Feeding and Quantifying Animal-Derived Blood and Artificial Meals in Aedes aegypti Mosquitoes.
Feeding and Quantifying Animal-Derived Blood and Artificial Meals in Aedes aegypti Mosquitoes.
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DOI：
10.3791/61835
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发表时间：
2020-10-22
期刊：
影响因子：
--
通讯作者：
Duvall LB
中科院分区：
文献类型：
--
作者：
Jové V;Venkataraman K;Gabel TM;Duvall LB
某些种类的雌性蚊子在叮咬脊椎动物宿主以获得卵发育所需的富含蛋白质的血餐时会传播疾病。在实验室中，研究人员可以通过膜喂食器向蚊子提供动物源性和人造血粉，这允许操纵膳食成分。在这里，我们提出的方法，用于喂养血液和人工血餐埃及伊蚊和量化的体积消耗个别女性。人工/血粉的定向喂养和定量具有广泛的用途，包括测试粉组分对蚊子行为和生理的影响，在不注射的情况下递送药理化合物，以及用特定病原体感染蚊子。在喂食前向膳食中添加荧光素染料允许随后的膳食量定量。蚊子消耗的食物体积可以通过重量来测量，如果雌性稍后用于行为实验，或者通过将个体雌性置于96孔板中并使用板读取器测量荧光水平作为终点测定来测量。膳食大小量化可用于确定改变膳食成分是否会改变摄入的膳食体积，或者不同蚊子品系之间的膳食消耗是否不同。精确的膳食量定量对于下游检测也至关重要，例如测量对宿主吸引力或繁殖力的影响。本文提出的方法可以进一步适用于在几天的过程中跟踪膳食消化，或者包括添加到不同膳食（如花蜜和血液）中的多个可区分的标记物，以量化单个蚊子对每餐的消耗。这些方法使研究人员能够单独进行高通量测量，以比较数百只蚊子的进食量。因此，这些工具将对蚊子研究人员回答各种生物学问题广泛有用。
Females of certain mosquito species can spread diseases while biting vertebrate hosts to obtain protein-rich blood meals required for egg development. In the laboratory, researchers can deliver animal-derived and artificial blood meals to mosquitoes via membrane feeders, which allow for manipulation of meal composition. Here, we present methods for feeding blood and artificial blood meals to Aedes aegypti mosquitoes and quantifying the volume consumed by individual females. Targeted feeding and quantification of artificial/blood meals have broad uses, including testing the effects of meal components on mosquito behavior and physiology, delivering pharmacological compounds without injection, and infecting mosquitoes with specific pathogens. Adding fluorescein dye to the meal prior to feeding allows for subsequent meal size quantification. The meal volume consumed by mosquitoes can be measured either by weight, if the females are to be used later for behavioral experiments, or by homogenizing individual females in 96-well plates and measuring fluorescence levels using a plate reader as an endpoint assay. Meal size quantification can be used to determine whether changing the meal components alters the meal volume ingested or if meal consumption differs between mosquito strains. Precise meal size quantification is also critical for downstream assays, such as those measuring effects on host attraction or fecundity. The methods presented here can be further adapted to track meal digestion over the course of days or to include multiple distinguishable markers added to different meals (like nectar and blood) to quantify the consumption of each meal by a single mosquito. These methods allow researchers to singlehandedly perform high-throughput measurements to compare the meal volume consumed by hundreds of individual mosquitoes. These tools will therefore be broadly useful to the community of mosquito researchers for answering diverse biological questions.
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影响因子：
4.6
作者：
Gonzales KK;Rodriguez SD;Chung HN;Kowalski M;Vulcan J;Moore EL;Li Y;Willette SM;Kandel Y;Van Voorhies WA;Holguin FO;Hanley KA;Hansen IA
通讯作者：
Hansen IA
影响因子：
16.2
作者：
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通讯作者：
Vosshall, Leslie B.
影响因子：
3.8
作者：
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通讯作者：
Scott TW
影响因子：
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作者：
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通讯作者：
Abeyewickreme, Wimaladharma
影响因子：
6.4
作者：
Guerbois, Mathilde;Fernandez-Salas, Ildefonso;Weaver, Scott C.
通讯作者：
Weaver, Scott C.