| Summary: | (1) Background: <i>Toxoplasma gondii</i> is a widespread zoonotic agent that greatly impacts Public Health, being responsible for one of the most important parasitic zoonosis worldwide. <i>T. gondii</i> has a heteroxenous life cycle, with cats being the definitive hosts and all warm-blooded animals, including humans, being intermediate hosts. Veterinary practitioners (VP) may be at a higher risk than the general population for <i>T. gondii</i> infection, as they have direct and daily contact with many animal species. The aim of the present study was to ascertain if VP were more likely to be anti-<i>T. gondii</i> IgG seropositive than the general population, as well as to understand if age, accidents with blood-contaminated sharps (cross-blood contamination), gender, working years, and geographic regions play a role as risk factors for <i>T. gondii</i> infection. For this purpose, a case–control study using archived samples was performed. (2) Methods: A total of 350 veterinary practitioners were tested using a commercial semiquantitative enzyme immunoassay for anti-<i>T. gondii</i> IgG. From the general population, 175 anonymous volunteers (matched with cases by region, age, and gender) were studied for anti-<i>T. gondii</i> IgG. (3) Results: There was no statistical difference found between the presence of anti-<i>T. gondii</i> IgG in practitioners (26%; CI = 21.40–30.60%) and the general population (33.14%; CI = 26.17–40.12%) (<i>p</i> = 0.108). Univariate and multivariate analysis showed that only age (older groups) was found to be associated with a higher prevalence of anti-<i>T. gondii</i> IgG, with significant <i>p</i> values (<i>p</i> < 0.05) for both univariate and multivariate analysis. (4) Conclusions: To the best of our knowledge, this is the first case–control study fully focused on the prevalence of anti-<i>T. gondii</i> IgG in VP in Portugal, showing that there was no significant risk for <i>T. gondii</i> infection in veterinarians exposed daily and repeatedly to different species of animals.
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