: Hepatitis E virus (HEV) can infect not only human but also several animals. This study has been conducted to evaluate the comprehensive anti-HEV seroprevalence in zoo animals in Korea. Anti-HEV antibodies were identified in 14 of 64 zoo animal species. HEV antibodies were detected for the first time in Eurasian Lynx, Setland Pony, Fallow Deer, Ezo Sika, Formosa Deer, East Wapitis, Barasingha, Corriedale, American Bison, Guanacos, Reticulated Giraffe, and Saanen. These results indicate that the several zoo animal species were exposed to HEV.Keywords : hepatitis E virus, seroprevalence, zoo animal species Hepatitis E virus (HEV) is an emerging zoonotic agent. HEV is the only member of the genus Hepevirus in the family Hepeviridae. HEV is a non-enveloped virus that has an approximately 7.2 kb single-stranded genome comprised of positive-sense RNA [7]. HEV strains have been classified into four major genotypes based on phylogenetic analysis of full genome sequences or highly conserved partial sequences [7]. HEV infections by genotypes 1 and 2 are restricted to humans and cause mainly waterborne outbreak of hepatitis in developing countries. However, HEV genotypes 3 and 4 are detected in both humans and animals, and induce sporadic cases of acute hepatitis in developed countries. Human infections by HEV genotypes 3 and 4 are possibly transmitted from animal species including pigs and wild boar [8]. Previous serological studies in human, pig, and cat populations in Korea reported seroprevalence of anti-HEV antibody as 18%, 15%, and 8.1%, respectively [3,12]. However, no reports have addressed the seroprevalence study in zoo animal species. This study was conducted to evaluate the comprehensive anti-HEV seroprevalence status in zoo animal species in Korea.This study was conducted to evaluate comprehensively the anti-HEV seroprevalence in zoo animal species. A total of 201 serum samples were obtained from zoo animals at a Seoul Zoo, Korea from 2005 to 2010. The samples were stored at -80 o C until analysis. The samples were all tested in duplicate for anti-HEV antibody with a direct sandwich enzyme immunoassay kit (Wantai Biopharmaceutical, China). The assay was carried out by following the manufacturer's instructions. Briefly, 50 µL of the serum sample was added to each well containing 50 µL of the diluent, and the microplate was incubated for 30 min at 37 o C. The microplate was washed five times with 350 µL of a wash solution. 100 µL of horseradish peroxidase-conjugated recombinant HEV antigen was added to each well, and the microplate was incubated for 30 min at 37 o C. The microplate was washed five times with 350 µL of a wash solution. 50 µL of chromogen A and chromogen B solution were added into each well, and the microplate was incubated at 37 o