Water, sewage, and soil are potential sources of infection for Toxoplasma gondii. Therefore, we sought to determine the prevalence of T. gondii infection and associated characteristics in 61 plumbers, 203 construction workers, and 168 gardeners in Durango City, Mexico. Participants were tested for T. gondii IgG and IgM antibodies with the use of enzyme-linked immunoassays. In addition, sociodemographic, clinical, and behavioral characteristics from each participant were obtained. IgG T. gondii antibodies were found in 4 (6.6%) plumbers, 17 (8.4%) construction workers, and 10 (6.0%) gardeners; T. gondii IgM antibodies were found in 3 (1.5%) construction workers and 4 (2.4%) gardeners, but in none of the plumbers. In the total population, the prevalence of T. gondii infection was significantly higher in workers living in suburban areas, without education, workers that consumed chorizo, and those who suffered from any disease (P < 0.05). In gardeners, prevalence of infection was significantly higher in those with blood transfusion, and memory impairment (P < 0.05). Multivariate analysis showed that T. gondii infection was positively associated with consumption of unwashed fruits (adjusted odds ratio [OR] = 2.41; 95% confidence interval [CI]: 1.13–5.13), and with raising animals (adjusted OR = 2.53; 95% CI: 1.00–6.37). This is the first report of contributing factors for T. gondii infection in workers occupationally exposed to water, sewage, and soil in a Mexican city, and results will contribute to the design of optimal preventive measures.