In vitro culture of the parasitic stage larvae of hematophagous parasitic nematode Haemonchus contortus

Lu Liu; Zongshan Zhang; Fuqiang Liu; Hui Liu; Lisha Ye; Feng Liu; Nishith Gupta; Chunqun Wang; Min Hu

doi:10.1016/j.ijpara.2025.01.007

In vitro culture of the parasitic stage larvae of hematophagous parasitic nematode Haemonchus contortus

Int J Parasitol. 2025 Jan 21:S0020-7519(25)00007-4. doi: 10.1016/j.ijpara.2025.01.007. Online ahead of print.

Authors

Lu Liu¹, Zongshan Zhang², Fuqiang Liu³, Hui Liu⁴, Lisha Ye⁵, Feng Liu⁶, Nishith Gupta⁷, Chunqun Wang⁸, Min Hu⁹

Affiliations

¹ National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: [email protected].
² National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: [email protected].
³ National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: [email protected].
⁴ National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: [email protected].
⁵ National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: [email protected].
⁶ National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: [email protected].
⁷ 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. Electronic address: [email protected].
⁸ National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: [email protected].
⁹ National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: [email protected].

PMID: 39848307
DOI: 10.1016/j.ijpara.2025.01.007

Abstract

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.

Keywords: Antioxidants; Defibrinated blood; Haemonchus contortus; Parasitic stage; in vitro culture.