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

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.

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

The Iraqi Journal of Veterinary Medicine ◽

10.30539/iraqijvm.v41i1.73 ◽

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.

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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 ◽

10.3390/ani11123567 ◽

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.

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Detection of the Newcastle disease virus and its effect on development of post-vaccination immunity in a commercial flock of laying hens

Acta Veterinaria Brno ◽

10.2754/avb201281010003 ◽

2012 ◽

Vol 81 (1) ◽

pp. 3-8

Author(s):

Jana Jeřábková ◽

Růžena Juranová ◽

Kateřina Rosenbergová ◽

Libuše Kulíková ◽

Alfred Hera ◽

...

Keyword(s):

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Laying Hens ◽

Haemagglutination Inhibition ◽

Pathogenic Strain ◽

Sequencing Analysis ◽

Tissue Samples ◽

Virus Rna ◽

Commercial Flock

The aim of this study was to monitor the concentration of antibodies against Newcastle disease after vaccination of laying hens at the beginning and in the end of the laying period. The study was carried out in one commercial flock of laying hens in Opatovice in the Czech Republic in the years 2008-2010. A total of 280 samples of blood sera were taken from laying hens coming from four poultry houses. The sera were tested by the haemagglutination inhibition test according to the OIE Manual. Virological testing was conducted as a consequence of atypical results of serological testing. Newcastle disease virus RNA was proved by the RT-nested PCR method in the pooled tissue samples of 5 hens, in the samples of intestines with ileocaecal tonsila, in trachea and also in one swab sample from the environment of one house. Based on sequencing analysis and subsequent phylogenetic analysis, the virus was identified as a low pathogenic strain of paramyxovirus (PMV-1). This low pathogenic strain did not have any impact on the health of laying hens.

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Detection of Newcastle Disease Virus by Immunohistochemistry on the Brains of Laying Birds with Clinical Signs Torticollis and Curled Toe Paralysis

KnE Life Sciences ◽

10.18502/kls.v3i6.1137 ◽

2017 ◽

Vol 3 (6) ◽

pp. 286

Author(s):

Ocie Harum Wulan ◽

Niken Yunita ◽

Hastari Wuryastuty ◽

Raden Wasito

Keyword(s):

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Environmental Changes ◽

Laying Hens ◽

Clinical Signs ◽

Economic Loss ◽

High Morbidity ◽

Fast Detection ◽

Rapid Spread

Poultry is one of animal protein sources that have been consumed much in Indonesia. Various of health disorders are caused by environmental changes, woof and infectious agents have caused decline in production and economic loss. One of diseases that have a big impact on poultry husbandry is Newcastle disease (ND). Newcastle disease virus (NDV) is an important disease in Indonesia, because this has spread around Indonesia and caused a big loss for poultry industry moreover, this has high morbidity and mortality and very rapid spread of the virus. The purpose of the research is to find out and determine a fast, exact and accurate, and efficient and effective application immunohistochemistry streptavidin biotin (IHK SB) test on NDV diagnosis affirmation and for NDV fast detection as a cause of disease on commercial laying hens with clinical torticollis and curled toe paralysis symptoms. The samples used in the form of tissues from 20 commercial laying hens with an indication of clinical torticollis and curled toe paralysis symptoms. Brains would be tested using streptavidin biotin. Inspection result data obtained by immunohistochemistry streptavidin biotin (IHK SB) method is analyzed in a descriptive and qualitative way. The result of the research proves that IHK SB coloration, NDV antigen could be detected on brains. Based on the result of the research, it could be concluded that IHK SB could be applied for NDV diagnosis affirmation and it proves that commercial laying hens with clinical torticollis and curled toe paralysis symptoms. Keywords: Newcastle disease virus, immunohistochemistry streptavidin biotin, torticollis, curled toe paralysis

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Detection of Newcastle Disease Virus (NDV) in Laughing Doves and the Risk of Spread to Backyard Poultry

Folia Veterinaria ◽

10.2478/fv-2020-0021 ◽

2020 ◽

Vol 64 (3) ◽

pp. 1-12

Author(s):

J. U. Okpanachi ◽

J. U. Umoh ◽

G. S. N. Kia ◽

A. A. Dzikwi

Keyword(s):

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Fusion Gene ◽

Haemagglutination Inhibition ◽

Viral Disease ◽

Poultry Production ◽

Rt Pcr ◽

Backyard Poultry ◽

Field Samples

AbstractNewcastle disease (ND) is a highly infectious viral disease of birds caused by the Newcastle disease virus (NDV) and doves have been incriminated in previous outbreaks of the disease that have discouraged backyard poultry productions. This survey was done to detect and characterize the NDV from 184 swabs from the cloacae and pharynxes of 67 trapped laughing doves and 25 backyard poultry birds. The study utilized haemagglutination assay (HA) followed by haemagglutination inhibition (HI) tests on HA positive samples to screen field samples. Conventional reverse transcriptase polymerase chain reaction (RT-PCR) was conducted on the HI positives to characterize the NDV. This study revealed that of 134 dove samples screened, 88 (65.7 %) were HA positive. Of these HA positives subjected to HI testing, 37 (42.1 %) were HI positive. Interestingly, 21 (56.8 %) of the HI positives were also RT-PCR positive: 8 lentogenic, 12 velogenic, while one had both lentogenic and velogenic NDV. Comparatively, of the 50 chicken samples screened, 23 (46 %) were HA positive; and of these, HA positives subjected to HI testing, 16 (69.6 %) were HI positive. Only 4 (25 %) of the HI positives were RTPCR positive: 3 lentogenic and a velogenic NDV. From this study it was concluded that laughing doves were demonstrated to be infected with either lentogenic or velogenic NDV or both. The use of red blood adsorption-de-adsorption concentration of NDV enhanced the RT-PCR detection using the fusion gene primers NDV-F 4829 and NDV-R 5031. The detection of not only lentogenic but velogenic NDV in laughing doves poses a great risk to backyard poultry production.

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Multi-epitope peptide vaccine prediction against Newcastle disease virus using immuno-informatics approaches

BULGARIAN JOURNAL OF VETERINARY MEDICINE ◽

10.15547/bjvm.2019-0074 ◽

2021 ◽

Vol 24 (1) ◽

pp. 136-143

Author(s):

N. Putri ◽

I. Wulandari ◽

R. Ernawati ◽

F. Abdul Rantam

Keyword(s):

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

3D Structure ◽

Peptide Vaccine ◽

Clinical Signs ◽

Economic Losses ◽

High Morbidity ◽

Epitope Peptide ◽

Worldwide Distribution

Newcastle disease is one of the most critical disease in poultry and wild birds, largely due to its high morbidity and mortality, as well as its worldwide distribution and threat of considerable economic losses to avian industries caused by Newcastle disease virus (NDV). The NDV can cause clinical signs varying from subclinical infections to 100% mortality, depending on the susceptibility of the host and the virulence of the virus. The virus is classified into velogenic (viscerotropic velogenic and neurotropic velogenic), mesogenic and lentogenic. The objectives of this study was to design a peptide vaccine using immunoinformatics approaches. In total, 12 NDV fusion proteins retrieved from NCBI database were aligned to determine the conservancy and candidate epitopes were analysed by predictions tools from Immune Epitope Database. Then the 3D structure of the conserved region was modelled using the Swiss Model and aligned using PyMol software. Two epitopes were predicted as a peptide vaccine for B cell (DKAVNVYTSSQT and NMPKDKEACAKAPLEA). This is a preliminary study of designing an epitope-based peptide vaccine against NDV, and we recommend further study to identify the interaction between these peptides with T cells and antibodies.

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

Nigerian Veterinary Journal ◽

10.4314/nvj.v41i1.8 ◽

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

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

Veterinary Pathology ◽

10.1354/vp.39-3-353 ◽

2002 ◽

Vol 39 (3) ◽

pp. 353-362 ◽

Cited By ~ 46

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.

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

Asian Journal of Medical and Biological Research ◽

10.3329/ajmbr.v1i3.26476 ◽

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

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Thymic transcriptome analysis after Newcastle disease virus inoculation in chickens and the influence of host small RNAs on NDV replication

Scientific Reports ◽

10.1038/s41598-021-89464-1 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Liangxing Guo ◽

Zhaokun Mu ◽

Furong Nie ◽

Xuanniu Chang ◽

Haitao Duan ◽

...

Keyword(s):

Innate Immunity ◽

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Viral Disease ◽

Immune Genes ◽

Comprehensive Information ◽

Virulent Newcastle Disease Virus ◽

Chicken Thymus ◽

Innate Immune Genes

AbstractNewcastle disease (ND), caused by virulent Newcastle disease virus (NDV), is a contagious viral disease affecting various birds and poultry worldwide. In this project, differentially expressed (DE) circRNAs, miRNAs and mRNAs were identified by high-throughput RNA sequencing (RNA-Seq) in chicken thymus at 24, 48, 72 or 96 h post LaSota NDV vaccine injection versus pre-inoculation group. The vital terms or pathways enriched by vaccine-influenced genes were tested through KEGG and GO analysis. DE genes implicated in innate immunity were preliminarily screened out through GO, InnateDB and Reactome Pathway databases. The interaction networks of DE innate immune genes were established by STRING website. Considering the high expression of gga-miR-6631-5p across all the four time points, DE circRNAs or mRNAs with the possibility to bind to gga-miR-6631-5p were screened out. Among DE genes that had the probability to interact with gga-miR-6631-5p, 7 genes were found to be related to innate immunity. Furthermore, gga-miR-6631-5p promoted LaSota NDV replication by targeting insulin induced gene 1 (INSIG1) in DF-1 chicken fibroblast cells. Taken together, our data provided the comprehensive information about molecular responses to NDV LaSota vaccine in Chinese Partridge Shank Chickens and elucidated the vital roles of gga-miR-6631-5p/INSIG1 axis in LaSota NDV replication.

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