Antibodies Against Hepatitis E Virus (HEV) in European Moose and White-Tailed Deer in Finland
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ORIGINAL PAPER
Antibodies Against Hepatitis E Virus (HEV) in European Moose and White‑Tailed Deer in Finland Emil Loikkanen1 · Satu Oristo1 · Natalia Hämäläinen1 · Pikka Jokelainen2,3 · Tuija Kantala1,4 · Antti Sukura3 · Leena Maunula1 Received: 2 June 2020 / Accepted: 27 August 2020 © The Author(s) 2020
Abstract The main animal reservoirs of zoonotic hepatitis E virus (HEV) are domestic pigs and wild boars, but HEV also infects cervids. In this study, we estimated the prevalence of HEV in Finnish cervid species that are commonly hunted for human consumption. We investigated sera from 342 European moose (Alces alces), 70 white-tailed deer (Odocoileus virginianus), and 12 European roe deer (Capreolus capreolus). The samples had been collected from legally hunted animals from different districts of Finland during 2008–2009. We analysed the samples for total anti-HEV antibodies using a double-sandwich ELISA assay. Seropositive sera were analysed with RT-qPCR for HEV RNA. HEV seroprevalence was 9.1% (31/342) in moose and 1.4% (1/70) in white-tailed deer. None of the European roe deer were HEV seropositive (0/12). No HEV RNA was detected from samples of seropositive animals. HEV seropositive moose were detected in all districts. Statistically, HEV seroprevalence in moose was significantly higher (p 1.1) were considered as positive in our study.
Detection of HEV RNA by RT‑qPCR We used quantitative reverse transcription polymerase chain reaction (RT-qPCR) to screen for HEV RNA from the HEV seropositive animals’ sera to detect possible acute HEV infections. From each HEV seropositive animals’ sample, 140 µl of undiluted sera was analysed. Before RNA extraction, we added 10 µl of whole mengovirus (strain MC0 grown in HeLa cells; kindly donated from Bosch A, University of Barcelona, Spain) to the samples to control the RNA extraction efficiency. RNA extraction was done using a commercial kit (E.Z.N.A.® Viral RNA Kit, Omega Biotek, United States) according to the manufacturer’s instructions without using Carrier RNA and with an elution volume of 70 µl. We cleaned the extracted RNA samples with a commercial inhibitor removal kit (OneStep™ PCR Inhibitor Removal, Zymo Research, United States) following the manufacturer’s instructions. In the presence of inhibition the sample was diluted. To measure the presence of HEV RNA, we used the method described earlier (Kantala et al. 2013) with slight modifications. Briefly, we used QuantiTect Probe RT-PCR
Table 1 Cervid samples included in the study according to the species, hunting districts, age groups, and sex, as well as moose densities in the studied hunting districts Species
European moose
District
Lapland, L North Ostrobothnia, NO Coastal Ostrobothnia, CO Central Finland, CF North Karelia, NK Southwest Finland, SW Southeast Finland, SE Total White-tailed deer Satakunta, S Southwest Finland, SW Total European roe deer Satakunta, S Southwest Finland, SW Total
Number of samples
Age group
Sex
Adult (%)
Calf (%)
Female (%) Male (%)
Moose density (numUnknown
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