| 摘要: | 本實驗開發新的多孔人工餵血系統，已成功做為埃及斑蚊繼代繁殖的餵血器，這個多孔人工餵血裝置使用牛膠原蛋白製備的人工腸衣，非常適合做為餵血器的膜系統。比較埃及斑蚊於多孔人工餵血系統對豬全血不添加或添加ATP的吸血率分別為89.7%及90.7%，與對活體小鼠的吸血率 (98.0%) 不具顯著差異，使用人工腸衣及石蠟膜對埃及斑蚊的吸血率不具顯著影響。本研究發現埃及斑蚊取食活體小鼠血液及豬全血，比較其吸血率，存活率、產卵率、子代卵孵化率、化蛹率及羽化率均不具顯著差異。建立人工餵血系統可減少實驗動物的耗損並符合動物保護的原則。本研究的多孔人工餵血系統也可應用做為防蚊液的檢測平台，開發一個含有鹽類、白蛋白、葡萄糖、腺嘌呤核?三磷酸即可誘引供試雌蚊吸血的人工替代血液，埃及斑蚊取食人工替代血液的吸血率與豬血間不具差異性，且吸血率顯著高於現在慣用的CDPA-1替代血液，以人工餵血系統進行檢測防蚊液的檢測，可避免實驗人員感染病媒傳播疾病的風險，也可減少實驗人員測試未知天然化合物造成過敏或中毒的風險。本系統可進行定量檢測，DEET 、香茅醛、香荊芥酚、香葉醇、丁子香酚及百里酚的 ID50 分別為 1.62 、 14.40 、 22.51 、23.29、 23.83 及 68.05 μg/cm2， DEET 的抑制效果明顯比各精油成分顯著。以多孔餵血裝置檢測植物精油的防蚊效果，以丁子香油的抑制吸血效果最佳，其次依序為百里香油、天龍香茅油及尤加利精油，貓薄荷精油抑制吸血的效果最差。以多孔餵血裝置檢測 DEET 的有效保護時間，一般市售防蚊液產品的 DEET含量為 7%，經檢測發現在第 3 個小時的吸血抑制率達 97.1%，第 5 小時降為 56.6%，此方法客觀準確，可降低進行人體皮膚測試時因手臂進出網籠或流汗造成的影響。關鍵字:埃及斑蚊、餵血器、in vitro 忌避劑生物檢測、植物精油。
A novel multiple membrane blood-feeding system for mosquitoes has been developed for the study and routine maintenance of Aedes aegypti L. for mosquitoes that normally require vertebrate blood meal to produce eggs. This blood-feeding system uses cattle collagen sausage casing membrane to facilitate feeding. The efficiency of this blood-feeding system was compared to a live mice blood source. We observed that Ae. aegypti that fed on pig whole blood had 89.7% (w/o ATP) and 90.7% (w/ ATP) blood feeding rates, which were not significantly different from the mice-fed ones (98.0%). The feeding rates did not differ between sausage casing membrane and Parafilm-M?. The survival rate, fecundity, pupation, and pupal emergence rates of Aedes females fed on pig whole blood were not significantly different from the mice-fed ones. The artificial blood feeder can be applied to replace live animals as blood sources. An artificial blood meal has been developed for feeding Aedes aegypti (L.), a mosquito that normally requires a vertebrate blood meal for engorgement. The artificial blood meal is a simple mixture with salt, albumin, dextrose, and adenosine triphosphate added to induce engorging. The feeding rates of Aedes aegypti on the artificial blood meal were not significantly different from those on pig blood, but the feeding rates were significantly higher than on CPDA-1 solutions. Because most assays currently rely on human subjects or vertebrate whole blood, an efficient and safe in vitro assay using a artificial blood meal and a multiple-membrane blood-feeding device could facilitate the development of a significantly simplified screening system for possible novel repellents. The dose-dependent biting inhibition rates were analyzed by probit analysis. The ID50 (the dose producing 50% inhibition of mosquito feeding) values of DEET, citronella, carvacrol, geraniol, eugenol and thymol were 1.62, 14.40, 22.51, 23.29, 23.83 and 68.05 μg/cm2, respectively.Keywords: Aedes agypti, blood feeder, in vitro repellent bioassay, essential oils。 |
