Abstract

A novel papillomavirus, Tursiops truncatus Papillomavirus type 2, TtPV-2, isolated from a genital lesion of a free-ranging bottlenose dolphin at the coast of Charleston, SC was identified by using the isothermal multiply primed rolling-circle-amplification technique. The complete nucleotide sequence was determined by sequencing transposon integration sites of the cloned viral DNA. A phylogenetic analysis revealed that TtPV-2 belongs to the close-to-root sea mammal genus together with TmPV-1 (Trichechus manatus latirostris PV-1), PsPV-1 (Phocoena spinipinnis PV-1) and TtPV-1 with the highest L1 nucleotide sequence similarity to TtPV-1 (70%). The L1 nucleotide sequences of TtPV-1 and -2 encoding the major component of the viral capsid were identified and cloned. To create VLPs as a vaccine which can be applied to captive animals, insect cells were transfected with a baculoviral vector containing TtPV-1 L1 and TtPV-2 L1, the coding sequence for the L1 proteins of the PV isolated from a captive Tursiops truncatus living in a Portuguese Aquarium and from the Tursiops truncatus in the Charleston Harbor. Antibodies against dolphin Ig were generated in mice, isolated and purified to serve as a control in future seroepidemiological studies to screen captive and free-ranging dolphins for active infections with TtPV-2 and -1. Therefore, antibodies against the VLPs will be induced in mice.

Acknowledgements

Dr. Rehtanz is a Postdoctoral fellow of Harbor Branch Oceanographic Institution, Fort Pierce, FL and works at the Brown Cancer Center, University of Louisville, KY.