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寄生虫的门Apicomplexa和他们的主机在历史上，有机体（主机寄生虫）和分子水平之间的相互作用。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）利用具有形态学和形态计量学特征的多基因组分子标记来研究顶复门的发生和宿主-寄生虫的协同进化：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），它们来自寄生于家养宿主和被忽视的野生宿主的复形寄生虫。我们新开发的PCR引物对将产生基于物种特异性COI的DNA条形码，其将用于特异性地鉴定球虫（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的原生生物寄生虫生物多样性和物种发现; 2）生成完整线粒体（mt）从生物多样性的apicomplexan寄生虫的基因组，探索mt基因组的结构进化，并产生必要的序列数据，推断进化历史，3）探讨家禽球虫的免疫防治。我们将确定感染国内火鸡的艾美耳球虫在国内和国际上的自然流行率，以确定抗火鸡球虫病的多价活球虫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源性免疫调节表位（HMGB 1）相结合。目前用于这种多价疫苗构建体的枯草杆菌表达载体将被转化为严格基于肽的（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。

Thelong term goal of my research program is to explore and understand the interactions betweenparasites of the phylum Apicomplexa and their hosts at the historical,organismal (host-parasite) and molecular levels. These parasites causeimportant animal and human diseases such as malaria, toxoplasmosis,cryptosporidiosis and coccidiosis. I propose to further our understanding ofthese relationships by pursuing three main areas of study:1) Investigate apicomplexan phylogeny and host-parasite coevolutionusing multi genome molecular markers with morphological and morphometriccharacters: We will determine the biodiversity of apicomplexan parasites inhumans, domesticated animals and wildlife (potential zoonotics) by generatingsequence data (nuclear 18S rDNA, plastid rDNA/polymerase B, and mt cytochrome c oxidase subunit I, COI) fromapicomplexan parasites that infect both domestic and, largely ignored, wildlifehosts. Our newly developed PCR primer pairs will generate species-specificCOI-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 NGSassay to both identify and enumerate mixed coccidia in biological samples suchas fecal or tissue samples; this method will permit rapid protistan parasitebiodiversity and species discovery;2)Generate complete mitochondrial (mt) genomes from biologically diverseapicomplexan parasites to explore the structural evolution of mt genomes andgenerate sequence data necessary for inferring the evolutionary history ofthese cosmopolitan parasites.3) Explore immunological control of coccidia of domestic poultry. Wewill determinethe natural prevalence of Eimeria that infect domestic turkeysnationally and internationally to determine the number of species required in amultivalent live coccidiosis vaccine against turkey coccidiosis. We will refine and further develop a recombinantmultivalent vaccine technology that combines two parasite antigenic epitopes with ahost-derived immune modulating epitope (HMGB1). The current Bacillus subtilis expression vector for this multivalentvaccine construct will be converted into a strictly peptide based (i.e.chemically synthesized) form and tested for biological activity in vivo whendelivered by various means.