Objectives: In an endemic area for leishmaniosis and filariasis, coinfection can occur and the immunomodulation triggered by Wolbachia infection might influence the clinical signs and progression of both diseases. The aims of this study were to determine the prevalence of Wolbachia in dogs infected with Dirofilaria immitis and other filarial nematodes, to evaluate the prevalence of coinfection of Leishmania infantum, filariae and Wolbachia and their association with clinical presentation. Methods: Polymerase chain reaction assays were performed to detect filarial species, Wolbachia species and Leishmania in 118 samples of dogs from southeastern Spain with leishmaniosis and/or filariasis. Results: Ninety-eight dogs were infected with Leishmania and 49 had filarial infection (29 were coinfected with both). Wolbachia DNA was detected in 30·6% of filariae-positive dogs (15/49). Dogs coinfected with Leishmania and filaria had more severe clinical signs. Wolbachia infection was significantly (P=0·026) more frequent in dogs that were not infected with Leishmania. There was no correlation between outcome and coinfection with these pathogens. Clinical Significance: This study highlights the increased sensitivity of polymerase chain reaction in the diagnosis of filariasis, confirms the presence of Wolbachia in dogs from the Mediterranean basin, shows the increased severity of clinical signs when Leishmania-filarial coinfection is present and suggests a protective role of Wolbachia in leishmaniosis. © 2013 British Small Animal Veterinary Association.