Lackovich J K, Brown D R, Homer B L, Garber R L, Mader D R, Moretti R H, Patterson A D, Herbst L H, Oros J, Jacobson E R, Curry S S, Klein P A
Department of Molecular Genetics and Microbiology, University of Florida, Gainesville 32610, USA.
Dis Aquat Organ. 1999 Jul 30;37(2):89-97. doi: 10.3354/dao037089.
Sea turtle fibropapillomatosis (FP) is a disease marked by proliferation of benign but debilitating cutaneous fibropapillomas and occasional visceral fibromas. Transmission experiments have implicated a chloroform-sensitive transforming agent present in filtered cell-free tumor homogenates in the etiology of FP. In this study, consensus primer PCR methodology was used to test the association of a chelonian herpesvirus with fibropapillomatosis. Fibropapilloma and skin samples were obtained from 17 green and 2 loggerhead turtles affected with FP stranded along the Florida coastline. Ninety-three cutaneous and visceral tumors from the 19 turtles, and 33 skin samples from 16 of the turtles, were tested. All turtles affected with FP had herpesvirus associated with their tumors as detected by PCR. Ninety-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 fibropapilloma. Also, 1 of 11 scar tissue samples from sites where fibropapillomas had been removed 2 to 51 wk earlier from 5 green turtles contained detectable herpesvirus. None of 18 normal skin samples from 2 green and 2 loggerhead turtles stranded without FP contained herpesvirus. The data indicated that herpesvirus was detectable only within or close to tumors. To determine if the same virus infected both turtle species, partial nucleotide sequences of the herpesvirus DNA polymerase gene were determined from 6 loggerhead and 2 green turtle samples. The sequences predicted that herpesvirus of loggerhead turtles differed from those of green turtles by only 1 of 60 amino acids in the sequence examined, indicating that a chelonian herpesvirus exhibiting minor intratypic variation was the only herpesvirus present in tumors of both green and loggerhead turtles. The FP-associated herpesvirus resisted cultivation on chelonian cell lines which support the replication of other chelonian herpesviruses. These results lead to the conclusion that a chelonian herpesvirus is regularly associated with fibropapillomatosis and is not merely an incidental finding in affected turtles.
海龟纤维乳头瘤病（FP）是一种以良性但使人衰弱的皮肤纤维乳头瘤以及偶尔出现的内脏纤维瘤增生为特征的疾病。传播实验表明，FP的病因与过滤后的无细胞肿瘤匀浆中存在的一种对氯仿敏感的转化因子有关。在本研究中，使用共识引物PCR方法来检测一种海龟疱疹病毒与纤维乳头瘤病的关联。从沿佛罗里达海岸线搁浅的17只患有FP的绿海龟和2只蠵龟身上获取了纤维乳头瘤和皮肤样本。对这19只海龟的93个皮肤和内脏肿瘤以及其中16只海龟的33个皮肤样本进行了检测。通过PCR检测发现，所有患有FP的海龟的肿瘤都与疱疹病毒有关。受影响海龟的肿瘤中有96%（89/93）含有可检测到的疱疹病毒，但皮肤样本中只有9%（3/33）含有。含有疱疹病毒的皮肤样本都在纤维乳头瘤的2厘米范围内。此外，在5只绿海龟身上，从2至51周前已切除纤维乳头瘤的部位采集的11个瘢痕组织样本中有1个含有可检测到的疱疹病毒。从2只绿海龟和2只蠵龟身上采集的18个未患FP而搁浅的正常皮肤样本中均未检测到疱疹病毒。数据表明，疱疹病毒仅在肿瘤内部或其附近可检测到。为了确定这两种海龟是否感染了同一种病毒，从6个蠵龟样本和2个绿海龟样本中测定了疱疹病毒DNA聚合酶基因的部分核苷酸序列。序列预测，蠵龟的疱疹病毒与绿海龟的疱疹病毒在所检测的60个氨基酸序列中仅有1个不同，这表明一种表现出微小型内变异的海龟疱疹病毒是绿海龟和蠵龟肿瘤中存在的唯一疱疹病毒。与FP相关的疱疹病毒在支持其他海龟疱疹病毒复制的海龟细胞系上无法培养。这些结果得出结论，一种海龟疱疹病毒经常与纤维乳头瘤病相关，而不仅仅是在受影响海龟中偶然发现。
