Global eukaryotic parasite diversity of platypus and echidna

The short beaked echidna (Tachyglossus aculeatus) and the platypus (Ornithorhynchus anatinus) are iconic egg-laying monotremes (Monotremata: Mammalia) in Australia. Recognition and understanding the importance of parasites such as apicomplexa (Eimeria, Theileria, Babesia, Hepatozoon) in monotreme’s health and disease is limited by the paucity of material to study. Using small subunit (18S) rDNA amplicon deep-sequencing we demonstrated presence of apicomplexan parasites confirmed by direct and cloned amplicon gene sequencing to characterise Theileria ornithorhynchi, Theileria tachyglossi, Eimeria echidnae and Cryptosporidium fayeri. Deep sequencing complements and supports traditional diagnostic methods including microscopy and histopathology to better define the status quo and is particularly suitable for wildlife disease investigation.短喙针鼹（Tachyglossus aculeatus）与鸭嘴兽（Ornithorhynchus anatinus）是澳大利亚极具标志性的卵生哺乳纲（Mammalia）单孔目（Monotremata）动物。目前学界对顶复门（Apicomplexa）寄生虫（包括艾美耳属（Eimeria）、泰勒虫属（Theileria）、巴贝斯虫属（Babesia）、肝簇虫属（Hepatozoon））在单孔目动物健康与疾病中的重要性的认知与理解，受限于研究材料的匮乏。本研究通过小亚基（18S）rDNA扩增子深度测序技术，并结合直接扩增子与克隆扩增子基因测序验证，确认了顶复门寄生虫的存在，成功鉴定出鸭嘴兽泰勒虫（Theileria ornithorhynchi）、针鼹泰勒虫（Theileria tachyglossi）、针鼹艾美耳球虫（Eimeria echidnae）以及法氏隐孢子虫（Cryptosporidium fayeri）。深度测序可作为包括显微镜检查与组织病理学在内的传统诊断方法的补充与支撑，能够更好地明确当前研究现状，尤其适用于野生动物疾病调查。