Molecular and morphological characterization of Glossina sp. and identification of associated Trypanosoma sp. in Nigeria including analysis for sialidase activity

These data sets include morphological and molecular data generated from a tsetse (Glossina sp.) collection carried out in Nigeria between 2014 and 2016. Traps were employed for one to three days in four different National Parks (Kainji Lake National Park, Old Oyo National Park, Cross River National Park, Gashaka Gumti National Park), one Game Reserve (Yankari Game Reserve) and two tsetse invested areas (Ijah Gwari, Tuongo). Each tsetse was identified morphologically and dissected in the field, all tissues preserved and used for further analysis. Selected tsetse wings were mounted and photographed to be used for morphometric analysis. The whole gut of each tsetse was homogenized in the field and partially preserved for DNA analysis and used to investigate for sialidase activity with a fluorescent substrate and a natural substrate. Analysis for sialidase activity was done by fluorescent measurement and reverse phase HPLC of processed gut samples. DNA was purified from preserved homogenized gut and used for phylogenetic analysis of tsetse by amplification of Cytochrome Oxidase C SU1 and Internal Transcribed Spacer-1 (ITS1). Presence of trypanosomes was investigated in gut and crudely extracted DNA from proboscis by amplification of ITS1. Identification of the respective trypanosome species was done based on lengths polymorphism of the ITS1 region as well as sequencing of selected ITS1 amplicons and additional glycosomal Glyceraldehyde-3-phosohate dehydrogenase.

这些数据集包含2014至2016年在尼日利亚开展的采采蝇（Glossina sp.）采集工作所产生的形态学与分子数据。在四个国家公园（卡因吉湖国家公园、旧奥约国家公园、克罗斯河国家公园、加沙卡古姆蒂国家公园）、一处野生动物保护区（扬卡里野生动物保护区）及两个采采蝇感染区域（伊贾格瓦里、图翁戈）内，陷阱的布设时长为1至3天。每只采采蝇均通过形态学鉴定后于野外解剖，所有组织均妥善保存以用于后续分析。选定的采采蝇翅膀经装片与拍摄处理，用于形态测量分析（morphometric analysis）。每只采采蝇的完整肠道于野外匀浆，部分样本留存供DNA分析使用，其余则通过荧光底物与天然底物研究唾液酸酶活性（sialidase activity）。唾液酸酶活性分析采用荧光测定法及处理后肠道样本的反相高效液相色谱（reverse phase HPLC）技术。从保存的匀浆肠道中纯化DNA，通过扩增细胞色素氧化酶C亚基1（Cytochrome Oxidase C SU1）与内部转录间隔区1（ITS1）进行采采蝇的系统发育分析（phylogenetic analysis）。通过扩增ITS1，检测肠道及口器粗提DNA中锥虫的存在情况。锥虫物种的鉴定基于ITS1区域的长度多态性（length polymorphism），以及选定ITS1扩增子与额外的糖体甘油醛-3-磷酸脱氢酶（glycosomal Glyceraldehyde-3-phosphate dehydrogenase）的测序结果。