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Xinhua Luo,1,* Jin Zhang,1,* Min Yuan,2 Sihua Mou,1 Mengqiao Xu,1 Dakang Hu,1 Qinfei Ma,1 Lingfen Sun,1 Piaopiao Li,1 Zhiwei Song,1 Lianhua Yu,1 Kai Mu3,4 1Department of Clinical Laboratory Medicine, Taizhou Municipal Hospital Affiliated with Taizhou University, Taizhou, 318000, People’s Republic of China; 2State Key Laboratory for Infectious Diseases Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, 102206, People’s Republic of China; 3Beijing Institute of Radiation Medicine, Beijing, 100850, People’s Republic of China; 4Beijing Key Laboratory of New Molecular Diagnosis Technologies for Infectious Diseases, Beijing, 100850, People’s Republic of China*These authors contributed equally to this workCorrespondence: Kai Mu, Beijing Key Laboratory of New Molecular Diagnosis Technologies for Infectious Diseases, Beijing, 100850, People’s Republic of China, Tel +86-010-66874794, Email kai_mu@outlook.com Lianhua Yu, Department of Clinical Laboratory Medicine, Taizhou Municipal Hospital affiliated with Taizhou University, Taizhou, 318000, People’s Republic of China, Email yulianhua64@126.comPurpose: This study aimed to explore the genomic characterization of multidrug-resistant IncHI5-carrying Klebsiella michiganensis strains and detailed genomic dissection of the IncHI5 plasmids.Materials and Methods: Through whole-genome sequencing, the IncHI5 plasmid pK92-qnrS was obtained from a single clinical K. michiganensis isolate K92. All complete genomes of K. michiganensis strains from the Genome database of NCBI were collected and used to construct a maximum likelihood (ML) phylogenetic tree. The epidemiology and geographic distribution of all the K. michiganensis strains were conducted. An extensive comparison of the seven IncHI5 plasmids of K. michiganensis (one from this study, six from GenBank) was applied.Results: This study revealed that all K. michiganensis strains carrying IncHI5 plasmids from different clonal groups were located in the southeast coastal area of China. The backbone regions of IncHI5 plasmids were composed of replicon (repHI5B and repFIB), partition (parABC), and conjugal transfer (tra1/tra2). The main accessory resistant regions of IncHI5 could be divided into two categories, Tn 1696-related region and Tn 6535-related region. These seven IncHI5 plasmids carried multiple drug-resistance genes which were all mediated by the mobile genetic elements (MGEs).Conclusion: Data presented here help to provide an overall in-depth understanding of epidemiology and geographic distribution of IncHI5-carrying K. michiganensis and the structure and evolutionary history of IncHI5 plasmids.Keywords: Klebsiella michiganensis, mobile genetic elements, Tn 1696, Tn 6535, antimicrobial resistance