Leptospirosis is a serious and potentially fatal zoonotic disease, but often neglected owing to lack of awareness. This study examined the knowledge, attitudes, and practices concerning leptospirosis among agricultural (n = 152) and non-agricultural (n = 115) workers in the lakeshore communities of Calamba and Los Baños, Laguna, Philippines. The findings showed no significant differences for the knowledge and attitude scores between agricultural and non-agricultural workers. However, agricultural workers had significantly lower prevention practice scores than non-agricultural workers. The ordinary least squares regression model identified gender, use of broadcast media as a source of health information, and knowledge and attitudes about leptospirosis as significant predictors of prevention practices common to both workers. Higher educational attainment was significantly associated with prevention practices among agricultural workers, while higher age and income level were significantly associated with prevention practices among non-agricultural workers. Public health interventions to improve leptospirosis knowledge and prevention practices should include health education and promotion programs, along with the strengthening of occupational health and safety programs in the agricultural sector.