Sex and culture duration differences in Ascaridia galli Extracellular Vesicles

To determine how to obtain the highest amount of EVs shed by A. galli, several kinetic experiments were conducted. Cleaned and sexed worms were grouped according to size. For both females and males, medium-sized worms (females: 5.7-6.8 cm; males: 3.7-4.8 cm) produced the most particles per mL, indicating that EV production depends on life stage.
The amount of EVs shed by A. galli was relatively stable across replicates, though the optimal culture duration for maximal EV yield differed between sexes. Female worms produced the highest average particle yield after 24 hours of culture, with no significant difference compared to 48 hours. Male worms shed the highest number of EVs after 40 hours, but again, no significant difference was found between timepoints.
No morphological differences were observed between EV fractions from different timepoints or worm sexes. Following size-exclusion chromatography, EVs from females were mainly detected in fractions 5-8, with the highest yield in fraction 6. In contrast, male worms produced significantly fewer EVs overall, and none were detected in fraction 8.
The size distribution of EVs did not differ between sexes, with most particles ranging from 100 to 200 nm.

为明确如何获取鸡蛔虫（A. galli）分泌的最多细胞外囊泡（Extracellular Vesicles，EVs），本研究开展了一系列动力学实验。将经过清洗并完成性别鉴定的虫体按体型大小分组。结果显示，雌雄两性的中型虫体（雌性：5.7~6.8 cm；雄性：3.7~4.8 cm）每毫升培养液中产生的囊泡数量最多，表明细胞外囊泡（EVs）的产生依赖于虫体发育阶段。
不同生物学重复中，鸡蛔虫分泌的细胞外囊泡总量相对稳定，但实现最大囊泡产量的最佳培养时长存在性别差异：雌性虫体在培养24小时后平均颗粒产量最高，且与培养48小时的结果无显著差异；雄性虫体在培养40小时时分泌的细胞外囊泡数量最多，但不同时间点的产量同样无显著差异。
不同培养时间点或不同性别的虫体所分泌的细胞外囊泡组分未观察到形态学差异。经尺寸排阻色谱分离后，雌性虫体的细胞外囊泡主要分布于组分5~8中，其中组分6的囊泡产量最高。与之相反，雄性虫体整体分泌的细胞外囊泡数量显著更少，且未在组分8中检测到囊泡。
不同性别的虫体所分泌的细胞外囊泡粒径分布无差异，绝大多数颗粒的粒径介于100~200 nm之间。