臺灣博碩士論文加值系統

吳郭魚為台灣重要的養殖漁業，屬於熱帶魚種，適於高溫養殖，近年來受到寒害和魚類病毒感染的影響，造成死亡率的提高。魚體內的血糖和乳酸濃度的變化是生理狀態異常的一個有用的指標，可以從濃度變化來確定魚體的生理狀態。藉此想出一套植入式魚體酵素電極之無線感測裝置。
本研究利用酵素法方式，製作了探針式酵素燃料電池和酵素感測針，主要針對吳郭魚體內的血糖和乳酸來進行發電和量測。針式酵素燃料電池利用葡萄糖去氫酶(Glucose dehydrogenase, GDH)與膽紅素氧化酶(Bilirubin oxidase, BOD)分別作為陽極與陰極的生物觸媒，建構出利用葡萄糖和氧氣為反應物之酵素燃料電池，植入吳郭魚體內進行發電量的測量與分析，量測出在22℃水溫下，最大電流密度約在41 (µA/cm^2)、開路電壓約為0.41V、最大功率密度為6.3(µW/cm^2)左右；改變水溫溫度至15℃，量測出最大電流密度上升到了52(µA/ cm^2)、最大功率密度則是8.6(µW/cm^2)，可以看出溫度的降低，吳郭魚的血糖會上升。
為了證實此無線感測針能夠確實應用於吳郭魚生理量測，本研究的葡萄糖酵素感測針是利用了葡萄糖去氫酶作為工作電極的催化酵素；乳酸酵素感測針則是乳酸氧化酶為催化酵素，使用GoSense無線恆電位儀做為無線接收器，進行降低水溫和施打鏈球菌的量測。在降低水溫(20℃至15℃)的量測下，可以看出血糖的變化從40mg/dl上升到90mg/dl，乳酸則是從20mg/dl下降至10mg/dl，反映在緊迫狀況下，是為了應付環境衝擊而生理調節所需能量之需求量增加所致。
施打足以致死的鏈球菌量(2×10^5cfu/g)，先進行24小時正常狀況下的量測，而後施打鏈球菌於腹腔中，在往後的24小時中觀察到吳郭魚的血糖由正常狀態下的41mg/dl上升到了92mg/dl，身體會產生對抗外界壓力的荷爾蒙，荷爾蒙會讓身體的胰島素需求增加；乳酸由正常狀態下的20mg/dl上升到了39mg/dl，在感染到高濃度的病菌時，使細胞內發生缺氧代謝，乳酸生成增加，肝臟的功能減弱，無法使乳酸回到正常值。

關鍵詞：吳郭魚、葡萄糖酵素、乳酸酵素、血液、鏈球菌、電化學分析測量儀、無線恆電位儀

Tilapia, an economically important cultured fish in Taiwan, plays a significant role in the fisheries development. It has less tolerable attributes of cold in winter and common bacterial infections in summer. Measurements of the variation of fish blood glucose and lactate concentration is a useful index to observe the change of physiological conditions.
In this study, we adopted enzyme method to developed enzymatic biofuel cells and mediator-type biosensor. Enzymatic biofuel cell that consists of a needle bioanode used for glucose dehydrogenase (GDH) and a gas-diffusion biocathode used for bilirubin oxidase (BOD). The assembled device for glucose oxidation was inserted into Tilapia, producing maximum current density 41 (µA/cm^2),open voltage 0.41V, maximum power density 6.3(µW/cm^2) at 22℃ and maximum current density 52 (µA/cm^2), maximum power density 8.6(µW/cm^2) at 15℃ in free-swimming fish
in the aquarium.
We developed a wireless biosensor system to monitor glucose and lactate concentration in Tilapia. The biosensor was used Pt-Ir wire as the working electrode and Ag/AgCl as the reference electrode. Glucose dehydrogenase and lactate oxidase were immobilized on the working electrode to be glucose biosensor and lactate biosensor respectively. The sensor was inserted into Tilapia in order to wirelessly monitor the glucose and lactate concentration in free-swimming fish.We confirmed that blood glucose levels increased from 40mg/dl to 90mg/dl and lactate levels decreased from 20mg/dl to 10mg/dl in the blood when the temperatures dropped from 20℃to 15℃ in the aquarium. We also monitored the concentrations of glucose and lactate in Tilapia when it was infected by Streptococcosis. The results confirmed that blood glucose concentration increased from 41mg/dl to 92mg/dl and lactate concentration increased from 20mg/dl to 39mg/dl in the blood. The concentration from the sensor gradually increased and decreased during the application of stress, which hinted that the stress was monitored by this system.



Keywords: Tilapia, Glucose enzyme, Lactate enzyme, Blood, Streptococcosis , Electrochemical analyzer , Wireless potential

摘要 I
Abstract II
目錄 III
圖目錄 V
表目錄 VII
第一章 緒論 1
1.1 前言 1
1.2研究背景 1
1.2-1酵素 1
1.2-2 生物燃料電池 2
1.2-3生物型感測器 3
1.2-4 吳郭魚(Tilapia) 4
1.2-5 細菌感染 5
第二章 文獻回顧 7
2.1 酵素燃料電池系統 7
2.2 酵素燃料電池充電之應用 12
2.3 酵素感測器之相關研究 17
2.4 研究動機與目的 23
第三章 實驗原理 25
3.1 酵素催化反應 25
3.2 電化學量測原理 25
第四章 實驗設計與架設 27
4.1感測系統之機台 27
4.1-1電化學CHI分析儀 27
4.1-2 GoSense無線恆電位儀 30
4.2探針式酵素燃料電池的製作 32
4.3酵素感測針的製作 33
4.3-1酵素感測針結構 33
4.3-2酵素感測針酵素固定 34
4.4 探針式酵素燃料電池植入魚體 36
4.5酵素感測針植入和鏈球菌施打 38
第五章 結果與討論 40
5.1 探針式酵素燃料電池效能分析 40
5.2 酵素感測針之效能分析 42
5.3量測環境溫度變化對吳郭魚的生理變化44
5.4量測感染鏈球菌之吳郭魚的生理變化 45
第六章 結論與未來展望 46
6.1 結論 46
6.2 未來展望 46
參考文獻 47

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