Three types of vaccine (1) a sonicated extract of killed larvae, (2) 5-fluorouracil-treated larvae and (3) 40 krad. X-irradiated larvae have been evaluated. Pups aged 3-4 months, weighing 2 +/- 0.5 kg and free of natural hookworm infection were selected for experimental studies. The sonicated extract was given either in a single dose or as 2 doses at weekly intervals, whereas the 5-fluorouracil-treated and X-irradiated larvae were given in a single dose only. After completion of the vaccination schedule, the pups were challenged with filariform larvae and were sacrificed 3 weeks after challenge. Non-vaccinated pups and those vaccinated with a single dose of sonicated extract lost weight and had diarrhoea or dysentery after challenge. Furthermore, in non-vaccinated pups, haemoglobin levels fell and at autopsy the intestinal mucosa was found to be grossly congested and free blood was present in the lumen of the intestine. Such findings were not obtained in pups vaccinated with the other types of vaccines. The percentage of infective larvae which developed to the adult stage in non-vaccinated pups was 82.7 +/- 9.53 compared with 45.1 +/- 12.33, 30.1 +/- 10.05, 20.6 +/- 10.60 and 8.0 +/- 7.46%, respectively in pups vaccinated with a single dose of sonicated extract, double dose of sonicated extract, 5-fluorouracil-treated and 40 krad. X-irradiated larval vaccines. While the irradiated larval vaccine provided maximum protection against clinical ancylostomiasis, 2 doses of the sonicated larval vaccine could also afford almost equal protection to pups.