In vitro culture of parasitic stages of Haemonchus contortus = Cultivo in vitro de estádios parasitários de Haemonchus contortus

Niciura, S. C. M., Minho, A. P., McIntyre, J. , Benavides, M. V., Okino, C. H., Esteves, S. N., Chagas, A. C. d. S. and Amarante, A. F. T. d. (2023) In vitro culture of parasitic stages of Haemonchus contortus = Cultivo in vitro de estádios parasitários de Haemonchus contortus. Revista Brasileira de Parasitologia Veterinaria, 32(1), e010122. (doi: 10.1590/s1984-29612023005) (PMID:36651422)

Abstract

Haemonchus contortus is a constraint to sheep production. Seeking to reduce the use of hosts and produce parasitic stages in large-scale, a 42-day in vitro culture protocol of H. contortus third-stage larvae was optimized using Dulbecco’s modified Eagle’s medium (DMEM). In cell-free culture, larvae were maintained at 39.6°C, in acidic media (pH 6.1) for 3 or 6 days with Δ4-dafachronic acid followed by DMEM pH 7.4 supplemented or not with Fildes’ reagent. In DMEM pH 7.4 at 37°C, supplementation with Caco-2 cells was compared to Fildes. On Day 14, fourth-stage larvae (L4) development rates in acidic media supplemented (86.8-88.4%) or not (74.4-77.8%) with Fildes and in Caco-2 cell co-culture (92.6%) were similar, and superior to DMEM pH 7.4 with Fildes (0.0%). On Day 21, Caco-2 cell co-culture resulted in higher larvae differentiation (25.0%) and lower degeneration (13.9%) compared to acidic media (1.5-8.1% and 48.6-69.9%, respectively). This is the first report of prolonged in vitro culture of H. contortus larvae using commercial media in co-culture with Caco-2 cells. Although no progression to the adult stage, Caco-2 cell co-culture resulted in morphological differentiation of H. contortus L4 and larval viability for up to 28 days.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: McIntyre, Dr Jennifer
• Authors: Niciura, S. C. M., Minho, A. P., McIntyre, J., Benavides, M. V., Okino, C. H., Esteves, S. N., Chagas, A. C. d. S., and Amarante, A. F. T. d.
• College/School: College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine
• Journal Name: Revista Brasileira de Parasitologia Veterinaria
• Publisher: Colegio Brasileiro de Parasitologia Veterinaria
• ISSN: 0103-846X
• ISSN (Online): 1984-2961
• Published Online: 13 January 2023
• Copyright Holders: Copyright © 2023 The Authors
• First Published: First published in Revista Brasileira de Parasitologia Veterinaria 32(1): e010122
• Publisher Policy: Reproduced under a Creative Commons License

Project Code	Award No	Project Name	Principal Investigator	Funder's Name	Funder Ref	Lead Dept
190824		The BUG consortium Building Upon the Genome: using H. contortus genomic resources to develop novel interventions to control endemic GI parasites	Eileen Devaney	Biotechnology and Biological Sciences Research Council (BBSRC)	BB/M003949/1	Institute of Biodiversity, Animal Health and Comparative Medicine