scholarly journals THE SEROPREVALENCE OF HEPATITIS E VIRUS INFECTION IN WILD BOARS IN SERBIA

2011 ◽  
Vol 4 (1) ◽  
pp. 19-29
Author(s):  
Diana Lupulović ◽  
Tamaš Petrović ◽  
Sava Lazić ◽  
Jasna Prodanov Radulović ◽  
Radoslav Došen ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Oral Route ◽  
Genotype 3 ◽  
Species Barrier ◽  
Serum Samples ◽  
Serological Tests ◽  
Wild Boars ◽  
Infected People ◽  
Contaminated Food

Hepatitis E (HEV) belongs to one of fi ve so far described types of viral hepatitis caused by human agents (hepatitis A, B, C, D and E). The disease is characterized by clinical and epidemiological signs of acute hepatitis and is transmitted primarily by fecal-oral route via contaminated food and water. The infection is mainly detected in the developing countries of the Middle East, Asia and Africa, especially in countries with poor sanitary conditions of life. HEV infection in pigs was fi rst recorded in 1990. Numerous studies that followed proved that HEV can skip species barrier and can be transmitted from pigs to humans. HEV has been demonstrated as a new zoonotic agent. Hepatitis E virus has infected people in Japan who consumed insuffi ciently cooked meat of deer, pig liver and meat of wild boar. In humans four genotypes have been determined: I, II, III and IV, while so far tested swine isolates belong to genotypes III and IV. It is also important to note that the swine HEV isolates from one geographical region are genetically closer to human isolates from the same area than to the other isolates from pigs in the world. Th e aim of this paper is to show whether and how much HEV infection is present in the population of wild boars in Serbia, which represent a reservoir of this disease caused by a signifi cant zoonotic agent. Preliminary serological tests included the examination of 92 blood serum samples of wild boars. In 32 animals, or 34.78%, the presence of specifi c antibodies against HEV genotype was detected. Th e blood samples were collected during 2009, 2010 and 2011 from 15 hunting sites in Serbia. Laboratory testing was performed by non-commercial ELISA (in-house ELISA), where the used antigen was recombinant capsid protein-HEV genotype 3 ΔORF 2, which was obtained by laboratory cloning procedure. The test results showed that the hepatitis E virus is present in wild boars in Vojvodina, that are a potential source of this infection, as well as for many other infections of diferrent etiology.

scholarly journals Liver Transudate, a Potential Alternative to Detect Anti-Hepatitis E Virus Antibodies in Pigs and Wild Boars (Sus scrofa)

2020 ◽  
Vol 8 (3) ◽  
pp. 450
Author(s):  
Alejandro Navarro ◽  
Carmen Bárcena ◽  
Pilar Pozo ◽  
Alberto Díez-Guerrier ◽  
Irene Martínez ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Intraclass Correlation ◽  
Kappa Statistic ◽  
Hepatitis E ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Wild Boars ◽  
Pork Products ◽  
Contaminated Food ◽  
Potential Alternative

In recent years, cases of hepatitis E virus (HEV) infection have increased in Europe in association with the consumption of contaminated food, mainly from pork products but also from wild boars. The animal’s serum is usually tested for the presence of anti-HEV antibodies and viral RNA but, in many cases such as during hunting, an adequate serum sample cannot be obtained. In the present study, liver transudate was evaluated as an alternative matrix to serum for HEV detection. A total of 125 sera and liver transudates were tested by enzyme-linked immunosorbent assay at different dilutions (1:2, 1:10, 1:20), while 58 samples of serum and liver transudate were checked for the presence of HEV RNA by RT-qPCR. Anti- HEV antibodies were detected by ELISA in 68.0% of the serum samples, and in 61.6% of the undiluted transudate, and in 70.4%, 56.8%, and 44.8% of 1:2, 1:10, or 1:20 diluted transudate, respectively. The best results were obtained for the liver transudate at 1:10 dilution, based on the Kappa statistic (0.630) and intraclass correlation coefficient (0.841). HEV RNA was detected by RT-qPCR in 22.4% of the serum samples and 6.9% of the transudate samples, all samples used for RT-qPCR were positive by ELISA. Our results indicate that liver transudate may be an alternative matrix to serum for the detection of anti-HEV antibodies.

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 Development of a novel immunoperoxidase monolayer assay for detection of swine Hepatitis E virus antibodies based on stable cell lines expressing the ORF3 protein

2014 ◽  
Vol 62 (2) ◽  
pp. 243-256 ◽  
Author(s):  
Huanbin Liang ◽  
Heng Wang ◽  
Liangquan Zhang ◽  
Honglang Gu ◽  
Guihong Zhang
Keyword(s):  
Cell Line ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Nucleotide Sequencing ◽  
Serum Samples ◽  
Orf3 Protein ◽  
Porcine Serum ◽  
Contaminated Food ◽  
Swine Hepatitis ◽  
Novel Method

Hepatitis E virus (HEV) strains are classified into 4 genotypes by nucleotide sequencing. Genotypes 3 and 4 infect humans and animals via HEV-contaminated food or water. HEV RNA was detected by PCR and antibodies were detected by ELISA. Since human studies showed that HEV IgG antibodies in sera can persist for extended periods, diagnosis of HEV infection in swine or humans is mainly based on serological detection using commercial ELISA kits. However, there is no supplemental method to verify ELISA results. Hence, we developed a novel method used for mutual correction of these common processes. Here, a modified stable HepG2 cell line was transfected with pcDNA3.1-ORF3 to express the swine HEV ORF3 protein. Based on this cell line, a novel immunoperoxidase monolayer assay (IPMA) was developed to detect antibodies against HEV. The results show that this method has good specificity, sensitivity and repeatability. When used to investigate 141 porcine serum samples, the IPMA had a coincidence rate of 92.2% with a commercial ELISA kit. The established IPMA described herein is valuable as a supplemental method to ELISA and can differentiate infections by HEV and other viruses.

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.

scholarly journals Hepatitis E virus in Italy: molecular analysis of travel-related and autochthonous cases

2011 ◽  
Vol 92 (7) ◽  
pp. 1617-1626 ◽  
Author(s):  
Giuseppina La Rosa ◽  
Michele Muscillo ◽  
Valentina Spuri Vennarucci ◽  
Anna Rosa Garbuglia ◽  
Patrizia La Scala ◽  
...  
Keyword(s):  
Acute Hepatitis ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
The Body ◽  
Industrialized Countries ◽  
Genotype 3 ◽  
Serum Samples ◽  
Specific Primers ◽  
Body Of Knowledge ◽  
Acute Hepatitis E

Human hepatitis E virus (HEV) is considered an emerging pathogen in industrialized countries. The aim of the present study was to contribute to the body of knowledge available on the molecular epidemiology of acute hepatitis E in Italy. Three sets of HEV-specific primers targeting the ORF1 and ORF2 were used to examine serum samples collected from acute hepatitis patients positive for anti-HEV IgG and/or IgM, between 2007 and 2010. Seventeen patients (39.5 %) tested HEV RNA-positive: 12 infections, due to genotype 1, were associated with travel to endemic areas (Bangladesh, India and Pakistan), while five infections, due to genotype 3, were presumably autochthonous. Risk factors identified in this group included exposure to raw seafood, pork liver sausages and wild boar. Results from the present study confirm that human HEV infection in Italy is caused by different genotypes, depending on whether the infection is travel-related or autochthonous.

Prevalence of hepatitis E virus in slaughter-age pigs in Scotland

2014 ◽  
Vol 143 (10) ◽  
pp. 2237-2240 ◽  
Author(s):  
C. CROSSAN ◽  
S. GRIERSON ◽  
J. THOMSON ◽  
A. WARD ◽  
J. NUNEZ-GARCIA ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Partial Sequence ◽  
Genotype 3 ◽  
Serum Samples ◽  
Specific Antibodies ◽  
First Report ◽  
Slaughter Age ◽  
The Uk

SUMMARYThe prevalence of anti-HEV isotype-specific antibodies and viraemia were investigated in serum samples collected from slaughter-age pigs (aged 22–24 weeks) from 23 farms in Scotland. Of 176 serum samples tested, 29·0% (n = 51) were anti-HEV IgG positive, 36·9% (n = 65) anti-HEV IgA positive and 29·0% (n = 51) anti-HEV IgM positive. Overall seroprevalence (anti-HEV IgG+ and/or IgA+ and/or IgM+) was 61·4% (n = 108). HEV RNA was detected in 72/162 serum samples (44·4%). Partial sequence of ORF2 (98 nt) was obtained from eight HEV RNA-positive samples and phylogenetic analysis confirmed that they were all of genotype 3. This is the first report on the prevalence of HEV in pigs in Scotland. Given the increasing incidence of locally acquired HEV infection in the UK, evidence that HEV is a foodborne zoonosis emphasizes the need for surveillance in pigs.

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.

scholarly journals Cross-Species Transmission of Swine Hepatitis E Virus Genotype 3 to Rabbits

Viruses ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 53 ◽  
Author(s):  
Sang-Hoon Han ◽  
Byung-Joo Park ◽  
Hee-Seop Ahn ◽  
Yong-Hyun Kim ◽  
Hyeon-Jeong Go ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Rna Virus ◽  
Hepatitis E ◽  
Genotype 3 ◽  
Serum Samples ◽  
Virus Genotype ◽  
Specific Pathogen ◽  
Swine Hepatitis ◽  
Negative Controls ◽  
Liver Tissues

Hepatitis E virus (HEV) is a quasi-enveloped, positive-sense single stranded RNA virus. HEV continually expands the host ranges across animal species. In this study, the possibility of cross-species infection with swine HEV-3 was investigated using rabbits. A total of fourteen 8-week old, specific pathogen-free rabbits were divided into three experimental groups. Four rabbits were used as negative controls, four rabbits were infected with rabbit HEV as positive controls, and six rabbits were inoculated with swine HEV-3. HEV RNA were detected from serum and fecal samples after viral challenge. The levels of anti-HEV antibodies, pro-inflammatory cytokines (IL-1, IL-6, TNF-α and IFN-α), and liver enzymes (alanine and aspartate aminotransferases) were determined in serum samples. Histopathological lesions were examined in liver tissues. Viral RNA and anti-HEV antibodies were identified in rabbits inoculated with swine HEV-3 demonstrating positive infectivity of the virus. However, pro-inflammatory cytokine and liver enzyme levels in serum were not significantly elevated, and only mild inflammatory lesions were detected in the liver tissues of rabbits infected with swine HEV-3. These results suggest that swine HEV-3 can engage in cross-species transmission to rabbits, but causes only mild inflammation of the liver.

scholarly journals Prevalence of hepatitis E virus (HEV) antibodies in Serbian blood donors

2014 ◽  
Vol 8 (10) ◽  
pp. 1322-1327 ◽  
Author(s):  
Tamaš Petrović ◽  
Diana Lupulović ◽  
Nereida Jiménez de Oya ◽  
Svetlana Vojvodić ◽  
Ana-Belén Blázquez ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Blood Donors ◽  
Hepatitis E ◽  
Enzyme Linked Immunosorbent Assay ◽  
Industrialized Countries ◽  
Limited Data ◽  
Genotype 3 ◽  
Serum Samples ◽  
High Prevalence ◽  
Polymerase Chain

Introduction: Hepatitis E virus (HEV) infection is rarely reported in industrialized countries, but recent studies have revealed quite variable seroprevalence rates among European populations, including blood donors. In Serbia, very limited data about HEV seroprevalence are available. This study aimed to determine the prevalence of anti-HEV IgG antibodies and HEV RNA in the sera of volunteer blood donors in Serbia. Methodology: Serum samples from 200 volunteer blood donors were tested for the presence of anti-HEV IgG by enzyme-linked immunosorbent assay (ELISA) using ORF-2 HEV genotype 3 recombinant proteins as antigen, and for the presence of HEV RNA by nested reverse transcriptase polymerase chain reaction (RT-PCR). Results: In total, 15% of the volunteer blood donors were seropositive. The prevalence increased with age; 21.5%, 14.2%, and 5.4% HEV seroprevalence rates were found in individuals older than 51 years, between 31 and 50 years, and in those younger than 30 years of age, respectively. However, no HEV RNA was detected in any of the individuals analyzed. Conclusions: The prevalence of anti-HEV IgG among blood donors as representatives of the general population is quite high in Serbia compared to data from many European countries. One of the reasons for this could be the high prevalence of HEV among Serbian pigs and the traditional consumption of piglet meat in the country. The relatively high HEV seroprevalence found among Serbian blood donors indicates the need for further investigation.

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