Epidemiology and molecular characterisation of duck hepatitis A virus from different duck breeds in Egypt

Erfan, Ahmed M.; Selim, Abdullah; Moursi, Mohamed; Nasef, Soad A.; Mostafa, Ahmed

doi:10.1016/j.vetmic.2015.03.020

Cited by 23 publications

(21 citation statements)

References 18 publications

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“…Peaked mortalities in examined flocks occurred at 2 nd and 3 rd day after showing signs, and its course take 6-7 days as high as (up to 70%). The obtained results were parallel to that previously recorded in DHV infection [6,10]. Liver of examined ducklings were enlarged with distinct petechial and ecchymotic haemorrhages.…”

Section: Discussionsupporting

confidence: 90%

“…The partially magnified region of the 3D gene (467bp) of DHAV of our samples using RT-PCR was detected in 22 samples (68.75%). Similarly, the presence of mentioned band indicates positive DHAV presence [9,10,12]. The RT-PCR confirmed the ECE inoculation results and consequently the RT-PCR could be a rapid and specific detection of the DHAV RNA as described by Liu et al [32] and Wei et al [33].…”

Section: Discussionsupporting

confidence: 54%

“…Enlarged embryonic liver with congestion necrotic patches were observed in inoculated ECE. The previously mentioned embryo lesions were reported as DHV pathological effect by many authors in previous studies [10] and [11].…”

Section: Discussionmentioning

confidence: 86%

“…Although all investigated birds were aroused from vaccinated breeders against DHAV, the positive virus infection was watched in our study in 68.75% of examined progeny and in consequence tremendous economic losses up to 70% were recorded. The failure in protection by maternal derived immunity (MDI) may be attributed to low MDI or misapplication as well as the possibility of vaccination of challenge virus among vaccinal strains [10].…”

Section: Discussionmentioning

confidence: 99%

“…In Egypt, Refaie [8] identified DHAV from Pekin, Balady and Rowan sick ducklings for the first time. Recently, Sultan and Talaat [9]; Erfan et al, [10]; Bayoumie and Abd ELSamie, [11] and EL-Samadony et al, [12] studied molecular characterization and epidemiology of DHAV from different duck breeds in Egypt. The DHAV is a nonenveloped single stranded RNA genome of approximately 7.8 kb [13].…”

Section: Introductionmentioning

confidence: 99%

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Isolation and Molecular Identification of Duck Hepatitis A Virus in Sharkia Governorate

Hassaan

Shahin

Eid

2018

Zagazig Veterinary Journal

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Thirty two flocks were examined to investigate the incrimination of DHAV in the nervous signs and massive mortalities recorded among ducklings during 2014-2016 in Sharkia Governorate. The examined birds aged (2-11 days) were suffering from nervous signs; ataxia followed by opisthotonos, kicking spasmodically then rapid death. Enlarged liver with marked petechial haemorrhages, enlarged spleen, nephritis and swelling of the kidneys were seen. Viral isolation in ECE revealed stunting, subcutaneous hemorrhages covering the body, necrotic areas and greenish discoloration of embryonic liver in 22 samples (68.75%). DHAV identification was succeeded by RT-PCR using RNA extracted from the livers of collected samples and specific primers based on 3D gene in all above positive samples. It could be concluded that RT-PCR could be carried out on hepatic tissues for rapid diagnosis of common DHAV. The subtyping of DHAV is useful for accurate diagnosis to avoid vaccination failure.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionsupporting

confidence: 54%

Section: Discussionmentioning

confidence: 86%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Isolation and Molecular Identification of Duck Hepatitis A Virus in Sharkia Governorate

Hassaan

Shahin

Eid

2018

Zagazig Veterinary Journal

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The 2A2 protein of Duck hepatitis A virus type 1 induces apoptosis in primary cell culture

Cao

Zhu

et al. 2016

Virus Genes

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Duck hepatitis A virus type 1, (DHAV-1) 2A2, is one of the most highly conserved viral proteins within the DHAV serotypes. However, its effect on host cells is unclear. We predicted that DHAV-1 2A2 was a GTPase-like protein based on the results of multiple sequence alignment and homologous modeling analysis. Upon transfection of a recombinant plasmid expressing DHAV-1 2A2, cells displayed fragmented nuclei, chromatin condensation, oligonucleosome-sized DNA ladder, and positive terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling staining; hence, cell death has the characteristics of apoptosis. By staining cells with fluorescein Annexin V-FITC and PI, it is possible to distinguish and quantitatively analyze nonapoptotic cells, early apoptotic cells, late apoptotic/necrotic cells, and dead cells through flow cytometry and fluorescence microscopy. The percentage of apoptotic cells gradually increased and reached a maximum after 48 h of transfection. In conclusion, apoptosis induced by this GTPase-like protein may contribute to DHAV-1 pathogenesis.

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The 3D protein of duck hepatitis A virus type 1 binds to a viral genomic 3′ UTR and shows RNA-dependent RNA polymerase activity

et al. 2017

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To explore the RNA-dependent RNA polymerase (RdRP) function of the 3D protein of duck hepatitis A virus type 1 (DHAV-1), the gene was cloned into the pET-32a(+) vector for prokaryotic expression. The 3' untranslated region (3' UTR) of DHAV-1 together with a T7 promoter was cloned into the pMD19-T vector for in vitro transcription of 3' UTR RNA, which was further used as a template in RNA-dependent RNA polymerization. In this study, three methods were applied to analyze the RdRP function of the 3D protein: (1) ammonium molybdate spectrophotometry to detect pyrophosphate produced during polymerization; (2) quantitative reverse transcription PCR (RT-qPCR) to investigate the changes in RNA quantity during polymerization; and (3) electrophoresis mobility shift assay to examine the interaction between the 3D protein and 3' UTR. The results showed the 3D protein was successfully expressed in bacteria culture supernatant in a soluble form, which could be purified by affinity chromatography. In 3D enzymatic activity assays, pyrophosphate and RNA were produced, the amounts of which increased based on approximative kinetics, and binding of the 3D protein to the 3' UTR was observed. These results indicate that prokaryotically expressed soluble DHAV-13D protein can bind to a viral genomic 3' UTR and exhibit RdRP activity.

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Epidemiology and molecular characterisation of duck hepatitis A virus from different duck breeds in Egypt

Cited by 23 publications

References 18 publications

Isolation and Molecular Identification of Duck Hepatitis A Virus in Sharkia Governorate

Isolation and Molecular Identification of Duck Hepatitis A Virus in Sharkia Governorate

The 2A2 protein of Duck hepatitis A virus type 1 induces apoptosis in primary cell culture

The 3D protein of duck hepatitis A virus type 1 binds to a viral genomic 3′ UTR and shows RNA-dependent RNA polymerase activity

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