Iodine deficiency can impair the reproductive performance of livestock and affect perinatal mortality of offspring, yet diagnosis of deficiency is complicated and guidelines for I supplementation are imprecise. We challenged pasture-grazing pregnant ewes with a long-acting I supplement and a goitrogenic forage, then monitored their I status during gestation and lactation and in their lambs from birth to weaning. Approximately 46 d into gestation, 376 ewes were assigned to 6 groups comprising 3 supplementation levels x 2 diet regimens. On d 0 the groups received an intramuscular injection of iodized oil providing 0, 300, or 400 mg of I. They grazed until d 23, then half of each supplementation group were fed brassica kale until d 85, then all groups returned to pasture for lambing (parturition approximately d 99) and remained there until weaning (d 192). Serum total I concentration (STIC) was measured repeatedly in 8 'monitor' ewes per group and in their lambs and in milk sampled postpartum. Severity of goiter was determined as the thyroid-weight:birth-weight (TW:BW) ratio in 82 newborn dead lambs. Mean +/- SE STIC for all ewes was initially 42 +/- 2 (range 24 to 105) mu g/L. Diet did not affect I concentrations in ewe serum or milk. Responses to iodized oil were proportional to dose level; STIC increased to approximately 150 and 240 mu g/L for the 300- and 400-mg I groups and remained greater than 0-mg I groups for 161 d (P < 0.05). Milk contained 26, 271, and 425 mu g I/L for the 0-, 300-, and 400-mg I groups, respectively. Mean STIC of lambs from supplemented ewes did not differ by diet; concentrations for the 300-and 400-mg I groups were 237 and 287 mu g I/L at birth, and by weaning all groups were similar (62 +/- 3 mu g/L). Lamb STIC measured at birth correlated with exposure to I in utero (R-2 = 0.59), which was estimated from the area under the curve (AUC) of ewe STIC measured during the last 99 d of gestation. Thyroid enlargement in lambs affecting the TW: BW ratio was a sensitive indicator of maternal nutrition, being greater with kale feeding (1.27 vs. 0.51 g/kg) and lesser with I supplementation (0.35 vs. 1.44 g/kg). Results support the use of STIC as a biochemical criterion. It was sensitive to the effects of I supplementation with responses in ewes and lambs proportional to dose level and it reflected the relationship between ewe and lamb I metabolism. However STIC did not discriminate between groups of ewes fed pasture vs. goitrogenic forage during pregnancy.