Whole-genome analysis of the potentially zoonotic Elizabethkingia miricola FL160902 with two new chromosomal MBL gene variants.

Objectives: Elizabethkingia is an emerging life-threatening pathogen in both humans and animals. We describe the whole-genome analysis of an Elizabethkingia miricola strain isolated from a diseased frog in China and investigate the molecular mechanism of carbapenem resistance in this pathogen. Methods: WGS of E. miricola FL160902 was performed using single-molecule, real-time technology. A phylogenetic tree was generated by SNP analysis, comparing the genome of our strain with other E. miricola isolates of amphibian and human origins. Antimicrobial resistance genes and virulence-related genes were identified using the Comprehensive Antibiotic Resistance Database (CARD) and the Virulence Factor Database (VFDB). Two putative carbapenemase genes were expressed in Escherichia coli to evaluate their contribution to antimicrobial resistance. Results: The genome of E. miricola FL160902 consists of a 4249586bp circular chromosome with 27 putative resistance genes and 38 predicted virulence-associated genes. Comparative genomic analysis demonstrated that the E. miricola strains of human and amphibian origins have similar virulence-associated gene profiles. In addition, all the amphibian isolates clustered together with one of the human isolates in the phylogenetic analysis. WGS revealed the presence of two novel MBL genes, designated bla(BlaB-16) and bla(GOB-19). Cloning of bla(BlaB-16) and bla(GOB-19) into E. coli DH5 alpha resulted in increased MICs of most beta-lactams, including imipenem, meropenem and ampicillin. Conclusions: We identified two chromosomal MBL gene variants, named bla(BlaB-16) and bla(GOB-19) in an amphibian E. miricola isolate, which was considered potentially zoonotic based on phylogenetic analysis and virulence-associated gene comparison. This study highlights the importance of E. miricola as a potential zoonotic pathogen and a reservoir of MDR genes.