Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 18 1 2026 02 14 48 54 EN Haifeng Mao Department of Clinical Laboratory, The First Affiliated Hospital of Kangda College of Nanjing Medical University, The First People's Hospital of Lianyungang, Lianyungang, China Xinzhe Wu Department of Clinical Laboratory, The First Affiliated Hospital of Kangda College of Nanjing Medical University, The First People's Hospital of Lianyungang, Lianyungang, China Danting Jin Department of Clinical Laboratory, The First Affiliated Hospital of Kangda College of Nanjing Medical University, The First People's Hospital of Lianyungang, Lianyungang, China 2025 03 02 2025 11 06 Background and Objectives: Non-O1/O139 Vibrio cholerae (NOVC) has been associated with extraintestinal infections; however, its isolation from bile remains exceedingly rare. This study reports the identification and comprehensive analysis of a NOVC strain isolated from bile, including its antimicrobial resistance profile, virulence gene content, and molecular characteristics. Materials and Methods: A NOVC isolate was obtained from the bile of a patient with a hepatobiliary tumor. The isolate was identified and subjected to antimicrobial susceptibility testing. Whole-genome sequencing was performed to characterize its molecular features, including antimicrobial resistance genes, virulence genes, and relevant genetic mutations. Results: The NOVC isolate presents a multi-drug resistance phenotype. Corresponding genomic analysis indicates that this strain belongs to a novel sequence type (ST1736), carrying various drug resistance genes and virulence factors. Moreover, the blaCTX-M-65 extended-spectrum β-lactamase (ESBL) gene was detected in its chromosomal genome. Conclusion: This study presents the first report of a multidrug-resistant NOVC strain isolated from bile in mainland China. Notably, the ESBL gene blaCTX-M-65 was identified chromosomally in NOVC for the first time. https://ijm.tums.ac.ir/index.php/ijm/article/view/5322 https://ijm.tums.ac.ir/index.php/ijm/article/download/5322/1861