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Hemosporidian parasites rarely infect aquatic birds. Few studies have been conducted in South America identifying some lineages of the genera Plasmodium, Leucocytozoon, and Haemoproteus, but none has been done in the Neotropic cormorant (Phalacrocorax brasilianus). This species is widely distributed through the American continent, from Southern USA to Tierra del Fuego, using a wide range of aquatic habitats. We molecularly studied the occurrence and diversity of hemosporidian lineages infecting individuals of Neotropic cormorant across a broad latitudinal gradient in Chile (Arica to Tierra del Fuego). As expected, a very low occurrence of individuals infected by Plasmodium sp. (4/123, 3.3%) and Leucocytozoon sp. (2/123, 1.6%) was detected. We found no evidence of Haemoproteus sp. We identified one lineage of Plasmodium (ZEMAC01) and one new lineage of Leucocytozoon (PHABRA01) infecting cormorants. Individuals infected by Plasmodium sp. were birds from only one site (i.e., Chillán), whereas Leucocytozoon sp. was found in one bird from Valdivia and another one from Tierra del Fuego. Our results expand the known range of hemosporidian parasite lineages in aquatic birds providing an essential baseline data that contribute to a better understanding of the geographic range of hemosporidian parasites infecting Phalacrocoracidae in South America.