BackgroundThe nematodes Dirofilaria immitis and D. repens are enzootic in Greece. In the light of evidence of dirofilariosis spreading to new areas around the world, the aim of the present study was to update and enrich the current knowledge on the prevalence of Dirofilaria infections in dogs in Greece, to assess the risk factors of heartworm infection, and to initiate the drawing of the epizootiological map of canine dirofilariosis, investigating Dirofilaria infections in five locations along the north-south axis of the country, i.e. municipalities of Thessaloniki, Larissa, Achaia, Attica and Heraklion, associated with the five largest urban centres of Greece.MethodsBlood samples collected from 750 dogs in total, were examined by the modified Knott’s method and by serology. A questionnaire including information about each examined dog was filled in and statistical analysis of the results was performed using the Chi-square test and a Binary Multiple Univariate Generalized Linear Model.ResultsIn total, 31 (4.1 %) out of 750 examined animals were found positive for D. immitis by any of the tests applied (Knott’s method and serological examination). Moreover, microfilariae of D. repens and Acanthocheilonema reconditum were detected by the Knott’s method in 17 (2.3 %) and 10 (1.3 %) of the animals, respectively. At the municipality level, the prevalence of infection was 14, 7, 5.3, 0.7 and 0 % for D. immitis, 1, 2, 8.7, 0.3 and 0 % for D. repens, and 0, 3, 2.7, 0.7 and 1 % for A. reconditum in Thessaloniki, Larissa, Achaia, Attica and Heraklion, respectively. In addition, in three dogs (one each in Thessaloniki, Achaia and Attica) mixed D. immitis - D. repens infections were detected by the Knott’s method. The area of the country, dog’s usage and age were determined as risk factors for heartworm infection.ConclusionsNorthern areas of Greece have higher Dirofilaria prevalence and the prevalence in a western province (Achaia) is recorded for the first time. The mosquito population dynamics recorded in the past is likely to play an important role in the distribution of Dirofilaria infections in Greece, and needs further investigation. Similarly, the role of wild reservoirs of filarial parasites in different areas needs to be clarified. Promotion by veterinarians of preventive treatment and compliance by pet owners is essential in all parts of Greece, regardless of the recorded prevalence of infection.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1797-6) contains supplementary material, which is available to authorized users.