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家蠅乙醯膽鹼酯酶活性及其對陶斯松、氧化態陶斯松與安丹之感受性
Activities of the Housefly Acetylcholinesterase and its Susceptibility to Chlorpyrifos, Chlorpyrifos Oxon and Propoxur
摘要
因應農產品農藥殘留快速篩檢需求，農業試驗所應用長期飼養於室內之感性品系家蠅（Musca domestica（Linnaeus, 1758））頭部所純化之乙醯膽鹼酯酶（acetylcholinesterase），於1985年推出可快篩有機磷（organophosphates）及胺基甲酸鹽類（carbamates）殺蟲劑之生化檢測技術。為增進家蠅乙醯膽鹼酯酶之產製效率及評估以其他蠅類生產乙醯膽鹼酯酶之可行性，本研究以長期飼養於室內之感性品系家蠅、東方果實蠅（Bactrocera dorsalis（Hendel, 1912））及瓜實蠅（Zeugodacus cucurbitae（Coquillett, 1899））頭部萃取之乙醯膽鹼酯酶進行測試。比較不同日齡家蠅頭部之乙醯膽鹼酯酶，可知家蠅在3日齡時乙醯膽鹼酯酶比活性（specific activity）最高。雄蠅乙醯膽鹼酯酶比活性顯著高於雌蠅，但兩性間的乙醯膽鹼酯酶總活性差距不大。另比較3種蠅類頭部之乙醯膽鹼酯酶，得知家蠅雄蠅之乙醯膽鹼酯酶比活性最高，為0.925μmol/min/mg protein，為東方果實蠅雄蠅的2.1倍。東方果實蠅雄蠅對陶斯松最敏感，陶斯松對其乙醯膽鹼酯酶活性之半抑制濃度（median inhibitory concentration, IC_(50)）為1.334μg/mL，家蠅雄蠅則略高為1.489μg/mL。顯示3種蠅類中，家蠅最適用來生產乙醯膽鹼酯酶。將家蠅之乙醯膽鹼酯酶進一步以管柱純化，其比活性增高為3.513μmol/min/mg protein，陶斯松（chlorpyrifos）、氧化態陶斯松（chlorpyrifos oxon）及安丹（propoxur）對此純化酵素之IC_(50)則分別下降至0.246、0.051及0.021μg/mL。相較於電鰻（Electrophorus electricus），此家蠅乙醯膽鹼酯酶比活性雖低於電鰻，但其對陶斯松、氧化態陶斯松與安丹之敏感性則遠高於電鰻。可見感性品系家蠅，對藥劑相對較為敏感，用來生產乙醯膽鹼酯酶檢測農產品農藥殘留最為適切。
並列摘要
In Taiwan, acetylcholinesterases (AChE) purified from the heads of a susceptible housefly strain (Musca domestica Linnaeus, 1758) has been used since 1985 for fast screening of cumulative toxicities of organophosphorus and carbamate insecticides in agricultural products by the Taiwan Agricultural Research Institute (TARI). To evaluate the efficiency of AChE of currently available insect colonies, comparative studies using housefly, oriental fruit fly (Bactrocera dorsalis (Hendel, 1912)) and melon fly (Zeugodacus cucurbitae (Coquillett, 1899)) were conducted. The highest AChE specific activity was recorded in three-day-old adult houseflies, and the specific activity of the male fly was higher than that of the female, although the total AChE activity of both male and female housefly was similar. The results of the AChE specific activity of three flies showed that the male housefly retained the highest activity (0.925 μmol/min/mg protein), the AChE from the male oriental fruit fly showed the highest sensitivity against chlorpyrifos, with an IC_(50) of 1.334 μg/mL, second was the male housefly with 1.489 μg/mL. Further purification of the housefly AChE, by column chromatography, increased the specific activity to 3.513 μmol/min/mg protein, and the IC_(50) to chlorpyrifos, chlorpyrifos oxon, and propoxur was decreased to 0.246 μg/mL, 0.051 ng/mL and 0.021 μg/mL, respectively. Although the specific activity of commercial AChE from the electric eel, Electrophorus electricus, is higher than that of the housefly colony in TARI, the AChE sensitivity of the housefly to chlorpyrifos and propoxur was much greater than that from the electric eel. In general, TARI's susceptible housefly strain retains a high sensitivity to insecticides, and is by far the best insect material that is adequate for effective residue control.
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