The objective of the present study was to characterize secretion of thymosin alpha 1 (A1) and thymosin beta 4 (B4) during different stages of ovarian function and pregnancy in bovine females. One hundred and thirty-five prepubertal heifers averaging 9 mo of age at the time the study was initiated were used. Estrous detection was conducted twice daily using intact bulls fitted with marking harnesses. Blood samples were collected at first behavioral estrus and at the time of rectal palpation for pregnancy. Heifers were weighed at the beginning of the study and every 56 days and additional blood samples were collected at those times. Plasma was harvested and radioimmunoassays were conducted to quantify concentrations of thymosin A1 and B4. Mean concentrations of thymosin A1 and thymosin B4 were determined during prepuberty, estrus during which conception did not occur, estrus during which conception did occur, pregnancy and in heifers that failed to attain puberty. Mean plasma concentrations of thymosin A1 and B4 were greater at estrus during which conception occurred than at any other period (P less than .01). Regressions of days prepubertal and days post conception on thymosin A1 and B4 were analyzed. During the 100 days prior to puberty, thymosin A1 and B4 did not change in the heifers that ultimately conceived. However, in the heifers that were never determined to be pregnant by rectal palpation, concentrations of both thymosin A1 and B4 increased linearly during prepuberty (P less than .01). A decline in thymosin A1 and B4 concentrations occurred in a quadratic fashion from conception through the first 100 d of pregnancy (P less than .01).(ABSTRACT TRUNCATED AT 250 WORDS)