Characterization of a hypervirulent fowl adenovirus 4 with the novel genotype newly prevalent in China and establishment of reproduction infection model of hydropericardium syndrome in chickens

AbstractFowl adenovirus serotype 4 (FAdV-4), the causative agent of hepatitis-hydropericardium syndrome (HHS), distributed widely in the poultry farms in China. Hexon is one of the major capsid proteins associated with viral species or serotypes. However, the epitopes of Hexon protein remain largely unknown. In this study, a monoclonal antibody (mAb) specific to Hexon protein of FAdV-4, designated as 3G8, was generated. Subsequently, the linear peptide recognized by 3G8 was mapped and identified as 213AYGAYVK219 using a series of overlapping peptides generated from Hexon protein. Amino acid sequence analysis revealed that the epitope recognized by 3G8 was highly conserved across all the FAdVs. The epitope was immunogenic and could be recognized by FAdV-4 positive chicken serum samples. These findings will enrich our knowledge regarding the epitope on Hexon and provide valuable information for further characterization of the antigenicity of Hexon protein.

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Development of a Novel Avian Vaccine Vector Derived From the Emerging Fowl Adenovirus 4

Frontiers in Microbiology ◽

10.3389/fmicb.2021.780978 ◽

2021 ◽

Vol 12 ◽

Author(s):

Qing Pan ◽

Yu Zhang ◽

Aijing Liu ◽

Hongyu Cui ◽

Yulong Gao ◽

...

Keyword(s):

Fluorescent Protein ◽

Virus Genome ◽

Open Reading Frames ◽

Vaccine Vector ◽

Economic Losses ◽

Foreign Gene ◽

The Novel ◽

Hydropericardium Syndrome ◽

Fowl Adenovirus ◽

Green Fluorescent

Severe hepatitis-hydropericardium syndrome (HHS) associated with a novel viral genotype, fowl adenovirus 4 (FAdV-4), has emerged and widely spread in China since 2015, causing severe economic losses to the poultry industry. We previously reported that the hexon gene is responsible for pathogenicity and obtained a non-pathogenic hexon-replacement rHN20 strain; however, the lack of information about the non-essential regions for virus replication limits the development of a FAdV-4 vector. This study first established an enhanced green fluorescent protein (EGFP)-indicator virus based on the FAdV-4 reverse genetic technique, effective for batch operations in the virus genome. Based on this, 10 open reading frames (ORFs) at the left end and 13 ORFs at the right end of the novel FAdV-4 genome were deleted separately and identified as non-essential genes for viral replication, providing preliminary insertion sites for foreign genes. To further improve its feasibility as a vaccine vector, seven combinations of ORFs were successfully replaced with EGFP without affecting the immunogenicity of the vector backbone. Finally, a recombinant rHN20-vvIBDV-VP2 strain, expressing the VP2 protein of very virulent infectious bursa disease virus (vvIBDV), was rescued and showed complete protection against FAdV-4 and vvIBDV. Thus, the novel FAdV-4 vector could provide sufficient protection for HHS and efficient exogenous gene delivery. Overall, our findings systemically identified 23 non-essential ORFs for FAdV-4 replication and seven foreign gene insertion regions, providing valuable information for an in-depth understanding of the novel FAdV-4 pathogenesis and development of multivalent vaccines.

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First detection and genotyping of Enterocytozoon bieneusi in pet fancy rats (Rattus norvegicus) and guinea pigs (Cavia porcellus) in China

Parasite ◽

10.1051/parasite/2020019 ◽

2020 ◽

Vol 27 ◽

pp. 21 ◽

Cited By ~ 1

Author(s):

Jingsong Wang ◽

Chaochao Lv ◽

Diandian Zhao ◽

Runan Zhu ◽

Chen Li ◽

...

Keyword(s):

Rattus Norvegicus ◽

Guinea Pigs ◽

Enterocytozoon Bieneusi ◽

Its Sequence ◽

The Novel ◽

Microsporidian Parasite ◽

Obligate Intracellular ◽

Reference Sequences ◽

Novel Genotype

Enterocytozoon bieneusi, an obligate intracellular microsporidian parasite, can infect humans and a wide variety of animals worldwide. However, information on the prevalence and molecular characterization of E. bieneusi in pet rats and guinea pigs is lacking. In this study, 325 fecal samples were collected from 152 pet fancy rats and 173 pet guinea pigs purchased from pet shops in Henan and Shandong provinces. The prevalence of E. bieneusi was 11.2% (17/152) in pet fancy rats and 20.2% (35/173) in pet guinea pigs. Genotypes D (n = 12), Peru11 (n = 3), S7 (n = 1) and SCC-2 (n = 1) were identified in pet fancy rats, and genotype S7 (n = 30) and a novel genotype PGP (n = 5) were identified in pet guinea pigs. The ITS sequence and its phylogenetic analysis showed that the novel genotype PGP was distinctly different; it exhibited less than 50% similarity to the reference sequences, and did not cluster with any of the known E. bieneusi genotype groups, forming a unique branch between groups 6 and 7. These data suggest that this is a new E. bieneusi genotype group. This is the first report of E. bieneusi infection in pet fancy rats and pet guinea pigs worldwide. The identification of zoonotic genotypes D, Peru11, and S7 suggests that pet fancy rats and guinea pigs can be potential sources of human microsporidiosis.

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Molecular cloning, expression and characterization of 100K gene of fowl adenovirus-4 for prevention and control of hydropericardium syndrome

Biologicals ◽

10.1016/j.biologicals.2015.10.002 ◽

2016 ◽

Vol 44 (1) ◽

pp. 19-23 ◽

Cited By ~ 10

Author(s):

M.S. Shah ◽

A. Ashraf ◽

M.I. Khan ◽

M. Rahman ◽

M. Habib ◽

...

Keyword(s):

Molecular Cloning ◽

Prevention And Control ◽

Hydropericardium Syndrome ◽

Fowl Adenovirus ◽

And Control

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Characterization of the novel genotype avian hepatitis E viruses from outbreaks of hepatic rupture haemorrhage syndrome in different geographical regions of China

Transboundary and Emerging Diseases ◽

10.1111/tbed.12987 ◽

2018 ◽

Vol 65 (6) ◽

pp. 2017-2026 ◽

Cited By ~ 6

Author(s):

Qi Su ◽

Yang Li ◽

Yawen Zhang ◽

Zhihui Zhang ◽

Fanfeng Meng ◽

...

Keyword(s):

Hepatitis E ◽

The Novel ◽

Hepatic Rupture ◽

Geographical Regions ◽

Novel Genotype

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Identification of novel B cell epitopes in the fiber protein of serotype 8 Fowl adenovirus

AMB Express ◽

10.1186/s13568-019-0895-1 ◽

2019 ◽

Vol 9 (1) ◽

Author(s):

Hao Lu ◽

Hongxia Shao ◽

Hongjun Chen ◽

Jianjun Zhang ◽

Weikang Wang ◽

...

Keyword(s):

B Cell ◽

Cell Epitope ◽

Economic Losses ◽

The Novel ◽

B Cell Epitopes ◽

Hydropericardium Syndrome ◽

Fowl Adenovirus ◽

Fiber Protein ◽

Inclusion Body Hepatitis ◽

B Cell Epitope

Abstract In recent years, hepatitis-hydropericardium syndrome (HHS) and inclusion body hepatitis (IBH) caused by fowl adenovirus (FAdV) infection have resulted in significant economic losses to the poultry industry worldwide. Epidemiological analysis revealed that serotype FAdV-8 is one of the major pathogenic FAdVs currently prevalent in domestic flocks. Although the fiber protein of FAdV plays vital roles in viral infection and pathogenesis, the B cell epitope in the fiber protein is less known. In this study, two monoclonal antibodies (mAbs) specific to fiber protein of FAdV-8, designated as 4D9 and 5F10, were prepared. Although the mAb 4D9 and 5F10 could not neutralize FAdV-8 infection, 4D9 and 5F10 showed good activities of indirect immunofluorescence, western blot and immunoprecipitation. Epitope analysis revealed that mAb 5F10 recognized 187–219aa in the fiber whereas mAb 4D9 recognized 113–149aa in the fiber. Sequence analysis showed that the epitope recognized by mAb 5F10 was conserve across serotypes FAdV-7, 8a and 8b whereas that for mAb 4D9 was only conserve in FAdV-8b. The generation of mAbs specific to fiber of FAdV-8 and the identification of the novel B cell epitopes here lay the foundation for further studying the antigenicity of the fiber and developing specific diagnosis for FAdV-8.

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Isolation and characterization of fowl adenovirus serotype 4 from chickens with hydropericardium syndrome in Korea

Korean Journal of Veterinary Research ◽

10.14405/kjvr.2011.51.3.209 ◽

2011 ◽

Vol 51 (3) ◽

pp. 209-216

Author(s):

Hong-Su Park ◽

Il-Soo Lim ◽

Sang-Kyu Kim ◽

Toh-Kyung Kim ◽

Sang-Geon Yeo

Keyword(s):

Adenovirus Serotype ◽

Hydropericardium Syndrome ◽

Isolation And Characterization ◽

Fowl Adenovirus ◽

Fowl Adenovirus Serotype 4

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The Natural Large Genomic Deletion Is Unrelated to the Increased Virulence of the Novel Genotype Fowl Adenovirus 4 Recently Emerged in China

Viruses ◽

10.3390/v10090494 ◽

2018 ◽

Vol 10 (9) ◽

pp. 494 ◽

Cited By ~ 5

Author(s):

Qing Pan ◽

Jing Wang ◽

Yulong Gao ◽

Hongyu Cui ◽

Changjun Liu ◽

...

Keyword(s):

The Novel ◽

Highly Pathogenic ◽

Reading Frame ◽

Fowl Adenovirus ◽

Nonpathogenic Strain ◽

High Virulence ◽

Specific Pathogen ◽

Novel Genotype ◽

First Time ◽

The Relationship

Since 2015, severe hydropericardium-hepatitis syndrome (HHS), caused by a highly pathogenic fowl adenovirus 4 (FAdV-4), emerged in China. In our previous study, the FAdV-4 has been identified as a novel genotype with a unique 1966-bp nucleotide deletion (1966Del) between open reading frame 42 and 43. In this study, the natural 1966Del was frequently identified among 17 clinical isolates and other reported Chinese clinical strains. To investigate the relationship between 1966Del and the increased virulence of the novel FAdV-4, a CRISPR/Cas9 operating platform for FAdV-4 was developed for the first time in this study. Based on this platform, a Re1966 strain was rescued, inserted the relative 1966Del sequence of a nonpathogenic strain KR5. In the pathogenicity study, the Re1966 strain retained high virulence for specific-pathogen-free chickens, similar to the parental wild-type HLJFAd15, although the survival time of chickens infected with Re1966 was much longer. Therefore, the natural 1966Del was identified as a non-essential site for the increased virulence of the emerged novel FAdV-4. Although further research on the virulence-determining region or point within the genome of the novel FAdV-4 is needed, the CRISPR/Cas9 operating platform for the novel FAdV-4 was developed and successfully applied to edit the genomic DNA for the first time, and it provides a novel powerful tool for both basic virology studies and vaccine vector development of FAdVs.

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