This short communication reports the impact of endometrial biopsies, uterine flushings and follicular fluid aspiration procedures at day 6 post artificial insemination (AI) on pregnancy rates. In Experiment 1, cows were timed AI (TAI) and assigned to the following treatment groups: control (n = 37), uterine flushing (n = 35) and endometrial biopsy (n = 38). On day 30 post AI, pregnancy rates were 40.5%, 33% and 28.5%, respectively (p > 0.1). Pregnancy rate on day 60 was lower (p < 0.004) in flushed cows than in the controls. In Experiment 2, oestrus was detected and cows were assigned to flushing (n = 32) or biopsy (n = 33) treatments 6 days after AI, which resulted in pregnancy rates of 31% and 36%, respectively (p > 0.1). In Experiment 3, cows were, 6 days after TAI, randomly assigned to the following treatments: control (n = 84) or aspiration of the largest follicle (n = 73). Pregnancy rates on day 30 post AI were 63.5% for the control group and 53% for the aspirated group (p > 0.1). In conclusion, uterine flushing and endometrial biopsy negatively affect pregnancy rates, but neither procedure can be considered to be incompatible with pregnancy maintenance. Follicular aspiration during pregnancy does not interact with pregnancy success. The amount and quality of samples obtained are compatible with the use of cellular and molecular analysis of uterine variables from cows that failed or succeeded on maintaining pregnancy.

The objective was to compare pharmacological strategies aiming to inhibit prostaglandin F2 alpha (PGF(2α)) synthesis (flunixin meglumine; FM), stimulate growth of the conceptus (recombinant bovine somatotropin; bST) and progesterone (P(4)) synthesis (human chorionic gonadotropin; hCG), as well as their combinations, regarding their ability to improve pregnancy rates in beef cattle. Lactating Nelore cows (N = 975), 35 to 70 days postpartum, were synchronized and inseminated by timed artificial insemination (TAI) on Day 0. On Day 7, cattle were allocated into eight groups and received one of the following treatments: saline (S) on Days 7 and 16 (Group Control); S on Day 7 and FM on Day 16 (Group FM); bST on Day 7 and S on Day 16 (Group bST); bST on Day 7 and FM on Day 16 (Group bST + FM); hCG on Day 7 and S on Day 16 (Group hCG); hCG on Day 7 and FM on Day 16 (Group hCG + FM); bST and hCG on Day 7 and S on Day 16 (Group bST + hCG), or bST and hCG on Day 7 and FM on Day 16 (Group bST + hCG + FM). The aforementioned treatments were administered at the following doses: 2.2 mg/kg FM (Banamine®; Intervet Schering-Plough, Cotia, SP, Brazil), 500 mg bST (Boostin®; Intervet Schering-Plough), and 2500 IU hCG (Chorulon®; Intervet Schering-Plough). Pregnancy diagnosis was performed 40 days after TAI by transrectal ultrasonography. Pregnancy rates were not significantly different among treatments. However, there was a main effect of hCG treatment to increase pregnancy rates (63.0 vs. 55.4%; P = 0.001). Concentrations of P(4) did not differ significantly among groups on Day 7 or on Day 16. However, consistent with the higher pregnancy rates, hCG increased P(4) concentrations on Day 16 (10.6 vs. 9.6 ng/mL, respectively; P = 0.05). We concluded that hCG treatment 7 days after TAI improved pregnancy rates of lactating Nelore cows, possibly via a mechanism leading to induction of higher P(4) concentrations, or by reducing the luteolytic stimulus during maternal recognition of pregnancy.

No abstract

The relevance of the uterine secretions during early embryonic development is due to its role as the only source of nutrients for embryo prior to implantation. Probing of the uterine secretions during early pregnancy without compromising gestation would be an important tool for the identification of fertility markers. Manipulations of the uterus could lead to a local increase in blood flow, which could result from the physical stimulation or tissue damage and inflammation. This study aimed to evaluate the effects of low-volume flushing of the uterine horn contralateral to the ovulation on uterine vascular perfusion and endometrial thickness 6 days after artificial insemination (AI). Transrectal B-mode and color Doppler ultrasonography (MyLab30 Vet Gold; Esaote Healthcare) were used to compare changes on endometrial thickness and uterine vascular perfusion, respectively. Examinations were carried out on 15 Nelore cows just prior to the flushing procedure (0 h), 6 and 24 h later. Cows were inseminated after oestrus detection and uterine flushings were performed 6 d after AI. The uterine horn, contralateral to the corpus luteum, was flushed with 20 mL of PBS using a Foley catheter. After massage of the uterine horn, flushing was collected in a syringe by suction. Vascular perfusion was estimated by scoring the extent of colored areas within the endometrium and mesometrium. Vascular perfusion scores indicated nil (1), minimal (2), intermediate (3), and maximal (4) vascular perfusion. Endometrial thickness was measured by taking the maximum diameter and its perpendicular diameter; these values were summed and then divided by four. Data that were not normally distributed were transformed to natural logarithms or ranks. For each variable, the main effect of side (flushed and nonflushed horns), hour, and their interaction were analyzed using the PROC MIXED procedure from the SAS software. The least significant difference method was used for the comparison of means among hours. An hour effect was detected for endometrial and mesometrial scores of vascular perfusion (Table 1), representing an increased vascular perfusion about 6 h after the flushing procedure and a return to basal perfusion at 24 h. Results indicate that the unilateral flushing procedure increases vascular perfusion on both uterine horns. For endometrial thickness, no effect of side, hour, or their interaction was detected. In conclusion, unilateral low-volume uterine flushing results in a transitory increase in uterine blood flow about 6 h post-flushing. Further studies are needed to evaluate other inflammatory characteristics and the potential effect on pregnancy rate in response to the uterine flushing procedure performed during early pregnancy. Table 1.Scores of endometrial and mesometrial vascular perfusion and endometrial thickness of both uterine horns at the time of uterine flushing (0 h), 6 and 24 h post-flushing (mean ± standard error of the mean)

Recent evidence indicates that the progesterone (P4) secretion by corpus luteum (CL) during early diestrus is affected by the size of ovulatory follicle and has a significant impact on embryo development and conception rates. Therefore, strategies to promote the growth of the dominant follicle and/or to stimulate the early development of the CL to increase P4 secretion become an alternative to improve conception rates in the beef cattle industry. Our aim was to study the effect of manipulations of the follicle growth on the diameter of the preovulatory follicle (POF) and subsequent size and function of the CL. Cyclic and non-lactating Nelore cows, pre-synchronized by 2 injections of prostaglandin F2α (PGF) 14 d apart, were manipulated to ovulate large or small follicles according to 3 experiments. In Experiment 1 (Exp. 1; n = 23), animals received a second-use intravaginal P4-releasing device along with an injection of oestradiol benzoate on Day –10 (Day 0 = GnRH injection). Cows were split to receive (large follicle group; LF) or not (small follicle group; SF) a PGF injection on Day –10. Progesterone devices were removed on Day –2.5 in the LF group and on Day –1.5 in the SF group. The PGF was injected at the removal of the P4 device. In Experiment 2 (Exp. 2; n = 38), cows in the LF group had the P4 device removed on Day –2.25 or Day –2, whereas in Experiment 3 (Exp. 3; n = 23), the device (first-use) was removed on Day –1.75 in the LF group and on Day –1.25 in the SF group; the other manipulations were similar to Exp. 1. Data analyses were done only on cows that had a functional CL on Day –10 (P4 > 1 ng mL–1) and that ovulated within 24 and 48 h post-GnRH (Exp. 1, n = 14; Exp. 2, n = 14; Exp. 3, n = 12). The three experiments were successful in inducing POF with different sizes, as indicated by the greater diameter of the POF in the LF group compared with SF in Exp. 1 (12.9 ± 0.5 mm v. 10.7 ± 0.6 mm; P < 0.03), Exp. 2 (14.1 ± 0.6 mm v. 11.7 ± 0.4 mm; P < 0.006), and Exp. 3 (13.8 ± 0.6 mm v. 11.7 ± 0.8 mm; P < 0.06). To evaluate the effect of POF size on size and function of the CL, a factorial analysis was performed by SAS software to test the effect of group, day, and their interaction. For CL volume, an effect of group was detected in Exp. 1 (P < 0.02) and in Exp. 3 (P < 0.06), but not in Exp. 2. The group effect represented greater average CL volume from Day 3 to Day 7 in LF (2.42 ± 0.27 and 2.5 ± 0.39 cm3) than in the SF group (1.39 ± 0.18 and 1.2 ± 0.15 cm3) for Exp. 1 and 3, respectively. For P4 concentrations, a group effect was detected only in Exp. 3 (P < 0.007), as indicated by greater average P4 concentrations from Day 3 to Day 7 in LF (2.31 ± 0.31 ng mL–1) than in the SF group (1.37 ± 0.19 ng mL–1). A day effect was detected in all experiments (P < 0.0001), as indicated by a progressive increase of CL volume and P4 concentrations from Day 3 to Day 7. Manipulation of follicle growth performed in Exp. 3 was the most efficient to modify the function and size of the CL. In conclusion, control of POF size by manipulation of P4 concentrations during growth of the dominant follicle alters the size and function of CL postovulation. CNPq, FAPESP, Ourofino, and PUSP-P.