scholarly journals Pathogenesis of Velogenic Genotype VII.1.1 Newcastle Disease Virus Isolated from Chicken in Egypt via Different Inoculation Routes: Molecular, Histopathological, and Immunohistochemical Study

Animals ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 3567
Author(s):  
Yassmin EL-Morshidy ◽  
Walied Abdo ◽  
Ehab Kotb Elmahallawy ◽  
Ghada Allam Abd EL-Dayem ◽  
Ahmed El sawak ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Time Course ◽  
Clinical Signs ◽  
Post Inoculation ◽  
Tissue Samples ◽  
Genetic Characteristics ◽  
Hemagglutination Inhibition Test ◽  
Death Time

Newcastle disease virus (NDV) remains a constant threat to the poultry industry. There is scarce information concerning the pathogenicity and genetic characteristics of the circulating velogenic Newcastle disease virus (NDV) in Egypt. In the present work, NDV was screened from tracheal swabs collected from several broiler chicken farms (N = 12) in Dakahlia Governorate, Egypt. Real-time reverse transcriptase polymerase chain reaction (RRT-PCR) was used for screening of velogenic and mesogenic NDV strains through targeting F gene fragment amplification, followed by sequencing of the resulting PCR products. The identified strain, namely, NDV-CH-EGYPT-F42-DAKAHLIA-2019, was isolated and titrated in the allantoic cavity of 10 day old specific pathogen-free (SPF) embryonated chicken eggs (ECEs), and then their virulence was determined by mean death time (MDT) and intracerebral pathogenicity index (ICPI). The pathogenicity of the identified velogenic NDV strain was also assessed in 28 day old chickens using different inoculation routes as follows: intraocular, choanal slit, intranasal routes, and a combination of both intranasal and intraocular routes. In addition, sera were collected 5 and 10 days post inoculation (pi) for the detection of NDV antibodies by hemagglutination inhibition test (HI), and tissue samples from different organs were collected for histopathological and immunohistochemical examination. A series of different clinical signs and postmortem lesions were recorded with the various routes. Interestingly, histopathology and immunohistochemistry for NDV nucleoprotein displayed widespread systemic distribution. The intensity of viral nucleoprotein immunolabeling was detected within different cells including the epithelial and endothelium lining, as well as macrophages. The onset, distribution, and severity of the observed lesions were remarkably different between various inoculation routes. Collectively, a time-course comparative pathogenesis study of NDV infection demonstrated the role of different routes in the pathogenicity of NDV. The intranasal challenge was associated with a prominent increase in NDV lesions, whereas the choanal slit route was the route least accompanied by severe NDV pathological findings. Clearly, the present findings might be helpful for implementation of proper vaccination strategies against NDV.

scholarly journals Isolation and pathotyping of Newcastle disease virus isolated from birds in Kerala

2021 ◽  
Vol 52 (3) ◽  
Author(s):  
Vijayakumar K ◽  
Vijayakumar K ◽  
Vijayakumar K ◽  
Vijayakumar K ◽  
Vijayakumar K
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Haemagglutination Inhibition ◽  
Clinical Signs ◽  
Viral Disease ◽  
High Morbidity ◽  
Isolation And Identification ◽  
Tissue Samples ◽  
Death Time

Newcastle disease (ND) is a pandemic viral disease of poultry. It is highly contagious and causes high morbidity and mortality in affected flocks. The disease is caused by Avian orthoavulavirus 1, commonly known as Newcastle disease virus (NDV) belongs to the family Paramyxoviridae. The virus affects almost 241 species of birds. Based on the pathogenicity, the virus is classified into five pathotypes viz., viscerotropic velogenic, neurotropic velogenic, mesogenic, lentogenic and asymptomatic enteric NDV. The severity of the disease varies with the viral pathotype. Isolation and identification along with pathotyping of the virus provides a basis for understanding the type of virus circulating in the region. In the present study, tissue samples from dead/ ailing birds showing lesions/clinical signs suggestive of ND were collected. They were subjected to virus isolation in embryonated chicken eggs and identified by haemagglutination test and confirmed by haemagglutination inhibition test. Eight NDV isolates were obtained out of 55 tissue samples and were classified into pathotypes by intracerebral pathogenicity index (ICPI) and mean death time (MDT). The ICPI values varied from 0.75 to 1.53 and MDT from 54 h. to 79.2 h. Out of eight isolates, three belonged to velogenic group and five were of mesogenic pathotype. The study revealed the circulation of virulent NDV in Kerala. The pathogenicity tests provide a basis for understanding the epidemiology of ND.

scholarly journals Molecular characterisation of Newcastle disease virus exotic isolate in East Java, Indonesia

2021 ◽  
Vol 24 (2) ◽  
pp. 191-199
Author(s):  
N. P. Kusumarahayu ◽  
N. Putri ◽  
R. Ernawati ◽  
J. Rahmahani ◽  
S. Suwarno ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Clinical Signs ◽  
Rt Pcr ◽  
Genetic Characteristics ◽  
Basic Work ◽  
Specific Pathogen ◽  
Baseline Information ◽  
Native Chickens

Newcastle disease virus (NDV) is ssRNA paramyxovirus causing clinical signs, varying from subclinical infections to 100% mortality in infected chickens. Haemagglutinin-neuraminidase (HN) protein has an important role related to infection and pathogenesis, therefore, the protein was characterised in this study. Samples were collected from 45 cloacal swabs of native chickens. They were isolated by inoculating in specific pathogen-free embryonated eggs. Molecular detection of NDV was done by reverse transcriptase polymerase chain reaction (RT-PCR) encoding HN protein. RT-PCR for HN gene of NDV generated DNA fragments sized 503 bp, which were then sequenced using ABI Prism. The results have shown that virus isolates were mostly lentogenic and might contribute to outbreak in East Java, Indonesia. Based on this fact, NDV infected native chickens can act as reservoir and contribute to outbreak in the poultry. Our study provides baseline information on genetic characteristics of NDV circulating in East Java and serves as a basic work for further research.

scholarly journals The Pathology of Vaccination of Chickens with Varying Doses of Lentogenic LaSota Strain of Newcastle Disease Virus

2021 ◽  
Vol 41 (1) ◽  
pp. 62-72
Author(s):  
A.O. Igwe ◽  
M.E. Sanda ◽  
U.E.I. Nnsewo ◽  
C.J. Okonkwo ◽  
O. Onyebgula
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Broiler Chickens ◽  
Haemagglutination Inhibition ◽  
Clinical Signs ◽  
Lymphoid Hyperplasia ◽  
Post Inoculation ◽  
Serum Samples ◽  
Antibody Titres

Recently, it was demonstrated under laboratory conditions that increased doses of LaSota vaccine increased ND antibody response significantly in chickens. In this study, we have used the same model to investigate whether vaccination with increased doses of  lentogenic LaSota strain of Newcastle disease virus are associated with pathological changes in chickens. Four-week-old broiler chickens (n=100) were randomly assigned into four groups of 25 each: ZD, each drenched with phosphate-buffered saline, SD, DD and TD broilers were each drenched with single, double and triple dose of LaSota vaccine, respectively. The chickens were observed for clinical signs and lesions. Serum samples were collected from the chickens in all the groups at weekly intervals post inoculation (PV) and assayed for haemagglutination inhibition (HI) antibodies. The vaccinated broilers showed no morbidity and mortality. Only the bursa of all the vaccinated groups appeared slightly reduced in size on day 10 PV. The histopathological changes were lymphoid  hyperplasia and formation of germinal centres in the spleen and caecal tonsils from days 3 to 6 PV and mild depletion of bursal lymphocytes on day 10 PV. Generally, the integrity of the lymphoid organs was intact. Groups DD and TD antibody titres were  significantly (P < 0.05) higher than that of the SD on day 21 PV. This suggests that increased doses of LaSota vaccine does not cause  pathologic impairment and may be considered in improving the performance of the vaccine in the control of velogenic ND. Key words: Newcastle disease, LaSota vaccine, pathology, broiler chickens

scholarly journals Adaptation of velogenic Newcastle disease virus in Vero cells: velogenicity of virus unaltered after adaptation

2017 ◽  
Vol 59 (4) ◽  
pp. 341 ◽  
Author(s):  
P. V. RAVINDRA ◽  
B. RATTA ◽  
U. CHATURVEDI ◽  
S. K. PALIA ◽  
P. K SUBUDHI ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Vero Cells ◽  
Post Inoculation ◽  
Death Time ◽  
Infected Cells ◽  
Velogenic Newcastle Disease Virus ◽  
Syncytia Formation ◽  
Chicken Eggs

In the present experiment, velogenic Newcastle disease virus (NDV) was adapted to Vero cells without the addition of exogenous trypsin. The cytopathic effect (CPE) in NDV-infected Vero cells was observed from passage 3 (P 3) onwards with concomitant increase in the haemagglutination (HA) and virus titres. The CPE was characterized by rounding of cells, their detachment from the surface and syncytia formation. The Vero-adapted NDV was confirmed by reverse transcription-polymerase chain reaction (RT-PCR), which detected the amplification of haemagglutinin neuraminidase (HN) gene from virus-infected cells. The mean death time (MDT) of 58 hours upon inoculation to embryonated chicken eggs and the intravenous pathogenicity index (IVPI) of 2.12 following inoculation to SPF chicken suggested that Vero adapted virus retained the velogenicity even after ten passages. Furthermore, the nervous signs and haemorrhages in the pro-ventriculus of birds that died at day 5 post inoculation with PIO virus confirmed that the Vero-adapted NDV was a neurotropic viscerotropic velogenic strain.

scholarly journals Awareness and antibody detection of Newcastle disease virus in a neglected society in Nigeria

2019 ◽  
Vol 12 (1) ◽  
pp. 112-118 ◽  
Author(s):  
Oluwafemi Babatunde Daodu ◽  
Julius Olaniyi Aiyedun ◽  
Rafiu Adebisi Kadir ◽  
Hauwa Motunrayo Ambali ◽  
Oladapo Oyedeji Oludairo ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Clinical Signs ◽  
Egg Yolk ◽  
Antibody Detection ◽  
Cross Sectional ◽  
Hemagglutination Inhibition Test ◽  
Egg Yolks ◽  
The Government

Aim: This study aimed to assess the level of awareness of rural poultry farmers on vaccination and to detect Newcastle disease virus (NDV) antibody in local birds (LB) and eggs in Kwara State, Nigeria. Materials and Methods: Data on farmers' attitude, knowledge, practices, and experiences on ND mortality were obtained through an interview using a structured cross-sectional checklist. NDV antibodies were detected in sera and egg yolks of local chickens (LC) and guinea fowls (GF) using hemagglutination inhibition test. Results: A total of 83 interviewees, 287 sera and 121 egg yolk extracts, were examined. The study revealed that 98.8% (82/83) of the interviewee had never vaccinated their flock before. 90% of the interviewee had reported high mortality in birds within 1-6 months old, while the major clinical signs were cold (40.4%) and torticollis (30.8%). Evidences of LB exposure to wild-type NDV were confirmed by the detection of NDV antibodies in 20.8% and 0% of LC and GF, respectively. The mortality differences experienced in <1 and 1-6 months old LB could be explained by the presence of maternally-derived NDV antibody (49.6%) in egg yolk. Conclusion: The study showed that LB suffers from NDV as a result of LB keepers' ignorance and neglect by the government. This has limited local investment and subsequent contribution to gross domestic product. This study suggests that the key factors to the prevention of ND remain awareness creation about poultry vaccination, production of affordable vaccines, and availability/accessibility to veterinarian (or trained personnel).

scholarly journals Pathogenesis of Six Pigeon-Origin Isolates of Newcastle Disease Virus for Domestic Chickens

2002 ◽  
Vol 39 (3) ◽  
pp. 353-362 ◽  
Author(s):  
G. D. Kommers ◽  
D. J. King ◽  
B. S. Seal ◽  
K. P. Carmichael ◽  
C. C. Brown
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Clinical Signs ◽  
Tunel Assay ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Apoptotic Cells ◽  
Viral Mrna ◽  
Tissue Samples ◽  
Matrix Gene

The pathogenesis of six pigeon-origin isolates of Newcastle disease virus (NDV) was investigated in chickens. Four isolates were previously defined as the variant pigeon paramyxovirus 1 (PPMV-1), and two isolates were classified as avian paramyxovirus 1 (APMV-1). Birds inoculated with PPMV-1 isolates were euthanatized, and tissue samples were collected at 2, 5, and 10 days postinoculation (DPI). Birds inoculated with APMV-1 isolates died or were euthanatized, and tissue samples were collected at 2, 4, and 5 DPI. Tissues were examined by histopathology, immunohistochemistry (IHC) for the presence of NDV nucleoprotein, and in situ hybridization (ISH) for the presence of viral mRNA for the matrix gene. Spleen sections were stained by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay and by IHC using an anti-active caspase-3 antibody (IHC-Casp) to detect apoptotic cells. Brain sections of PPMV-1-infected birds were examined by IHC to detect T and B lymphocytes and glial fibrillary acidic protein (GFAP). Histologically, birds inoculated with PPMV-1 isolates had marked lesions in the heart and brain. Presence of viral nucleoprotein and viral mRNA in the affected tissues was confirmed by IHC and ISH, respectively. Numerous reactive astrocytes were observed in brain sections stained for GFAP. Among all the isolates, the IHC-Casp demonstrated that apoptosis was very prominent in the ellipsoid-associated cells of the spleen at 2 DPI. Results of the TUNEL assay indicated that apoptotic cells were prominent at 5 DPI and were more randomly distributed. The clinical signs and gross and histopathologic changes observed in the APMV-1-infected birds were characteristic of an extensive infection with highly virulent NDV evident by IHC.

scholarly journals Isolation and identification of the Newcastle disease virus from field outbreaks in broiler and layer flocks in Iraq

2017 ◽  
Vol 41 (1) ◽  
pp. 23-27
Author(s):  
Mushtaq T. B. AL-Zuhariy
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Sequence Similarity ◽  
Haemagglutination Inhibition ◽  
Clinical Signs ◽  
Economic Losses ◽  
Isolation And Identification ◽  
Death Time ◽  
Polymerase Chain

     Newcastle disease is one of serious pathological problems and causes of vast economic losses during 2011-2016 in Iraq. The disease caused high mortalities in all types of poultry nevertheless of vaccination. In this study all samples were collected from infected flocks with clinical signs of the disease. Inoculation of chicken embryonated eggs was carried out for virus isolation, identification, Haemagglutination and Haemagglutination Inhibition assay. Using Reverse Transcriptase-Polymerase Chain Reaction to confirm the presence of the virus, Intra Cerebral Pathogenicity Index and Mean Death Time were used to confirm all the isolates that were velogenic. The important determinant of Newcastle disease virus pathogenicity is fusion protein that has been used for phylogenetic analysis. sequencing and compared genetically of Newcastle disease virus Iraqi isolate to publish sequences acquired from GenBank showed 99% sequence similarity to the Iran isolate IRI 1392k (KJ176996.1). It can concluded from these data that introduction new virus was occurred in Iraq.

scholarly journals Determination of in-vivo growth kinetics of virulent Newcastle disease virus in layer chicken

2016 ◽  
Vol 1 (3) ◽  
pp. 526-536
Author(s):  
Md Mostofa Kamal ◽  
Mohammad Aynul Haque ◽  
Shuvho Chakra Borty ◽  
AKM Khashruzzaman ◽  
Mohammad Nizam Uddin Chowdhury ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Research Work ◽  
Clinical Signs ◽  
Avian Embryo ◽  
Death Time ◽  
Virulent Newcastle Disease Virus ◽  
Almost All

The research work was conducted on 105 layer chicks with a view to determine the rate of distribution of neurotropic virulent Newcastle disease virus (NVNDV) in various organs following infection through natural (intranasal, intraocular and oral) and parenteral (intravenous, intramuscular and subcutaneous) routes of inoculation at different ages (7, 15 and 28 days of old). Each bird received a dose of 0.2 ml contained 300 ELD50 of reference NVNDV. The highest body temperature (?1080F) was recorded in the birds of almost all the experimental groups within 48 to 72 hours of PI. Appearance of clinical signs was observed earlier (48 to 72 hours of PI) in parenterally infected birds than those of inoculated through natural routes. The shortest duration (24-48 hours of PI) and longest duration (74-138 hours of PI) of death time were recorded in birds those inoculated through IV and oral routes of infection respectively. Isolation of NDV was positive from day 2 of PI and onward in all the groups with some minor variations in some cases. The CEF system was found more sensitive for the isolation of viruses compare to that of avian embryo. The highest HA titre of NDV was found in the brain tissue followed by lungs and kidney. Significantly (p<0.01) higher HA titre of NDV isolate was recorded in the birds of all the experimental groups inoculated through IV route. Following infection, the MDA titre decreased day by day in the birds with the increase of HA titres of NDV.Asian J. Med. Biol. Res. December 2015, 1(3): 526-536

scholarly journals Effect of Immunosuppression on Newcastle Disease Virus Persistence in Ducks with Different Immune Status

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Lucy W. Njagi ◽  
Phillip N. Nyaga ◽  
Lilly C. Bebora ◽  
Paul G. Mbuthia ◽  
Uswege M. Minga
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Immune Status ◽  
Experimental Period ◽  
Hemagglutination Inhibition Test ◽  
Viral Excretion ◽  
Antibody Titres ◽  
Antibody Levels ◽  
Chicken Embryo Fibroblasts

This study was carried out to verify the possibility that ducks are sources of Newcastle disease (ND) virus infection for chickens in mixed flocks. Immunosuppressed (IS) and non immunosuppressed (NIS) birds, at three different antibody levels (medium, low and absent) were used; the titres having been induced through vaccination, and Immunosuppression done using dexamethazone. Each of the 3 respective groups was further divided into 2 groups of about 12 ducks each: one challenged with velogenic ND virus; the other not challenged. Selected ducks from all groups had their antibody titres monitored serially using hemagglutination inhibition test, while two birds from each of the challenged groups were killed and respective tissues processed for ND viral recovery, using chicken embryo fibroblasts. In general, antibody titres of IS and NIS challenged ducks were significantly higher than their unchallenged counterparts (P<0.05). Non-challenged pre-immunised ducks had a progressive decrease in antibody levels; non-immunised ducks did not seroconvert. Newcastle disease virus was isolated from livers and kidneys of the challenged ducks throughout the experimental period; indicating a possibility of viral excretion, especially when the birds are stressed. It, therefore, provides another possible model of viral circulation within mixed flocks.

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