Cultivation of Haemonchus contortus (Nematoda: Trichostrongylidae) from infective larvae to the adult male and the egg-laying female

The parasitic nematode Haemonchus contortus was cultured in vitro to the adult male and the egg-laying female. Gastric contents from 2 cannulated calves and 2 sheep as well as extracts of stomach mucosa from both hosts were added to a mixture of API-1 culture medium and Fildes' reagent (a pepsin digest of defibrinated bovine blood) adjusted to pH 6.4 for the first week and pH 6.8 thereafter. The gas phase was 85% N2:5% O2:10% CO2. Best results were obtained when API-1 culture medium plus Fildes' reagent was supplemented with ovine gastric contents. Adult males up to 10 mm were obtained, and they produced and stored sperm at about 28 days in culture. Spermatozoa were never seen in the uteri of females grown in vitro. After 36 days in culture, female worms up to 16 mm were obtained which produced and layed eggs. Some of the eggs underwent division up to the 8-cell stage but did not develop further. No live larvae were observed at a stage earlier than the fourth. Presumably, this was because no matings were observed, although large clumps of worms did occur in the culture medium and mating might have occurred. The growth of adults from young to mature phases of this stage included: for the female, oogenesis and laying of eggs; for the male, development of paired sclerotized spicules, and spermatogenesis with the production of spermatozoa.