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Bovine tuberculosis (bTB), a chronic zoonotic disease in cattle, has a substantial socio-economic and public health impact. This study was conducted to estimate the prevalence and geographical distribution of bTB in the Sylhet district of Bangladesh. A cross-sectional study was conducted at all 12 upazilas of Sylhet district, which included 512 randomly selected cattle from 48 farms. Selected animals were tested with the Caudal Fold Tuberculin (CFT) test to identify bTB-positive cattle. Out of 512 cattle, only one animal was identified as a reactor, providing an estimated prevalence of 0.19% (95% Confidence Interval; 0-0.58%). The only positive reactor was found in Zakiganj upazila. As the prevalence of bTB in cattle in Sylhet appears to be low, it indicates that most of the upazilas of Sylhet district are free of the bTB infection. This prevalence is lower than the reported prevalence in other parts of Bangladesh. Thus, attempts should be made to maintain the current situation of bTB infection in cattle of Sylhet district., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)