Introduction method

Trichostrongylus colubriformis is a common, widely distributed, nematode parasite of the small intestine in ruminants. In previous attempts to grow advanced stages of T. colubriformis from infective larvae in vitro a few became young adult males and females (Douvres, 1980). This occurred in a two-step, roller culture system that consisted of complex cell-free media, RFN-2 (Step 1) and API-16, with appropriate additions of a reducing agent and pepsin (Step 2).

Nelson & Douvres (1984) reported that Trichostrongylus colubriformis from guinea pigs and sheep: survival and yield of advanced stages that developed from larvae in fourth stage and fourth molt in a roller culture system using medium RPMI 1640.In vivo-grown 7-8-day-old young adult males and females did not grow or develop further in vitro.

From previous attempts to grow advanced stages of Trichostrongylus colubriformis in vitro , from infective larvae which had varying degrees of success . We hypothesize that when Trichostrongylus colubriformis L3 larvae are cultured in an in vitro system that mimics the intestinal niche of the sheep that the parasite will develop into L4 and adult stages.

The primary objective of the present study was to improve the existing cultivation system that enhance the growth and development of the nematode (Haemonchus Contortus and Trichostrogylus colubriformis ).

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