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Toxoplasmosis is a parasitic disease with worldwide prevalence. Despite the relatively similar effects of toxoplasmosis and smoking on alteration in neurotransmitters, especially dopamine, little is known about the relation of <i>Toxoplasma gondii</i> infection and addiction to cigarette smoking. Therefore, the main objective of this study was to assess the relationship between latent toxoplasmosis and smoking. Through a case-control study, 216 regular cigarette smokers and 324 nonsmoker age- and gender-matched subjects were evaluated for anti-<i>T.gondii</i> IgG antibodies with enzyme-linked immunosorbent assay (ELISA). During the sampling, a structured questionnaire was used to obtain the demographic information of participants and the risk factors of acquired <i>Toxoplasma</i>. The median ages of case and control groups were 51.04 ± 18.1 (22–97 years) and 51.03 ± 16.5 (21–89 years), respectively (<i>p</i> = 0.99). Anti-<i>T.gondii</i> IgG antibodies were detected in 44 (20.37%) cases and in 135 (41.67%) controls. There was a statistically significant difference for the positivity rate between the smokers and the control group (OR = 0.35; 95%CI: 0.19–0.65; and <i>p</i> = 0.001). The overall prevalence was 33.14%. This study indicated the inverse association between seropositivity to <i>Toxoplasma</i> infection and cigarette smoking. This relationship could be due to the changes that latent toxoplasmosis has on the neurotransmitters, especially dopamine, which needs more research.