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The optimization of Fixed-Time Artificial Insemination (FTAI) protocols is key for profitability in tropical livestock farming. This study compared the effect of two FTAI timings (48 vs. 56 hours post-PGF2a) within an Ovsynch protocol on reproductive parameters in crossbred dairy cows (Bos taurus x Bos indicus). Forty-six lactating cows were randomly assigned to one of two groups: Ovsynch-48 (n=24) and Ovsynch-56 (n=22). The pregnancy rate was not significantly different between the 48-hour group (37.5%) and the 56-hour group (18.2%) (x²=1.267, p=0.260). However, the Ovsynch-56 protocol significantly reduced the total days open compared to the Ovsynch-48 protocol (p=0.039). Additionally, no significant effect of the number of parities or body condition on the probability of pregnancy was found. It is concluded that, while both protocols yield statistically similar pregnancy rates, the 56-hour protocol offers a strategic advantage by shortening the days open, thereby improving a key indicator of reproductive efficiency.