Sea turtle fibropapillomatos~s (FP) is a disease marked by proliferat~on of b e n~g n but debilitating cutaneous fibropapillomas and occasional visceral fibromas Transmission experiments have implicated a chloroform-sensltlve transforming agent present in filtered cell-free tumor homogenates in the etiology of FP In t h~s study, consensus pnmer PCR methodology was used to test the a s s o c~a t~o n of a chelonian herpesvirus with f~bropapillomatosis Fibiopap~lloma and skin samples were obtalned f~o m 17 green and 2 loggerhead turtles affected iwth FP stranded along the Flor~da coastline Ninety-three cutaneous and vlsceral tumors fiom the 19 turtles, and 33 skln samples from 16 of the turtles, were tested All turtles affected with FP had herpesvlrus associated w t h thelr tumors as detected by PCR N~nety-six percent (89/93) of the tumors but only 9 % (3/33) of the skin samples from affected turtles contained detectable herpesvirus The skin samples that contained herpesvirus were all within 2 cm of a flbropapillo~na Also. 1 of 11 scar tissue samples from sites where fibropapillomas had been removed 2 to 51 wk earlier from 5 green turtles contalned detectable herpesvirus None of 18 normal skln samples from 2 green and 2 loggerhead turtles stranded without FP contained herpesvirus The data indicated that heipesvlrus was detectable only withln or close to tumors To determine if the same vlrus infected both turtle specles, partial nucleotide sequences of the herpesvlrus DNA polymerase gene were determined from 6 loggerhead and 2 green turtle samples The sequences predicted that herpesviius of loggerhead turtles dlffered from those of green turtles by only 1 of 60 a m~n o acids In the sequence examined, Indicating that a chelonlan herpesvlrus exhibltlng minor intratyplc vanation was the only helpesvlrus present in tumors of both green and loggerhead turtles The FP-assoc~ated herpesvlrus reslsted cultlvat~on on chelonian cell hnes whlch support the replicat~on of other chelonian herpesvlruses These results lead to the conclus~on that a chelonian herpesvirus IS regularly associated with f~bropap~llomatosis and 1s not merely an ~ncldental flndlng in affected turtles

FIBROPAPILLOMATOSIS (FP) of sea turtles is a debilitating disease which is characterised by multiple benign cutaneous fibropapillomas and occasional visceral fibromas (Herbst 1994). Fibroepithelial tumours, which are commonly found on the turtles' conjunctivae and skin, interfere with vision, feeding and locomotion, and visceral nodules can fatally disrupt normal organ function (Herbst 1994, Herbst and others 1999). The aetiology of FP is unproven, but the disease has been experimentally induced in tumour-free turtles in captivity by inoculation of cell-free filtrates of tumour homogenates, consistent with a viral aetiology (Herbst and others 1995). The transforming agent is chloroform-sensitive, which suggests that if the agent is viral, it is most likely to be an enveloped virus (Herbst and others 1996a, b). However, similar fibropapillomas are caused in many vertebrates by papillomaviruses, that is, naked DNA viruses of the family Papovaviridae. Although papillomaviruses were absent from fibropapillomas of green turtles from Florida and Hawaii examined by using electron microscopy, immunohistochemistry and nucleic acid blot probing (Jacobson and others 1989), subsequent development of sensitive polymerase chain reaction (PCR) diagnostics for papillomaviruses has prompted reinvestigation of a possible association between papillomaviruses and sea turtle FP. Lu (1998) used degenerate PCR primers to amplify a DNA fragment which included a short nucleotide sequence motif identical to a sequence in the late structural capsid protein 1 (Li) gene of mammalian papillomaviruses. The fragment was amplified from fibropapillomas, lung, kidney, heart, skin, and cultured cells derived from fibropapillomas, testis and lung of FP-affected green turtles from Hawaii. The data were interpreted to represent detection of papillomaviruses in tumour tissues of green turtles.This short communication reports the results of an assay for papillomaviruses in nucleic acid samples purified from 26 fibropapillomas from 10 green turtles and one loggerhead turtle from Florida. Four pairs of PCR primers with known efficiency of detection were used.The nucleic acid samples used for the PCR templates were obtained by using standard methods of digestion (0.5 g sample plus 50 mg trypsin in 500 VI of 10mM Tris-HCl pH 8, 0-4M sodium chloride, 0-6 per cent sodium dodecyl sulphate [SDS], and 50mM ethylenediaminetetraacetic acid [EDTA], incubated at 37°C until liquified) and phenol:chloroform extraction of 26 cutaneous fibropapillomas from 10 green turtles and one loggerhead turtle stranded along the east coast of Florida (Lackovich and others 1999).The PCR was performed by using a Perkin-Elmer Gene Amp 2400 thermal cycler. Reactions included 10mM Tris-HCl pH 9, 50mM potassium chloride, 0.1 per cent Triton X-100, 2-5 mg template nucleic acid or 500 ng positive control plasmid DNA, 0-2mM deoxynucleoside triphosphates, 50 pmol each primer, and 2-5 U Taq DNA polymerase in 100 1I volumes. For consensus degenerate primer combination MYO9 + MY11, complementa...