Localization of fibropapilloma-associated turtle herpesvirus in green turtles (Chelonia mydas) by in-situ hybridization.

Fibropapilloma-associated turtle herpesvirus (FPTHV) is the presumed aetiological agent of sea turtle fibropapillomatosis (FP). Intralesional DNA and RNA of the virus have been detected by polymerase chain reaction (PCR) and reverse transcriptase-PCR (RT-PCR), respectively, but the exact location and distribution of the virus within the tumours have not been addressed. In this study, in-situ hybridization (ISH) was used to investigate viral transcriptional activity and localization of FPTHV. Twenty-five tumours were obtained from the skin or conjunctiva of 105 green turtles (Chelonia mydas) examined on two islands in Puerto Rico (Culebra and Culebrita). These lesions comprised 19 fibropapillomas and six fibromas. FPTHV mRNA transcripts were detected by ISH in three fibropapillomas, with positive reactions confined to the nuclei of clusters of epithelial cells. Viral DNA was detected by riboprobe ISH combined with denaturation in 14 tumours, including both fibropapillomas and fibromas. Signals were confined to the nuclei of acanthotic epithelial cells and were not seen in the subepithelial fibrous areas of the tumours. These results suggest that FPTHV is present in epithelial cells and transcriptionally active in fibropapillomas.