In vitro culture of parasitic stages of Haemonchus contortus

Author(s)

Niciura, Simone Cristina Meo ; Minho, Alessandro Pelegrine ; McIntyre, Jennifer ; Benavides, Magda Vieira ; Okino, Cintia Hiromi ; Esteves, Sergio Novita ; Chagas, Ana Carolina de Souza ; do Amarante, Alessandro Francisco Talamini

• Document type: Journal article
• Source: REVISTA BRASILEIRA DE PARASITOLOGIA VETERINARIA; v. 32, n. 1, p. 12-pg., 2023-01-01.

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 degrees C, in acidic media (pH 6.1) for 3 or 6 days with Delta 4-dafachronic acid followed by DMEM pH 7.4 supplemented or not with Fildes' reagent. In DMEM pH 7.4 at 37 degrees 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. (AU)

• FAPESP's process: 19/02967-2 - Development of methods to study Haemonchus contortus and anthelmintic resistance to monepantel
• Grantee: Simone Cristina Méo Niciura
• Support Opportunities: Regular Research Grants