Microbial interplay between ZIKA virus and the native microbiome in mosquitoes

ZIKA 病毒与蚊子体内微生物组之间的微生物相互作用

• 批准号: 9726225
• 负责人: Grant Leslie Hughes
• 金额: $ 13.5万
• 依托单位: TMLIVERPOOL SCHOOL OF TROPICAL MEDICINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9726225
• 关键词: Aedes; Affect; African; Alphavirus; Antibiotics; Antiviral Agents; Arbovirus Infections; Arboviruses; Area; Arthropods; Asians; Attention; Bacteria; Biological Assay; Biology; Chikungunya virus; Competence; Complex; Culicidae; Data; Dengue Virus; Disease; Flavivirus; Foundations; Future; Generations; Gnotobiotic; High-Throughput Nucleotide Sequencing; Individual; Inherited; Invaded; Investigation; Investigative Reports; Knowledge; Laboratories; Mediating; Medical; Microbe; Modeling; Mosquito-borne infectious disease; Oral; Phenotype; Property; Role; Seminal; Shapes; Supplementation; Techniques; Technology; Variant; Viral; Viral Physiology; Virus Diseases; West Nile virus; Work; Yellow fever virus; ZIKV infection; Zika Virus; arbovirus disease; bacteriome; base; experimental study; fungus; gut microbes; insight; microbial; microbiome; microbiome composition; microbiota; microorganism; microorganism interaction; mycobiome; novel; offspring; pathogen; symbiont; vector; viral transmission

Project summary. The native microbiota of mosquitoes profoundly influences many aspects of mosquito biology, including the ability of mosquitoes to transmit arboviral pathogens. Perturbation of the mosquito microbiome, either by supplementation of antibiotics or by oral inoculation of bacterial isolates into the mosquito gut can interfere with flaviruses such as Dengue virus (DENV) and West Nile Virus as well as alphaviruses like Chikungyunya (CHIKV). Most studies indicate that gut microbes are antagonistic to pathogens, but particular bacteria facilitate viral infection in mosquitoes. Additionally, we are beginning to appreciate that arbovirus infection in mosquitoes can alter the bacterial microbiome. Taken together, it is clear that there is complex interplay, either by direct or indirect mechanisms, between native gut microbes of mosquitoes and invading arbovirus, and these interactions have a profound effect on vector competence. Here we will characterize the interplay between gut microbes of Aedes mosquitoes and Zika virus to determine how these interactions influence ZIKV vector competence in mosquitoes. Our work will focus on both bacterial and fungal taxa of Aedes aegypti and Aedes albopictus. While it is emerging that bacterial symbionts alter vector competence, we have a poor understanding of the role of fungal microbes on mosquito biology and vector competence. Our work will exploit high throughput sequencing technologies to obtain a culture-independent quantitative characterization of the bacterial and fungal microbiome. Importantly, experiments will use both lab-reared and field-collected mosquitoes as our appreciation of the role of the microbiome of on vector competence of mosquitoes in the field is poorly understood. In specific aim 1, we will identify bacterial and fungal taxa that alter vector competence by high throughput sequencing comparing perturbed and conventionally reared mosquitoes. Using field-collected mosquitoes, ZIKV vector competence will be correlated to microbial abundance and composition. As the composition of the microbiome is variable, the presence or abundance of specific symbionts may account for variation in arbovirus vector competence. ZIKV vector competence assays will be completed on gnotobiotic mosquito lines that have had isolates reinfected, functionally validating candidate taxa identified from our sequencing experiments. In specific aim 2, we will examine how ZIKV infection alters the bacterial and fungal microbiome using high throughput sequence. We will determine if microbes that ZIKV inhibits have anti-viral properties and examine if ZIKV-mediated modulation of the microbiome is transferred between generations. Lastly, we will determine if such transgeneration alteration of the micobiome affects vector competence of subsequent generations to ZIKV. This work will provide insights in microbial interactions within mosquito, a poorly understudied but important area of vector biology that has promising applications to control ZIKV and other mosquito-borne viral disease. This project will lay the foundation for future work in the Hughes laboratory, which is to develop applied novel microbial-based strategies suitable for simultaneously controlling multiple arboviruses such as ZIKV, DENV, CHIKV, and yellow fever virus.

项目摘要。 蚊子的天然微生物群深刻影响蚊子生物学的许多方面，包括 蚊子传播丁香病毒病原体。通过补充蚊子微生物组的扰动 抗生素或通过口服接种细菌分离株进入蚊子肠道会干扰登革热等黄病毒 病毒（DENV）和西尼罗河病毒以及Chikungyunya（Chikv）等α。大多数研究表明肠道 微生物对病原体是拮抗的，但是特定的细菌促进了蚊子中的病毒感染。另外，我们是 开始欣赏蚊子中的arbovirus感染会改变细菌微生物组。两者一起，是 清楚地表明，通过直接或间接机制存在复杂的相互作用，在天然肠道微生物之间 和入侵的Arbovirus，这些相互作用对向量能力产生了深远的影响。在这里，我们将表征 埃德斯蚊子和寨卡病毒的肠道微生物之间的相互作用，以确定这些相互作用如何影响zikv 蚊子的矢量能力。我们的工作将重点放在埃及伊德斯和艾德斯的细菌和真菌分类群上 白化病。尽管细菌共生体改变了向量能力，但我们对 真菌微生物在蚊子生物学和载体能力上的作用。我们的工作将利用高通量测序 获得细菌和真菌微生物组的独立定量表征的技术。 重要的是，实验将使用实验室奔跑和现场收集的蚊子作为我们对该作用的欣赏 蚊子在田间的载体能力的微生物组知之甚少。在特定目标1中，我们将确定 细菌和真菌分类群，通过比较扰动和 常规饲养的蚊子。使用田间收集的蚊子，ZIKV矢量能力将与 微生物丰度和成分。由于微生物组的组成是可变的，因此存在或丰度 特定的共生体可以解释Arbovirus矢量能力的变化。 zikv矢量能力分析将是 在已重新感染分离株的gnotobiotic蚊子线上完成，在功能上验证了候选类群 从我们的测序实验。在特定目标2中，我们将研究ZIKV感染如何改变细菌和真菌 使用高吞吐量序列的微生物组。我们将确定ZIKV抑制的微生物是否具有抗病毒特性 并检查微生物组的ZIKV介导的调制是否在世代之间传递。最后，我们会的 确定MICOBIOME的这种转变改变是否影响后代的向量能力 zikv。这项工作将为蚊子内的微生物相互作用提供见解，这是一个研究不足但重要的领域 载体生物学的应用，可用于控制ZIKV和其他蚊子传播病毒疾病。这个项目 将为休斯实验室的未来工作奠定基础，该实验室将开发应用新颖的基于微生物 适用于同时控制多个Arbovirus的策略，例如ZIKV，DENV，CHIKV和黄热病 病毒。