Background: Although the Coxiella burnetii infection has been investigated in dogs, its role in human transmission remains to be fully established, particularly in close and daily human–dog contact settings, such as in Police K-9 Units. Methods: Accordingly, this study aimed to assess anti-C. burnetii antibodies in clinically healthy police officers by an in-house indirect immunofluorescence assay (IFA), and working dogs by a commercial IFA Kit, from the State Special Operations Battalion, Paraná, Southern Brazil. Results: Overall, 1/18 (5.5%) police officers and 9/30 (30.0%; CI 95% 16.66–47.88) dogs tested seropositive to anti-C. burnetii IgG antibodies. Conclusions: To date, this is the highest prevalence of Q fever seropositivity among military dogs worldwide. Despite the low sampling rate, a statistically significant association was found between seropositivity and female dogs (p = 0.0492). Further studies with larger sample sizes should be conducted to establish the prevalence of Q Fever in other Brazilian K-9 Units. In summary, this study is the first to conduct a concomitant serosurvey of police officers and working dogs, and its findings should be considered a warning for cross-exposure and transmission of Coxiella burnetii among Police K-9 Units in Brazil and worldwide.