Objective：To reveal the serum，biotype distribution and molecular characteristics of isolates from different hosts in Jiangsu Province，2016-2019. Methods：Different kinds of specimens were collected for isolation strains，the isolated strains of Yersinia enterocolitic were evaluated by serotyping and biotyping. The five virulence genes（ail，virF，yadA，ystA and ystB）and multilocus sequence typing were analysed by using next-generation sequencing technology. Results：In this study，6 398 specimens were collected，and 186 strains of Yersinia enterocolitic were isolated. The serotype of 109 strains could not be identified，the dominant serotype that can be identified are O：8 and O：3. There are 3 biotypes of all isolates，most of which are biotype 1A. Based on sequencing analysis，type Ⅲ and type Ⅳ are the mainly virulence gene type in this area. All strains were divided into 81 STs by MLST. Conclusion：The serotype of Yersinia enterocolitic isolated from different hosts are complex，the biotype is relatively concentrated. Virulence gene type Ⅳ strains can be isolated from diarrhea patients. ST genotypes are distributed divergently. The hosts of pig，chick and dog should be supervision and surveillance.