Dataset Information

Quantitative evaluation of viral interference among Egyptian isolates of highly pathogenic avian influenza viruses (H5N1 and H5N8) with the lentogenic and velogenic Newcastle disease virus genotype VII in specific pathogen-free embryonated chicken eggs model.

ABSTRACT:

Background and aim

Mixed infections of the highly pathogenic avian influenza virus (HPAIV) and Newcastle disease virus (NDV) are considered the most distressing problem of the poultry industry. The problem arises due to the influence of a hidden virus on the replication of another suspected virus. Consequently, misdiagnosis of the real cause of disease may become a source of infection for other healthy stock by transmission and dissemination of the hidden virus. This study aimed to determine the impact of HPAIV and NDV on each other in a specific pathogen-free embryonated chicken egg (SPF-ECE) model.

Materials and methods

HPAIVs (H5N1 and H5N8) and NDVs [avirulent NDV [avNDV] and velogenic NDV [vNDV]) were inoculated into the allantois cavity of SPF-ECE with graded titers (2, 3, and 4 log₁₀ EID50) at 24 and 48 h of incubation, followed by the collection of allantoic fluid. A quantitative reverse transcription real-time polymerase chain reaction was used to determine the viral RNA copies of both viruses.

Results

Obvious interference was reported on the growth of NDVs when co-inoculated with AIVs. NDV RNA titers reduction ranged from <3 to 5 log₁₀ to complete suppression, but slight interference with the growth of AIVs occurred. H5N1 RNA titers showed <1-2 log₁₀ reduction when co-inoculated with vNDV compared with the H5N1 control. The interference impact of H5N8 was more powerful than that of H5N1, while vNDV showed more resistance for interference than the avNDV strain. On the other hand, interference of AIVs was not observed except when vNDV was inoculated before H5N1. The interfering impact was increased after 48 h of inoculation, whereas no titer of avNDV was detectable.

Conclusion

AIV strains had a powerful effect on NDV growth, regardless of which infection occurred first.

SUBMITTER: Soliman MA

PROVIDER: S-EPMC6925047 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature

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Publications

Quantitative evaluation of viral interference among Egyptian isolates of highly pathogenic avian influenza viruses (H5N1 and H5N8) with the lentogenic and velogenic Newcastle disease virus genotype VII in specific pathogen-free embryonated chicken eggs model.

Soliman Mohamed A MA Nour Ahmed A AA Erfan Ahmed M AM

Veterinary world 20191123 11

<h4>Background and aim</h4>Mixed infections of the highly pathogenic avian influenza virus (HPAIV) and Newcastle disease virus (NDV) are considered the most distressing problem of the poultry industry. The problem arises due to the influence of a hidden virus on the replication of another suspected virus. Consequently, misdiagnosis of the real cause of disease may become a source of infection for other healthy stock by transmission and dissemination of the hidden virus. This study aimed to deter ...[more]

PMID: 32009763

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Project description:Newcastle disease (ND) is considered to be one of the most economically significant avian viral diseases. It has a worldwide distribution and a continuous diversity of genotypes. Despite its limited zoonotic potential, Newcastle disease virus (NDV) outbreaks in Egypt occur frequently and result in serious economic losses in the poultry industry. In this study, we investigated and characterized NDV in wild cattle egrets and house sparrows. Fifty cattle egrets and fifty house sparrows were collected from the vicinity of chicken farms in Kafrelsheikh Governorate, Egypt, which has a history of NDV infection. Lung, spleen, and brain tissue samples were pooled from each bird and screened for NDV by real-time reverse transcriptase polymerase chain reaction (RRT-PCR) and reverse transcriptase polymerase chain reaction (RT-PCR) to amplify the 370 bp NDV F gene fragment. NDV was detected by RRT-PCR in 22 of 50 (44%) cattle egrets and 13 of 50 (26%) house sparrows, while the conventional RT-PCR detected NDV in 18 of 50 (36%) cattle egrets and 10 of 50 (20%) of house sparrows. Phylogenic analysis revealed that the NDV strains identified in the present study are closely related to other Egyptian class II, sub-genotype VII.1.1 NDV strains from GenBank, having 99.7-98.5% identity. The pathogenicity of the wild-bird-origin NDV sub-genotype VII.1.1 NDV strains were assessed by experimental inoculation of identified strains (KFS-Motobas-2, KFS-Elhamoul-1, and KFS-Elhamoul-3) in 28-day-old specific-pathogen-free (SPF) Cobb chickens. The clinical signs and post-mortem changes of velogenic NDV genotype VII (GVII) were observed in inoculated chickens 3 to 7 days post-inoculation, with 67.5-70% mortality rates. NDV was detected in all NDV-inoculated chickens by RRT-PCR and RT-PCR at 3, 7, and 10 days post-inoculation. The histopathological findings of the experimentally infected chickens showed marked pulmonary congestion and pneumonia associated with complete bronchial stenosis. The spleen showed histocytic cell proliferation with marked lymphoid depletion, while the brain had malacia and diffuse gliosis. These findings provide interesting data about the characterization of NDV in wild birds from Egypt and add to our understanding of their possible role in the transmission dynamics of the disease in Egypt. Further research is needed to explore the role of other species of wild birds in the epidemiology of this disease and to compare the strains circulating in wild birds with those found in poultry.

Project description: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.

Project description:Newcastle disease (ND) is caused by virulent strains of avian paramyxovirus type 1, also known as Newcastle disease virus (NDV). Despite vaccination, the frequency of reported outbreaks in Ethiopia has increased. From January to June 2022, an active outbreak investigation was conducted in six commercial chicken farms across areas of central Ethiopia to identify the circulating NDV strains. Thirty pooled tissue specimens were collected from chickens suspected of being infected with NDV. A questionnaire survey of farm owners and veterinarians was also carried out to collect information on the farms and the outbreak status. NDV was isolated using specific-pathogen-free (SPF)-embryonated chicken eggs and detected using haemagglutination and the reverse transcriptase-polymerase chain reaction (RT-PCR). The genotype and virulence of field NDV isolates were determined using phylogenetic analysis of fusion (F) protein gene sequences and the mean death time (MDT) test in SPF-embryonated chicken eggs. The questionnaire results revealed that ND caused morbidity (23.1%), mortality (16.3%), case fatality (70.8%), and significant economic losses. Eleven of thirty tissue specimens tested positive for NDV using haemagglutination and RT-PCR. The MDT testing and sequence analysis revealed the presence of virulent NDV classified as genotype VII of class II velogenic pathotype and distinct from locally used vaccine strains (genotype II). The amino acid sequences of the current virulent NDV fusion protein cleavage site motif revealed 112RRQKR↓F117, unlike the locally used avirulent vaccine strains (112GRQGR↓L117). The epidemiological data, MDT results, cleavage site sequence, and phylogenetic analysis all indicated that the present NDV isolates were virulent. The four NDV sequences were deposited in GenBank with accession numbers F gene (PP726912-15) and M gene (PP726916-19). The genetic difference between avirulent vaccine strains and circulating virulent NDV could explain the low level of protection provided by locally used vaccines. Further studies are needed to better understand the circulating NDV genotypes in different production systems.

Dataset Information

Quantitative evaluation of viral interference among Egyptian isolates of highly pathogenic avian influenza viruses (H5N1 and H5N8) with the lentogenic and velogenic Newcastle disease virus genotype VII in specific pathogen-free embryonated chicken eggs model.

Background and aim

Materials and methods

Results

Conclusion

Publications

Quantitative evaluation of viral interference among Egyptian isolates of highly pathogenic avian influenza viruses (H5N1 and H5N8) with the lentogenic and velogenic Newcastle disease virus genotype VII in specific pathogen-free embryonated chicken eggs model.

Similar Datasets

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