Abstract
The sex ratio of mammals has previously been shown to be affected by maternal stress. In our previous study, the proportion of female embryos collected from superovulated and artificially inseminated Holstein heifers that were frequently placed in stanchions and subjected to transrectal examinations of the ovaries during the follicular phase tended to be higher than the expected 50%. The goal of the present study was to test the validity of this observation using a greater number of heifers. Superovulated heifers were artificially inseminated at 56 and 72 h after PGF2α treatment using a single batch of frozen semen. Frequent capture (FC), transrectal examination and/or blood sampling were performed at 4-h intervals from 36 to 76 h after PGF2α treatment (n=13). Nine heifers were used as the Control (non-treatment). Seven-day-embryos were recovered by uterine flushing. Male and female embryos were separated using the loop-mediated isothermal amplification procedure. The proportion of female transferable embryos in the FC group (67.8%, 78/115) was significantly higher than that in the Control group (51.2%, 43/84, P<0.05). The peak concentration of plasma cortisol during the follicular phase following superovulatory treatment was 20.6 ng/ml in the FC group. These results suggest that subjecting heifers to stress during the follicular phase following superovulatory treatment may increase the female sex ratio of embryos.