食蛇龜正常健康體態比例、血液及生化參考值之研究

Abstract

食蛇龜（Cuora flavomarginata）為台灣原生種烏龜，且為台灣四種原生龜中唯一陸棲型淡水龜，且由於長年棲地的破壞以及走私捕捉，面臨瀕臨絕種的危機，但現今爬蟲類文獻中對於食蛇龜的研究仍相當缺乏，尤其基礎生理及血液學資料皆尚未建立，因此本研究目的在於建立食蛇龜的正常體態參考指標及血液生化參考值，以期應用於獸醫臨床醫學及生態保育研究。 本實驗研究食蛇龜以圈養且臨床理學檢查正常之成年食蛇龜53隻，其中雄龜17隻、雌龜36隻作為觀測對象，健康體態參考指標以Jackson’s ratio、體重除以背甲長的對數回歸方程式進行分析。結果顯示食蛇龜的體重與背甲長存在正相關性，體重除以背甲長的比例會隨著背甲長增加；同時建立了食蛇龜的體重背甲長回歸方程式，可根據季節和性別的不同用來預測正常的體重範圍。 在血液生化參考值之建立上，以同一群圈養之食蛇龜於1月、4月、7月、10月為台灣四季代表月份，以物理保定方式進行頸靜脈抽血，檢驗血液中項目包含血容比、血紅素、紅白血球計數及血球細胞染色。血液生化學包含總蛋白、白蛋白、尿酸、尿素氮、肌胺酸酐、葡萄糖、天門冬酸鹽轉氨酶、氨基丙酸轉氨酶、膽固醇、三酸甘油脂、鹼磷酶、血鈣、血磷、肌酸磷激脢共14項，以國際臨床病理聯盟（International Federation of Clinical Chemistry, IFCC）法則建立正常參考值的範圍，並分析性別及季節單獨對於血液生化值的影響和性別與季節的交互作用。 血液學和血液生化值的參考值分析，雄龜和雌龜間的血紅素、血容比、紅血球總數、平均血球血紅素、淋巴球比例、嗜酸性球比例、單核球比例、鹼磷酶、天門冬酸鹽轉氨酶、尿素氮、血鈣、尿酸、膽固醇、肌酐激脢、血糖、血磷、總蛋白、三酸甘油脂具有差異存在。季節因子方面，除了平均血球血紅素、白血球總數、淋巴球比例、嗜鹼性球比例、氨基丙酸轉胺酶、血鈣、膽固醇、總蛋白不受影響外，其餘均有差異。繁殖行為及度冬是主要造成性別與季節間的差異原因。食蛇龜的血球在型態外觀上和其他爬蟲相似，在Liu’s Stain可得到良好的染色效果。此外和台灣另兩種原生烏龜班龜和柴棺龜比較，食蛇龜具有較高的嗜鹼性球比例和總蛋白，較低氨基丙酸轉胺酶濃度，而雌龜的三酸甘油脂則較高。 本實驗所完成的健康體態參考指標和血液生化參考值，可直接應用於獸醫臨床和生態保育相關工作上，根據實驗觀察此兩項檢查皆具有低侵入性的優點，實驗模式及資料未來可成為其他爬蟲類動物臨床或研究之參考。

Yellow-margined box turtle (Cuora flavomarginata) is the only largely terrestrial geoemydide turtle species in Taiwan. Because of habitat destruction and smuggling, yellow-margined box turtle is endangered in the island. So far scientific research are lacking especially in basic physiology and hematological information. For this purpose, the objective of this study are establishing normal body ratio, hematological and biochemical reference values of yellow-margined box turtle. The results will be applied to clinical veterinary medicine and wildlife conservation. Fifty three captive adult yellow-margined box turtles including 17 males and 36 females were used in this study. Clinical physical examinations were normal in all turtles. Jackson’s ratio and the equation of logarithm body weight regressed against logarithm carapace length were adopted to assess the health condition. There was the high correlation between body weight and carapace length, Jackson’s ratio increased with increasing carapace length. Equations of logarithm body weight regressed against logarithm carapace length were established. Body weight can be predicted by the equation based on season and sex. In hematological and biochemical reference values establishment, blood samples were obtained from jugular vein in January, April, July, and October. Complete blood count (CBC) was performed and included PCV, Hb, RBC count, WBC count and WBC differential count. Chemistry analysis included albumin, total protein, uric acid, urea (BUN), creatinine, glucose, aspartate aminotransferase (AST), alanine aminotransferase (ALT), Alkaline phosphatase (ALKP), cholesterol, triglycerides, calcium, phosphorus and creatine kinase (CK). The principle form International Federation of Clinical Chemistry (IFCC) was utilized for reference range. Data were also analyzed on the basis of sex, season and interaction between sex and season, using repeated measures analysis of variance. Significant sex differences were observed for PCV, Hb, RBC, MCHC, lymphocyte percentage, eosinophil percentage, monocyte percentage, ALKP, AST, BUN, calcium, uric acid, cholesterol, CK, glucose, phosphorus, total protein and triglycerides. Marked seasonal variation was noted in most parameters except MCH, WBC, lymphocyte percentage, basophil percentage, ALT, calcium, cholesterol and total protein. The differences between sex and seasons were primarily associated with reproduction and lower temperature in winter. Morphology of blood cells stain well with Liu’s Stain, and was similar to other reptiles. In addition, when compared other native turtles, Chinese striped-necked turtle (Ocadia sinensis) and Asian yellow pond turtle (Mauremys mutica), yellow-margined box turtles had higher basophil percentage, higher total protein and lower ALT values. But only female turtles had higher triglycerides values. The results present in this study can contribute to clinical veterinary medicine and wildlife conservation directly. According to experimental observation, the two procedures used in this study have the advantages of being fast and required minimal invasion to the subjects. The results and methods will become a reference for future research.