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PubMed2005

[Generation of newcastle disease virus strain ZJI isolated from an outbreak in the goose using reverse genetics technique].

The full-length cDNA clone, NDV3GM122, and the three helperplasmids pCI-NP, pCI-P and pCI-L of Newcastle disease virus strain ZJI isolated from an outbreak in the goose were cotransfected into BSR-T7/5 cell expressing T7 RNA polymerase. Meanwhile, the full-length cDNA clone NDV3GM122 and the three helperplasmids, pCIneoNP, pCIneoP and pCIneoL which were derived from NDV strain La Sota, were also cotransfected into the cell, respectively. Indiect immunofluorescence assay (IFA) was performed 48 to 96 hours post-transfection using NDV HN-specific monoclonal anbtibody (McAb) 6B1 and bright stainings were found in the transfectants, indicating that the full-length clone was functional and the HN protein was expressed. The transfected cell and the supernatant were mixed well and thereafter the mixture was inoculated into specific pathogen free (SPF) chicken eggs. The allanotoic fluid of the injected eggs gave a positive hemagglutinin( HA) titer ranging from 16 to 32 in the secondary passage and increased to 128 in the third passage, which was same to the level of parent wild-type virus. The allantoic fluid containing the recovered NDV was analyzed in hemagglutination inhibition( HI) test by using McAb 6B1 and the specific inhibition was found. The typical morphology of the produced NDV was detected in the electronic microscope. The results mentioned above demonstrated that infectious NDV of strain ZJI was successfully generated, which laid good foundation for the further related research.

Animals↗
PubMedDate not supplied

Detection of a macrophage-specific antigen and the production of interferon gamma in chickens infected with Newcastle disease virus.

Formalin-fixed, paraffin-embedded spleen and intestinal tissues were harvested at 2 days postinfection from 4-wk-old white rock chickens infected with five different strains of Newcastle disease virus (NDV). These tissues were examined for the presence of macrophage antigen expression, virus replication, and interferon gamma (IFN gamma) production. The five strains represented all three NDV pathotypes. Viral replication and IFN gamma, as determined by riboprobe in situ hybridization, were detected only in those chickens infected with velogenic viscerotropic NDV (VVNDV) strains. Macrophage antigen expression, an indicator of macrophage activation, was determined by immunohistochemistry with a macrophage-specific antibody, CVI-ChNL-68.1. Presence of macrophage antigen was most prominent in VVNDV-infected chickens. The distribution of this antigen within tissues was far more diffuse than the staining for viral mRNA. The presence of IFN gamma mRNA was detected in the spleen and intestinal lymphoid tissue of VVNDV-infected chickens. There was also increased macrophage antigen expression in the mesogen-infected birds, but it was less dramatic than in tissues from VVNDV-infected chickens. One of two lentogen-infected birds had evidence of increased macrophage antigen expression only in the spleen.

Animals↗
PubMedDate not supplied

Virus distribution and histopathologic changes in organs of chickens inoculated with Newcastle disease virus (avian paramyxovirus-1) isolated from racing pigeons.

The virus distribution and histopathologic changes in organs of 1-week-old chickens inoculated with three representative isolates of Newcastle disease virus isolated from racing pigeons in Japan were examined. All three isolates were recovered from various organs, including brain, for several days, but not from the blood. Results were highly correlated with their high intracerebral pathogenicity indices (ICPI), in spite of their long mean death time of minimum lethal dose (MDT/MLD).

Animals↗
PubMed1987

Immunity to Newcastle disease in fowl of different breeds, primarily vaccinated with commercial inactivated oil-emulsion vaccines: a laboratory experiment.

Fowl were primarily vaccinated with commercial inactivated oil emulsion (IOE) Newcastle disease (ND) vaccines and immunity was established by both determination of haemagglutination inhibiting (HI) antibodies and by challenge, over periods similar to the economic life span of the birds. The effect of level of maternal antibodies, route of vaccine administration, and vaccine dose on the vaccination results were studied. In one experiment, immunity provoked by vaccination with IOE vaccine was compared with that obtained by vaccination with live vaccines. Vaccination by the subcutaneous route induced a significantly stronger HI response than intramuscular vaccination. Also the vaccine dose per bird and the level of maternal antibodies influenced the vaccination results. In both brown layer and White Leghorn (WL) hens a strong immunity was induced up to at least 45 and 78 weeks of age respectively, by vaccination with IOE vaccine at 6 weeks of age. Also WL hens primarily vaccinated with IOE vaccine at 70 weeks of age, responded with high levels of HI antibodies, which persisted for at least 39 weeks. However, immunity in meat type fowl vaccinated at 6 weeks of age was poor. Following subcutaneous vaccination of one-day-old broilers with high maternal antibody titres with one dose of IOE vaccine per bird, immunity during the fattening period was at an insignificantly lower level than in birds vaccinated with live vaccine at 7 days of age. Twofold vaccination with IOE vaccine at one day old and 18 weeks of age of meat type fowl with high maternal antibody titres provoked a moderate immunity up to at least 55 weeks of age.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1984

[Effect of levamisole on the level of antihemagglutinins, serum sialic acid and lysozyme in the aerosol vaccination of chickens against Newcastle disease].

Tested was the effect of the levamisole antihelminthic on the dynamic of anti-hemagglutinins, serum sialic acid, and lysozyme in the process of building active immunity against Newcastle disease following aerosol vaccination. It was found that there was unidirectional correlation of the active immunologic process with the level of serum sialic acid and lysozyme. The use of levamisole in adequate doses, applied opportunely enhanced the unspecific humoral defense of the body, having no direct influence on the quality of actively built specific immunity.

Aerosols↗
PubMed2001

Rapid detection and differentiation of Newcastle disease virus isolates by a triple one-step RT-PCR.

A triple one-step RT-PCR was developed to screen and differentiate virulent from avirulent Newcastle disease virus (NDV) isolates. Three sets of oligonucleotides were designed, each specific for amplifying NDV fusion protein gene-specific RNA from virulent, avirulent or all isolates respectively. The sensitivity of one-step RT-PCR was determined using viral RNA extracted from serially diluted NDV-infected allantoic fluid and found to be 10(-5) HA units. Application of one-step RT-PCR to various NDV samples, including wild-type virulent isolates and avirulent vaccine strains, demonstrated the potential for rapid identification (3-4 h) of NDV isolates as well as the differentiation of virulent from avirulent strains.

Animals↗
PubMed2004

Genomic sequence of an isolate of Newcastle disease virus isolated from an outbreak in geese: a novel six nucleotide insertion in the non-coding region of the nucleoprotein gene. Brief Report.

We have completely sequenced the genome of an isolate of Newcastle disease virus (NDV) obtained following an outbreak in geese. The genomic sequence consists of 15192 nt, which is six nt longer than the published full length genome of the NDV strains La Sota, B1, Clone-30, Beaudette C and HB V4. The six nt insertion was located in the non-coding region of the nucleoprotein (NP) gene between nt 1646 and nt 1647 of the NDV genome (numbered according to the genomic sequence of the La Sota strain). An additional 22 NDV strains were searched for the existence of this six nt insertion. NDVs in genotypes VI, VII, VIII and IX had this insertion while NDV's in genotypes I, II, III, IV, and V did not. The significance of this insertion in NDV evolution is discussed.

Animals↗
PubMedDate not supplied

An oligonucleotide probe that distinguishes isolates of low virulence from the more pathogenic strains of Newcastle disease virus.

A synthesized oligonucleotide, termed cleavage probe (NDV-CL), has been designed to complement the cleavage-activation site of the fusion gene of the Texas GB isolate of Newcastle disease virus (NDV). This oligonucleotide probe, 21 bases in length, bound with RNA from velogenic strains of NDV tested in a slot-blot hybridization assay. The probe also recognized RNA from the mesogenic strains used in this assay, although no signal was observed with RNA isolated from lentogenic NDVs or with that from other common avian viruses used as controls. This probe did not recognize RNA from isolates of other paramyxovirus serotypes (PMV-2 or PMV-3) included in this study. The ability of this probe to distinguish lentogenic NDVs, which cause little or no clinical disease, from those strains that may produce severe morbidity and/or mortality suggests a potential use for the probe in a molecular diagnostic assay.

Animals↗
PubMed2005

Virulence of Newcastle disease virus is determined by the cleavage site of the fusion protein and by both the stem region and globular head of the haemagglutinin-neuraminidase protein.

Virulence of Newcastle disease virus (NDV) is mainly determined by the amino acid sequence surrounding the fusion (F) protein cleavage site, since host proteases that cleave the F protein of virulent strains are present in more tissues than those that cleave the F protein of non-virulent strains. Nevertheless, comparison of NDV strains that carry exactly the same F protein cleavage site shows that significant differences in virulence still exist. For instance, virulent field strain Herts/33 with the F cleavage site 112RRQRRF117 had an intracerebral pathogenicity index of 1.88 compared with 1.28 for strain NDFLtag, which has the same cleavage site. This implies that additional factors contribute to virulence. After generating an infectious clone of Herts/33 (FL-Herts), we were able to map the location of additional virulence factors by exchanging sequences between FL-Herts and NDFLtag. The results showed that, in addition to the F protein cleavage site, the haemagglutinin-neuraminidase (HN) protein also contributed to virulence. The effect of the HN protein on virulence was most prominent after intravenous inoculation. Interestingly, both the stem region and the globular head of the HN protein seem to be involved in determining virulence.

Animals↗
PubMed2005

Genomic sequences of low-virulence avian paramyxovirus-1 (Newcastle disease virus) isolates obtained from live-bird markets in North America not related to commonly utilized commercial vaccine strains.

Avian paramyxovirus 1 (APMV-1), also referred to as Newcastle disease virus (NDV), variants of low virulence were isolated from chickens, ducks and other unidentified species found in live-bird markets of the northeastern United States. These isolates were characterized as APMV-1 by the hemagglutination-inhibition (HI) assay utilizing NDV-specific polyclonal antisera. However, the isolates failed to react with a monoclonal antibody that has specificity for a wide variety of APMV-1 isolates. Although only highly virulent isolates require reporting to international regulatory agencies, the ability to correctly identify APMV-1 types is important for control and regulatory purposes. Protein gel patterns of the purified isolates resembled previously reported APMV-1 and anti-NDV polyclonal sera recognized the viral proteins. For three isolates oligonucleotide primers specific for the nucleoprotein, fusion protein and polymerase genes of NDV were utilized to synthesize cDNA using viral RNA as a template. Approximately 12kb of the genome was subsequently sequenced for the three isolates that included the nucleoprotein, phosphoprotein, matrix protein, fusion (F) protein, hemagglutinin-neuraminidase protein genes and a 5' portion of the polymerase gene. The isolates had an F protein cleavage site sequence of ERQER/LVG indicating low-virulence viruses that phylogenetically separated with other unique NDV isolates designated as a lineage 6 genotype. Additionally, a four amino acid insert was detected in the predicted phosphoprotein which complies with the "rule of six" among paramyxoviruses. These APMV-1 genotypes have not been previously reported in North America and further substantiate the heterogeneous genetic nature of these commercially important pathogens found worldwide.

Amino Acid Sequence↗
PubMed1980

Collection and processing of blood samples dried on paper for microassay of Newcastle disease virus and avian influenza virus antibodies.

A practical method for collection and processing of dried whole blood samples on filter paper was developed to facilitate large-scale testing programs for Newcastle disease virus and avian influenza virus antibodies. A modified paper punch was used to cut and place dried blood samples simultaneously in a standard 96-well microlate for elution of antibody. Twelve eluted samples were simultaneously transferred to another microplate for the hemagglutination-inhibition (HI) microtest. Similar HI titers were obtained with simultaneously collected serum and dried blood samples. Minor differences were not considered of practical importance in diagnostic serologic studies. Dried blood titers were not markedly affected by method of drying (37 C for 2 hours or 26 C for 4 hours), by storage for 24 hours before drying, or by storage of dried samples at 4 C for 28 days or 30 C for 14 days. Blood dried on paper was a satisfactory sample for assay of HI antibodies to Newcastle disease virus and avian influenza virus.

Animals↗
PubMed1964

STUDIES ON PERSISTENT INFECTIONS OF TISSUE CULTURES. V. THE INITIAL STAGES OF INFECTION OF L(MCN) CELLS BY NEWCASTLE DISEASE VIRUS.

The initial stages of infection of L(MCN) cell populations with standard Newcastle disease virus (NDV(ST)) were analyzed in an effort to elucidate the steps leading to survival of the cultures and to the indefinite persistence of the infectious process at a low level. Cells were exposed in suspension to NDV at varying multiplicities and the monolayer cultures derived from such cells assayed at intervals for cellular growth rates, percentage of infected cells as determined by immunofluorescence, yields of viral progeny and of interferon, and, on occasion, resistance to superinfection with vesicular stomatitis virus. The percentage of cells calculated to be initially infected on the basis of adsorption data was found to match closely the percentage of immunofluorescent cells resulting from the first infectious cycle (up to 24 hours). Cells initially infected with NDV(ST) produced a mixed progeny of infectious virus (from 15 to 40 pfu/cell) and about 10 times as many non-infectious particles in 24 hours [NDV(L(MCN))], but little or no interferon. If all cells were infected the cultures ultimately died. At multiplicities of infection (m) of 2 or less the cultures survived with increasing ease as the percentage of infected cells was reduced. The number of pfu per infected cell was of the above order during the first 3 days; it declined thereafter. Limited secondary spread of the infection was noted by 48 hours and no further cycling was noted thereafter. As m decreased from 2.0 to 0.1 there was an increase in the yields of interferon and the time at which peak titers were reached. Addition of anti-NDV serum 2 hours after infection prevented measurable production of interferon. In contrast, following exposure of cells to NDV(L(MCN)) at multiplicities ranging from 20.0 to 0.2 (based on infectious virus) all cultures survived, no secondary spread was noted, the number of pfu per infected cells was reduced at the higher multiplicities, and the yields of interferon were similar and maximal by 24 hours and not affected by anti-NDV serum added after an adsorption period of 2 hours. It is concluded that the non-infectious virus particles in the progeny released from NDV(ST)-infected cells induce resistance in remaining cells or, if adsorbed simultaneously with infectious virus, abort the intracellular infectious process. In both instances interferon is produced which may then render additional cells resistant. The non-infectious component is considered an incomplete or defective product of viral replication and not merely thermally inactivated virus. NDV(ST) partially or completely inactivated at 37 degrees C induced neither cellular resistance nor synthesis of interferon. The incomplete viral component behaved in all respects like ultraviolet-inactivated NDV(ST) except that it was significantly more efficient in inducing interferon synthesis. On the basis of the presented data a scheme has been devised and discussed which appears to explain satisfactorily the events which take place on initial infection of L(MCN) cells with NDV and which lead to the persistence of the infectious process.

Animals↗
PubMed1964

USE-DILUTION TEST AND NEWCASTLE DISEASE VIRUS.

The use-dilution test for evaluating the effectiveness of disinfectants against bacteria was modified to determine the effectiveness of disinfectants against a group of viruses. Modifications were kept to a minimum to retain the general principles of the test and thereby retain the test's familiarity among testing laboratory personnel. Modifications included the use of a standard allantoic fluid suspension of Newcastle disease virus instead of a standard bacterial culture. The only other modification was the inoculation of six embryonated chicken eggs (10 to 12 days old) with 0.1 ml of nutrient broth into which a carrier ring was transferred after a standard period in diluted disinfectant. The death or survival of 60 embryos, then, is the criterion by which a disinfectant can be judged effective at use-dilution. Experiments are described which establish the validity of the modified test procedure. The effectiveness of nine common disinfectants against Newcastle disease virus as judged by this test procedure is reported.

Animals↗
PubMed2000

Determination of the disulfide bond arrangement of Newcastle disease virus hemagglutinin neuraminidase. Correlation with a beta-sheet propeller structural fold predicted for paramyxoviridae attachment proteins.

Disulfide bonds stabilize the structure and functions of the hemagglutinin neuraminidase attachment glycoprotein (HN) of Newcastle disease virus. Until this study, the disulfide linkages of this HN and structurally similar attachment proteins of other members of the paramyxoviridae family were undefined. To define these linkages, disulfide-linked peptides were produced by peptic digestion of purified HN ectodomains of the Queensland strain of Newcastle disease virus, isolated by reverse phase high performance liquid chromatography, and analyzed by mass spectrometry. Analysis of peptides containing a single disulfide bond revealed Cys(531)-Cys(542) and Cys(172)-Cys(196) linkages and that HN ectodomains dimerize via Cys(123). Another peptide, with a chain containing Cys(186) linked to a chain containing Cys(238), Cys(247), and Cys(251), was cleaved at Met(249) with cyanogen bromide. Subsequent tandem mass spectrometry established Cys(186)-Cys(247) and Cys(238)-Cys(251) linkages. A glycopeptide with a chain containing Cys(344) linked to a chain containing Cys(455), Cys(461), and Cys(465) was treated sequentially with peptide-N-glycosidase F and trypsin. Further treatment of this peptide by one round of manual Edman degradation or tandem mass spectrometry established Cys(344)-Cys(461) and Cys(455)-Cys(465) linkages. These data, establishing the disulfide linkages of all thirteen cysteines of this protein, are consistent with published predictions that the paramyxoviridae HN forms a beta-propeller structural fold.

Amidohydrolases↗