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Post-mortem inspection in slaughterhouses plays a key role in the epidemiological surveillance of infectious diseases, including bTB. This study assessed the prevalence of bTB in cattle in Sicily during two different three-year periods (2010–2012; 2017–2019), using data from meat inspections and histopathological methods. Out of 100,196 cattle, 5221 (5.21%) were diagnosed with tuberculous lesions. Higher prevalence of bTB was recorded during the triennium 2010–2012 (6.74%; <i>n</i> = 3692) compared to the triennium 2017–2019 (3.36%; <i>n</i> = 1529), with a decreasing trend in annual occurrence throughout the study period and a heterogenous proportion of infected cattle among the Sicilian provinces (<i>p</i> < 0.01). Lower rates of infection were recorded in animals aged <12 months and >84 months (<i>p</i> < 0.0001). Pearson’s chi square analysis revealed a higher localization of lesions in the thoracic cavity (<i>p</i> < 0.0001). Gathered findings show that even though bTB occurrence in cattle was greatly reduced over the years, the disease still remains widespread in Sicily, also posing severe implications for public health.