比較生物檢測法及生化分析法在埃及斑蚊抗藥性之研究

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標題:	比較生物檢測法及生化分析法在埃及斑蚊抗藥性之研究 Bioassays and Biochemical Analyses of Insecticide Resistance in Southern Taiwan Aedes aegypti (L.)
作者:	曾宏傑
貢獻者:	嘉南藥理科技大學：生物科技系暨研究所羅怡珮
關鍵字:	埃及斑蚊抗藥性生化分析高百滅寧耐受性生物檢測 Aedes aegypti detoxification enzymes biochemical assays bioassay insecticide resistance
日期:	2009
上傳時間:	2011-10-27 14:43:09 (UTC+8)
摘要:	本實驗以苓雅、左營、前鎮、鼓山、鹽埕、旗津、台南市東區、台南市中西區、台南市南區、台南市北區、台南市安平區、永康等12個地區及NS、Bora Bora 等2個敏感性品系埃及斑蚊，進行百滅寧、賽滅寧、第滅寧、安丹及撲滅松等殺蟲劑對埃及斑蚊雌蚊的生物檢測。結果顯示供試12個地區品系埃及斑蚊對於除蟲菊酯殺蟲劑具高度抗藥性，其中苓雅品系對百滅寧的抗性比為100.11，鼓山品系對賽滅寧的抗性比為229.99，鼓山品系對第滅寧的抗性比為73.89。各品系對氨基甲酸鹽劑及有機磷劑的抗性情形較不明顯，苓雅品系對安丹抗性比為5.61，台南市安平區對撲滅松的抗性比為4.12。以供試12個地區及2個敏感性品系之單隻雌蚊測定??#37238;、β酯酶、GST、單加氧酶、乙醯膽鹼酯酶等五種酵素活性，並進行生化分析與生物檢測之相關性分析。結果顯示??#37238;、β酯酶及單加氧酶3種酵素活性與百滅寧及安丹生物檢測之半致死濃度有關。??#37238;、β酯酶、GST及乙醯膽鹼酯酶4種酵素活性與賽滅寧及撲滅松生物檢測之半致死濃度有關。??#37238;、β酯酶、GST、單加氧酶及乙醯膽鹼酯酶等5種酵素活性與第滅寧生物檢測之半致死濃度有關。將各殺蟲劑之半致死濃度與相關之酵素活性進行多元迴歸分析，判定係數r2介於0.086～0.322之間。以多元迴歸計算式預估各地區埃及斑蚊對殺蟲劑的半致死濃度，計算預估值與實際值的差異百分比。各地區品系埃及斑蚊對百滅寧半致死濃度的平均差異百分比為40.23%，賽滅寧半致死濃度的平均差異百分比為69.91%，第滅寧半致死濃度的平均差異百分比為30.54 %，安丹半致死濃度的平均差異百分比為37.38%，撲滅松半致死濃度的平均差異百分比為45.34%。在統計上是屬於符合預期的範圍。以不同濃度百滅寧藥膜處理苓雅、左營、前鎮、鼓山等四個品系埃及斑蚊，並測定24小時後存活蚊體的??#37238;、β酯酶、GST、單加氧酶等4種酵素活性。結果顯示在測試四個品系中，隨著百滅寧藥膜濃度的增加，存活蚊體的酵素活性有增加的趨勢。以酵素活性與百滅寧藥膜處理濃度進行相關性分析，顯示供試四個品系中β酯酶、GST及單加氧酶等三種解毒酵素與埃及斑蚊對百滅寧的抗性有關。苓雅品系以單加氧酶相關性最高為0.544，左營品系及鼓山品系以GST的相關性最高，分別為0.443及0.448。前鎮品系以??#37238;的相關性最高為0.362。因此推測在不同的品系間，由於殺蟲劑對各品系的選汰壓力不同，埃及斑蚊對百滅寧產生的抗性機制可能有所差異。 Aedes aegypti, one of the primary vectors of dengue fever in the southern of Taiwan, was evaluated for the susceptibility of the 3 major insecticides (organophosphates, pyrethroids, and carbamate) by bioassay and biochemical assay. The susceptibilities of field strains of Adeds aegypti adult females to permethrin, cypermethrin, deltamethrin, propoxur and fenitrothion were investigated and compared with two susceptible laboratory strains (NS strain and Bora Bora strain) by using a filter paper (12 X 15 cm) impregnation method. The hightest resistance ratio between the 50% lethal concentration value (RR) of the field strains and the NS strain shows that the RR for permethrin was 100.11(Lingya strain), the RR for cypermethrin was 229.99(Gushan strain), the RR for deltamethrin was 73.89(Gushan strain), the RR for propoxur was 5.61(Lingya strain), and the RR for fenitrothion was 4.12(Anping Dist strain). Adult bioassays showed widespread, significant resistance to pyrethroid insecticides. Microplate assays were performed to measure levels of insensitive acetylcholinesterase, ?esterase, β-esterase, glutathione-S-transferase, monooxygenase enzymes. Comparison between bioassays and biochemical assays, the higher levels ?esterase activity, β-esterase activity and monooxygenase activity were significant correlation with the 50% lethal concentration value of permethrin and propoxur, the higher levels ?esterase activity, β-esterase activity, glutathione-S-transferase activity and insensitive acetylcholinesterase activity were significant correlation with the 50% lethal concentration value of cypermethrin and fenitrothion, the higher levels ?esterase activity, β-esterase activity, glutathione-S-transferase activity, monooxygenase activity and insensitive acetylcholinesterase activity were significant correlation with the 50% lethal concentration value of deltamethrin. The simultaneous multiple regression analysis was to establish an ideal formula for bioassays and biochemical assays to estimate the 50% lethal concentration value of insecticides. The formulas of permethrin, propoxur and deltamethrin were satisfied, and the formulas of cypermethrin and fenitrothion had to modified to increase the accuracy. The effect of exposure of Aedes aegypti adult to different concentration of permethrin impregnation filter paper, the higher concentration exposure treatment survival adults had the elevated levels of detoxification enzyme activity. Mechanism of permethrin resistance of Aedes aegypti involving β-esterase activity, glutathione-S-transferase activity, monooxygenase activity were apparent.
關聯:	校內校外均不公開，學年度：97， 104 頁
顯示於類別:	[生物科技系(所)] 博碩士論文

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