We evaluated the prevalence of selected vector-borne diseases in 131 dogs in an animal shelter in central Spain using point-of-care assays (SNAP 4DX and SNAP Leishmania; IDEXX Laboratories, Westbrook, ME). The SNAP 4DX detects Dirofilaria immitis (Di) antigen and antibodies against Ehrlichia canis (Ec), Borrelia burgdorferi (Bb), and Anaplasma phagocytophylum (Aph); the SNAP Leishmania kit detects antibodies against Leishmania infantum (Li). Dogs were classified as healthy or sick based on physical examination, complete blood counts, and serum chemistry profiles. The prevalence of positive test results was as follows: Ec, 5.3% (n = 7); Aph, 19.0% (n = 25); Bb, 0%; Di, 0%; and Li, 5.3% (n = 7). Four dogs (3%) were coexposed to Ec and Aph, and three dogs (2.3%) were coexposed to Aph and Li. There was no statistically significant correlation between positive serology and clinical status (sick vs. healthy) or hematologic/biochemical abnormalities. The prevalence of Aph was the highest and is in agreement with a recent report in a dog shelter in northwestern Spain. These point-of-care assays may be more valuable as epidemiologic than as clinical tools.