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Polyomaviruses (PyVs) are small double-stranded DNA viruses able to infect species across all vertebrate taxa. In cetaceans, PyVs have been reported only in short-beaked common dolphin (Delphinus delphis), common bottlenose dolphin (Tursiops truncatus) and killer whale (Orcinus orca). Herein, we surveyed PyV in 119 cetaceans (29 mysticetes and 90 odontocetes) stranded along the Brazilian coast, from 2002 to 2022, comprising 18 species. DNA extracted from the lungs was tested using a nested PCR targeting the major capsid protein gene of PyV. Polyomavirus was detected in lung samples of 1.7% (2/119) cetaceans: two juvenile female Guiana dolphins (Sotalia guianensis) stranded in Rio de Janeiro (Rio de Janeiro state) and Guriri (Espírito Santo state), in 2018. Both retrieved sequences were identical and presented 93.3% amino acid identity with Zetapolyomavirus delphini, suggesting a novel species. On histopathology, one of the PyV-positive individuals presented basophilic intranuclear inclusion bodies morphologically consistent with polyomavirus in the lungs. Other available tissues from both cases were PyV-PCR-negative; however, both individuals tested positive for Guiana dolphin morbillivirus. To our knowledge, this is the first report of PyV infection in cetaceans of the Southern Hemisphere and the first description of a co-infection with morbillivirus.