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冈比亚按蚊对常用杀虫剂的抗性及细胞色素 P450 单加氧酶在拟除虫菊酯抗性中的作用。

Resistance of Anopheles gambiae s.s. against commonly used insecticides and implication of cytochrome P450 monooxygenase in resistance to pyrethroids in Lambaréné (Gabon).

机构信息

Centre de Recherches Médicales de Lambaréné, Lambaréné, P.O. Box 242, Gabon.

Institut für Tropenmedizin, Eberhard-Karls-Universität, Wilhelmstrasse 27, Tübingen, P.O. Box 72076, Germany.

出版信息

BMC Infect Dis. 2024 Oct 30;24(1):1221. doi: 10.1186/s12879-024-10021-y.

Abstract

BACKGROUND

Insecticides are a crucial component of vector control. However, resistance constitute a threat on their efficacy and the gains obtained over the years through malaria vector control. In Gabon, little data on phenotypic insecticide resistance in Anopheles vectors are published, compromising the rational implementation of resistance management strategies. We assessed the susceptibility to pyrethroids, carbamates and organophosphates of Anopheles gambiae sensu lato (s.l.) and discuss the mechanisms involved in the pyrethroid resistance-phenotype.

METHODS

A. gambiae s.l. larvae were collected from breeding sites in Lambaréné. Emerging adults were used in WHO tube assays at an insecticide concentration that defines resistance (diagnostic concentration). Subsequently, deltamethrin and permethrin were used at 5x and 10x diagnostic concentrations and after preexposure with the cytochrome p450 (and glutathione S-transferase) inhibitor piperonyl butoxide (PBO). A subset of mosquitoes was typed by molecular methods and screened using Taqman assays for mutations conferring target site resistance at the Voltage-gated sodium channel 1014 (Vgsc-1014) locus and the acetylcholinesterase (Ace-1) gene.

RESULTS

All mosquitoes were A. gambiae sensu stricto (s.s.) and resistant to permethrin, deltamethrin and alphacypermethrin (mortality less than 98%). However, mosquitoes were susceptible to malathion but resistant to bendiocarb. The level of resistance was high for permethrin and at least moderate for deltamethrin. Pre-exposure to PBO significantly increased the mortality of resistant mosquitoes (P < 0.0001). They became fully susceptible to deltamethrin and permethrin-induced mortality increased 4-fold. The G119S Ace-1 resistance allele, which confers resistance to both organophosphates and carbamates, was not present. All sampled mosquitoes were either homozygous for the Vgsc-L1014F or heterozygous for Vgsc-L1014F/L1014S, a marker for resistance to pyrethroids and organochlorides.

CONCLUSION

These findings demonstrate a role of cytochrome P450 monooxygenases in the pyrethroid-resistance of A. gambiae s.s. from Lambaréné. Combining PBO with pyrethroids, as done in second generation bednets, may be used to revert resistance. In addition, malathion could also be used in combination with pyrethroids-based methods for resistance management.

摘要

背景

杀虫剂是病媒控制的重要组成部分。然而,它们的功效和多年来在疟疾病媒控制方面取得的成果受到了抗药性的威胁。在加蓬,有关冈比亚按蚊媒介的表型杀虫剂抗药性的数据很少,这影响了抗药性管理策略的合理实施。我们评估了冈比亚按蚊对拟除虫菊酯、氨基甲酸酯和有机磷的敏感性,并讨论了与拟除虫菊酯抗药性表型相关的机制。

方法

从兰巴雷内的滋生地收集冈比亚按蚊 s.l. 幼虫。将新出现的成虫用于世界卫生组织管试验,使用定义抗药性的杀虫剂浓度(诊断浓度)。随后,用 5 倍和 10 倍诊断浓度的溴氰菊酯和氯菊酯以及预先用细胞色素 P450(和谷胱甘肽 S-转移酶)抑制剂增效醚(PBO)处理后,对其进行测试。一小部分蚊子通过分子方法进行分型,并使用 Taqman 检测针对电压门控钠通道 1014(Vgsc-1014)基因和乙酰胆碱酯酶(Ace-1)基因的靶位抗性的突变。

结果

所有蚊子均为冈比亚按蚊 s.s.,对氯菊酯、溴氰菊酯和高效氯氰菊酯具有抗药性(死亡率低于 98%)。然而,它们对马拉硫磷敏感,但对丙硫克百威有抗药性。对氯菊酯的抗药性水平很高,对溴氰菊酯的抗药性至少是中度的。预先用 PBO 处理显著增加了抗性蚊子的死亡率(P<0.0001)。它们对溴氰菊酯和氯菊酯诱导的死亡率完全敏感,死亡率增加了 4 倍。未发现同时对有机磷和氨基甲酸酯具有抗药性的 G119S Ace-1 抗性等位基因。所有采样的蚊子要么是 Vgsc-L1014F 纯合子,要么是 Vgsc-L1014F/L1014S 杂合子,这是对拟除虫菊酯和有机氯杀虫剂产生抗药性的标志。

结论

这些发现表明细胞色素 P450 单加氧酶在兰巴雷内冈比亚按蚊 s.s. 的拟除虫菊酯抗药性中发挥作用。在第二代蚊帐中,将 PBO 与拟除虫菊酯结合使用,可能有助于逆转抗药性。此外,马拉硫磷也可与基于拟除虫菊酯的方法结合用于抗药性管理。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1e4/11523776/8f9beb13c602/12879_2024_10021_Fig1_HTML.jpg

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