Abstract

Skin lesions suggestive of papillomatosis were sampled from a free-ranging male calf Florida manatee (Trichechus manatus latirostris) (Tml) in Homosassa Springs, Florida, in February 2003. Total DNA was extracted from these lesions and amplified by polymerase chain reaction (PCR) using primers MY09 and MY11 routinely used in humans for the diagnosis of papilloma virus (HPV) infection in cervical carcinomas. These primers target sequences located within the L1 capsid protein gene and amplify a DNA fragment of 458-bp that translates into a peptide of 152 amino acid residues. GenBank searches identified this peptide as a highly conserved domain within the late L1 capsid protein of Tml papilloma virus (TmlPV). Comparison of the L1 nucleotide and amino acid sequences derived from this TmlPV with those from HPV showed a deletion of three nucleotides within the TmlPV L1 fragment. However, fragments were, on average, about 76 percent similar at the amino acid level. A second DNA fragment of approximately 1.5 kb was recently obtained, using the same set of primers, from a skin lesion that had been conserved in DMSO for at least one year. After sequencing, this fragment was shown to contain the open reading frame (ORF) that codes for the E6 oncogenic early protein known to bind the p53 tumor suppressor protein in addition to the L1 capsid protein fragment. These preliminary results provide molecular evidence of the association of a papilloma virus with skin lesions in the Florida manatee.

Acknowledgements

A Training Grant for the Care of Marine Mammals from the Florida Fish and Wildlife Conservation Commission supported this work.