AIM:To investigate the seroprevalence and evolutionary dynamics of hepatitis E virus(HEV)and assess the ancestor of HEVs in China’s Shandong Province.METHODS:A total of 2028 serum,60 fecal and 82 bile samples were collected from the general human population,patients and swine,respectively.This seroepidemiological study was conducted using an immunnosorbent assay and HEV RNA was detected by the reverse transcription-nested polymerase chain reaction(RTnPCR)method.Complete genome sequences of the prevalent strains(CH-YT-HEV01,CH-YT-HEV02 and CHYT-sHEV01)were determined,and the sequences were analyzed phylogenetically.In addition,the evolutionary dynamics of three HEV isolates were determined using the framework of coalescent analysis in the program package BEAST,and the time of the most recent common ancestors(TMRCAs)of China-indigenous genotype4 HEV isolates was calculated.RESULTS:The overall viral burden in the general human population was 0.1%,and the positive rates of anti-HEV IgG and IgM in the serum specimens were25.1%(509/2028)and 2.3%(51/2028),respectively.In addition,IgG positivity increased with age.The phylogenetic analysis based on the full-length nucleotide sequences showed that the strain CH-YT-HEV02was directly related to CH-YT-sHEV01 with a 94%identity,suggesting that they were involved in crossspecies transmission.The isolate CH-YT-HEV01 was close to HB-3 and CHN-SD-sHEV with a bootstrap value of 100%,sharing a 96.1%-96.4%identity with each other.Surprisingly,the HB-3 strain was a representative strain prevalent in swine in Hubei,and the isolate CHNSD-sHEV was obtained from swine in Shandong in a previous report.TMRCA for the clade of CH-YT-HEV01and HB-3 was 2003,which was consistent with the TMRCA for the clade of CHN-SD-sHEV and HB-3,and they were both earlier than the TMRCA for the clade of CHYT-HEV01 and CHN-SD-sHEV(2004).CONCLUSION:The strains CH-YT-HEV01,CHN-SDsHEV and HB-3 are involved in trans-regional transmission,and the ancestors of HEVs in Shandong come from Hubei Province. AIM: To investigate the seroprevalence and evolutionary dynamics of hepatitis E virus (HEV) and assess the ancestor of HEVs in China’s Shandong Province. METHODS: A total of 2028 serum, 60 fecal and 82 bile samples were collected from the general human population, patients and swine, respectively. This seroepidemiological study was conducted using an immunosorbent assay and HEV RNA was detected by the reverse transcription-nested polymerase chain reaction (RTnPCR) method. Complete genome sequences of the prevalent strains (CH-YT-HEV01, CH- YT -HEV02 and CHYT-sHEV01) were determined, and the sequences were analyzed phylogenetically. In addition, the evolutionary dynamics of three HEV isolates were determined using the framework of coalescent analysis in the program package BEAST, and the time of the most recent common ancestors (TMRCAs) of China-indigenous genotype 4 HEV isolates was calculated .RESULTS: The overall viral burden in the general human population was 0.1%, and the positive rates of anti-HEV IgG a nd IgM in the serum specimens were 25.1% (509/2028) and 2.3% (51/2028), respectively. In addition, IgG positivity increased with age. The phylogenetic analysis based on the full-length nucleotide sequences showed that the strain CH-YT-HEV02 was directly related to CH-YT-sHEV01 with a 94% identity, suggesting that they were involved in crossspecies transmission. The isolate CH-YT-HEV01 was close to HB-3 and CHN-SD-sHEV with a bootstrap value of 100%, sharing a 96.1% -96.4% identity with each other. Surprisingly, the HB-3 strain was a representative strain prevalent in swine in Hubei, and the isolate CHNSD-sHEV was obtained from swine in Shandong in a previous report . TMRCA for the clade of CH-YT-HEV01 and HB-3 was 2003, which was consistent with the TMRCA for the clade of CHN-SD-sHEV and HB-3, and they were both earlier than the TMRCA for the clade of CHYT -HEV01 and CHN-SD-sHEV (2004) .CONCLUSION: The Sections CH-YT-HEV01, CHN-SDsHEV and HB-3 are involved in trans-regional transmission, and the ancestors of HEVs i nShandong come from Hubei Province.