Non-invasive approaches to mosquito-borne pathogen surveillance using excreta

项目摘要 蚊子传播的疾病正在全球迅速蔓延。新近爆发的西方疫情 尼罗河、登革热、基孔肯雅和寨卡病毒已使数亿人患病，而 疟疾等寄生虫每年导致40多万人死亡。有能力的媒介及其传播 病原体传播到新的大陆和气候，加上最近在全球范围内的快速扩张 其他感染人类的新型RNA病毒强调了快速和可靠的必要性 病原体监测技术比以往任何时候都要先进。及早发现可确保病媒 控制努力将在正确的时间指向正确的地点，以避免潜在的流行病。 监测和检测受感染的蚊子通常是即将到来的早期预测因素 然而，人类感染伴随着挑战，包括需要时间来分类、识别和 从大量蚊子中提取RNA，同时保持冷链。这需要 在病毒检测之前对蚊子进行监测、鉴定和汇集。它最近一直在 显示传播感染的蚊子排泄(作为排泄物/粪便)虫媒病毒和 在实验室采样时适合检测的浓度的寄生虫。这允许快速 和无损采样和病原体检测，然而，电流场捕捉设备是 无法有效收集随机分布的约1.5微米L排泄物液滴。要解决这个问题 针对这一问题，我们建议设计两个新的收集排泄物的蚊子捕捉器来寻找寄主 和产卵(或怀孕)的蚊子，分别利用糖喂食站和 由纳米结构的超疏水表面组成的定制的收集系统 过滤并浓缩排泄物样本。适当的疏水表面构象将 首先在最初的田间试验中进行筛选和药效测试。然后这个原型将被按比例调整 设置为诱捕野生蚊子种群，同时疏导和聚集排泄物 在诱捕器内生产到外部连接的标准微离心管，以快速和 轻松收集。为了测试这两种设备有效提取潜在病原体的能力 诱捕蚊子，诱捕器将部署在新泽西州的五个县，跨越一个 与县蚊子控制机构合作，建立大量蚊子栖息地。这个 捕获的排泄物将接受变态基因组测序，以识别和组装基因组 来自样本中已知的和潜在的新的RNA病毒的全部数据， 代表了新泽西蚊子的广泛多样性。这个项目代表着一个基础的 迈向对病媒传播病原体的哨兵“无蚊子”监测，这将有助于 在地方性疾病暴发发生前作出快速反应以减轻其影响。

Project Summary Mosquito-borne diseases are expanding rapidly across the globe. Recent outbreaks of West Nile, dengue, chikungunya and Zika viruses have sickened hundreds of millions of people, while parasites such as malaria kill over 400,000 annually. The spread of competent vectors and their pathogens to new continents and climates, coupled with the recent rapid global expansion of other novel RNA viruses that infect humans, emphasizes the need for rapid and reliable pathogen surveillance techniques now more than ever. Early detection ensures that vector control efforts are directed to the right location at the right time to avert a potential epidemic. Surveillance and detection of infected mosquitoes is often an early predictor of impending human infection however comes with challenges, including the time involved to sort, identify and extract RNA from large numbers of mosquitoes while maintaining a cold chain. This requires that mosquito surveillance, identification, and pooling precede virus testing. It has recently been shown that mosquitoes with a disseminated infection excrete (as excreta/feces) arbovirus and parasites in concentrations suitable for detection when sampled in the lab. This allows for rapid and non-destructive sampling and pathogen testing, however current field trapping devices are unable to efficiently collect the randomly distributed ca. 1.5 µL excreta droplets. To address this issue, we propose here to design two novel excreta-collecting mosquito traps for host-seeking and ovipositing (or gravid) mosquitoes, respectively, that utilize sugar feeding stations and a custom fabricated collection system composed of a nanostructured superhydrophobic surface to funnel and condense the excreta sample. An appropriate hydrophobic surface conformation will first be selected and tested for efficacy during initial field trials. This prototype will then be scaled up and configured to trap wild mosquito populations while channeling and aggregating excreta produced within the trap to a standard microcentrifuge tube attached externally for quick and easy collection. To test the ability of both devices to effectively sample potential pathogens from trapped mosquitoes, the traps will be deployed across five New Jersey counties spanning a multitude of mosquito habitat in collaboration with county mosquito control agencies. The captured excreta will be subject to metavirome sequencing to identify and assemble genome data from the full breadth of both known and potentially novel RNA viruses in the samples, representing a broad diversity of New Jersey mosquitoes. This project represents a foundational step towards sentinel “mosquito free” surveillance of vector-borne pathogens that will facilitate rapid response to mitigate enzootic outbreaks before they occur.