Is it a Fluke or is it Reproducible: Understanding Isolate Variation in Fasciola hepatica for Diagnostic Potential

大约55%的家畜疾病是由寄生虫引起的，对全球粮食安全和食源性疾病构成重大威胁，由于缺乏疫苗，只能通过驱虫药物进行控制。特别是，由肝吸虫引起的人畜共患片吸虫病对牲畜生产和福利产生深远的负面影响，造成死亡或慢性消耗性疾病和易患细菌性疾病，每年在全世界造成约30亿美元的损失。由于依赖三氯苯达唑（TCBZ）作为一线驱虫药，控制工作因TCBZ耐药性的出现而受到阻碍。为了促进改进控制，现在迫切需要新的诊断方法，能够区分对TCBZ耐药的肝吸虫和对TCBZ敏感的肝吸虫。新的肝吸虫分离物对TCBZ既敏感又耐药，现在可以利用后基因组技术探索产生下一代肝吸虫诊断方法。因此，本项目旨在采用多组学方法揭示肝吸虫分离物的特异性特征，这些特征可用于TCBZ分化诊断，并支持分离物TCBZ状态的确认。本项目的目标是：1)利用蛋白质组学技术对肝吸虫分离物的躯体蛋白、排泄分泌蛋白和细胞外囊泡蛋白进行指纹图谱分析；2)对肝吸虫分离物的转录组学图谱进行序列变异分析；3)优化液体AP (MA)LDI以支持分离物的快速分化；4)开发一种实验性笔侧诊断测试。这项工作将为支持未来肝吸虫TCBZ耐药/易感性诊断奠定重要基础。学生将获得广泛的分子和寄生虫学方法的经验和培训，包括生物化学，分子生物学和体外寄生虫维持。这也将包括细胞外囊泡的纯化和分析。此外，将提供多组学技术和生物信息学方面的培训，以分析大规模组学数据集。此外，学生将获得分析质谱和生物分子纯化的经验。这名学生还将在英国兽医服务公司Ridgeway Research Ltd工作，在那里他们将接受寄生虫生命周期管理方面的培训。学生还将在与利益相关者（当地农民和农民工会等）沟通项目目标和进展方面发挥核心作用。

Helminth parasites are responsible for >55% of livestock diseases representing a major threat to global food security and food borne disease, with control being through anthelmintic drugs due to an absence of vaccines. In particular, zoonotic fascioliasis, caused by liver fluke, has a profound, negative impact on livestock production and welfare, causing death or chronic wasting disease and predisposition to bacterial diseases with losses of ~$US3 billion/annum worldwide. With a reliance on triclabendazole (TCBZ) as the frontline anthelmintic drug, control has been hampered by the emergence of TCBZ resistance. To facilitate improved control, there is now an urgent need for new diagnostics capable of differentiating between liver fluke that are TCBZ resistant and those that are TCBZ susceptible. New liver fluke isolates, that are both susceptible and resistant to TCBZ, are now available to explore using post genomic technologies to generate the next generation of liver fluke diagnostics. Therefore, this project aims to employ a multiomic approach to reveal liver fluke isolate specific profiles that can be exploited for TCBZ differentiation diagnostics and to support the confirmation of isolate TCBZ status. This project will aim to 1) Utilise proteomics to fingerprint liver fluke isolate somatic1, excretory-secretory and extracellular vesicle2 proteins 2) Transcriptome profile liver fluke isolates for sequence variations 3) Optimize Liquid AP (MA)LDI to Support Rapid Isolate Differentiation and 4) develop a pilot pen-side diagnostic test. This work will importantly lay the foundations to support future TCBZ resistance/susceptibility diagnostics for the liver fluke.The student will gain experience and training in a wide range of molecular and parasitological methods including biochemistry, molecular biology and in vitro parasite maintenance. This will also include purification and analysis of extracellular vesicles. In addition, training will be provided in multiomics technologies and bioinformatics for the analysis of large-scale omic datasets. Furthermore, the student will gain experience in analytical mass spectrometry and biomolecule purification. The student will also spend time with Ridgeway Research Ltd, a UK-based veterinary service firm, where they will receive training in parasite life cycle management. The student will also play a central role in communicating project goals and progress with stakeholders (local farmers and farmers unions etc).