Trypanosoma cruzi is an emerging zoonotic vector-borne parasite infecting dogs and other mammals in the United States. In this study we evaluated shelter dogs in one northeastern and one southeastern county in Oklahoma for prevalence of exposure to T. cruzi. Dogs were tested for antibodies against T. cruzi using the Chagas STAT PAK® assay and for T. cruzi in circulation by PCR. In addition, dogs were tested for evidence of infection with other vector-borne organisms using the SNAP® 4Dx® Plus Test and PCR. Overall, 26 of 197 (13.2%) shelter dogs had detectable antibodies against T. cruzi and 3 of 189 (1.6%) dogs were PCR positive. In addition, we found that 42 of 197 (21.3%) shelter dogs had evidence of exposure to or were infected with at least one vector-borne agent other than T. cruzi based on serology and/or PCR; 9 of 42 (21.4%) of these dogs were also positive for T. cruzi antibodies. Other infections identified in dogs included Anaplasma phagocytophilum, Anaplasma platys, Babesia sp. (Coco), Dirofilaria immitis, Ehrlichia canis, Ehrlichia chaffeensis, Ehrlichia ewingii, and Hepatozoon americanum. This study serves to boost state-wide veterinary and public health awareness of T. cruzi and other vector-borne pathogens infecting shelter dogs in Oklahoma. Results indicate the need for more comprehensive screening of shelter dogs in Oklahoma for exposure to vector-borne agents to enhance surveillance and to identify dogs in need of additional specific veterinary care.