This study aimed to compare three ways of FSH administration [single-dose subcutaneous (sc) using an adjuvant, single-dose epidural without an adjuvant and the conventional intramuscular (i.m) repeated decreasing doses] on blood FSH levels and superovulation response. Holstein cows (n:18) were divided into three groups for the superovulation procedure (n=6). The animals in group (1) received intramuscular (im) decreasing doses of FSH (Stimufol, 500 μg porcine FSH and 100 μg porcine LH, Ulg FMV, Liège, Belgium) at 12-h intervals for 4 d. The animals in group (2) received FSH (500 μg) into the epidural space (e). The animals in group (3) received a single subcutaneous (SC) dose of FSH (500 μg) added in 10 mL of Montanide ISA 206 adjuvant. CL count in the ovary was measured by ultrasonography and superovulation was determined based on the presence of?>4 CL. For FSH measurement, blood samples were collected from the V. jugularis of all the animals. Serum FSH (pFSH) concentration was determined by ELISA. After synchronisation and superovulation procedures, all animals had estrus (100%). Superovulation response (≥4 CL) was detected in six animals in group 1 (im), one animal in group 2 (e), and four animals in group 3 (sc). Superovulation responses in e group were lower than those in im group (P<0,05), but they were like those in sc group (P>0.05). Serum FSH concentrations were similar between groups for all sampling times (P>0.05).