Cysteine proteinase enriched fractions obtained by thiol-sepharose chromatography of Ostertagia ostertagi membrane-bound protein extract (S3-thiol) or total adult excretory-secretory (ES-thiol) products were tested in a vaccination experiment to evaluate their protective efficacy against O. ostertagi in cattle. Calves were vaccinated three times and subsequently challenged with a trickled infection of 25,000 infective larvae in total over 25 days (1000 L3/day, 5 days/week). Geometric mean cumulative egg counts in the ES-thiol group were reduced by 60% during the 2-month period between the first challenge infection and necropsy, compared to the control group (P < 0.002). No reduction in egg output was observed in the S3-thiol group. At necropsy, calves immunized with ES-thiol had a significantly higher percentage of inhibited L4 larvae (9.8%) and had in total 18% less worms than the control calves, but this reduction was not statistically significant. Both the female and male adult worms were significantly smaller in the ES-thiol group than in the control group. Although no significant difference was observed in the number of eggs per female worm between the groups, there was a trend to less eggs per female worm in the ES-thiol group. Number of worms, size of adult worms and number of eggs per female worm were not significantly different between the S3-thiol group and the control group. Systemic immunization with QuilA as adjuvant induced a significant rise in Ostertagia-specific antibody levels in the abomasal mucosa. Ostertagia-specific local antibody levels showed a significant negative correlation with the size of the adult worms, the number of eggs per female worm and the cumulative faecal egg counts. However, these correlations were quite weak and did not appear to be isotype-specific.