scholarly journals Cell Culture Isolation and Whole Genome Characterization of Hepatitis E Virus Strains from Wild Boars in Germany

2021 ◽  
Vol 9 (11) ◽  
pp. 2302
Author(s):  
Katja Schilling-Loeffler ◽  
Oliver Viera-Segura ◽  
Victor Max Corman ◽  
Julia Schneider ◽  
Ashish K. Gadicherla ◽  
...  
Keyword(s):  
Cell Culture ◽  
Wild Boar ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Enriched Library ◽  
Genome Copy ◽  
Genotype 3 ◽  
Wild Boars ◽  
Copy Numbers ◽  
Cell Culture Isolation

Infection with hepatitis E virus (HEV) can cause acute and chronic hepatitis in humans. The HEV genotype 3 can be zoonotically transmitted from animals to humans, with wild boars representing an important reservoir species. Cell culture isolation of HEV is generally difficult and mainly described for human isolates so far. Here, five sera and five liver samples from HEV-RNA-positive wild boar samples were inoculated onto PLC/PRF/5 cells, incubated for 3 months and thereafter passaged for additional 6 weeks. As demonstrated by RT-qPCR, immunofluorescence and immune electron microscopy, virus was successfully isolated from two liver samples, which originally contained high HEV genome copy numbers. Both isolates showed slower growth than the culture-adapted HEV strain 47832c. In contrast to this strain, the isolated strains had no insertions in their hypervariable genome region. Next generation sequencing using an HEV sequence-enriched library enabled full genome sequencing. Strain Wb108/17 belongs to subtype 3f and strain Wb257/17 to a tentative novel subtype recently described in Italian wild boars. The results indicate that HEV can be successfully isolated in cell culture from wild boar samples containing high HEV genome copy numbers. The isolates may be used further to study the zoonotic potential of wild boar-derived HEV subtypes.

scholarly journals Geographical restriction of Hepatitis E virus circulation in wild boars (Sus scrofa) in Emilia-Romagna region, Northern Italy

2020 ◽  
Vol 9 (1) ◽  
Author(s):  
Silvia Bonardi ◽  
Virginia Filipello ◽  
Enrico Pavoni ◽  
Valentina Carta ◽  
Luca Bolzoni ◽  
...  
Keyword(s):  
Wild Boar ◽  
Cervus Elaphus ◽  
Hepatitis E Virus ◽  
Red Deer ◽  
Hepatitis E ◽  
Northern Italy ◽  
Genotype 3 ◽  
Hunting Season ◽  
Wild Boars ◽  
Emilia Romagna Region

Hepatitis E virus (HEV) is a singlestrand RNA virus that causes an acute viral hepatitis in humans. Among its eight recognized genotypes, HEV-3 and HEV-4 are zoonotic, infecting humans, pigs and wild boars. Recently, HEV-3 has been also detected in red deer, which represents another reservoir of HEV. Consumption of raw pork products (mainly liver sausages), undercooked wild boar meat, raw wild boar liver and deer meat has been responsible for foodborne HEV human worldwide. From November 2018 to March 2019, liver samples collected from 97 wild boars hunted in Emilia-Romagna region (Northern Italy) were tested for HEV RNA. The hunting area included two territories for an extension of 33 km2, named A (about 13 km2, natural park, deciduous wood) and B (about 20 km2, cultivated fields in proximity of a river) areas. Distance between the two areas ranged between 8 to 10 km. A total of 73 wild boars were hunted in area A, and 24 in area B. HEV RNA was detected by Realtime RT–PCR in 23/73 liver samples of wild boars living in area A only (31.5% - 95% CI: 22.0-42.8%). The HEV sequences (n=13) clustered within genotype 3. The majority of positives belonged to animals < 12 months (12/25; 48%), followed by subadults (13-24 months) (7/16; 43.8%) and adults (4/32; 12.5%). This difference was found to be statistically significant (p=0.0024). In absence of pig farms, the restriction of HEV-positive animals to a well-defined territory of 13 km2 (Boschi di Carrega Regional Park) could hypothetically be related to the presence of red deer (Cervus elaphus), which lived in area A at the beginning of the hunting season. Further studies are needed to confirm or deny our hypothesis.

DETECTION AND PHYLOGENETIC CHARACTERIZATION OF HEPATITIS E VIRUS GENOTYPE 3 IN A CAPTIVE WILD BOAR IN THAILAND

2012 ◽  
Vol 43 (3) ◽  
pp. 640-644 ◽  
Author(s):  
Anuwat Wiratsudakul ◽  
Ladawan Sariya ◽  
Phirom Prompiram ◽  
Siriporn Tantawet ◽  
Duangkhamol Suraruangchai ◽  
...  
Keyword(s):  
Wild Boar ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Genotype 3 ◽  
Virus Genotype ◽  
Phylogenetic Characterization

scholarly journals Robust hepatitis E virus infection and transcriptional response in human hepatocytes

2020 ◽  
Vol 117 (3) ◽  
pp. 1731-1741 ◽  
Author(s):  
Daniel Todt ◽  
Martina Friesland ◽  
Nora Moeller ◽  
Dimas Praditya ◽  
Volker Kinast ◽  
...  
Keyword(s):  
Cell Culture ◽  
Hepatitis E Virus ◽  
Hepatoma Cell ◽  
Hepatitis E ◽  
Human Hepatocytes ◽  
Cell Culture Model ◽  
Genotype 3 ◽  
Human Hepatoma Cell ◽  
Culture Model ◽  
Viral Titers

Hepatitis E virus (HEV) is the causative agent of hepatitis E in humans and the leading cause for acute viral hepatitis worldwide. The virus is classified as a member of the genus Orthohepevirus A within the Hepeviridae family. Due to the absence of a robust cell culture model for HEV infection, the analysis of the viral life cycle, the development of effective antivirals and a vaccine is severely limited. In this study, we established a protocol based on the HEV genotype 3 p6 (Kernow C-1) and the human hepatoma cell lines HepG2 and HepG2/C3A with different media conditions to produce intracellular HEV cell culture-derived particles (HEVcc) with viral titers between 105 and 106 FFU/mL. Viral titers could be further enhanced by an HEV variant harboring a mutation in the RNA-dependent RNA polymerase. These HEVcc particles were characterized in density gradients and allowed the trans-complementation of subgenomic reporter HEV replicons. In addition, in vitro produced intracellular-derived particles were infectious in liver-humanized mice with high RNA copy numbers detectable in serum and feces. Efficient infection of primary human and swine hepatocytes using the developed protocol could be observed and was inhibited by ribavirin. Finally, RNA sequencing studies of HEV-infected primary human hepatocytes demonstrated a temporally structured transcriptional defense response. In conclusion, this robust cell culture model of HEV infection provides a powerful tool for studying viral–host interactions that should facilitate the discovery of antiviral drugs for this important zoonotic pathogen.

scholarly journals Wild Boar as a Sylvatic Reservoir of Hepatitis E Virus in Poland: A Cross-Sectional Population Study

Viruses ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 1113
Author(s):  
Iwona Kozyra ◽  
Artur Jabłoński ◽  
Ewelina Bigoraj ◽  
Artur Rzeżutka
Keyword(s):  
Wild Boar ◽  
Regional Differences ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Igg Antibodies ◽  
Cross Sectional ◽  
Wild Boars ◽  
Wildlife Species ◽  
Faecal Samples ◽  
Molecular Assessment

The most important wildlife species in the epidemiology of hepatitis E virus (HEV) infections are wild boars, which are also the main reservoir of the virus in a sylvatic environment. The aim of the study was a serological and molecular assessment of the prevalence of HEV infections in wild boars in Poland. In total, 470 pairs of samples (wild boar blood and livers) and 433 samples of faeces were tested. An ELISA (ID.vet, France) was used for serological analysis. For the detection of HEV RNA, real-time (RT)-qPCR was employed. The presence of specific anti-HEV IgG antibodies was found in 232 (49.4%; 95%CI: 44.7–54%) sera, with regional differences observed in the seroprevalence of infections. HEV RNA was detected in 57 (12.1%, 95%CI: 9.3–15.4%) livers and in 27 (6.2%, 95%CI: 4.1–8.9%) faecal samples, with the viral load ranging from 1.4 to 1.7 × 1011 G.C./g and 38 to 9.3 × 107 G.C./mL, respectively. A correlation between serological and molecular results of testing of wild boars infected with HEV was shown. HEV infections in wild boars appeared to be common in Poland.

Susceptibility of Pigs to Zoonotic Hepatitis E Virus Genotype 3 Isolated from a Wild Boar

2016 ◽  
Vol 64 (5) ◽  
pp. 1589-1597 ◽  
Author(s):  
D. Thiry ◽  
N. Rose ◽  
A. Mauroy ◽  
F. Paboeuf ◽  
L. Dams ◽  
...  
Keyword(s):  
Wild Boar ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Genotype 3 ◽  
Virus Genotype

Chronically infected wild boar can transmit genotype 3 hepatitis E virus to domestic pigs

2015 ◽  
Vol 180 (1-2) ◽  
pp. 15-21 ◽  
Author(s):  
Josephine Schlosser ◽  
Ariel Vina-Rodriguez ◽  
Christine Fast ◽  
Martin H. Groschup ◽  
Martin Eiden
Keyword(s):  
Wild Boar ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Genotype 3 ◽  
Domestic Pigs

scholarly journals Wild boars as sources for infectious diseases in livestock and humans

2009 ◽  
Vol 364 (1530) ◽  
pp. 2697-2707 ◽  
Author(s):  
X. J. Meng ◽  
D. S. Lindsay ◽  
N. Sriranganathan
Keyword(s):  
Infectious Diseases ◽  
Wild Boar ◽  
Hepatitis E Virus ◽  
Classical Swine Fever ◽  
Hepatitis E ◽  
Domestic Animals ◽  
Wild Boars ◽  
Suburban Areas ◽  
Wild Boar Meat ◽  
Human Habitation

Wild boars ( Sus scrofa ) are indigenous in many countries in the world. These free-living swine are known reservoirs for a number of viruses, bacteria and parasites that are transmissible to domestic animals and humans. Changes of human habitation to suburban areas, increased use of lands for agricultural purposes, increased hunting activities and consumption of wild boar meat have increased the chances of exposure of wild boars to domestic animals and humans. Wild boars can act as reservoirs for many important infectious diseases in domestic animals, such as classical swine fever, brucellosis and trichinellosis, and in humans, diseases such as hepatitis E, tuberculosis, leptospirosis and trichinellosis. For examples, wild boars are reservoirs for hepatitis E virus, and cluster cases of hepatitis E have been reported in Japan of humans who consumed wild boar meat. In Canada, an outbreak of trichinellosis was linked to the consumption of wild boar meat. The incidence of tuberculosis owing to Mycobacterium bovis has increased in wild boars, thus posing a potential concern for infections in livestock and humans. It has also been documented that six hunters contracted Brucella suis infections from wild swine in Florida. This article discusses the prevalence and risk of infectious agents in wild boars and their potential transmission to livestock and humans.

scholarly journals Construction of an infectious cDNA clone of hepatitis E virus strain JE03-1760F that can propagate efficiently in cultured cells

2009 ◽  
Vol 90 (2) ◽  
pp. 457-462 ◽  
Author(s):  
Kentaro Yamada ◽  
Masaharu Takahashi ◽  
Yu Hoshino ◽  
Hideyuki Takahashi ◽  
Koji Ichiyama ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cdna Clone ◽  
Hepatitis E Virus ◽  
Culture Supernatant ◽  
Cultured Cells ◽  
A549 Cells ◽  
Hepatitis E ◽  
Infectious Cdna Clone ◽  
Post Inoculation ◽  
Genotype 3

A full-length infectious cDNA clone (pJE03-1760F/wt) of a genotype 3 hepatitis E virus (HEV) (strain JE03-1760F) obtained from a faecal specimen was constructed in this study. Upon transfection of the capped in vitro transcripts of pJE03-1760F/wt into PLC/PRF/5 cells, the viral RNA levels in the culture supernatant started to increase on day 6 post-transfection (p.t.) and reached 107 copies ml−1 on day 28 p.t. Detection of increasing numbers of cells with ORF2 protein expression by immunofluorescence assay at 5, 7, 11 and 15 days p.t. indicated the spread of HEV infection in cell culture. When the cDNA-derived virus in culture supernatant was inoculated into PLC/PRF/5 or A549 cells, it grew as efficiently as the faeces-derived virus in both cells, reaching 106 copies ml−1 at 30 days post-inoculation. Our reverse genetics system for HEV that is usable in a robust cell-culture system will be useful for elucidation of the mechanism of HEV replication and functional roles of HEV proteins.

scholarly journals Analysis of the full-length genome of hepatitis E virus isolates obtained from wild boars in Japan

2005 ◽  
Vol 86 (12) ◽  
pp. 3321-3326 ◽  
Author(s):  
Tsutomu Nishizawa ◽  
Masaharu Takahashi ◽  
Kazunori Endo ◽  
Shinji Fujiwara ◽  
Nobuo Sakuma ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Full Length ◽  
Partial Sequence ◽  
Genotype 3 ◽  
Entire Genome ◽  
Wild Boars ◽  
Full Length Sequence ◽  
Full Length Genome ◽  
Virus Isolates

Two (2·3 %) of 87 wild-caught boars in Japan had detectable hepatitis E virus (HEV) RNA. The two boar HEV isolates (wbJTS1 and wbJYG1) obtained in the present study and a previously reported isolate (wbJSG1) whose partial sequence had been determined were sequenced over the entire genome. The wbJSG1, wbJTS1 and wbJYG1 isolates comprised 7225 or 7226 nt, excluding the poly(A) tail, and segregated into genotype 3. They differed by 8·5–11·2 % from each other and by 8·6–18·4 % from 17 reported genotype 3 HEV isolates, including one boar isolate, in the full-length sequence. When compared with 191 reported genotype 3 HEV isolates whose partial sequences were known, these three boar isolates were closer to Japanese isolates than to isolates of non-Japanese origin (89·2±2·6 vs 85·9±2·2 %; P<0·0001). A proportion of wild boars in Japan are infected with markedly heterogeneous HEV strains that are indigenous to Japan and may serve as reservoirs of HEV.

Sign in / Sign up

Export Citation Format

Share Document