建立健康斑龜及柴棺龜血液蛋白質電泳參考值

Abstract

蛋白質電泳是將血中蛋白質區分及定量的檢驗，血中各種蛋白質是維持動物正常生理功能所必須，其含量會隨不同的生理和病理情況而改變，故用來評估動物的健康狀況，為相當有用的診斷工具。此外，爬蟲類血中白蛋白必須用蛋白質電泳法，方能得到正確之結果。這項檢驗在人類、哺乳類動物及鳥類已應用多年，但在爬蟲類缺乏參考值可提供診斷依據，其應用並不積極。因此本研究擬分別建立斑龜及柴棺龜（Ocadia sinensis and Mauremys mutica）之血液蛋白質電泳參考值，期望藉由蛋白質電泳的分析，配合其它臨床檢查及診斷試驗，提供治療及預後的指標。 研究對象為臨床理學檢查、血液學皆檢驗正常之成年斑龜30隻，其中雄龜15隻、雌龜15隻，及成年柴棺龜35隻，其中雄龜11隻、雌龜24隻。於民國100年至101年春夏秋冬四季各採一次血，以醋酸纖維素法進行蛋白質電泳（Cellulose acetate electrophoresis），檢體為血漿（plasma）。 結果顯示斑龜之血漿蛋白質可以分成白蛋白、α球蛋白、β球蛋白和γ球蛋白；而柴棺龜之血漿蛋白質則可分成白蛋白、α1球蛋白、α2球蛋白、β球蛋白和γ球蛋白。斑龜皆有雙白蛋白血症（bisalbuminemia）的現象，應和此品種烏龜的基因相關。 結果經統計分析可得到，斑龜雄雌龜間，白蛋白百分比、α球蛋白、白蛋白/球蛋白比例有顯著差異；季節方面出現差異的項目包括α球蛋白百分比、α球蛋白、β球蛋白、γ球蛋白百分比和球蛋白。柴棺龜雄雌龜間，白蛋白、α球蛋白百分比、β球蛋白、γ球蛋白百分比、γ球蛋白、球蛋白皆出現差別；季節這部分則所有分析項目皆達顯著差異。檢視血漿蛋白質在性別與季節的變化，可發現造成差異的主要原因是生殖行為和季節溫度變化。 由上結果得知斑龜及柴棺龜血漿蛋白質電泳參考值的建立，對往後診斷其健康狀態應極有助益，並可將此參考值提供為臨床檢查及治療預後的指標。 除了建立斑龜及柴棺龜之蛋白質電泳參考值外，由於斑龜血液學檢驗項目尚未齊全，包括血液生化值血鈣、血磷、肌氨酸激酶、乳酸鹽脫氫酶（Ca, Phosphorus, CK, LDH）和血液電解質鈉、鉀、氯離子（Na+, K+, Cl-）。本研究在進行斑龜蛋白質電泳之餘，額外補足這些項目，使斑龜之血液學檢驗資料能夠更臻完整。 關鍵字：斑龜、柴棺龜、蛋白質電泳、雙白蛋白血症、血液生化值、電解質

Protein electrophoresis is the method for fractionation and quantification of plasma proteins. Proteins are responsible for maintaining normal body functions, and change markedly in physiological and pathological conditions. Protein electrophoresis can be used to evaluate the health status of animals, and it is a valuable diagnostic tool. In reptiles, accurate albumin concentrations should be measured with protein electrophoresis. This diagnostic tool is widely applied in human, mammalian and avian medicine, but reference values have not been establish for all reptiles species, reducing the utility of protein electrophoresis in reptile medicine. The purpose of this study is to determine the protein electrophoresis reference values of Ocadia sinensis and Mauremys mutica, which can be used with other clinical and laboratory findings to make more complete diagnosis, disease monitoring and prognosis. Study animals were 30 adult Ocadia sinensis and 35 adult Mauremys mutica, including 15 males and 15 females, 11 males and 24 females respectively. Physical examinations, hematology and biochemistry tests were normal in all turtles. Plasma samples were obtained in four seasons from 2011 to 2012. Cellulose acetate electrophoresis method was used. The results showed that 4 protein fractions were identified in Ocadia sinensis: albumin, α-globulin, β-globulin,γ-globulin, and 5 fractions were found in Mauremys mutica, including albumin, α1-globulin, α2-globulin, β-globluin,γ-globulin. All Ocadia sinensis has bisalbuminemia. We suggest in this species, bisalbuminemia has a genetic origin, and it is a normal phenomenon for Ocadia sinensis. In Ocadia sinensis, significant sex differences of albumin (%), α-globulin (g/dL) and A/G ratio were observed; seasonal variations were noted in α-globulin (%),α-globulin (g/dL), β-globulin (g/dL), γ-globulin (%) and globulin (g/dL). In Mauremys mutica, sex differences were found in albumin, α-globulin (%), β-globulin (g/dL), γ-globulin (%), γ-globulin (g/dL), globulin (g/dL); all parameters has seasonal variations. The differences between sex and seasons were primarily associated with female reproductive stage and temperature changes of four seasons. In conclusion, protein electrophoresis reference intervals can be used in combination with other diagnostic tools to assess health status of Ocadia sinensis and Mauremys mutica and indicate disease prognosis. Besides of protein electrophoresis reference values, this study also establish the biochemistry and electrolyte values of Ocadia sinensis, including Ca, Phosphorus, CK, LDH and Na+, K+, Cl-, hoping to make more complete hematology database for this turtle. Key words: Ocadia sinensis, Mauremys mutica, protein electrophoresis, bisalbuminemia, biochemistry, electrolyte