is a genus of nematodes, which infects a variety of hosts, principally dogs and cats, with potential zoonotic risks to humans. spp. larvae are capable of migrating throughout the host tissues, eliciting eosinophilic and granulomatous reactions, while surviving for extended periods of time, unchanged, in the host. It is postulated that larvae are capable of altering the host's immune response through the release of excretory-secretory products, containing both proteins and extracellular vesicles (EVs). The study of EVs has increased exponentially in recent years, largely due to their potential use as a diagnostic tool, and in molecular therapy. To this end, there have been multiple isolation methods described for the study of EVs. Here, we use nanoparticle tracking to compare the yield, size distribution, and % labelling of EV samples acquired through various reported methods, from larval cultures of and containing excretory-secretory products (TES). The methods tested include ultracentrifugation, polymer precipitation, magnetic immunoprecipitation, size exclusion chromatography, and ultrafiltration. Based on these findings, ultrafiltration produces the best results in terms of yield, expected particle size, and % labelling of sample. Transmission electron microscopy confirmed the presence of EVs with characteristic cup-shaped morphology. These findings can serve as a guide for those investigating EVs, particularly those released from multicellular organisms, such as helminths, for which few comparative analyses have been performed.

中文翻译：

---

用于研究犬弓蛔虫和猫弓蛔虫排泄分泌 (TES) 产品内外泌体货物的细胞外囊泡分离方法的比较


是一种线虫属，可感染多种宿主，主要是狗和猫，对人类具有潜在的人畜共患风险。种。幼虫能够在宿主组织中迁移，引起嗜酸性和肉芽肿反应，同时在宿主体内存活较长时间，保持不变。据推测，幼虫能够通过释放含有蛋白质和细胞外囊泡（EV）的排泄分泌产物来改变宿主的免疫反应。近年来，对 EV 的研究呈指数级增长，这主要是由于它们作为诊断工具和分子治疗的潜在用途。为此，已经描述了多种用于电动汽车研究的隔离方法。在这里，我们使用纳米颗粒追踪来比较通过各种报道的方法从排泄分泌产物 (TES) 的幼虫培养物中获取的 EV 样品的产量、尺寸分布和标记百分比。测试的方法包括超速离心、聚合物沉淀、磁免疫沉淀、尺寸排阻色谱和超滤。根据这些发现，超滤在产量、预期粒径和样品标记百分比方面产生最佳结果。透射电子显微镜证实了具有特征杯形形态的 EV 的存在。这些发现可以为那些研究电动汽车的人提供指导，特别是那些从多细胞生物（例如蠕虫）释放的电动汽车，对此很少进行比较分析。