Background Current mark release recapture methodologies are limited in their ability to address complex problems in vector biology, such as studying multiple groups overlapping in space and time. Additionally, limited mark retention, reduced post-marking survival, and the large effort in marking, collection and recapture, all complicate effective insect tracking. Method We have developed and evaluated a marking method using a fluorescent dye (SmartWater) combined with synthetic DNA tags to informatively and efficiently mark adult mosquitoes using an airbrush pump and nebulizer. Using handheld UV flashlights, the fluorescent marking enabled quick and simple initial detection of recaptures in a simple, field-ready, and non-destructive approach that when combined with a simple, extraction-free PCR on individual mosquito legs provides potentially unlimited marking information. Results This marking, first tested in the laboratory with Anopheles gambiae s.l. mosquitoes, did not affect survival (median ages 24-28 days, p-adj > 0.25), oviposition (median eggs/female of 28.8, 32.5, 33.3 for water, green, red dyes, respectively, p-adj > 0.44), or Plasmodium competence (mean oocysts 5.56 to 10.6, p-adj > 0.95). DNA and fluorescence had 100% retention up to 3 weeks (longest time point tested) with high intensity, indicating marks would persist longer. Conclusions We describe a novel, simple, no/low-impact, and long-lasting marking method that allows separation of multiple insect sub-populations by combining unlimited length and sequence variation in the synthetic DNA tags. This method can be readily deployed in the field for marking multiple groups of mosquitoes or other insects.

中文翻译：

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一种新颖的荧光和DNA组合，可长期复杂地标记蚊子

背景技术当前的标记释放重新捕获方法论解决媒介生物学中复杂问题的能力有限，例如研究时空重叠的多个群体。此外，有限的标记保留，减少的标记后存活以及标记，收集和重新捕获的大量工作，都使有效的昆虫追踪变得复杂。方法我们已经开发并评估了一种使用荧光染料（SmartWater）结合合成DNA标签的标记方法，以使用喷枪泵和雾化器有效，有效地标记成年蚊子。使用手持式紫外线手电筒，荧光标记可以通过一种简单，现场就绪且无损的方法快速，简单地对捕获物进行初始检测，将其与简单，单个蚊子腿上的无萃取PCR提供了潜在的无限标记信息。结果该标记首次在实验室中用冈比亚按蚊蚊子测试，不影响存活（中位年龄24-28天，p-adj> 0.25），产卵（卵中位数/雌性卵为28.8、32.5、33.3，水，绿色） ，红色染料分别表示p-adj> 0.44）或疟原虫能力（平均卵囊5.56至10.6，p-adj> 0.95）。DNA和荧光具有高强度的长达3周（测试的最长时间点）的100％保留，表明标记将持续更长的时间。结论我们描述了一种新颖，简单，无/低影响和持久的标记方法，该方法可以通过在合成DNA标签中结合无限的长度和序列变异来分离多个昆虫亚群。