Brucellosis, a contagious bacterial disease affecting animals globally presents a substantial zoonotic risk that is frequently underestimated, hinders animal trade, and endangers livestock and human health. The present study was conducted from November 2023 to June 2024 in Central and North Gondar Zone, Ethiopia. The current cross-sectional study aimed to evaluate the prevalence of brucellosis and to explore the related knowledge, attitudes, and practices within the specified region. A total of 384 serum samples were collected via random sampling from 20 dairy farms located in Ethiopia. Both local and cross-breed samples screened using the Rose Bengal Plate test and confirmed through an enzyme-linked immunosorbent assay. The seroprevalence of bovine brucellosis for both tests was 17.71% overall, with 9.62% for Central Gondar and 27.27% for North Gondar. Extensive farms exhibited notably higher odds of brucellosis compared to intensive farms, with unadjusted crude odds ratios of 3.01 and adjusted odds ratios of 2.37, respectively. Medium-sized herds also demonstrated increased odds in the multivariate analysis compared with small herds. Young respondents displayed the highest awareness levels, followed by adults and older individuals, with statistically significant differences observed across all categories. Regarding a semi-structured survey from 150 farmers on the association between sociodemographic data and knowledge, females exhibited higher awareness levels, with 117 (80.14%) responding positively. Young respondents showed a higher positive response rate of 58% compared to adults (52.7%) and the elderly (36.54%). In conclusion, these results emphasize the need for comprehensive strategies to address the factors influencing bovine brucellosis prevalence and respondent awareness.