优化直接膜喂食法中斑须按蚊种群蚊子的喂食率。
Optimization of the feeding rate of Anopheles farauti s.s. colony mosquitoes in direct membrane feeding assays.
机构信息
Papua New Guinea Institute of Medical Research, Madang, Papua New Guinea.
Australian Institute of Tropical Health and Medicine, James Cook University, Smithfield, QLD, 4870, Australia.
出版信息
Parasit Vectors. 2021 Jul 7;14(1):356. doi: 10.1186/s13071-021-04842-y.
BACKGROUND
Direct membrane feeding assays (DMFA) are an important tool to study parasite transmission to mosquitoes. Mosquito feeding rates in these artificial systems require optimization, as there are a number of factors that potentially influence the feeding rates and there are no standardized methods that apply to all anopheline species.
METHODS
A range of parameters prior to and during direct membrane feeding (DMF) were evaluated for their impact on Anopheles farauti sensu stricto feeding rates, including the starving conditions and duration of starving prior to feeding, membrane type, DMF exposure time, mosquito age, feeding in the light versus the dark, blood volume, mosquito density and temperature of water bath.
RESULTS
The average successful DMFA feeding rate for An. farauti s.s. colony mosquitoes increased from 50 to 85% when assay parameters were varied. Overnight starvation and Baudruche membrane yielded the highest feeding rates but rates were also affected by blood volume in the feeder and the mosquito density in the feeding cups. Availability of water during the pre-feed starvation period did not significantly impact feeding rates, nor did the exposure duration to blood in membrane feeders, the age of mosquitoes (3, 5 and 7 days post-emergence), feeding in the light versus the dark, or the temperature (34 °C, 38 °C, 42 °C and 46 °C) of the water bath.
CONCLUSION
Optimal feeding conditions in An. farauti s.s. DMFA were to offer 50 female mosquitoes in a cup (with a total surface area of ~ 340 cm with 1 mosquito/6.8 cm) that were starved overnight 350-500 µL of blood (collected in heparin-coated Vacutainer tubes) per feeder in feeders with a surface area ~ 5 cm (with a maximum capacity of 1.5 mL of blood) via a Baudruche membrane, for at least 10-20 min.
背景
直接膜喂食法（DMFA）是研究寄生虫传播给蚊子的重要工具。在这些人工系统中，需要优化蚊子的喂食率，因为有许多因素可能影响喂食率，而且没有适用于所有按蚊种的标准化方法。
方法
在直接膜喂食（DMF）之前和期间，评估了一系列参数对按蚊远缘亚种（Anopheles farauti sensu stricto）喂食率的影响，包括饥饿条件和饥饿前的持续时间、膜的类型、DMF 暴露时间、蚊子的年龄、在黑暗中和在光线下喂食、血量、蚊子密度和水浴温度。
结果
当改变实验参数时，按蚊远缘亚种（An. farauti s.s.）群体蚊子的平均成功 DMFA 喂食率从 50%增加到 85%。过夜饥饿和 Baudruche 膜产生了最高的喂食率，但喂食率也受到喂食器中血量和喂食杯中蚊子密度的影响。在预喂食饥饿期间水的可用性对喂食率没有显著影响，膜喂食器中血液暴露时间、蚊子的年龄（孵化后 3、5 和 7 天）、在黑暗中和在光线下喂食、或水浴温度（34°C、38°C、42°C 和 46°C）也没有影响。
结论
在按蚊远缘亚种 DMFA 中，最佳的喂食条件是在一个杯子中提供 50 只雌性蚊子（总表面积约为 340 cm²，每只蚊子 6.8 cm²），在饥饿 350-500 µL 血液（收集在肝素涂层的 Vacutainer 管中）的情况下，饥饿过夜，每台喂食器的表面积约为 5 cm²（最大容量为 1.5 mL 血液），通过 Baudruche 膜，至少喂食 10-20 分钟。
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