Introduction: Brucellosis is one of the most common zoonosis diseases in developing and undeveloped countries, with adverse socio-economic status and animal and human health. The essential element for effective prevention and control of brucellosis is to improve the community’s Knowledge, Attitude, and Practice (KAP) through Health Education Intervention (HEI). Aim: To assess health education intervention’s effect on promoting knowledge, beliefs, and preventive behaviors on brucellosis among rural populations in Nagpur district, Maharashtra, India. Methods and Materials: 382 subjects over 18 years with a history of animal contact or consuming animal products were randomly selected. Data were collected through questionnaires and checklists. The pre-test was implemented and followed by HEI. The post-test was conducted after 45 to 60 days. For the attitude five-point Likert scale and knowledge and practice, a two-point assessment scale [yes, no] was applied. SPSS was used to analyze paired t -test, and P < 0.05 was considered significant. Result: Of 382 subjects, 300 (78.5%) were male, and the mean age of 42.15 ± 13.72. Before HEI, 18 (5%) subjects heard about brucellosis. After HEI, reduction in the risk behaviors practices like raw milk consumption ( P < 0.0001), assisted animal delivery without gown ( P = 0.002), throwing animal birth products in the dustbin ( P < 0.0001) were statistically significant. After implementing HEI, subjects were more aware of animal and human brucellosis signs/symptoms ( P < 0.0001). Awareness of disease transmission route ( P < 0.0001) and up-gradation in knowledge ( P < 0.0001) were statistically significant among subjects after HEI. Conclusions: HEI substantially affects KAP and changes community behaviors to prevent brucellosis transmission. The authors recommend implementing HEI in the community to prevent brucellosis.