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无患子提取物对埃及伊蚊(双翅目:蚊科)酯酶和磷酸酶的影响。

Effects of extract of soapnut Sapindus emarginatus on esterases and phosphatases of the vector mosquito, Aedes aegypti (Diptera: Culicidae).

机构信息

Laboratory of Pathobiology, Department of Zoology, University of Madras, Guindy Campus, Chennai, India.

出版信息

Acta Trop. 2011 Apr;118(1):27-36. doi: 10.1016/j.actatropica.2011.01.003. Epub 2011 Jan 18.

Abstract

Our earlier investigations with kernels from the soapnut Sapindus emarginatus revealed it as a new source of botanical biocide with potent antimosquito activity, as evident from the proven unique ability of the aqueous kernel extract to kill all the developmental stages of three important vector mosquito species, Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus. This extract was also found to be safe for two non-target aquatic insects. As a sequel to these findings, we have further examined quantitative and qualitative changes in total proteins, esterases, and phosphatases in whole body homogenates of fourth instar larvae and pupae of A. aegypti exposed to this extract at an appropriate threshold time for its lethal effect to gain insights into the impact of the botanical biocide on biochemical characteristics of the target vector mosquito at two distinct developmental stages. The profiles of proteins, esterases (acetylcholinesterse, α- and β-carboxylesterases), and phosphatases (acid and alkaline) exhibited distinct patterns of variation during normal development of fourth instar larvae and pupae, indicating intrinsic difference in biochemical features between these two developmental stages of A. aegypti. Upon exposure of the larvae to the extract, significant reduction in the activities of acetylcholinesterse, β-carboxylesterase, and acid phosphatases were recorded, whereas the total proteins, α-carboxylesterase and alkaline phosphatase activities were unaffected. By contrast, only alkaline phosphatase activity was significantly affected in pupae exposed to the extract. Analysis of these enzymes in native PAGE revealed that they exist in isoforms in both the larvae and pupae. The alterations in the levels of enzymatic activities observed from the quantitative assays of various enzymes were reflected by the respective zymograms with perceptible differences in the intensity and the number of bands detected especially with β-carboxylesterase, acid and alkaline phosphatase activity between the control and exposed test organisms. Despite the fact that the soapnut kernel extract causes mortality of both the larvae and pupae of A. aegypti, the findings of this study demonstrate that the impact of this extract is most pronounced in various enzyme profiles of the larvae rather than the pupae. Such discrepancy implicates the presence of unique biochemical mechanisms in the pupae of mosquito for detoxification of botanical biocides.

摘要

我们之前对肥皂果(Sapindus emarginatus)果仁的研究表明,它是一种具有强大抗蚊活性的新型植物源杀生物剂,这从水提果仁提取物杀死三种重要病媒蚊(埃及伊蚊、致倦库蚊和淡色库蚊)所有发育阶段的独特能力中显而易见。该提取物还被发现对两种非靶标水生昆虫是安全的。作为这些发现的后续,我们进一步研究了暴露于该提取物时埃及伊蚊四龄幼虫和蛹的全身体匀浆中总蛋白、酯酶和磷酸酶的定量和定性变化,以深入了解该植物源杀生物剂对两个不同发育阶段靶标病媒蚊生化特征的影响。在埃及伊蚊四龄幼虫和蛹的正常发育过程中,蛋白质、酯酶(乙酰胆碱酯酶、α-和β-羧酸酯酶)和磷酸酶(酸性和碱性)的图谱显示出明显的变化模式,表明这两个发育阶段的生化特征存在内在差异。幼虫暴露于提取物后,记录到乙酰胆碱酯酶、β-羧酸酯酶和酸性磷酸酶的活性显著降低,而总蛋白、α-羧酸酯酶和碱性磷酸酶的活性不受影响。相比之下,仅暴露于提取物的蛹的碱性磷酸酶活性受到显著影响。在天然 PAGE 分析这些酶时,发现它们在幼虫和蛹中都以同工酶的形式存在。定量分析各种酶的酶活性变化反映在各自的同工酶图谱中,特别是在β-羧酸酯酶、酸性和碱性磷酸酶活性方面,与对照和暴露于测试物的生物体内检测到的带强度和数量存在明显差异。尽管肥皂果仁提取物会导致埃及伊蚊的幼虫和蛹死亡,但本研究的结果表明,该提取物对幼虫各种酶谱的影响比蛹更为明显。这种差异暗示了蚊子蛹中存在独特的生化解毒机制。

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