Velogenic Newcastle Disease Virus Strain Research Articles

Isolation and Characterization of a Pathogenic Newcastle Disease Virus from a Natural Case in Indonesia

This study was performed to isolate a velogenic Newcastle disease virus (NDV) strain currently found in Indonesia for establishing a domestic reference virus for future pathological and molecular epidemiological studies. A chicken suspected to have contracted Newcastle disease (ND) in a local outbreak in Bali was selected for NDV isolation. Atrophy of lymphoid tissues such as the bursa of Fabricius, thymus, and spleen; intestinal haemorrhage; and oedema of the brain were observed in the chicken. Histopathological examination revealed severe non-suppurative meningoencephalomyelitis characterised by neuronal necrosis, multifocal to diffuse gliosis, and perivascular cuffing of mononuclear cells, hemorrhagic necrosis of the trachea, intestines and bursa of Fabricius, and various degree of lymphoid depletion and necrosis of the lymphoid tissues. After ND was confirmed immunohistochemically, the NDV was propagated by inoculating tissue homogenate of the diseased chicken in embryonated eggs. Phylogenetic analysis based on the F gene nucleotide sequence revealed that this isolate belonged to genotype VII. The deduced amino acid sequence of the isolated NDV F protein at the cleavage site was (112)RRQKRF(117), which is typically found in virulent NDV isolates. Pathogenicity indexes such as the mean death time (MDT) and intracerebral pathogenicity index (ICPI) were 54 hr and 1.77, respectively. Pathological findings, phylogenetic analysis, amino acid sequence of the F protein cleavage site, and pathogenicity index test results revealed the NDV isolate, designated as NDV/Bali-1/07, to be a novel Indonesian velogenic NDV strain belonging to group VII.

Newcastle Disease Virus Infection Promotes Bax Redistribution to Mitochondria and Cell Death in HeLa Cells

Background/Aims: Newcastle disease virus (NDV) is an avian paramyxovirus that has gained a lot of interest in cancer viro-therapeutic applications because of its ability to selectively induce apoptosis in human cancer cells. However, the underlying mechanisms by which NDV induces apoptosis in human cancer cells are still not entirely understood. Methods: In this study we examined the effect of a Malaysian velogenic strain of NDV, known as AF2240, on some elements of the intrinsic pathway of apoptosis. Results: We show that NDV infection leads to conformational change of Bax protein. This is associated with the translocation of Bax from the cytoplasm to mitochondria and the release of cytochrome c into the cytoplasm. Interestingly, the level of Bcl-2 protein was not affected by NDV treatment. Conclusion: We have shown that Bax conformational change and subcellular distribution is involved in the intrinsic pathway of apoptosis induced by NDV.

Response of Village Chickens to Newcastle Disease (ND) Vaccine with Broken Sorghum as the Vehicle

The suitability of sorghum as carrier for Newcastle Disease (ND) V4-UPM virus vaccine for vaccination of freerange chickens was assessed by standard methods. The grain was ground rough or broken, soaked in water for three days, washed, sun-dried, coated with the virus and finally dried at room temperature. The food vaccine which was supplied to chicken owners in two different locations was fed to the birds in their natural habitat. Blood for serum was collected two weeks after each feeding event for haemagglutination inhibition (HI) assays. Samples of vaccinated and unvaccinated chickens were bought back from owners and exposed to velogenic strain of ND virus. Results showed that out of 129 birdsvaccinated, 87 (67.4%) propduced detectable HI antibody with only 13 (10.1%) attaining antibody level of log2 > 3.0 regarded as protective and geometric mean titre (GMT) of 3.1 after primary vaccination. After a booster dose on 116 birds in the same flock, 95 (81.9%) seroconverted with 67 (57.8%) attaining log2 > 3.0 and GMT of 11.8. Exposure of 34 vaccinated buy-back and 25 unvaccinated (control) chickens to velogenic challenge virus led to 26 (76.5%) chickens and 4 control birds (16.0%) surviving respectively. Sorghum, if adequately treated could be a suitable carrier for thermostable ND vaccines, and as dried food vaccine, it could have good shelf life at room temperature.

Determination of Minimum Hemagglutinin Units in an Inactivated Newcastle Disease Virus Vaccine for Clinical Protection of Chickens from Exotic Newcastle Disease Virus Challenge

The potency of inactivated Newcastle disease virus (NDV) vaccines in the United States is currently determined using vaccination and challenge of experimental animals against a velogenic strain of NDV. Because velogenic strains of NDV are now classified as select agents in the United States, all vaccine potency testing must be performed in live animals under biosafety level 3 agriculture conditions. If the minimum amount of inactivated viral antigen required for clinical protection can be determined using other methods, vaccines meeting these criteria might be considered of adequate potency. The linearity of correlation between the hemagglutination (HA) assay measurement and the 50% embryo infectious dose titer ofNDV Hitchner B1 vaccine virus was determined. Correlation between hemagglutinin units (HAU) per vaccine dose, clinical protection, and antibody response was then determined using a vaccinate-and-challenge model similar to Chapter 9 of the U.S. code of federal regulations approved method for vaccine potency testing. The dose providing 50% protection of an in-house water-in-oil emulsion vaccine formulated with inactivated NDV B1 was determined to be between 400 and 600 HAU from two separate trials. A positive correlation (R2 = 0.97) was observed between antibody response and HAU per vaccine dose. Serum antibody responses from vaccinated birds indicate HA inhibition titers >2(5) log2 would provide 100% protection from morbidity and mortality and require a minimum protective dose of 1000 HAU per bird. These are the first studies to examine establishing both a minimum protective HAU content for inactivated ND vaccines and a minimum serologic response necessary to ensure potency.

Genetic and antigenic analysis of Newcastle disease viruses from recent outbreaks in Taiwan

Portions of the haemagglutinin-neuraminidase (HN) and fusion protein (F) genes of Newcastle disease virus (NDV) isolated from recent outbreaks in Taiwan were amplified and sequenced. These isolates were velogenic, based on the amino acid sequences of the F protein cleavage site and the mean death time in chicken embryos. All the recent viruses contained the amino acid sequences 112RRQKR116 for the C-terminus of the F2 protein. The serological relatedness of recent isolates was determined using a serum neutralization (SN) test. Relatedness values, determined by a cross-SN test, revealed that all belonged to a single serotype but could be classified into distinct subtypes, suggesting that antigenic variations occurred in these isolates. Phylogenetic trees based on the nucleotide sequences of the HN and F genes revealed that recent Taiwanese isolates had evolved into two groups. Antigenic analysis also suggested that there are at least two groups of NDVs involved in recent outbreaks and that these outbreaks in Taiwan might have been caused by co-circulation of multiple velogenic NDV strains.

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.

Occurrence of genotypes IV, V, VI and VIIa in Newcastle disease outbreaks in Germany between1939 and 1995

Forty-five velogenic Newcastle disease virus strains isolated in Germany between 1939 and 1995 were analysed by restriction enzyme digestion and sequencing to shed light on the relationships of past epizootics. Viruses derived from the period prior to 1970 belonged to a clade (IVea) of genotype IV comprising the earliest isolates from Europe, and could be isolated until the late seventies from poultry. Essex'70-like viruses, the prototype of genotype V, were already present at the beginning of the 1970-74 epizootic and in sporadic cases thereafter, indicating that these Newcastle disease outbreaks started in Western Europe. A genotype VI (subtype VIc)isolate was obtained in the early 1980s from a single outbreak in poultry. Outbreaks between 1993-95 were again part of a Western European epizootic caused by a genotype VIIa virus that was prevalent in the Far East.

Protective Immunity against Newcastle Disease: The Role of Cell-Mediated Immunity

The role of cell-mediated immunity (CMI) in protection of birds from Newcastle disease was investigated by two different strategies in which only Newcastle disease virus (NDV)-specific CMI was conveyed without neutralizing antibodies. In the first strategy, selected 3-wk-old specific-pathogen-free (SPF) birds were vaccinated with either live NDV (LNDV), ultraviolet-inactivated NDV (UVNDV), sodium dodecyl sulfate-treated NDV (SDSNDV), or phosphate-buffered saline (PBS) (negative control) by the subcutaneous route. Birds were booster vaccinated 2 wk later and challenged with the velogenic Texas GB strain of NDV 1 wk after booster. All vaccinated birds had specific CMI responses to NDV as measured by a blastogenesis microassay. NDV neutralizing (VN) and hemagglutination inhibition (HI) antibody responses were detected in birds vaccinated with LNDV and UVNDV. However, birds vaccinated with SDSNDV developed antibodies that were detected by western blot analysis but not by the VN or HI test. Protection from challenge was observed only in those birds that had VN or HI antibody response. That is, birds with demonstrable CMI and VN or HI antibody response were protected, whereas birds with demonstrable CMI but no VN or HI antibody response were not protected. In the second strategy, birds from SPF embryos were treated in ovo with cyclophosphamide (CY) to deplete immune cells. The birds were monitored and, at 2 wk of age, were selected for the presence of T-cell activity and the absence of B-cell activity. Birds that had a significant T-cell response, but not a B-cell response, were vaccinated with either LNDV, UVNDV, or PBS at 3 wk of age along with the corresponding CY-untreated control birds. The birds were booster vaccinated at 5 wk of age and were challenged with Texas GB strain of NDV at 6 wk of age. All birds vaccinated with LNDV or UVNDV had a specific CMI response to NDV, VN or HI NDV antibodies were detected in all CY-nontreated vaccinated birds and some of the CY-treated vaccinated birds that were found to have regenerated their B-cell function at 1 wk postbooster. The challenge results clearly revealed that CY-treated birds that had NDV-specific CMI and VN or HI antibody responses to LNDV or UVNDV were protected, as were the CY-nontreated vaccinated birds. However, birds that had NDV-specific CMI response but did not have VN or HI antibodies were not protected from challenge. The results from both strategies indicate that specific CMI to NDV by itself is not protective against virulent NDV challenge. The presence of VN or HI antibodies is necessary in providing protection from Newcastle disease.

Evaluation of the efficacy of the crude extracts of Capsicum frutescens, Citrus limon and Opuntia vulgaris against Newcastle disease in domestic fowl in Tanzania

Prophylactic and therapeutic efficacy of a combination of Capsicum frutescens (red pepper), Citrus limon (lemon) and Opuntia vulgaris (prickly pear) against Newcastle disease (ND) in domestic fowl were evaluated. Eighty-eight broiler chickens were divided into five groups. Birds from three groups were inoculated with velogenic ND virus strain, whereas birds from two groups were left as controls. Two groups received a mixture of the plant extract three days prior to inoculation and birds from one group were given the plant extract for two days following development of clinical signs. Blood samples were collected for haemaglutination inhibition tests (HI) for detection of ND virus antibodies. Body weights were monitored during the experiment. Three birds died from the group that was inoculated with ND virus and treated with the plant extract; two died from the group that received the plant extract as a prophylaxis and inoculated with ND virus; and one bird died from the group that was inoculated with ND virus but not given the plant extract. No death was observed in any of the birds in the control groups. Antibody titers for ND virus rose four-fold in the inoculated birds but remained low in the un-inoculated groups. Mean body weights of birds in group B declined markedly compared to the other groups. The results indicated that there was no prophylactic or therapeutic value of the plant extract against ND. The plant extract showed a negative effect on body weights in birds with ND.

Embryo vaccination of turkeys against Newcastle disease infection with recombinant fowlpox virus constructs containing interferons as adjuvants

Recombinant fowlpox viruses (rFPV) expressing the fusion and hemagglutinin-neuraminidase glycoproteins of Newcastle disease virus (NDV) as well as chicken type I interferon (IFN) or type II IFN were used to vaccinate specific pathogen-free (SPF) turkeys in ovo. No significant changes in the hatchability, survival rate, performance and weight gain were observed after vaccination with the rFPV vaccines in comparison to diluent-inoculated embryos. The rFPV-NDV-IFN-II construct induced the onset of anti-NDV antibody production in SPF birds at one week post hatch, one week earlier than other vaccine constructs. Three to five weeks post hatch, the turkeys were challenged with the neurotropic velogenic NDV strain Texas GB (NDV-GB-Tx). The rFPV-NDV-IFN-II construct was the most protective vaccine against NDV. rFPV vaccines significantly ( p<0.05) suppressed the mitogenic response of peripheral blood leukocytes in vaccinated turkeys in comparison to placebo inoculated controls at 25 days post vaccination. Birds vaccinated with rFPV-NDV-IFN-I construct did not have an inhibition in the mitogenic response.

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.

Newcastle Disease Virus Surveillance in Argentina: Use of Reverse Transcription-Polymerase Chain Reaction and Sequencing for Molecular Typification

Newcastle disease virus (NDV) remains a major pathogen of poultry where highly virulent strains require reporting to the Office of International Epizootes. NDV is a paramyxovirus existing as different strains classified on the basis of severity of the disease they cause. The present study was conducted in Argentina to determine the prevalence of highly virulent velogenic NDV strains in commercial poultry farms. Tracheal and cloacal swabs from 693 flocks, representing 14% of the broiler production, were collected and pooled. A pool amplified twice in embryonated eggs presented a limited hemagglutination titer. We performed reverse transcription coupled to polymerase chain reaction to amplify fusion and matrix protein gene sequences of the isolate and the strain Trenque Lauquen, isolated in Argentina during an outbreak in 1970-71 and previously characterized as velogenic viscerotropic by biological methods. The amino acid sequences were deduced from nucleotide sequences of the amplification products and the pathotype predicted according to the sequences obtained. From the samples analyzed, we found only one type of NDV, being the isolate identified as lentogenic NDV. This strain is probably the one used in vaccination of flocks where that sample was obtained. These data have allowed us to consider a velogenic NDV-free status in Argentina's commercial poultry.

Protective antibody titre against Newcastle disease in ostriches (Struthio camelus)

The aim of this study was to define an estimated antibody titre which was considered to be protective against Newcastle disease (ND) virus infection in ostriches. Eighteen young ostriches of 4 days of age were divided into two groups each containing 9 birds. The first group was unvaccinated and the second group was vaccinated against ND virus twice at 4 and 14 days of age. Antibody titres were monitored at 1, 14, 28, 42, 56, 70 and 85 days of age by haemagglutination inhibition (HI) test. All birds were then challenged with a velogenic strain of ND virus, Ita strain, at 42 days of age. The excretion of the challenge virus were monitored daily after challenge up to the end of this experiment. Several organs such as brain, trachea, lungs and spleen were collected from died birds for re-isolation of the challenged virus. Results indicated that all unvaccinated birds succumbed to the challenged virus, except one bird that survived challenged. In contrast to the unvaccinated birds, all vaccinated birds survived challenged, except two birds with low antibody titres succumbed challenged. All birds with antibody titres of 4 (HI-log2) or greater survived challenged. All challenged birds excreted the challenged virus through out their oropharyngs. Moreover, challenged virus can be successfully re-isolated from most organs of the died birds. This study concludes that : (a) the estimated protective titre against ND in ostriches is 4 (HI-log2), (b) the immune status for ostrich with antibody titre less the 4 (HI-log2) could not be defined, and (c) vaccination against Newcastle disease in ostriches could successfully prevent birds from sick and died of ND, but unable to prevent virus infection and unable to stop carrier status after infection. Key words : Newcastle disease, ostrich, antibody, protective titre

Onset of Protective Immunity in Chicks after Vaccination with a Recombinant Herpesvirus of Turkeys Vaccine Expressing Newcastle Disease Virus Fusion and Hemagglutinin-Neuraminidase Antigens

The onset of protective immunity from lethal Newcastle disease virus (NDV) challenge of chicks was determined after vaccination with a recombinant herpes virus of turkeys (HVT) expressing the fusion and hemagglutinin-neuraminidase proteins of NDV. One-day-old specific-pathogen-free chicks devoid of maternal antibodies to NDV were vaccinated with 130 to 3300 plaque forming units of HVT (depending on the trial) and then challenged at 4, 7, 10, and 14 days postvaccination (DPV) with a neurotropic velogenic strain of NDV (GB Texas). The recombinant vaccine afforded 0%, 35-75%, 85%, and 94-100% protection when the vaccinated birds were challenged at 4, 7, 10, and 14 DPV, respectively. In all trials, challenge caused 100% mortality in unvaccinated control chicks. Newcastle disease virus was reisolated from the lung, liver, spleen, and brain of birds dying in all trials regardless of vaccine dosage or time of challenge, except when challenge occurred at 14 DPV.

Protective efficacy of a recombinant herpesvirus of turkeys as an in ovo vaccine against Newcastle and Marek's diseases in specific-pathogen-free chickens

We investigated the potential of a herpesvirus of turkey (HVT)-based recombinant virus (rHVT) as an in in ovo vaccine to protect specific-pathogen-free chickens against Newcastle disease (ND) and Marek's disease (MD). The rHVT, designed to express fusion (F) and hemagglutinin-neuraminidase (HN) glycoproteins of the lentogenic Hitchner B1 strain of ND virus (NDV), as well as glycoproteins A and B of the GA strain of serotype 1 MD virus (MDV) was efficacious in protecting chickens against ND and MD. No adverse effects on hatchability or the survival of chickens were observed following in ovo vaccination with rHVT. A single administration at embryonation day 18 (ED18) or at hatch protected chickens against challenge-exposures with virulent MDV strain RB-1B and velogenic NDV strain GB-Texas (NDV-GB-TX). Vaccinated chickens developed antibodies against both viruses as detected by serological tests, namely, hemagglutination inhibition, virus neutralization and western immunoblotting for NDV, and immunofluorescence and radioimmunoprecipitation assays for MDV. PCR analysis showed that in ovo vaccination with rHVT resulted in a persistent infection leading to systemic immunity against ND for up to 8 weeks of age, the longest period of time tested in this study. However, virus isolation tests indicated that rHVT-vaccinated chickens were only partially protected from the replication of NDV-GB-TX in the trachea. The results of the study indicate that rHVT is safe for both ED18 and posthatch vaccination for ND and MD, and because the vaccine persists, it may induce longer lasting immunity than conventional live NDV vaccines.

The epidemiology of Newcastle disease in rural poultry: A review

Rural poultry are the dominant form of poultry kept in the developing world. They are a natural resource whose potential is not fully exploited for the welfare of rural populations. The most devastating disease of rural poultry is Newcastle disease (ND). ND is an epidemic disease in intensive poultry and is responsible for high economic losses. Its epidemiology in intensive poultry is well understood, but little has been published on its behaviour in village poultry populations. Some research has been carried out during the past few years and it now appears that rural poultry are receiving increasing support for research and development from many government and international funding agencies throughout the world. All strains of Newcastle disease virus (NDV) occur in rural poultry, but velogenic strains are reported to be more common. Serological surveys in conjunction with isolation studies have shown that velogenic NDV strains are endemic in rural poultry populations even in isolated villages and possibly in isolated flocks. Although NDV is endemic in village poultry, the clinical disease usually follows an epidemic pattern. ND outbreaks often occur once or twice a year at regular intervals affirming the endemicity of the virus, however, 'mini' outbreaks in individual flocks and sporadic cases in individual birds may occur. Epidemics usually occur at times of climatic stress, leading to seasonal occurrence. The spread of NDV within and between village poultry populations is relatively slow due to a low contact rate. The major mode of transmission appears to be by the faecal-oral route. The respiratory route may also play a role in flocks where close bird-to-bird associations exist. Other poultry species, wild and feral birds, wild animals, communal water reservoirs and domestic animals may play a role in transmission; however, their role has not been properly investigated. In non-immune, intensively reared poultry, introduction of a sufficient quantity of virulent NDV is sufficient to cause severe outbreaks, but in rural poultry epidemic disease depends upon a number of factors. These include NDV pathotype, age structure and immunity of the host population, breed susceptibility, concurrent disease and seasonal influences. Although ND is endemic in rural poultry, many aspects of the epidemiology and economics of ND have not been fully understood. Well-designed, longitudinal studies made within a socio-economic framework are required to improve our understanding of the main problems of rural poultry systems.

Efficacy in Chickens of a Herpesvirus of Turkeys Recombinant Vaccine Containing the Fusion Gene of Newcastle Disease Virus: Onset of Protection and Effect of Maternal Antibodies

The onset of protection against Newcastle disease and the effect of maternal antibodies to Newcastle disease virus (NDV) and Marek's disease virus (MDV) on vaccine efficacy were determined following vaccination of chickens with a recombinant herpesvirus of turkeys (HVT) vaccine expressing the fusion (F) glycoprotein gene of NDV. Onset of protection following intra-abdominal administration of the recombinant HVT/F vaccine at 1 day of age and subsequent ocular challenge with the neurotropic velogenic Texas GB strain of NDV was determined to occur between days 14 and 21 post-vaccination (PV). Vaccination with the Hitchner B1 strain of NDV resulted in protection by day 6 PV, and vaccination with an inactivated NDV oil-emulsion vaccine induced protection by day 14 PV. One-day-old broiler-type chickens with maternal antibodies to both NDV and MDV and 1-day-old specific-pathogen-free (SPF) white leghorn chickens lacking maternal antibodies were vaccinated with the recombinant HVT/F vaccine or with control vaccines, challenged intra-abdominally with the very virulent RB1B strain of MDV on day 8 PV, and challenged with the Texas GB strain of NDV on day 29 PV. The HVT/F and NDV strain Hitchner B1 vaccines provided 73% and 80% protection, respectively, against NDV in broilers, whereas both vaccines resulted in 100% protection in SPF leghorns.

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.

Protection of Chickens from Newcastle and Marek's Diseases with a Recombinant Herpesvirus of Turkeys Vaccine Expressing the Newcastle Disease Virus Fusion Protein

Recombinant strains of herpesvirus of turkeys (HVT) were constructed that contain either the fusion protein gene or the hemagglutinin-neuraminidase gene of Newcastle disease virus (NDV) inserted into a nonessential gene of HVT. Expression of the NDV antigens was regulated from a strong promoter element derived from the Rous sarcoma virus long terminal repeat. Recombinant HVT strains were stable and fully infectious in cell culture and in chickens. Chickens receiving a single intra-abdominal inoculation at 1 day of age with recombinant HVT expressing the NDV fusion protein had an immunological response and were protected (> 90%) against lethal intramuscular challenge at 28 days of age with the neurotropic velogenic NDV strain Texas GB. Recombinant HVT expressing the NDV hemagglutinin-neuraminidase provided partial protection (47%) against the same challenge. Chickens vaccinated with recombinant HVT vaccines had low levels of protection against NDV replication in the trachea when challenged ocularly. Recombinant HVT vaccines and the parent HVT strain provided similar levels of protection to chickens challenged with the very virulent RB1B strain of Marek's disease virus, indicating that insertion of foreign sequences into the HVT genome did not compromise the ability of HVT to protect against Marek's disease.

Vaccination against Newcastle Disease with a Recombinant Baculovirus Hemagglutinin-Neuraminidase Subunit Vaccine

Vaccination of chickens with an oil-emulsion vaccine containing a recombinant baculovirus that expressed the hemagglutinin-neuraminidase (HN) of Newcastle disease virus (NDV)-induced hemagglutination-inhibition (HI) and virus-neutralizing antibodies against NDV. HI antibody titers obtained in response to vaccination with the live recombinant virus were higher than those obtained when the recombinant was inactivated with beta-propiolactone, and the titers were lower than those obtained in response to the same HN concentrations in live or beta-propiolactone-inactivated NDV strain B1. The serological response to the recombinant baculovirus was differentiated from the response to NDV by an enzyme-linked immunosorbent assay in which purified NDV nucleoprotein was used as antigen. Chickens vaccinated with the live recombinant or with inactivated NDV resisted an oculonasal challenge with the neurotropic velogenic Texas GB strain of NDV, which was lethal in unvaccinated controls. It was concluded that the HN protein of NDV expressed as a subunit by a recombinant baculovirus was protective against Newcastle disease.