應用形態病理學、穿透式電子顯微鏡及聚合酶鏈反應診斷陸龜疱疹病毒感染症

The Diagnosis of Herpesvirus Infection in Tortoises by Pathology, Transmission Electron Microscopy and Polymerase Chain Reaction

張益誠(Yi-Cheng Chang)

指導教授 ： 劉振軒

國立臺灣大學/生物資源暨農學院/獸醫學研究所/碩士(2005年)

https://doi.org/10.6342/NTU.2005.00485

摘要

自1999年1月至2005年4月，收集因消化道感染死亡之圈飼陸龜共56隻及採集144隻圈飼陸龜之口腔拭子進行病理學檢查、穿透式電子顯微鏡檢查及巢式聚合酶鏈反應檢測。罹病陸龜常見之臨床症狀為黃白色分泌物分別由口腔與鼻腔流出、食慾不振、沈鬱及眼瞼腫脹。由肉眼病理學與組織病理學檢查，有18隻（18/56, 32.1﹪）呈現壞死性口炎、胃炎與腸炎，其中有3隻（3/18, 16.6﹪），於胃（1隻）與小腸（2隻）之壞死病灶區伴隨出現嗜酸性核內包涵體，感染之品系分別為1隻四爪陸龜、1隻紅腿象龜與1隻蘇卡達象龜。以穿透式電子顯微鏡觀察口腔與胃組織中之包涵體，可見典型疱疹病毒之病毒顆粒分佈在黏膜上皮細胞之細胞核與細胞質內，病毒顆粒之直徑大約100 nm、具有封套、核蛋白衣、與電子緻密小體。採集144隻陸龜之口腔拭子及罹病陸龜之口腔與胃組織，經由巢式聚合酶鏈反應檢測疱疹病毒核酸，有3個口腔拭子與1個胃組織呈現陽性反應，感染品系分別為2隻四爪陸龜、1隻輻射龜與1隻緬甸星龜。由病理學、穿透式電子顯微鏡與巢式聚合酶鏈反應，證實國內圈飼陸龜已有疱疹病毒感染病例而死亡。此為台灣地區首次報告陸龜疱疹病毒感染症。

關鍵字

陸龜疱疹病毒感染症 ； 巢式聚合酶鏈反應 ； 陸龜

並列摘要

Between January in 1999 and April in 2005, a total of 56 necropsies of tortoises with clinical signs of oral and nasal discharge, anorexia, lethargy, and palpebral edema and 144 oral swabs were collected. On gross and microscopic examination, 18 of 56 necropsies (32.1%) showed varying degrees of necrotic lesions distributed in the oral cavity and gastrointestinal tract. Three of 18 necropsies (16.6%) with digestive lesions showed marked necrotizing gastroenteritis associated with intralesional eosinophilic intranuclear inclusion bodies (INIB), in which 1 case in the stomach and 2 cases in the small intestine were observed in Geochelone carbonaria, Geochelone sulcata, and Testudo horsfieldi. Electron microscopic examination of INIB in the oral tissue and stomach showed numerous herpesviral particles about 100 nm in diameter in the nuclei and cytoplasm of mucosal epithelium. The viral particles contained envelope, nucleocapsids, and an electron-dense body. Nested polymerase chain reaction analysis using a herpesvirus consensus primer method confirmed the presence of herpesvirus DNA in three oral swabs and stomach. Based on the clinical signs, and the results of pathological examination, electron microscopy and nest polymerase chain reaction, herpesvirus infection was diagnosed. This is the first report to verify herpesvirus infection of tortoises in Taiwan, which induced digestive lesions and caused fatal outcome in some cases.

並列關鍵字

Herpesvirus infection ； nested polymerase chain reaction ； tortoises

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