Felids are important in the epidemiology of Toxoplasma gondii infection because they are the only hosts that shed resistant oocysts in the environment. A total of 242 serum samples and 80 fecal samples from domestic cats in Latvia was tested for T. gondii infection. Serum samples were tested for T. gondii antibodies by an indirect in-house ELISA; antibodies were found in 125 (51.6%) of 242 cats; seroprevalence increased with age, indicating postnatal infection. Using multiple logistic regression analyses, age and outdoor access were found to be the most significant (F = 21.70, P < 0.05) factors associated with T. gondii infection in Latvia. Toxoplasma gondii-like oocysts were detected in 2 of the cats examined microscopically using the salt flotation method, but definitive diagnosis could not be made because a bioassay was not performed.