Temperature dependent impacts of RNA virus population evolution

摘要 节肢动物传播病毒(虫媒病毒)的出现是一个持续的公共卫生问题。这些条件 导致新病毒基因型别出现的原因目前还知之甚少。在蚊子身上，温度 通过复制率和病毒从 并最终传播到中肠。然而，关于外源性孵化的影响，文献中还存在一个空白。 (EI)温度对蚊子体内病毒的多样化有影响。因此，我们计划研究这一角色 温度在虫媒病毒EI期间的进化中起作用。首先，我们将确定EI的程度 温度对寨卡病毒(ZIKV)和西尼罗河病毒(WNV)在伊蚊和库蚊中载体能力的影响 然后使用rna-seq来确定一致的基因组是如何变化的。 对EI温度的响应。接下来，我们将评估温度驱动的单核苷酸变体和 进行种群遗传学分析以确定EI温度对病毒种群的影响程度 多样性。这可能提供了病毒进化的特定温度模型。最后，我们将验证我们的 通过与本地传播的西尼罗河病毒现场分离株进行比较，建立了温度驱动的病毒进化模型。这个 这些研究的结果将为虫媒病毒的进化提供洞察力，并可能被用于预测 可能导致进化和选择新的病毒基因型的环境条件，这些新的病毒基因型会导致严重的 疫情爆发。 假设：我们假设WNV和ZIKV等虫媒病毒在体内经历外源孵化 它们各自的载体、高温孵化驱动了病毒种群的多样性，导致了突变 单倍型正在传递。这一假设将在三个具体目标中得到检验 目的1.确定外源性孵化温度对ZIKV和WNV载体的影响程度 蚊子的能力，并识别任何温度驱动的一致和可预测的共识基因组 改变。 目的2.确定外在孵化温度对ZIKV和WNV种群多样性的影响 在蚊子体内。 目的3.描述2013-2017年间温度对柯林斯堡西尼罗河病毒演变的影响。

Abstract The emergence of arthropod-borne viruses (arboviruses) is an ongoing public health concern. The conditions that lead to the emergence of new virus genotypes are poorly understood. In mosquitoes, temperature profoundly impacts virus transmission via replication rate and the likelihood that virus will disseminate from the midgut and ultimately be transmitted. Yet, there is a gap in the literature on the effect that extrinsic incubation (EI) temperature has on virus diversification in the mosquito. Therefore, we plan to study the role that temperature plays in arbovirus evolution during the EI period. First, we will determine the extent to which EI temperature impacts Zika virus (ZIKV) and West Nile virus (WNV) vector competence in Aedes and Culex mosquitoes respectively, and following this use RNA-seq to identify how the consensus genome changes in response to the temperature of EI. Next, we will assess temperature driven single-nucleotide variants and conduct population genetics analysis to determine the extent of the EI temperature’s effect on viral population diversity. This may provide a temperature specific model of viral evolution. Last, we will validate our temperature driven, viral evolution model by comparing it to locally transmitted WNV field isolates. The outcome of these studies will provide insight to arbovirus evolution and potentially be used to predict environmental conditions that may lead to evolution and selection of novel virus genotypes that cause severe outbreaks. Hypothesis: We hypothesis that when arboviruses such as WNV and ZIKV undergo extrinsic incubation in their respective vector, a high temperature of incubation drives viral population diversity, leading to mutant haplotypes being transmitted. This hypothesis will be tested in three specific aims Aim 1. Determine the extent in which the temperature of extrinsic incubation impacts ZIKV and WNV vector competence in mosquitoes and identify any temperature driven consistent and predictable consensus genome changes. Aim 2. Determine how the temperature of extrinsic incubation influences ZIKV and WNV population diversity within mosquitoes. Aim 3. Characterize the effects of temperature on WNV evolution in Fort Collins from 2013-2017.