropic virus. The velogenic viscerotropic strain is less pathogenic for turkeys than for hens. It causes slighter haemorrhagic necrotic modification in turkeys viscera than in hens. Nonsuppurative encephalitis established histologically in both turkeys and hens infected with neurotropic and viscerotropic Newcastle disease virus is a constant finding and may be used as a diagnostic sign of the disease.

Animals↗
PubMed2005

High-throughput real-time RT-PCR assay to detect the exotic Newcastle Disease Virus during the California 2002--2003 outbreak.

During the 2002--2003 Exotic Newcastle Disease (END) outbreak in Southern California, a high-throughput real-time reverse transcriptase-polymerase chain reaction (RRT-PCR) system was developed to respond to the large diagnostic and surveillance sample workload. A 96-well RNA extraction method, using magnetic bead technology, combined with a 96-well RRT-PCR assay, allowed 1 technician to process and test more than 400 samples per day. A 3-technician team could complete testing on approximately 1,900 samples per day. The diagnostic sensitivity of the high-throughput RRT-PCR assay was 0.9967 (95% CI 0.9937-0.9997) based on 926 virus isolation confirmed positive samples. Diagnostic specificity using an initial 434 virus isolation confirmed negative samples was 100%. A diagnostic specificity of 0.9999 (95% CI 0.9999, >0.9999) was subsequently calculated on the basis of 2 false-positive results among 65,343 surveillance samples collected after the final END-positive case was confirmed in May 2003. Assay performance over 500 replicates, including reproducibility of the combined extraction and RRT-PCR amplification steps yielded a standard deviation of 0.70 RRT-PCR cycle thresholds (Ct) and a standard deviation of 0.59 Ct for the RRT-PCR steps alone. The high-throughput RRT-PCR developed for END contributed significantly to the 2002--2003 END control effort, reducing the predicted timeline for eradication from 3 years to just 11 months, primarily because of the large number of samples that could be rapidly tested. The 96-well approach described for high-throughput END RRT-PCR could be adapted to other rapid, high-volume testing needs, as required for potential foreign animal disease responses or intensive surveillance efforts.

Animals↗
PubMed1999

Tumor stimulator cell modification by infection with Newcastle Disease Virus: analysis of effects and mechanism in MLTC-CML cultures.

Effects of tumor stimulator cell modification by infection with Newcastle Disease Virus (NDV) are described as analysed in vitro in mixed lymphocyte tumor cell cultures (MLTC). Direct antitumor effects were seen with human melanoma or colon-carcinoma cells in a dose- and time-dependent manner when using live but not UV inactivated virus. When T cell stimulation was measured by [3H]-thymidine uptake, NDV infected tumor stimulator cells did not show an augmentation but rather an inhibitory effect in comparison to non-infected stimulator cells. Virus infected tumor stimulator cells were, however, capable of augmenting the induction of tumor specific cytotoxic T cells in MLTC-CML assays when using murine ESb lymphoma immune cells and syngeneic NDV modified ESb cells as stimulators. A CML stimulatory effect was also shown for NDV modified third party cells and thereof derived conditioned medium. These effects are most likely explained by interferon- which is induced in tumor cells by NDV infection and by interferon-á which is induced in responder cells when stimulated with NDV infected stimulator cells.

Animals↗
PubMed1981

The isolation of salmonellae, Newcastle disease virus and other infectious agents from quarantined imported birds in Canada.

Necropsy and culture results are presented for 269 consignments of imported birds (mainly psittacine and passerine species) examined between January 1977 and August 1980. Consignments were submitted for diagnosis of clinical illness or deaths occurring among these birds while they were in quarantine before entry into Canada. Enteritis and injury were the most frequent diagnoses. Pathogens or potential pathogens were isolated from 77% of consignments. Newcastle disease virus was isolated nine times, and Chlamydia psittaci was isolated once. Escherichia coli (from 113 consignments) and salmonellae (from 49) were the most common bacteria isolated, and reoviruses (from 22) and paramyxoviruses other than Newcastle disease virus (from 22) were the most common viruses. Salmonella typhimurium was the most common Salmonella serovar. Salmonella hadar was isolated from turkey poults imported from Great Britain. The possible public health significance of the role of imported birds in the introduction of exotic Salmonella serovars, or of serovars resistant to several antimicrobials is discussed.

Animals↗