Newcastle Disease Virus Outbreaks Research Articles

MRNA Profiling and Transcriptomics Analysis of Chickens Received Newcastle Disease Virus Genotype II and Genotype VII Vaccines.

Newcastle Disease Virus (NDV) genotype VII (GVII) is becoming the predominant strain of NDV in the poultry industry. It causes high mortality even in vaccinated chickens with a common NDV genotype II vaccine (GII-vacc). To overcome this, the killed GVII vaccine has been used to prevent NDV outbreaks. However, the debate about vaccine differences remains ongoing. Hence, this study investigated the difference in chickens' responses to the two vaccines at the molecular level. The spleen transcriptomes from vaccinated chickens reveal that GVII-vacc affected the immune response by downregulating neuroinflammation. It also enhanced a synaptogenesis pathway that operates typically in the nervous system, suggesting a mechanism for the neurotrophic effect of this strain. We speculated that the down-regulated immune system regulation correlated with protecting the nervous system from excess leukocytes and cytokine activity. In contrast, GII-vacc inhibited apoptosis by downregulating PERK/ATF4/CHOP as part of the unfolded protein response pathway but did not affect the expression of the same synaptogenesis pathway. Thus, the application of GVII-vacc needs to be considered in countries where GVII is the leading cause of NDV outbreaks. The predicted molecular signatures may also be used in developing new vaccines that trigger specific genes in the immune system in combating NDV outbreaks.

Newcastle viral disease causation web correlations with laying hen productivity

Environmental conditions profoundly impact the health, welfare, and productivity of laying hens in commercial poultry farming. We investigated the association between microclimate variations, production indices, and histopathological responses to accidental Newcastle disease virus (NDV) infection within a controlled closed-house system. The study was conducted over seven months in a laying hen facility in Cairo, Egypt. Microclimate measurements included temperature, relative humidity (RH%), air velocity (AV), and the temperature humidity index (THI) that were obtained from specific locations on the front and back sides of the facility. Productivity indices, including the egg production percentage (EPP), egg weight (EW), average daily feed intake, and feed conversion ratio, were assessed monthly. During an NDV outbreak, humoral immune responses, gross pathology, and histopathological changes were evaluated. The results demonstrated significant (p < 0.05) variations in EPP and EW between the front and back sides except in April and May. AV had a significant (p = 0.006) positive effect (Beta = 0.346) on EW on the front side. On the back side, AV had a significant (p = 0.001) positive effect (Beta = 0.474) on EW, while it negatively influenced (p = 0.027) EPP (Beta = − 0.281). However, temperature, RH%, and THI had no impact and could not serve as predictors for EPP or EW on either farm side. The humoral immune response to NDV was consistent across microclimates, highlighting the resilience of hens. Histopathological examination revealed characteristic NDV-associated lesions, with no significant differences between the microclimates. This study underscores the significance of optimizing microclimate conditions to enhance laying performance by providing tailored environmental management strategies based on seasonal variations, ensuring consistent airflow, particularly near cooling pads and exhaust fans, and reinforcing the importance of biosecurity measures under field challenges with continuous monitoring and adjustment.

Genomic Diversity and Geographic Distribution of Newcastle Disease Virus Genotypes in Africa: Implications for Diagnosis, Vaccination, and Regional Collaboration

The emergence of new virulent genotypes and the continued genetic drift of Newcastle disease virus (NDV) implies that distinct genotypes of NDV are simultaneously evolving in different geographic locations across the globe, including throughout Africa, where NDV is an important veterinary pathogen. Expanding the genomic diversity of NDV increases the possibility of diagnostic and vaccine failures. In this review, we systematically analyzed the genetic diversity of NDV genotypes in Africa using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Information published between 1999 and 2022 were used to obtain the genetic background of different genotypes of NDV and their geographic distributions in Africa. The following genotypes were reported in Africa: I, II, III, IV, V, VI, VII, VIII, XI, XIII, XIV, XVII, XVIII, XX, and XXI. A new putative genotype has been detected in the Democratic Republic of the Congo. However, of 54 African countries, only 26 countries regularly report information on NDV outbreaks, suggesting that this number may be vastly underestimated. With eight different genotypes, Nigeria is the country with the greatest genotypic diversity of NDV among African countries. Genotype VII is the most prevalent group of NDV in Africa, which was reported in 15 countries. A phylogeographic analysis of NDV sequences revealed transboundary transmission of the virus in Eastern Africa, Western and Central Africa, and in Southern Africa. A regional and continental collaboration is recommended for improved NDV risk management in Africa.

Fate and Transport of Coronavirus Surrogate through Compacted Clays for Pathogenic Waste Disposal.

An increased pathogenic waste post-COVID-19 pandemic forced policymakers to treat biomedical waste (BMW) similar to municipal solid waste (MSW) to dispose into dumpsites and MSW landfills across the globe. The granular bentonite of geosynthetic clay liners (GCLs) does not completely seal the macro-voids upon saturation due to the loss of osmotic potential in the salt environment from the leachate. Such behavior of GCLs can lead to advection-dominant virus migration through the liner system. A knowledge of the fate and transport of coronavirus and other viral pathogens in compacted clays is essential for safe disposal of the viral pathogens in MSW landfills. Although the attenuation and transport parameters for coronavirus have been recently evaluated theoretically, experimental backup is currently lacking. The present work uses Newcastle disease virus (NDV) as a surrogate to coronavirus due to structural similarities for studying the fate and transport in the compacted natural clays. This study also implicitly addresses the waste management facilities for waste generated from NDV outbreaks through poultry litter and carcasses. The interaction of bentonite and kaolin clays with the NDV was studied by varying the virus concentration, interaction time, and clay dose using batch sorption tests. The studied clays showed excellent attenuation efficiency for the NDV. Design parameters, viz., the diffusion coefficient and retardation factor, were evaluated, affirming the suitability of these clays for exclusive pathogenic waste disposal protocols that are discussed in this article.

BotCl, the First Chlorotoxin-like Peptide Inhibiting Newcastle Disease Virus: The Emergence of a New Scorpion Venom AMPs Family.

Newcastle disease virus (NDV) is one of the most serious contagions affecting domestic poultry and other avian species. It causes high morbidity and mortality, resulting in huge economic losses to the poultry industry worldwide. Despite vaccination, NDV outbreaks increase the need for alternative prevention and control means. In this study, we have screened fractions of Buthus occitanus tunetanus (Bot) scorpion venom and isolated the first scorpion peptide inhibiting the NDV multiplication. It showed a dose dependent effect on NDV growth in vitro, with an IC50 of 0.69 µM, and a low cytotoxicity on cultured Vero cells (CC50 > 55 µM). Furthermore, tests carried out in specific pathogen-free embryonated chicken eggs demonstrated that the isolated peptide has a protective effect on chicken embryos against NDV, and reduced by 73% the virus titer in allantoic fluid. The N-terminal sequence, as well as the number of cysteine residues of the isolated peptide, showed that it belongs to the scorpion venom Chlorotoxin-like peptides family, which led us to designate it "BotCl". Interestingly, at 10 µg/mL, BotCl showed an inhibiting effect three times higher than its analogue AaCtx, from Androctonus australis (Aa) scorpion venom, on NDV development. Altogether, our results highlight the chlorotoxin-like peptides as a new scorpion venom AMPs family.

EVALUATION OF DIFFERENT NEWCASTLE DISEASE VIRUS VACCINATION REGIMES AGAINST CHALLENGE WITH RECENTLY ISOLATED GENOTYPE VII VIRUS FROM EGYPT

Newcastle disease virus (NDV) genotype VII is incriminated in the currently circulating NDV outbreaks in the Middle East region. In this study, evaluation of different vaccination regimes including genetically-matched or mismatched vaccines to the currently circulating field virulent NDV (vNDV) genotype VII was performed. One-day-old Arbor Acres broiler chicks were divided into nine groups; groups 1 to 3 were vaccinated with live or inactivated genetic-mismatched vaccines (genotype II) or both of them. Groups 4 to 6 were vaccinated with either live or inactivated genetic-matched vaccine to vNDV genotype VII or combination of them. Group (Gp) 7 was vaccinated with a combination of inactivated genetic-matched and live genetic-mismatched vaccines to vNDV genotype VII while groups 8 and 9 were kept as control non-vaccinated. The groups that received a combination of live and inactivated vaccines from either genetically-matched or mismatched origins had the highest serological responses and protection against mortality which was 100%. The two groups received a combination of inactivated genetic matched vaccine and live vaccines of either genetic-matched or mismatched origins had the lowest clinical index and were nearly completely protected against vNDV clinical signs. The virus tracheal and cloacal shedding titers and number of shedders were significantly reduced or nearly neglicable in the instance of application of inactivated genetic-matched vaccine to the challenge virus either alone or boosted with live genetic-matched or mismatched vaccine. In consistent inactivated genetic-matched vaccine inhibited the transmissibility of the challenged virus to contacted birds. We concluded from our results that application of NDV vaccination regimes included a combination of inactivated NDV genotype VII vaccine and live vaccine regardless of its genotype provides better clinical protection and minimize virus shedding and subsequently decrease transmissibility and virus load to the surrounding environment.

A study of risk factors associated with Newcastle disease and molecular characterization of genotype XIII Newcastle disease virus in backyard and commercial poultry in Assam, India

The continuous emergence of Newcastle disease virus (NDV) poses a persistent threat to the poultry industry. Recent increasing outbreaks of NDV in the North East region of India have highlighted the need to closely monitor and analyze the potential risk factors for Newcastle disease (ND) outbreaks. In the present study, an attempt was made to genotype the circulating Newcastle disease virus (NDV) in the backyard and commercial poultry flocks in Assam, India. Sera samples from unvaccinated backyard poultry flocks and tissue samples of ND suspected cases were collected and tested for the presence of NDV antibodies using the Haemagglutination inhibition (HI) test. A total of seven NDV isolates were analyzed from different districts of Assam, India, both genotypically and pathotypically. All isolates were characterized as virulent, carrying 112RRKQRF117 amino acid residues at the cleavage site. As determined by phylogenetic analysis, the isolates clustered with members of genotype XIII of class II NDV. Further analysis of risk factors of ND occurrence was conducted through a questionnaire survey. All the results indicated an occurrence of genotype XIII of NDV in the farms with inadequate biosecurity and farming practices.

Performance comparison of homologous and heterologous Newcastle disease virus in vaccines and antibody tests

Antigenic differences between commercial Newcastle Disease Virus (NDV) vaccine and circulating field virus reduce vaccine efficacy. Fifty-layer chickens were divided into five groups: three vaccinated chicken groups using killed LaSota (Genotype II/GII), Mega, or VD (Genotype VII/GVII) viral strains, negative, and positive control groups. On day 28, Hemagglutination Inhibition (HI) serology of vaccinated chickens was performed using whole virus antigens of RIVS, LaSota, Mega, and VD strains. Sera were also tested with an alternative antigen, using an ELISA to detect antibody for the cleavage site F protein peptide from GII and GVII NDV strains. Vaccinated and unvaccinated positive control birds underwent infectious challenges using VD and Mega strains. HI testing showed that antibody titers were higher when tested using homologous antigens than heterologous antigens. ELISA performed with alternative antigens did not perform as well as the established HI test using homologous strains. Viral shedding was reduced by vaccination that was homologous to the infectious challenge in comparison with vaccination using the LaSota strain virus. We conclude that superior results are obtained when serological testing, vaccinations, and vaccine challenge experiments all use circulating strains of ND virus. Implementation of this recommendation would likely reduce viral shedding by vaccinated chickens and be more effective in preventing outbreaks of virulent NDV.

Development of flock immunity against Newcastle disease in native chicken in a locality

Preventive measures to combat with Newcastle disease virus (NDV) can be achieved by vaccination programs that are practiced in many countries of the world. Even after vaccination outbreaks of NDV occurred in the flock indicate that vaccination against NDV cannot provide effective immunity that may cause disease in partially vaccinated flock. Hence, the objectives of this study was to development of flock immunity in chicken against Newcastle disease in a local community. In this study a vaccination programme for the native flock in a locality (Dhamrai, Savar, Dhaka, Bangladesh) was conducted and effectiveness of the Ranikhet disease vaccine (RDV) was identified with detection of antibody titers that may protect the native flocks against the disease but mortality or the infection was present. The study showed that after routine vaccination with BCRDV (Baby Chicks Ranikhet Disease Vaccine) and RDV (Ranikhet Disease Vaccine) vaccine the native chicken showed high antibody titre in native chicken. Overall herd immunity of native chickens can be achieved if the maximum (&gt;80%) chicken obtains high antibody titers (log2 haemagglutination inhibition titer ≥3) after vaccinations. So, it can be concluded that successful vaccinations programme is the key point to control NDV with higher herd health immunity. Asian J. Med. Biol. Res. 2022, 8 (1), 37-41

Cormorants as a Potentially Important Reservoir and Carrier of Newcastle Disease Virus on the Asian Continent.

Despite numerous disease prevention measures and control programs, Newcastle disease (ND) remains one of the most significant infections in poultry worldwide, especially in developing countries. It is known that wild birds, mainly of the Anseriformes order, are the main carrier of lentogenic (non-pathogenic) variants of Newcastle disease virus (NDV) in nature. But the question of the reservoir of velogenic (highly pathogenic) NDV in nature still remains open. In the 1970s, 1990s, and 2000s in North America during epizootics among cormorants, velogenic NDV strains were isolated. It was later concluded that cormorants play an important role in the maintenance and circulation of NDV in North America. New data have been obtained on the circulation of velogenic NDV strains in wild birds in Central Asia: VIIb and XIII genotype strains were isolated from cormorants for the first time in Kazakhstan. Interestingly, outbreaks of NDV registered in poultry in Central and Southern Asia were phylogenetically close to the viruses from cormorants that support the idea that cormorants can serve as the potential reservoir of velogenic NDV in developing countries of Asia. The seasonal migrations of cormorants may contribute to the distribution of the virus throughout Asia but more evidence must be obtained to confirm this hypothesis. There is increasing evidence of the introduction of NDV into the poultry farms from wild nature worldwide. This article continues the discussion on the likelihood of cormorants to serve as a reservoir and carrier of NDV on the Asian continent.

Presence of Newcastle Disease Virus in Vaccinated Indigenous Chicken in Selected Regions in Kenya —A Cross-Sectional Study

Vaccination of flocks against Newcastle disease virus (NDV) outbreaks is the main approach for controlling the spread of Newcastle disease (ND). Nevertheless, NDV outbreaks have been reported in vaccinated chickens. In this study, we determined the prevalence of NDV among vaccinated indigenous chickens (ICs) and examined the relationship of the disease with the weather (temperature, rainfall, humidity, and wind speed) at the time of sample collection, production system, and the presence of other species. The genetic diversity of the NDV matrix and fusion genes was also inferred. A total of 1,210 swabs were collected between 2017 and 2018 from ICs that were vaccinated or unvaccinated against NDV in free-range and semi-free-range production systems. We collected 650 swabs each from the oropharynx and cloaca of ICs in 68 households within the Bomet, Baringo, Kilifi, Nakuru, Kakamega, and Machakos counties in Kenya. NDV matrix genes were detected using reverse transcription-polymerase chain reaction, and amplicons of matrix and fusion genes were sequenced using a capillary sequencer from the pooled samples. Among the vaccinated ICs, the prevalence of NDV was 78.5% (p=0.045). There were significant relationships between the presence of NDV and vaccination history of the ICs (p=0.034), the type of production system for ICs (p=0.004) and the months of sample collection (p < 0.0001). However, no significant relationship was found between the presence of NDV and the interaction between ICs and other birds. The presence of matrix and fusion genes in samples from vaccinated flocks indicated the presence of both virulent and low-virulence strains of NDV. These findings highlight the significant presence of NDV among vaccinated ICs and suggest the possibility of inadequate vaccination and viral shedding post-vaccination as the drivers of infections.

Clinical, molecular and cytopathological characterization of a Newcastle disease virus from an outbreak in Baghdad, Iraq

BackgroundThe frequent outbreaks of Newcastle disease virus (NDV) in Iraq pose a constant threat to commercial poultry, despite the introduction of routine vaccination programmes. Several factors, particularly stress factors and coinfections, might play a role in increasing NDV outbreaks in poultry species.ObjectivesThe current study was aimed to characterize an NDV isolate from an outbreak in North Baghdad, Iraq.MethodsClinical pathogenicity of the isolate was determined experimentally in chickens. In vitro studies included cytopathological examination, as well as molecular and phylogenetic analyses.ResultsBased on the clinical studies and pathogenicity indices (mean death time and intracerebral and intravenous pathogenicity indices), the isolate was characterized as velogenic (highly virulent). Reverse transcriptase polymerase chain reaction targeting the partial fusion protein gene of the NDV genome confirmed the detection. Partial sequencing of the hypervariable region of the fusion gene identified the presence of an avirulent (lentogenic) fusion protein motif (GRQGRL). Phylogenetic analysis of the new isolate along with previously known regional isolates revealed that the new isolate was related to genotype II strains. Additionally, sequence analysis indicated a distinct genetic lineage of the new isolate, which was related to some of the lineages identified in previous outbreaks in the Middle East.ConclusionThe current study offers essential information on the epidemiology, characteristics and diagnosis of NDV for disease control in Iraq. The isolate was found to belong to genotype II and possess an avirulent fusion protein motif.

First Isolation and Rapid Identification of Newcastle Disease Virus from Aborted Fetus of Dromedary Camel Using Next-Generation Sequencing

Newcastle disease virus (NDV) causes morbidities and mortalities in wild and domestic birds globally. For humans, exposure to infected birds can cause conjunctivitis and influenza-like symptoms. NDV infections in mammals are rarely reported. In this study, using next-generation sequencing, an NDV was identified and isolated from Vero cells inoculated with the nasal swab of an aborted dromedary fetus in Dubai, during the time when an NDV outbreak occurred in a pigeon farm located in close proximity to the dairy camel farm where the mother of the aborted dromedary fetus resided, and there were a lot of pigeons in the camel farm. Genome analysis revealed that the structurally and functionally important features of other NDVs were also present in this dromedary NDV genome. Phylogenetic analysis based on the nucleotide sequences of fusion protein (F), hemagglutinin-neuraminidase protein (HN) and complete polyprotein showed that the virus belonged to sub-genotype VIg of class II NDV and is most closely related to pigeon NDVs in Egypt in the same year. The present study is the first that demonstrated isolation of NDV in dromedaries. Further study is warranted to investigate the relationship between NDV infection and abortion.

Recombinant duck enteritis viruses expressing the Newcastle disease virus (NDV) F gene protects chickens from lethal NDV challenge

Newcastle disease virus (NDV) is a major threat to poultry worldwide. Virulent Newcastle disease virus infection can cause 100% morbidity and mortality in chickens. Vaccination is the most effective way to prevent and control NDV outbreaks in poultry. Previously, we demonstrated that a duck enteritis virus (DEV) vaccine strain is a promising vector to generate recombinant vaccines in chickens. Here, we constructed two recombinant DEVs expressing the F protein (rDEV-F) or HN protein (rDEV-HN) of NDV. We then evaluated the protective efficacy of these recombinant DEVs in specific-pathogen-free chickens. rDEV-F induced 100% protection of chickens from lethal NDV challenge after a single dose of 104 TCID50, whereas rDEV-HN did not induce effective protection. rDEV-F may therefore serve as a promising vaccine candidate for chickens. This is the first report of a DEV-vectored vaccine providing robust protection against lethal NDV infection in chickens.

Pathological and molecular investigation of velogenic viscerotropic Newcastle disease outbreak in a vaccinated chicken flocks.

Newcastle disease virus (NDV) belongs to genus Avulavirus and family Paramyxoviridae. There are thirteen serotypes named APMV-I (Avian Paramyxovirus-I) to APMV-13 of which NDV has been designated as APMV-1. The disease has been reported worldwide affecting both domestic and wild avian species. Morbidity and mortality rates up to 100% have been reported in cases of unvaccinated flocks. Stringent vaccination schedule is practiced in endemic/disease prone areas in order to prevent the disease. Despite this, NDV outbreaks have been reported even in cases of vaccinated populations. In this study we describe detailed pathological and molecular investigation that were undertaken in an organized poultry farm from Bareilly region, Uttar Pradesh, India, involving layer flocks which succumbed to ND outbreak in spite of following strict vaccination protocol. The mortality rate ranged from 76.80 to 84.41% in different flocks with an average mortality of 79.50%. Necropsied birds had gross lesions suggestive of viscerotropic ND including petechial hemorrhages on the proventricular tips, intestinal lumen with necrotic areas covered with hemorrhages, hemorrhagic cecal tonsils, para-tracheal edema and mottling of spleen. The characteristic histopathological lesions were mainly seen in the blood vessels and lymphoid tissues. Vascular changes characterized by congestion, edema, and hemorrhage were found in majority of the organs. Lymphocytolysis in spleen and cecal tonsils was evident. Immunohistochemical studies revealed positive signals mostly in macrophage and lymphocytes. PCR assay was done to confirm the NDV genome, which revealed an amplicon size of 356bp. The phylogenetic analysis revealed the resemblance of the present isolate (ADS01) with class II genotype NDV XIIIA. The isolate belonged to velogenic NDV as the Minimum Lethal Dose (MLD) and Mean Death Time (MDT) for the present isolate were 10-8 and 41h, respectively. Thus this study clearly demonstrates that in spite of strict vaccination regime and biosecurity procedures, ND continues to be rampant. Hence it is important to effectively administer the present vaccine in addition to strains matching to the field isolates to provide longer and optimal protection against spreading of virus by means of reducing the extent of viral shedding.

Spillover of Newcastle disease viruses from poultry to wild birds in Guangdong province, southern China

Despite intensive vaccination programs in many countries, including China, Newcastle disease has been reported sporadically and is still a significant threat to the poultry industry in China. Newcastle disease virus (NDV) is infectious for at least 250 bird species, but the role of wild birds in virus epidemiology remains largely unknown. Fourteen NDV isolates were obtained from 2040 samples collected from wild birds or the environment in Guangdong province, southern China, from 2013 to 2015. The isolation rate was the highest in the period of wintering and lowest during the periods of spring migration, nesting, and postnesting. A maximum clade credibility phylogenetic analysis revealed that at least four genotypes circulate in southern China: three class II genotypes (II, VI, and IX) and one class I (1b). We also demonstrated that most isolates from wild birds were highly similar to isolates from poultry, and two isolates were linked to viruses from wild birds in northern China. These data suggested that wild birds could disseminate NDV and poultry-derived viruses may spillover to wild birds. Accordingly, vaccine development and poultry management strategies should be considered to prevent future NDV outbreaks, particularly given the strength of the poultry industry in developing countries, such as China.

Monitoring of Newcastle disease virus in environmental samples.

Newcastle disease virus (NDV) is an important pathogen in poultry. Waterfowl and a number of other avian species serve as the host for NDV. Severity of the disease is variable and infected animals mainly develop respiratory and neurological symptoms. Outbreaks of NDV in poultry are recorded regularly in the Republic of Kazakhstan despite the widespread use of vaccines. Here we present evidence that nucleic acid found in open water bodies in Kazakhstan can be detected by means of next-generation sequencing and belongs to at least three distinct genotypes of NDV.

Evaluation of fusion protein cleavage site sequences of Newcastle disease virus in genotype matched vaccines.

Newcastle disease virus (NDV) causes a devastating poultry disease worldwide. Frequent outbreaks of NDV in chickens vaccinated with conventional live vaccines suggest a need to develop new vaccines that are genetically matched against circulating NDV strains, such as the genotype V virulent strains currently circulating in Mexico and Central America. In this study, a reverse genetics system was developed for the virulent NDV strain Mexico/01/10 strain and used to generate highly attenuated vaccine candidates by individually modifying the cleavage site sequence of fusion (F) protein. The cleavage site sequence of parental virus was individually changed to those of the avirulent NDV strain LaSota and other serotypes of avian paramyxoviruses (APMV serotype-2, -3, -4, -6, -7, -8, and -9). In general, these mutations affected cell-to-cell fusion activity in vitro and the efficiency of the F protein cleavage and made recombinant Mexico/01/10 (rMex) virus highly attenuated in chickens. When chickens were immunized with the rMex mutant viruses and challenged with the virulent parent virus, there was reduced challenge virus shedding compared to birds immunized with the heterologous vaccine strain LaSota. Among the vaccine candidates, rMex containing the cleavage site sequence of APMV-2 induced the highest neutralizing antibody titer and completely protected chickens from challenge virus shedding. These results show the role of the F protein cleavage site sequence of each APMV type in generating genotype V-matched vaccines and the efficacy of matched vaccine strains to provide better protection against NDV strains currently circulating in Mexico.

Molecular screening and isolation of Newcastle disease virus from live poultry markets and chickens from commercial poultry farms in Zaria, Kaduna state, Nigeria

Newcastle disease is one of the major economic threats to poultry population because of its high morbidity and mortality varying from 90-100%. It is caused by Avian Paramyxovirus-1 (APMV-1). This research work was carried out to identify Newcastle disease virus (NDV) by using reverse transcription-polymerase chain reaction (RT-PCR) assay and further isolate the virus in embryonated chicken eggs. A total of 127 cloacal swabs were collected from local chickens in live bird market and exotic chickens in commercial poultry farms in Zaria and environs, Nigeria between November, 2014 and January, 2015. Five commercial poultry farms and four live bird markets were purposively sampled. Molecular screening of NDV Matrix-gene (M-gene) was performed on all the samples using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). Newcastle disease positive samples were further inoculated into embryonated chicken eggs for isolation of Newcastle disease virus. Isolates were confirmed as Newcastle disease virus by haemaggulitination inhibition (HI) test. Newcastle disease virus Matrix-gene was detected in 16 (12.5%) out of 127 cloacal swabs; 13 (10.2%) from live bird markets and 3 (2.3%) from commercial poultry farms. However, only 10 Newcastle disease viruses were isolated in embryonated chicken eggs as confirmed by Haemagglutination inhibition (HI) test. Due to the higher detection rate recorded by reverse transcriptase-polymerase chain reaction (RT-PCR), it is therefore important that molecular technique be made easily accessible so that samples from each suspected outbreaks of NDV be screened so that rapid and confirmatory diagnosis can be achieved.Keywords: Embryonated chicken eggs, Haemagglutination inhibition test, Newcastle disease virus, RT-PCR

Genetic Modification of Oncolytic Newcastle Disease Virus for Cancer Therapy.

Clinical development of a mesogenic strain of Newcastle disease virus (NDV) as an oncolytic agent for cancer therapy has been hampered by its select agent status due to its pathogenicity in avian species. Using reverse genetics, we have generated a lead candidate oncolytic NDV based on the mesogenic NDV-73T strain that is no longer classified as a select agent for clinical development. This recombinant NDV has a modification at the fusion protein (F) cleavage site to reduce the efficiency of F protein cleavage and an insertion of a 198-nucleotide sequence into the HN-L intergenic region, resulting in reduced viral gene expression and replication in avian cells but not in mammalian cells. In mammalian cells, except for viral polymerase (L) gene expression, viral gene expression is not negatively impacted or increased by the HN-L intergenic insertion. Furthermore, the virus can be engineered to express a foreign gene while still retaining the ability to grow to high titers in cell culture. The recombinant NDV selectively replicates in and kills tumor cells and is able to drive potent tumor growth inhibition following intratumoral or intravenous administration in a mouse tumor model. The candidate is well positioned for clinical development as an oncolytic virus. Avian paramyxovirus type 1, NDV, has been an attractive oncolytic agent for cancer virotherapy. However, this virus can cause epidemic disease in poultry, and concerns about the potential environmental and economic impact of an NDV outbreak have precluded its clinical development. Here we describe generation and characterization of a highly potent oncolytic NDV variant that is unlikely to cause Newcastle disease in its avian host, representing an essential step toward moving NDV forward as an oncolytic agent. Several attenuation mechanisms have been genetically engineered into the recombinant NDV that reduce chicken pathogenicity to a level that is acceptable worldwide without impacting viral production in cell culture. The selective tumor replication of this recombinant NDV, both in vitro and in vivo, along with efficient tumor cell killing makes it an attractive oncolytic virus candidate that may provide clinical benefit to patients.