Molecular Phylogenetics, Diagnostics and Control of Apicomplexan Parasites

我的研究计划的长期目标是探索和理解顶复门的寄生虫和它们的宿主之间的相互作用在历史上，生物体（宿主-寄生虫）和分子水平。这些寄生虫引起重要的动物和人类疾病，如疟疾、弓形虫病、隐孢子虫病和球虫病。我建议通过以下三个主要研究领域来进一步理解这些关系：*1）使用具有形态和形态测量 * 特征的多基因组分子标记来研究顶复门的发育和宿主-寄生虫的共同进化 *：我们将确定人类、家养动物和野生动物中顶复门寄生虫的生物多样性（潜在的人畜共患病）通过生成 * 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）从顶复门寄生虫，感染国内和很大程度上被忽视，野生动物宿主。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，该条形码将用于特异性鉴定球虫 *（艾美球虫属），包括肠球虫（例如艾美球虫）* 和组织球虫（例如弓形虫）*。我们将修改我们的DNA条形码引物，用于多重NGS* 检测，以识别和计数生物样品（如粪便或组织样品）中的混合球虫;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;*2）* 从生物多样性 * 顶复门寄生虫中生成完整的线粒体（mt）基因组，以探索mt基因组的结构进化，以及 * 生成推断这些世界性寄生虫进化历史所需的序列数据。3)家禽球虫免疫控制的探讨。我们将在国内和国际上确定感染火鸡的艾美耳球虫的自然流行率，以确定针对火鸡球虫病的多价活球虫病疫苗所需的物种数量。我们将改进和进一步开发一种重组多价疫苗技术，该技术将两种寄生虫抗原表位与一种宿主衍生的免疫调节表位（HMGB 1）结合起来。* 用于该多价疫苗构建体的当前枯草芽孢杆菌表达载体将被转化为严格基于肽的表达载体（即，化学合成的）形式，并在通过各种方式递送时测试体内生物活性。 使用非活疫苗构建体成功 * 接种艾美球虫属物种（* 全球第一）将解决疫苗生产、运输和保质期 * 问题，并避免在商业家禽中使用活球虫病疫苗 * 的健康影响。

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.**