title: Development of a versatile in vitro method for understanding the migration of Fasciola hepatica newly excysted juveniles.
abstract: Fasciola hepatica is a parasitic trematode that causes serious losses to livestock producers, and also zoonotic disease. The limitations of chemotherapy for the control of fasciolosis have led to significant interest in the development of vaccines to protect cattle and sheep from infection. However, relatively few studies have concentrated on the mechanisms of invasion of the gut by newly excysted juvenile liver flukes (NEJ) and the host response triggered by this event. The aim of this work was to develop an in vitro model to study invasion by NEJ, while also reducing the requirement for challenge infections of experimental animals. Fasciola hepatica metacercariae were excysted in vitro and placed into compartments containing rat distal jejunal sheets. Variations in incubation medium, chamber size and incubation temperature were used to identify optimal conditions for NEJ migration across the gut. Histological examination showed increased migration until 120 min post-incubation. The use of RPMI, without gassing at 39 °C, as the incubation medium was found to be optimal, with 40·5% of NEJ migrating after 150 min. This study describes a readily-reproducible method for studying the migration of F. hepatica NEJ within the definitive host. It will be useful for identifying potential drug and vaccine targets.
authors.affiliation: School of Veterinary Medicine,University College Dublin,Veterinary Sciences Centre,Belfield,Dublin 4,Ireland.
authors.forename: Andres
authors.initials: A
authors.lastname: Garcia-Campos
authors.affiliation: School of Veterinary Medicine,University College Dublin,Veterinary Sciences Centre,Belfield,Dublin 4,Ireland.
authors.forename: Alan W
authors.initials: AW
authors.lastname: Baird
authors.affiliation: School of Veterinary Medicine,University College Dublin,Veterinary Sciences Centre,Belfield,Dublin 4,Ireland.
authors.forename: Grace
authors.initials: G
authors.lastname: Mulcahy
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000469:parasitology*; D004195:Disease Models, Animal; D005210:Fasciola hepatica / Q000502:physiology*; D005211:Fascioliasis / Q000469:parasitology / Q000662:veterinary*; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D012756:Sheep; D012757:Sheep Diseases / Q000469:parasitology*
keywords: Fasciola hepatica; NEJ; gut; horizontal diffusion chamber; jejunum; method optimization