Two experiments were conducted to determine the effects of immunoneutralization of growth hormone-releasing factor [GRF(1-29)-NH 2 ] on concentrations of somatotropin (ST) and insulin-like growth factor I (IGF-I) in lactating beef cows. In Experiment 1, mUltiparous Hereford cows were immunized against 2 mg GRF(l-29)-(GlY)4-Cys-NH2 conjugated to human serum albumin (GRFi, n=3) or 2 mg human serum albumin (HSAi, n=3) at 52 ± 1 d prior to parturition. Boosters (1 mg) were administered on days 12,40 and 114 postpartum (pp). Serum samples were collected at IS-min intervals for 5 hr on days 18,46 and 120 pp, followed by administration CIV) of an opioid agonist (FK33-824; 10 J..lg/kg) and an antagonist (naloxone; .5 mg/kg) at hours 5 and 7, respectively. A GRF analog ([desamino-Tyrl, D-Ala 2 , Ala l5 ] GRF (l-29)-NH 2 ; 3.5 ~lkg) and arginine (.5 glkg) were administered at hour 10 on days 47 and 121, respectively. Percentage binding of [ 125 I]GRF (1:100 dilution of serum) 28 d after primary immunization was greater in GRFi (14.3 ± 4.9) than in HSAi (.7 ± .3) cows. Binding increased to 29.3 ± 6.5% after first booster in GRFi cows. Episodic release of ST was abolished by immunization against GRF; concentration and frequency of release of ST were lower (P<.05) in GRFi than in HSAi cows on all days pp. Concentrations of IGF-I were lower in GRFi than in HSAi cows throughout lactation. Serum ST failed to increase following FK33-824 or arginine in GRFi; however, ST increased after both compounds in HSAi cows. Concentrations of ST following GRF-analog were greater (P<.05) in HSAi than in GRFi cows. Experiment 2 was con ducted to determine if a lower dose of antigen and a single booster would be sufficient to lower ST and IGF-I in lactating cows. Multiparous Hereford and Angus cows were assigned to GRFi (n=6) or HSAi (n=6). Primary (1.2 mg) and booster (.5 mg) immunizations were administered -14 and 8 d from calving, respectively. Cows were restricted to 60% of recommended intake of energy dur ing lactation in order to elevate concentrations of ST. Serum samples were collected at IS-min in tervals for 6 hr on days 26, 50, 73, 90 and 109 pp. Two of six GRFi cows had binding less than 10% (1 : 1 ,000 dilution of serum) and were omitted from further analyses. Percentage binding at a I: 1,000 dilution was greater in GRFi (17.0 ± 4.7) than in HSAi (1.4 ± .4) cows at 89 d pp. Consequently, frequency of release, and concentration of ST and IGF-I were lower (P < .05) in GRFi than in HSAi cows. Concentrations of ST increased following FK33-824 in HSAi, but not in GRFi cows. Across experiments, interval from calving to ovulation (estimated from weekly progesterone concentrations) was greater in HSAi than in GRFi cows. In conclusion, gestating-lactating beef cows were effec , tively immunized against GRF as evidenced by antibody binding of [ 125 I]GRF, absence of pulsatile release of ST, low concentrations of ST and IGF-I and failure of ST to increase after IV opioid ago nist or arginine.