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Introduction: India is a developing country with many poor sanitation areas. Cholera, a water borne disease is rampant in areas of poor sanitation and is mainly due to Vibrio cholera of O1 and O139 serogroups causing acute cases of rice watery diarrhoea and high mortality. Infection due to Multidrug Resistant (MDR) Vibrio is on the rise. It is endemic in over 50 countries including India leading to a number of epidemics and pandemics. Till date, about seven pandemics have been identified due to cholera infection. Aim: To isolate Vibrio cholerae from acute diarrhoea cases with their antibiotic susceptibility and identify MDR strains, if any. Materials and Methods: A hospital-based, cross-sectional study was conducted in the Department of Microbiology, Burdwan Medical College, Purba Burdwan, West Bengal, India, from January 2021 to December 2021. Rectal swabs/faeces collected in Cary-Blair Transport Media were brought to the laboratory and incubated at 37°C for six hours; a loopful was inoculated in nutrient agar, MacConkey agar and selective media Thiosulphate-Citrate-Bile Salts-Sucrose agar (TCBS). Gram stain, motility, colony characteristics, oxidase test, cholera-red reaction and slide agglutination test was done. Diagnosis was confirmed; antibiotic susceptibility done and interpreted as per Clinical and Laboratory Standard Institute (CLSI) guidelines. The data was collected and entered into Microsoft Excel software and presented as frequency and percentages. Results: Total 60 samples were collected and tested, out of which 24 (40%) were positive for Vibrio cholerae and nine in 24 (37.5%) samples were MDR strains. Twelve samples were from femlaes and 12 were from males, majority {5 (41.67%) females, 9 (75%) males} belonged to the age group of 0-5 years. Serotyping revealed that Vibrio cholerae O1 serogroup was identified in 8 (33.33%) cases and O-139 serotype in 16 (66.67%) samples. Rest were Escherichia coli (n=10, 25%), Klebsiella pneumoniae (n=5, 12.5%) and Staphylococcus aureus (n=1, 25%). Conclusion: There is a rise of infections due to MDR strains of Vibrio O139 sero group in the community which needs early diagnosis and treatment for control. Vibrio cholerae strains were more resistant to fluoroquinolones, macrolides, tetracycline and Ampicillin. [ABSTRACT FROM AUTHOR]