The objective of this randomized controlled experiment was to evaluate the effect of delaying induction of ovulation and timed artificial insemination (TAI) on expression of estrus before AI and first service reproductive outcomes. A secondary objective was to evaluate the effects of delaying induction of ovulation in a Double-Ovsynch protocol on ovarian function. Lactating Holstein cows (n = 4,672) from 2 commercial dairy farms fitted with sensors for automated detection of estrus were synchronized with a Double-Ovsynch protocol up to the first PGF2α (PGF-L) of the Breeding-Ovsynch portion of the protocol (Pre-Ovsynch: GnRH, 7 d later PGF2α, 3 d later GnRH, 7 d later Breeding-Ovsynch: GnRH, 7 d later PGF2α, 1 d later PGF2α). At PGF-L, cows blocked by parity (primiparous vs. multiparous) and semen used for first service (sex-sorted dairy vs. conventional beef) were randomly assigned to the G56 (n = 2,338) or G80 (n = 2,334) treatments. Cows in G56 had 56 h whereas cows in G80 had 80 h from PGF-L to induction of ovulation with the last GnRH (GnRH2) before AI. For both treatments, TAI occurred ∼16 h after GnRH2. All cows with automated estrus alerts between PGF-L and TAI were inseminated at detected estrus (AIE) without GnRH. Ovarian function and responses to synchronization were monitored based on circulating concentrations of progesterone and examination of the ovaries by ultrasonography. Data for binary outcomes were analyzed by logistic and continuous outcomes with lineal regression. More cows in G80 received AIE and had estrus before AI. Overall, pregnancies per AI (P/AI) did not differ for the G80 and G56 treatments. Cows in G80 that received TAI and had no estrus had fewer P/AI than cows with estrus that received AIE or TAI in G80, and fewer P/AI than cows AIE and cows that received TAI and had or did not have estrus in the G56 treatment. No differences were observed between treatments or for cows with and without estrus for pregnancy loss. Unlike some minor differences between treatments for concentrations of progesterone at GnRH2, the most notable differences in ovarian function were for cows in both treatments with or without estrus that received TAI. Cows with estrus, were more likely to have follicles > 16 mm, had larger follicles before ovulation, and had a greater ovulation risk after AI. Likewise, within the G80 treatment only, cows with estrus that received AIE or TAI had larger follicles, were more likely to have complete luteal regression, had greater ovulation risk, were more likely to have a functional corpus luteum, and had more circulating progesterone after AI. We concluded that delaying induction of ovulation and TAI was effective for allowing more cows to express estrus before AI which had different ovarian function outcomes and greater P/AI than cows that did not express estrus. However, the greater P/AI of cows that expressed estrus was insufficient to compensate for the reduced P/AI of cows that did not express estrus, and thus increase overall P/AI compared with the treatment without delayed induction of ovulation. Detection of estrus before AI in cows undergoing synchronization of ovulation could help identify cows with different likelihoods of pregnancy after insemination.
Keywords: automated detection of estrus; delayed induction of ovulation; fertility; timed artificial insemination.
© 2025, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).