文獻來源: 王馨蓮. 2006. 台灣中部地區豢養陸龜上呼吸道疾病之黴漿菌分離與鑑定 (Isolation and Identification of Mycoplasma spp. of Upper Respiratory Tract Disease in Pet Tortoises in Central Taiwan). 國立中興大學獸醫學系暨研究所碩士論文. [線上摘要]摘要
上呼吸道疾病為台灣豢養龜類之常見疾病，致病病原包含Mycoplasma agassizii, M. testudnium 及 Herpesvirus等。本研究目的為瞭解台灣中部地區罹患上呼吸道疾病之豢養陸龜黴漿菌感染情形，並提供臨床獸醫作為診斷治療之參考依據。自2005 年3月至2006年5月於中興大學獸醫教學醫院野生動物科門診採集罹患上呼吸道疾病陸龜的鼻腔灌洗液或拭子樣本共110個，分別以黴漿菌分離培養及聚合酶 鍊鎖反應檢測黴漿菌之發生率。黴漿菌分離使用SP4培養基，於30℃；5％ CO2 環境中培養1-6週。聚合酶鍊鎖反應則配合黴漿菌屬別特異性引子對P1-MGSO進行16S rDNA 核酸序列增幅，陽性產物經純化後以核酸定序比對進行確認。並分析自2004年8月至2006年5月門診罹患上呼吸道疾病陸龜病例與氣候因子之相關性。結果 顯示，黴漿菌屬別特異性聚合酶鍊鎖反應陽性率為16.3％。然而僅自這些檢體中分離出一株黴漿菌分離株，經核酸定序比對後確認為M. agassizii，證實台灣地區豢養陸龜確有黴漿菌感染。同時病例之發生除與氣溫改變具相關趨勢外，與其他氣候因子則無相關。和黴漿菌分離培養比較，以 聚合酶鍊鎖反應配合核酸序列分析比對進行黴漿菌檢測較分離培養可迅速、確實地診斷陸龜黴漿菌症，不但可用於上呼吸道疾病的病因診斷，也可作為帶菌陸龜的篩 檢方法，對於陸龜臨床疾病診治及預防本病傳播應當有所助益。
Abstract
Upper respiratory tract disease (URTD) was one of the common diseases in captive chelonian in Taiwan. It has been reported that Mycoplasma agassizii, M. testudnium and Herpesvirus can become the etiological agents and cause URTD in tortoise. The purpose of this study was to investigate the prevalence of mycoplasma infection in captive tortoises with URTD in central Taiwan that may provide the reference to clinical veterinarian for diagnosis of URTD in tortoises. We collected nasal discharge specimens of 110 tortoises with URTD from March, 2005 to May, 2006 at the wildlife section of Veterinary Teaching Hospital, National Chung Hsing University. Specimens were cultured in SP4 medium at 30℃ with 5％ CO2 for 1 to 6 weeks and examined by polymerase chain reaction (PCR). Mycoplasma genus-specific primer pair (P1-MGSO) based upon the sequence information of 16S rDNA was used in PCR detection. The positive PCR product was sequenced and then these sequences were compared with NCBI data bank. In addition, we analyzed the relationship between prevalence of URTD and climate factors (temperature difference and relative humidity) from August, 2004 to May, 2006. The result showed that the prevalence of Mycoplasma spp. was 16.3％ (18/110) detected by PCR. However, only one isolated specimens was identified as M. agassizii by 16S rDNA sequencing. In addition, URTD was only related to temperature change, but not to other climate factors. Compared to traditional mycoplasma culture PCR may be a good and rapid method for diagnosis of mycoplasmosis in tortoises with URTD. The method not only can use for clinically etiological diagnosis but also be a good screen method for carrier.
