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Resistance to velogenic Newcastle disease virus in leghorn chickens by use of prophylactic lymphokines.

A group of 1-day-old commercial leghorn chickens was prophylactically treated with lymphokines obtained from lymphocyte cultures of chickens previously infected with Salmonella enteritidis (S. enteritidis-immune lymphokines [SE-ILK]) with the objective to investigate the effect of SE-ILK on development of Newcastle disease (ND) infection caused by Chimalguacan strain, a Mexican velogenic ND virus (vNDV). Clinical signs, histologic lesions, and hemagglutination-inhibition (HI) serum titers were compared with four other groups, namely, chickens without SE-ILK treatment with virus challenge; with SE-ILK without virus challenge; with nonimmune lymphokine (NILK) treatment and virus challenge; with lymphokine treatment and no virus challenge. SE-ILK was administered intraperitoneally in a dose of 0.5 ml/chicken and was followed 30 min later with the challenge of vNDV in a dose of 10(7.6) 50% embryo lethal dose/ml per bird. Birds were observed during 21 days of postchallenge. Detection of histologic changes and virus isolation procedures were carried out on the third, seventh, 14th, and 21st postinoculation days. HI tests were performed first before treatment and later on the days of histologic sample collection except on the third postinoculation day. Results showed that SE-ILK administration conferred resistance to the chickens because: 1) it significantly diminished the severity of ND infection by inhibiting appearance of clinical signs (P < 0.001), lesions (P < 0.005), and histopathologic changes (P < 0.005); 2) it decreased vNDV isolation rate from the organs (P < 0.001), and 3) it potentialized and even accelerated (P < 0.005) primary immune response by antibodies in the presence of vNDV.

Animals↗

Experimental infection of turkeys with avian pneumovirus and either Newcastle disease virus or Escherichia coli.

Avian pneumoviruses (APVs) are RNA viruses responsible for upper respiratory disease in poultry. Experimental infections are typically less severe than those observed in field cases. Previous studies with APV and Escherichia coli suggest this discrepancy is due to secondary agents. Field observations indicate APV infections are more severe with concurrent infection by Newcastle disease virus (NDV). In the current study, we examined the role of lentogenic NDV in the APV disease process. Two-week-old commercial turkey poults were infected with the Colorado strain of APV. Three days later, these poults received an additional inoculation of either NDV or E. coli. Dual infection of APV with either NDV or E. coli resulted in increased morbidity rates, with poults receiving APV/NDV having the highest morbidity rates and displaying lesions of swollen infraorbital sinuses. These lesions were not present in the single APV, NDV, or E coli groups. These results demonstrate that coinfection with APV and NDV can result in clinical signs and lesions similar to those in field outbreaks of APV.

Animals↗

Use of enzyme linked immunosorbent assay for estimation of antibody to Newcastle disease virus.

Serum antibody assay of 1045 serum samples from birds vaccinated with Newcastle Disease vaccine by HI test and ELISA was carried out. Five hundred and twenty seven birds were challenged with virulent virus and data on HI test, ELISA and challenge test results were used for comparison. A good correlation between HI titres, ELISA absorbance and potency was observed indicating the usefulness of ELISA for potency estimation of vaccines and determination of immunization level in vaccinated flocks.

Animals↗

The correlation of fatty acid content of infected cells and virions with Newcastle disease virus (NDV) virulence.

Chick embryo fibroblasts (CEF) infected with virulent strains of Newcastle disease virus (NDV) showed a dramatic increase in total unsaturated fatty acids (UFA). This increase was not seen in cells infected with the avirulent strains of NDV, Sendai virus or influenza A (PR8). The virions of the virulent strains of NDV harvested from the chorioallantoic cavity also had a higher UFA content compared to the avirulent ones. The kinetics of UFA increase in the virulent strains could be correlated with published data on the kinetics of RNA and protein synthesis inhibition, polykaryon formation and membrane permeability changes.

Animals↗

Experimental infection of rosellas (Platycercus eximius) with velogenic viscerotropic Newcastle disease virus (VVNDV).

Plaque-purified and non-plaque-purified velogenic viscerotropic Newcastle disease viruses (VVNDVs) were inoculated into golden-mantled rosellas (Platycercus eximius). VVNDV produced acute clinical disease in this species: all birds died within 6 days postexposure. There was no difference between the two inoculation groups in clinical signs. Seven tissues and five tissue swabs were collected from each of 15 birds. The VVNDV concentration in each specimen was titrated, and the concentrations were compared. The lung and trachea had the highest concentrations of virus in both the tissue suspensions and the swab suspensions. The average virus concentrations of the lung were 10(5.9) 50% embryo lethal doses (ELD50) per 0.1 ml for the tissue suspension and 10(4.9) for the swab. The average virus concentrations of the trachea were 10(5.6) ELD50 per 0.1 ml of tissue suspension and 10(4.6) for the swab.

Animals↗

[Characteristics of 24S and 35S virus-specific RNA in cells infected with Newcastle disease virus].

The properties of 24S and 35S virus-specific RNAs of Newcastle disease virus were studied after denaturation. It was shown that 60-80% of 24S and 35SRNA consisted of agglomerates of molecules of virus-specific 18S RNA. Undissociable upon denaturation, 24S RNA contained mainly the same components as undenaturated RNA and consisted completely of sequences of virus-specifis 18S RNA. Undissociable 35S RNA contained no "heavy" heterogenous material present in the intact undenaturated RNA and 75% of it consisted os sequences of 18S virus-specific RNA. Some sequences present in undissociable 24S RNA were absent in undissociable 35S RNA. The aggregation of virus-specific na-transcripts and subsequent formation of covalent bonds between transcripts as well as possible formation of "solid" transcripts of the adjacent genes are discussed.

Newcastle disease virus↗

Cloning and expression of the phosphoprotein gene of Newcastle disease virus in Escherichia coli.

The phosphoprotein (P) gene of a heat stable Newcastle disease virus (NDV) was cloned, sequenced and expressed in Escherichia coli. SDS-PAGE analysis of the recombinant P protein (395 amino acids) and a C-terminal extension derivative (424 amino acids), gave rise to two distinct protein bands with molecular masses of approximately 53-55 and 56-58 kDa, respectively, which are approximately 26-30% heavier than those calculated from the deduced amino acid sequences. The differences in molecular mass on SDS-PAGE are thought to be attributed to the acidic nature of the P protein (pI=6.27) and also the different degrees of phosphorylation in the prokaryotic cell. Amino acid sequence comparison of the P protein among the published NDV strains showed that they were highly conserved particularly at the putative phosphorylation sites.

Amino Acid Sequence↗

The influence of test conditions on Newcastle disease hemagglutination-inhibition titers.

Replicate samples of serum from chickens immune to Newcastle disease were titrated to determine the influence of certain test conditions on hemagglutination-inhibition (hi) titers. The test conditions studied were those most likely to vary in normal laboratory operations. The most marked effect on magnitude of HI titers was incubation time of twofold serum dilutions in antigen-saline; the average titer increase after incubation of the serum-antigen mixture for 1 hr at 37 C was log2 2.3 (fivefold). Twofold increases in virus concentration of the antigen-saline diluent caused an average titer reduction of log2 0.8. Shifts in HI titers were only minor with twofold changes in erythrocyte concentration (log2 0.3), with variations of test reading times from 0.5 to 2.0 hr (log2 0.1), and with variations in the period between preparation of the initial 1:10 serum dilution in antigen-saline and the subsequent serum dilutions (log2 0.3).

Animals↗

Identification of field isolates of infectious bronchitis virus by interference with the La Sota of strain of Newcastle disease virus.

The interference phenomenon of infectious bronchitis virus (IBV) with growth of Newcastle disease virus (NDV) in embryonating chicken eggs (ECE) was used as a diagnostic method. Fifteen field isolates obtained from presumptively infectious-bronchitis-affected chickens were analyzed by the IBV-NDV interference test. Eight isolates were capable of interfering with the growth of the La Sota strain of NDV, as measured by hemagglutination (HA) activity when IBV was inoculated 10 hr before NDV into ECE. The interference was considered specific for IBV, because it could be eliminated by adding homologous anti-IBV serum. The sensibility of this method could be demonstrated, because in some cases low-passage levels of IBV isolates showing HA interference ability were not capable of producing lesions in ECE. Furthermore, serologically negative IBV samples did not interfere with NDV growth. From these results, the IBV-NDV interference test appears to be a potential diagnostic alternative for identifying IBV field isolates.

Animals↗

Modulation of adjuvant-enhanced delayed-type hypersensitivity by the interferon inducers poly I:C and Newcastle disease virus.

The modulation by the interferon (IFN) inducers poly I:C and Newcastle disease virus (NDV) of the effector phase of adjuvant-enhanced delayed-type hypersensitivity (DH) was studied in mice. A strongly enhanced DH was induced in mice to ultraviolet (UV) light inactivated Semliki forest virus (SFV) by the use of the adjuvant dimethyldioctadecylammonium bromide. At day 6 after intracutaneous immunization, DH was elicited with SFV and measured 24 and 48 h later as increase in footpad thickness (footpad swelling test). Systemic, intravenous administration of either poly I:C, UV-inactivated NDV, or NDV-induced IFN prior to elicitation of DH with antigen resulted in a temporarily suppressed DH reaction. Both the poor swelling at 3 h and strong swelling at 24 h were suppressed, while the swelling at 48 h was enhanced. The model described provides a sensitive in vivo method to study modulating effects of drugs and microbial agents on the effector phase of DH.

Animals↗

Strain variation and nuclear association of Newcastle disease virus matrix protein.

Five monoclonal antibodies to the matrix (M) protein of Newcastle disease virus (NDV) Australia-Victoria (AV) strain were generated and characterized. In competitive antibody-binding assays, the antibodies fell into three discrete groups. The antigenic sites described by these antibody groups were designated M1, M2, and M3. Each antibody reacted with a panel of NDV strains in a manner unique to its group, confirming the grouping by competitive antibody binding. Only site M1 was found on all 12 of the strains tested and may be a "pan-NDV" epitope. A large portion of the M protein of strain AV was detected in the nuclei of infected cells by all five monoclonal antibodies. In addition, the antibodies only stained the nuclei of cells infected with NDV strains expressing M protein containing the corresponding antigenic site. These results confirm that the immunoreactivity in the nucleus is actually caused by the M protein and not by a cross-reacting host protein induced by viral infection.

Animals↗

Cytopathic effect of lentogenic strains of Newcastle disease virus in cultured chicken muscle cells.

The ability of lentogenic strains of Newcastle disease virus (NDV) to inhibit the development of embryonic chicken muscle cells in vitro is described. In contrast to the uninfected control, which displayed a thick network of branched multinucleated muscle fibers, cultures infected with NDV exhibited a severely reduced potential to differentiate. The few differentiated muscle fibers that formed in the presence of NDV were poorly developed, displayed an attenuated morphology, and detached from the substrate 2 to 3 days before uninfected cultures showed signs of cell aging. When NDV-infected cultures were stained before muscle fiber detachment with a fluorescent-labeled NDV-specific antibody, the infected muscle fibers reacted strongly relative to neighboring myoblasts and fibroblasts. All six strains of NDV induced a similar inhibitory effect, and no strain-specific differences were detectable. These results indicate that differentiating muscle fibers are highly susceptible to infection by NDV in vitro and that this tissue exhibits an early NDV-induced cytopathic effect.

Animals↗