Zambia experienced the largest cholera epidemic in the country's history in 2023-2024; however, the antimicrobial susceptibility profile of Vibrio cholerae during the epidemic is unknown. A total of 2,384 stool samples were collected from suspected cholera cases in Eastern, Lusaka, and Luapula provinces in Zambia from January 2023 to March 2024. Among them, 549 (23.5%, n = 549 of 2,341) were culture positive for V. cholerae O1, and antimicrobial susceptibility results were available for 431 (78.5%, n = 431 of 549). Sensitivity for tetracycline was 84.5% (n = 316 of 374) in Lusaka, whereas it was 100% in Eastern and Luapula provinces. Isolates resistant to azithromycin were found only in Lusaka (1.6%, n = 1 of 61). Sensitivity for ciprofloxacin was 81.8% (n = 260 of 318) in Lusaka province, whereas it was 100% in other provinces. Our results suggested an increased prevalence of antimicrobial resistance in V. cholerae against the first- and second-line antibiotic treatments, particularly in the capital. Careful monitoring of the regional antibiogram is warranted.
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