Levamisole (LEV) was tested in four experiments to compare efficacy values against Ostertagia ostertagi when larval maturation was occurring (September), following inhibition and also when populations were expected to be largely adult (winter). A primary objective was to determine the importance of developing fourth-stage larvae (DL4) and inhibited, early fourth-stage larvae (EL4) in replacing adult worms lost through treatment and the effect of this on reduced efficacy against adult worms. Young crossbred beef calves ranging in weight from 150 to 230 kg were used in the first (September 1981), second (September 1983) and third experiments (January 1987). Jersey calves of 110 kg average weight were used in the fourth experiment (December 1988). Calves were randomized to groups according to weight and group sizes ranged from three to five calves. All parasite infections were naturally acquired, but a mixture of nematode third-stage larvae (L3) (22 500 per calf), including 20% Ostertagia ostertagi, was inoculated into Jersey calves of Experiment 4 following a 2 week exposure to natural infection. All LEV treatments were by subcutaneous injection at dosages of 6 and 8 mg kg-1. Treatment with ivermectin was used only in Experiment 3-as an efficacy reference. All calves were killed at 8-10 days after treatment. The efficacy of LEV against all developmental stages of Ostertagia ostertagi was consistently low in all experiments and a dose-dependent response was not evident. Large numbers of all Ostertagia ostertagi developmental stages were present in non-treated calves in both September experiments. Percent reduction of adults, DL4 and EL4 at the 6 mg kg-1 and 8 mg kg-1 dosages, respectively, were adults, 51.7 and 23.6 (1981), 8.7 and 51.3 (1983); DL4 40.3 and 13.2 (1981), 37.9 and 33.1 (1983); EL4, 19.6 and 0 (1981), 59.6 and 42.9 (1983). Smaller numbers of Ostertagia ostertagi were present in winter experiments and adult worms greatly outnumbered larval stages. Percent reductions of adults, DL4 and EL4, respectively, were (1987) LEV 6 mg kg-1, 40.2, 0 and 0; ivermectin 200-mu-g kg-1, 98.7, 97.7 and 100.0; (1988) LEV 6 mg kg-1, 62.4, 100.0 and 100.0; LEV 8 mg kg-1, 49.1 65.0 and 74.1. Too few larval stages were present in the latter experiment for valid efficacy values. Data from September experiments suggested that DL4 and EL4 could have replaced adults lost through LEV treatment based on progressive maturation of larval stages in September, slightly reduced numbers of larval stages in treated calves compared to controls, particularly in 1981, and the increased proportion of young female worms without eggs in LEV-treated calves in Experiment 2. The consistent low activity of LEV against all developmental stages in both September and winter suggested some degree of LEV resistance across all stages, but this was not proven in the present experiments.