347 ARTIFICIAL INSEMINATION OF LACTATING HOLSTEIN COWS WITH SEXED SPERM

The recommended use of sexed sperm has been limited to heifers. The objective of this field trial was to determine pregnancy rates in lactating dairy cows following artificial insemination of 2 or 10 � 106 sexed or 10 � 106 unsexed frozen/thawed sperm. Sperm were separated into X chromosome-bearing populations at rates of 3000-5000/s, on the basis of DNA content, with a flow cytometer/sperm sorter operating at 40 psi and using 150 mW of laser intensity. Sperm were sorted, collected and cryopreserved in a Tris-based medium. Holstein cows, averaging 2.4 lactations (range: 1-9), were presynchronized with prostaglandin F2� (PGF)2�; (25 mg) i.m. at 40-58 days in milk (DIM) (average, 44 days). A second dose of PGF2� (25 mg) i.m. was administered 14 days later. All cows were screened with ultrasound 14 days later, and those with normal ovarian and uterine status were used for this study. Qualifying cows received 100 �g gonadotropin-releasing hormone (GnRH) i.m. (average, 72 DIM) and 25 mg PGF2� i.m. 7 days later, and then 100 �g GnRH i.m. 2 days after the prostaglandin. Cows (n = 172) were fixed-timed inseminated 16-19 h after the last GnRH treatment, balanced over three sperm insemination treatments, two Holstein bulls, and two inseminators. Both 32-day pregnancy and 60-day fetal sex were determined using ultrasound. Cows carrying fetuses identified as dying at the 32-day ultrasound check were considered not pregnant. Data were subjected to ANOVA. Effects of sperm treatment, bulls, inseminators, and subsequent interactions on pregnancy rate were not significant (P > 0.05). The 32-day pregnancy rate and 95% confidence interval for 10 � 106 unsexed, 10 � 106 sexed, and 2 � 106 sexed inseminates were 55.6 (42.5-68.6), 43.9 (31.6-57.0), and 40.4% (28.4-53.6), respectively. Additionally, DIM or cow parity did not affect pregnancy rate (P > 0.05). Fetal death (<32 days) for unsexed inseminates was 5.5% (two pregnancies) compared to 9.5% (four pregnancies) for pooled sexed inseminates (P > 0.05). There were four confirmed pregnancies of sexed sperm that also aborted between Days 32-60. Fetal sex ratio was significantly altered (P < 0.05) for sexed (93% female fetuses) compared to unsexed (52% female fetuses) inseminates. Pregnancy rate in these selected lactating cows inseminated with sexed sperm was not significantly lower (P > 0.05) than that for unsexed sperm. However, these results should be viewed cautiously due to the limited numbers of cows used. Successful use of sexed sperm in dairy cows, which approached an elective waiting period of 80 DIM, was achieved with cows that were selected for reproductive status prior to insemination.