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Objectives were to evaluate risk factors affecting ovulatory responses and conception rate to the Ovsynch protocol. Holstein cows, 466, were submitted to the Ovsynch protocol [day 0, GnRH-1; day 7, prostaglandin (PG) F; day 9, GnRH-2] and 103 cows were inseminated 12 h after GnRH-2. Information on parity, days in milk at GnRH-1, body condition, milk yield, exposure to heat stress, pre-synchronization with PGF and the use of progesterone insert from GnRH-1 to PGF was collected. Ovaries were scanned to determine responses to treatments. Overall, 54.7%, 10.6%, 2.2%, 81.1%, 9.0%, 91.5% and 36.9% of the cows ovulated to GnRH-1, multiple ovulated to GnRH-1, ovulated before GnRH-2, ovulated to GnRH-2, multiple ovulated to GnRH-2, experienced corpus luteum (CL) regression and conceived, respectively. Ovulation to GnRH-1 was greater in cows without a CL at GnRH-1, cows with follicles >19 mm and cows not pre-synchronized with PGF 14 days before GnRH-1. Multiple ovulations to GnRH-1 increased in cows without CL at GnRH-1 and cows with follicles ≤19 mm at GnRH-1. Ovulation before GnRH-2 was greater in cows without CL at PGF. Ovulation to GnRH-2 increased in cows that received a progesterone insert, cows with a CL at GnRH-1, cows with follicles not regressing from the PGF to GnRH-2, cows with larger follicles at GnRH-2, cows that ovulated to GnRH-1 and cows not pre-synchronized. Multiple ovulations after GnRH-2 increased in cows with no CL at GnRH-1, multiparous cows and cows that multiple ovulated to GnRH-1. Conception rate at 42 days after AI increased in cows with body condition score > 2.75 and cows that ovulated to GnRH-2. Strategies that optimize ovulation to GnRH-2, such as increased ovulation to GnRH-1, should improve response to the Ovsynch protocol.