Abstract BACKGROUND: Cholera remains a significant public health concern, particularly in endemic regions like Bangladesh. This study aimed to assess the prevalence and characteristics of Vibrio cholerae contamination in pani-puri plate wash water in Dhaka city, employing biochemical, microscopic, and antibiotic resistance analyses. METHODS: A total of 20 pani-puri plate wash water samples were collected from various locations in Dhaka between August 2023 and October 2023. The samples were analyzed at the Department of Microbiology, Stamford University Bangladesh. RESULTS: V. cholerae was found to be highly prevalent, especially in crowded urban areas. Biochemical tests confirmed the identity of the isolates as V. cholerae. Antibiotic susceptibility testing revealed concerning resistance patterns, with 100% resistance to commonly used drugs such as meropenem and ampicillin. In addition, resistance to cefixime, nalidixic acid, and chloramphenicol was observed, emphasizing the need for alternative treatment options. CONCLUSION: These findings highlight the significance of ongoing monitoring of V. cholerae populations for virulence factors and changes in antibiotic resistance to inform public health interventions and cholera control measures. The study underscores the importance of strict hygienic standards in the preparation of street food, particularly in endemic locations such as Dhaka, Bangladesh, to reduce the risk of waterborne diseases. Continuous surveillance for drug resistance and microbiological contamination is crucial for effective cholera treatment and prevention.