Randomized controlled trial of reinsemination strategies in dairy cows diagnosed nonpregnant using color flow Doppler ultrasonography on d 21 after insemination

The objective of this study was to quantify the reproductive performance of 4 reinsemination strategies in cows diagnosed nonpregnant using corpus luteum color flow Doppler ultrasonography on d 21 after last insemination. A total of 2,140 color flow Doppler ultrasonography exams from 845 Holstein cows from 10 commercial dairy herds were used in this study. Farms were visited every 2 wk by the research team. On d 7 after insemination, cows were enrolled in a randomized controlled trial to be assigned 1 of 4 treatments if they were subsequently diagnosed nonpregnant on d 21. All cows were then examined on d 21 using Doppler ultrasonography to diagnose nonpregnancy. Treatment A (CON) was the control group: nonpregnant cows were programmed to receive a standard Ovsynch protocol starting on d 32. Nonpregnant cows in treatment B (GnRH) were injected i.m. with GnRH on d 21 after insemination and reinseminated immediately. Cows in treatment C (2×GnRH) received an i.m. injection of GnRH on d 11 after insemination. If diagnosed nonpregnant on d 21 after insemination, they were injected i.m. with GnRH on d 21 after insemination and inseminated immediately. Cows in treatment D (Resynch) received an i.m. injection of GnRH on d 14 after insemination. If diagnosed nonpregnant on d 21 after insemination, they were injected i.m. with PGF(2α) on d 21 after insemination and injected i.m. with GnRH on d 24. Then, a standard Ovsynch protocol was started on d 32. Statistical analyses were performed using mixed logistic regression models accounting for cow clustering and herd effect. A theoretical simulation was performed for each treatment to quantify the proportion of cows remaining nonpregnant 42 d after initial insemination. Of the 2,140 color flow Doppler ultrasonography exams in the study, 870 (40.6%) had a Doppler score of D0 (n = 444) or D1 (n = 426), which were indicative of nonpregnancy and used for data analysis. Overall, the number of exams assigned to each treatment was as follows: CON = 223 (25.6%), GnRH = 214 (24.6%), 2×GnRH = 220 (25.3%), and Resynch = 213 (24.5%). The final mixed multivariable logistic regression model included treatment, parity, days in milk at enrollment, and herd. Conception risk at first insemination following enrollment was 31.4, 20.6, 31.9, and 48.7% for treatments CON, GnRH, 2×GnRH, and Resynch, respectively. Based on the simulation, if 100 cows were diagnosed nonpregnant at their color flow Doppler ultrasonography exam on d 21 after last insemination, the total number of cows remaining nonpregnant 42 d after the initial insemination would be 69, 72, 58, and 51 for treatments CON, GnRH, 2×GnRH, and Resynch, respectively. Thus, the treatments used in groups 2×GnRH and Resynch for cows diagnosed nonpregnant on d 21 after insemination yielded better results than those used in the other groups.