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Manipulation of the reproductive tract can cause inflammatory processes in the endometrium and release of cytokines and prostaglandins. It has been shown that PGF2α has direct negative effects on embryonic survival and development. Treatment with nonsteroidal antiinflammatory drugs (e.g., ibuprofen lysinate, flunixin meglumine) might improve pregnancy rates after embryo transfer in recipient heifers. The primary objective of this study was to evaluate the effect of a nonsteroidal antiinflammatory drug on reproductive performance in lactating dairy cows when administered at the time of first-service artificial insemination (AI) based on the hypothesis that uterine manipulation during AI might be similarly intense compared with embryo transfer in its effect on prostaglandin release. A total of 970 cows (333 primiparous and 637 multiparous) from 17 Holstein dairy farms were enrolled. On the day of first AI, cows were randomly allocated to 1 of 3 treatment groups. Cows of group 1 received 1.4 mg/kg of body weight (BW) of carprofen subcutaneously immediately after AI (SC group). In group 2, 1.4 mg/kg of BW of carprofen was administered into the uterus using a sterile disposable catheter 12 to 24 h after AI (IU group). Animals of group 3 remained as untreated controls. First AI conception rate was similar for the SC group (42.2%) compared with the untreated control group (45.1%). A binary logistic regression model for the odds of conception at first AI revealed a negative effect of an intrauterine administration of carprofen on conception rate (38.3%). Cows allocated to the IU group had a lower likelihood of being pregnant within 200 d in milk than cows in the control group. In summary, subcutaneous treatment with the nonsteroidal antiinflammatory drug carprofen at the time of AI did not influence conception rate, whereas an intrauterine administration of carprofen 12 to 24 h after first AI had a negative effect on first-service conception rate in lactating dairy cows.