Liu Lu, Zhang Zongshan, Liu Fuqiang, Liu Hui, Ye Lisha, Liu Feng, Gupta Nishith, Wang Chunqun, Hu Min
National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070 China.
Intracellular Parasite Education and Research Labs (iPEARL), Department of Biological Sciences, Birla Institute of Technology and Science, Pilani (BITS-Pilani), Hyderabad, India; Department of Molecular Parasitology, Faculty of Life Sciences, Humboldt University, Berlin, Germany.
Int J Parasitol. 2025 Apr;55(5):263-271. doi: 10.1016/j.ijpara.2025.01.007. Epub 2025 Jan 21.
Current research on common parasitic nematodes is limited because their infective stages cannot be propagated in vitro. Here, we report a culture system for developing L4s of Haemonchus contortus, a blood-feeding nematode of ruminants. Our results demonstrated that a proportionate mixture of NCTC-109 to Luria-Bertini (1:2) media promoted the formation of early L4s and then into late L4s upon inclusion of 12.5% (v/v) defibrinated blood, albeit with a decline in survival. Adding antioxidants (0.3 mg/mL of L-glutathione or 200 nmol of vitamin C) improved survival of L4s, with approximately 90% developing to late L4s by 22 days. These L4s showed parallel morphological features (such as digestive and reproduction systems) compared with in vivo L4s at day 7 (following challenge infection), although with delayed development. Our work optimized the in vitro culture system for L4s while providing an important platform for in-depth molecular research on Haemonchus and other related parasitic nematodes.
目前对常见寄生线虫的研究有限，因为它们的感染阶段无法在体外繁殖。在此，我们报告了一种用于培养捻转血矛线虫（一种反刍动物的吸血线虫）L4期幼虫的培养系统。我们的结果表明，NCTC - 109培养基与吕氏培养基按比例混合（1:2），加入12.5%（v/v）去纤维蛋白血后可促进早期L4期幼虫的形成，进而发育为晚期L4期幼虫，尽管存活率有所下降。添加抗氧化剂（0.3 mg/mL的L - 谷胱甘肽或200 nmol的维生素C）可提高L4期幼虫的存活率，约90%的幼虫在22天内发育为晚期L4期幼虫。这些L4期幼虫与感染后第7天的体内L4期幼虫相比，具有相似的形态特征（如消化系统和生殖系统），尽管发育有所延迟。我们的工作优化了L4期幼虫的体外培养系统，同时为深入开展血矛线虫及其他相关寄生线虫的分子研究提供了重要平台。
