RESUMO
Fibropapillomatosis (FP), a transmissible neoplastic disease of marine turtles characterized by a likely herpesviral primary etiology, has emerged as an important disease in green sea turtles (Chelonia mydas) over the past three decades. The objectives of this study were to determine the suitability of three different chelonid fibropapilloma-associated herpesvirus (CFPHV) gene targets in polymerase chain reaction (PCR) assays of affected tissues; to explore the presence of CFPHV in non-affected skin from turtles with and without tumors; and to better understand tissue localization of the CFPHV genome in a tumor-free turtle by evaluating CFPHV presence in microanatomic tissue sites. Two aggregations of green sea turtles (Chelonia mydas) in Puerto Rico were evaluated, with six sampling intervals over the three-year period 2004-2007. Primary and nested PCR for three different herpesviral gene targets- DNA polymerase, capsid maturation protease, and membrane glycoprotein B- were performed on 201 skin biopsies taken from 126 turtles with and without external tumors. Laser capture microdissection and nested PCR were used to identify tissue localizations of CFPHV in skin from a normal turtle. Of the turtles sampled in Manglar Bay, 30.5% had tumors; at the relatively more pristine Culebrita, 5.3% of turtles sampled had tumors. All three PCR primer combinations successfully amplified CFPHV from tumors, and from normal skin of both tumored and tumor-free turtles. Via nested PCR, the polymerase gene target proved superior to the other two gene targets in the positive detection of CFPHV DNA. CFPHV infection may be common relative to disease incidence, supporting the idea that extrinsic and/or host factors could play a transforming role in tumor expression. Laser capture microdissection revealed CFPHV in skin from a tumor-free turtle, harbored in both epidermal and dermal tissues. Identification of CFPHV harbored in a non-epidermal site (dermis) of a tumor-free turtle indicates that virus is latent in a non-tumored host.
RESUMO
Seven species of helminths and six species of arthropods are reported from 23 of 40 brown pelicans, Pelecanus occidentalis, collected from various localities in Puerto Rico. Helminth parasites include three nematodes (Contracaecum multipapillatum, Contracaecum mexicanum, and Eustrongylides sp.), three trematodes (Galactosomum darbyi, Mesostephanus appendiculatoides, and Ribeiroia ondatrae), and one cestode (Tetrabothrium sulae). Arthropod parasites include Colpocephalum occidentalis, Neottialges apunctatus, Ornithodoros capensis, Phalacrodectus pelecani, Phalacrodectus punctatissimus, and Phalacrodectus sp. The presence of R. ondatrae in the brown pelican is a new species host record, and P. pelecani, P. punctatissimus and N. apunctatus are new subspecies host records. C. multipapillatum, C. mexicanum, G. darbyi and M. appendiculatoides are new locality records for Puerto Rico, and N. apunctatus, P. pelecani, P. punctatissimus and T. sulae are new locality records for the Caribbean. Necrosis produced by C. multipapillatum, C. mexicanum, and R. ondatrae may have contributed to the emaciation and death of the brown pelicans examined in the present study.