Heterogeneity and Seroprevalence of a Newly Identified Avian Hepatitis E Virus from Chickens in the United States

Huang, F. F.; Haqshenas, Gholamreza; Shivaprasad, H. L.; Guenette, D. K.; Woolcock, P. R.; Larsen, C. T.; Pierson, F. W.; Elvinger, François; Toth, Thomas E.; Meng, Xiang‐Jin

doi:10.1128/jcm.40.11.4197-4202.2002

Cited by 130 publications

(187 citation statements)

References 46 publications

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“…For RT-PCR, the degenerate primers helicase F and helicase R were used together with the parameters described by Huang et al (2002). Amplification products (25 ml) were analysed by agarose gel electrophoresis after ethidium bromide staining and visualized under UV light.…”

Section: Methodsmentioning

confidence: 99%

Avian hepatitis E virus infection and possible associated clinical disease in broiler breeder flocks in Hungary

Morrow¹,

Samu

Mátrai

et al. 2008

Avian Pathology

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In broiler breeder flocks in one broiler integration in Hungary, a new syndrome appeared in January 2005 with initially four successive post-peak flocks experiencing significant decreases in egg production. Clinically birds became depressed and there was a small increase in the mortality rate. Postmortem examinations revealed enlarged livers in up to 19% of birds dying, and enlarged spleens in some. Also observed were birds with either clotted blood or serosanguineous fluid in the abdomen and subcapsular haemorrhages of the liver. Histopathology and polymerase chain reaction excluded tumours and the presence of common tumour-associated viruses. Chronic bacterial infections (especially causing hepatitis, peritonitis and airsacculitis) were common but many enlarged livers had no obvious bacterial involvement. After a 9-month period during which a majority of flocks became affected, no newly affected flocks occurred. Investigations showed that all tested affected flocks were seropositive in the big liver and spleen (BLS) Agar Gel Immuno Diffusion (AGID) test. Subsequent flocks without post-peak egg-production drops were shown to be seronegative in the BLS AGID test, as were all the parent flocks contributing to the affected flocks. Liver samples and cloacal swabs were positive by polymerase chain reaction (aHEV helicase target), and calicivirus-like particles were demonstrated in bile samples from affected birds. These observations are similar to hepatitis-splenomegaly syndrome as described in North America and BLS syndrome as described in Australia. Histopathological features were a non-specific chronic hepatitis similar to those described in BLS and hepatitis-splenomegaly syndrome. Immunohistochemistry using a BLS-specific monoclonal antibody confirmed the presence of avian hepatitis E virus antigen in livers and spleen.

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Section: Methodsmentioning

confidence: 99%

Avian hepatitis E virus infection and possible associated clinical disease in broiler breeder flocks in Hungary

Morrow¹,

Samu

Mátrai

et al. 2008

Avian Pathology

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“…Horseradish peroxidase (HRP)-conjugated rabbit anti-chicken immunoglobulin G (IgG; Sigma) or HRP-conjugated goat anti-turkey IgG (Kirkegaard & Perry Laboratories, Inc.) was used as the secondary antibody for chicken and turkey sera, respectively. The cutoff values for these assays were set conservatively at 0.30, as described previously (10,28). Sera from serologically negative SPF chickens and SPF turkeys were used as negative controls, and convalescent-phase sera from SPF chickens experimentally infected with avian HEV were included as positive controls.…”

Section: Virusmentioning

confidence: 99%

“…The PCR primers used for the detection of avian HEV from fecal, serum, and bile samples from the experimentally inoculated SPF chickens and turkeys were based on the ORF1 helicase gene region (8,10,27). Primers AHEV F-1/S (5Ј-GAGCTTGTGAAGGGCGTTGAGG-3Ј) and AHEV R-1/S (5Ј-ACCCAAGATCAACGGCGCTC-3Ј) were used as the external primer set, and primers AHEV F-2/S (5Ј-CGGCAAGTCGTCGTCTGTTGACCAT-3Ј) and AHEV R-2/S (5Ј-CCCCCAGCATAACAACATCGCGC-3Ј) were used as the internal primer set.…”

Section: Virusmentioning

confidence: 99%

“…A truncated recombinant ORF2 capsid protein of avian HEV was expressed in Escherichia coli strain BL21Star (DE3)pLysS (Invitrogen) and purified with a BugBuster His-Bind Purification kit (Novagen) (9,10,28). The purified avian HEV ORF2 protein was used as the antigen to standardize an enzyme-linked immunosorbent assay (ELISA) for the detection of avian HEV antibodies in chickens and turkeys as described previously (10). Briefly, the purified antigen was coated onto 96-well flat-bottom microtiter plates (Thermo Labsystems).…”

Section: Virusmentioning

confidence: 99%

“…Regressive ovaries, red fluid in the abdomen, and an enlarged liver and spleen were often seen in infected chickens with histological changes of hepatic necrosis and hemorrhage (25). Avian HEV has been genetically identified from chickens with HS syndrome as well as from healthy chickens (8,10,28). Avian HEV shares approximately 50 to 60% nucleotide sequence identities with known human and swine HEVs and approximately 80% sequence identity with the Australian chicken big liver and spleen disease virus (8)(9)(10)21).…”

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confidence: 99%

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Generation and Infectivity Titration of an Infectious Stock of Avian Hepatitis E Virus (HEV) in Chickens and Cross-Species Infection of Turkeys with Avian HEV

et al. 2004

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Avian hepatitis E virus (HEV), a novel virus identified from chickens with hepatitis-splenomegaly syndrome in the United States, is genetically and antigenically related to human HEV. In order to further characterize avian HEV, an infectious viral stock with a known infectious titer must be generated, as HEV cannot be propagated in vitro. Bile and feces collected from specific-pathogen-free (SPF) chickens experimentally infected with avian HEV were used to prepare an avian HEV infectious stock as a 10% suspension of positive fecal and bile samples in phosphate-buffered saline. The infectivity titer of this infectious stock was determined by inoculating 1-week-old SPF chickens intravenously with 200 l of each of serial 10-fold dilutions (10 ؊2 to 10 ؊6 ) of the avian HEV stock (two chickens were inoculated with each dilution). All chickens inoculated with the 10 ؊2 to 10 ؊4 dilutions of the infectious stock and one of the two chickens inoculated with the 10 ؊5 dilution, but neither of the chickens inoculated with the 10 ؊6 dilution, became seropositive for anti-avian HEV antibody at 4 weeks postinoculation (wpi). Two serologically negative contact control chickens housed together with chickens inoculated with the 10 ؊2 dilution also seroconverted at 8 wpi. Viremia and shedding of virus in feces were variable in chickens inoculated with the 10 ؊2 to 10 ؊5 dilutions but were not detectable in those inoculated with the 10 ؊6 dilution. The infectivity titer of the infectious avian HEV stock was determined to be 5 ؋ 10 5 50% chicken infectious doses (CID 50 ) per ml. Eight 1-week-old turkeys were intravenously inoculated with 10 5 CID 50 of avian HEV, and another group of nine turkeys were not inoculated and were used as controls. The inoculated turkeys seroconverted at 4 to 8 wpi. In the inoculated turkeys, viremia was detected at 2 to 6 wpi and shedding of virus in feces was detected at 4 to 7 wpi. A serologically negative contact control turkey housed together with the inoculated ones also became infected through direct contact. This is the first demonstration of cross-species infection by avian HEV.Hepatitis E virus (HEV), the causative agent of hepatitis E, is an important human pathogen (1-2, 23-24, 26, 34-35). HEV is a positive-sense, single-stranded, nonenveloped RNA virus. The genome of HEV is about 7.2 kb and contains three open reading frames (ORFs) (23)(24)26). Hepatitis E is primarily transmitted through the fecal-oral route, with an incubation period of about 15 to 60 days. The mortality rate is generally low (about 1%); however, it can reach up to 15 to 25% among infected pregnant women (7,23,24

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Hepatitis E as a Zoonotic Disease

Meng¹

2005

Viral Hepatitis

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Heterogeneity and Seroprevalence of a Newly Identified Avian Hepatitis E Virus from Chickens in the United States

Cited by 130 publications

References 46 publications

Avian hepatitis E virus infection and possible associated clinical disease in broiler breeder flocks in Hungary

Avian hepatitis E virus infection and possible associated clinical disease in broiler breeder flocks in Hungary

Generation and Infectivity Titration of an Infectious Stock of Avian Hepatitis E Virus (HEV) in Chickens and Cross-Species Infection of Turkeys with Avian HEV

Hepatitis E as a Zoonotic Disease

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