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.

项目总结。 蚊子的本地微生物区系深刻地影响着蚊子生物学的许多方面，包括 蚊子传播虫媒病毒病原体。对蚊子微生物群的干扰，要么是通过补充 抗生素或向蚊子肠道内口服接种细菌分离株可干扰登革热等闪光病毒 病毒(DENV)和西尼罗河病毒(West Nile Virus)以及基孔庆雅(Chikunya)等甲型病毒(CHIKV)。大多数研究表明，肠道 微生物对病原体具有拮抗作用，但特定的细菌促进了蚊子的病毒感染。此外，我们正在 开始意识到蚊子中的虫媒病毒感染可以改变细菌的微生物群。综上所述，它是 很明显，蚊子的肠道微生物之间存在着复杂的相互作用，无论是直接的还是间接的机制。 和入侵的虫媒病毒，这些相互作用对媒介能力有深远的影响。在这里，我们将描述 伊蚊肠道微生物与寨卡病毒的相互作用以确定这些相互作用如何影响寨卡病毒 蚊子的媒介能力。我们的工作将集中在埃及伊蚊和伊蚊的细菌和真菌分类群上 白纹伊蚊。虽然出现了细菌共生体改变媒介能力的现象，但我们对 真菌微生物在蚊子生物学和媒介能力中的作用。我们的工作将利用高通量测序 获得细菌和真菌微生物群的独立于培养的定量特征的技术。 重要的是，实验将使用实验室饲养和野外采集的蚊子作为我们对 关于蚊子在野外传播媒介能力的微生物群还知之甚少。在具体目标1中，我们将确定 通过比较扰动和高通量测序改变载体能力的细菌和真菌分类 按常规饲养蚊子。利用现场采集的蚊子，ZIKV媒介能力将与 微生物丰度和组成。由于微生物组的组成是可变的，所以存在或丰富的 特定的共生体可能解释了虫媒病毒载体能力的变化。ZIKV载体能力分析将是 在已有分离物再次感染的灵知生物蚊子品系上完成，从功能上验证已确定的候选分类群 从我们的测序实验中。在特定的目标2中，我们将研究ZIKV感染如何改变细菌和真菌 使用高通量序列的微生物组。我们将确定ZIKV抑制的微生物是否具有抗病毒特性 并检查ZIKV介导的微生物组的调制是否在世代之间转移。最后，我们将 确定这种微生物体的转化改变是否会影响后续世代的载体能力 齐科夫。这项工作将对蚊子内的微生物相互作用提供洞察力，这是一个研究较少但很重要的领域 媒介生物学在控制寨卡病毒和其他蚊媒传播的病毒疾病方面具有很好的应用前景。这个项目 将为休斯实验室未来的工作奠定基础，即开发适用的新型微生物基础 适用于同时控制多种虫媒病毒的策略，如ZIKV、DENV、CHIKV和黄热病 病毒。