Molecular Phylogenetics, Diagnostics and Control of Apicomplexan Parasites

顶复门寄生虫的分子系统发育、诊断和控制

• 批准号: RGPIN-2016-05826
• 负责人: Barta, John
• 金额: $ 2.4万
• 依托单位: University of Guelph
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2016
• 资助国家: 加拿大
• 起止时间: 2016-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=615925
• 关键词: Molecular; Phylogenetics; Diagnostics; Control; Apicomplexan

The long term goal of my research program is to explore and understand the interactions between parasites of the phylum Apicomplexa and their hosts at the historical, organismal (host-parasite) and molecular levels. These parasites cause important animal and human diseases such as malaria, toxoplasmosis, cryptosporidiosis and coccidiosis. I propose to further our understanding of these relationships by pursuing three main areas of study: 1) Investigate apicomplexan phylogeny and host-parasite coevolution using multi genome molecular markers with morphological and morphometric characters: We will determine the biodiversity of apicomplexan parasites in humans, domesticated animals and wildlife (potential zoonotics) by generating sequence data (nuclear 18S rDNA, plastid rDNA/polymerase B, and mt cytochrome c oxidase subunit I, COI) from apicomplexan parasites that infect both domestic and, largely ignored, wildlife hosts. Our newly developed PCR primer pairs will generate species-specific COI-based DNA barcodes that will be used to identify specifically coccidia (Eimeriorina) including enteric (e.g. Eimeria) and tissue (e.g. Toxoplasma) coccidia. We will modify our DNA barcode primers for use in a multiplexed NGS assay to both identify and enumerate mixed coccidia in biological samples such as fecal or tissue samples; this method will permit rapid protistan parasite biodiversity and species discovery; 2) Generate complete mitochondrial (mt) genomes from biologically diverse apicomplexan parasites to explore the structural evolution of mt genomes and generate sequence data necessary for inferring the evolutionary history of these cosmopolitan parasites. 3) Explore immunological control of coccidia of domestic poultry. We will determine the natural prevalence of Eimeria that infect domestic turkeys nationally and internationally to determine the number of species required in a multivalent live coccidiosis vaccine against turkey coccidiosis. We will refine and further develop a recombinant multivalent vaccine technology that combines two parasite antigenic epitopes with a host-derived immune modulating epitope (HMGB1). The current Bacillus subtilis expression vector for this multivalent vaccine construct will be converted into a strictly peptide based (i.e. chemically synthesized) form and tested for biological activity in vivo when delivered by various means. Successful vaccination against Eimeria species using a non-viable vaccine construct (a global first) would address vaccine production, transportation and shelf life issues as well as avoid health implications of using live coccidiosis vaccines in commercial poultry.

的 我的研究计划的长期目标是探索和理解之间的相互作用 在历史上， 生物体（宿主-寄生虫）和分子水平。这些寄生虫导致 重要的动物和人类疾病如疟疾，弓形体病， 隐孢子虫病和球虫病。我建议进一步了解 这些关系通过追求三个主要研究领域： 1)研究顶复门的进化和寄主-寄生虫的协同进化 使用多基因组分子标记， 特征：我们将确定顶复门寄生虫的生物多样性， 人类、家养动物和野生动物（潜在的人畜共患病）， 序列数据（核18 S rDNA、质体rDNA/聚合酶B和mt细胞色素c氧化酶亚基I，COI）来自 感染家养动物和野生动物的顶复体寄生虫， hosts.我们新开发的PCR引物对将产生物种特异性 基于COI的DNA条形码将用于特异性识别球虫 （艾美球虫属），包括肠道（例如艾美球虫属） 和组织（例如弓形虫） 球虫我们将修改我们的DNA条形码引物用于多重NGS 鉴定和计数生物样品中混合球虫的测定 作为粪便或组织样本;这种方法将允许快速原生生物寄生虫 生物多样性和物种发现; （二） 从生物多样性中生成完整的线粒体（mt）基因组 apicomplexan寄生虫，以探索mt基因组的结构进化， 生成推断进化历史所需的序列数据， 这些世界性的寄生虫 3)家禽球虫免疫控制的探讨。我们 将决定 感染火鸡艾美耳球虫自然流行 在国家和国际范围内， 抗火鸡球虫病多价活疫苗。我们将改进并进一步开发一种 多价疫苗技术，其将两种寄生虫抗原表位与 宿主来源的免疫调节表位（HMGB 1）。 目前用于该多价表达载体的枯草芽孢杆菌表达载体是 疫苗构建体将被转化为严格基于肽的（即， 化学合成的）形式，并在体内测试生物活性， 通过各种方式交付。成功 使用非活疫苗构建体（a）接种抗艾美耳球虫属物种的疫苗 全球第一）将解决疫苗生产、运输和保质期问题 以及避免使用活球虫疫苗对健康的影响 商业家禽。