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Bovine twin birth is associated with detriments, including increased embryo/fetal losses, malpresentation, and dystocia. Incidence of these is lessened in bilateral compared with unilateral twin pregnancy. This study was undertaken to assess the use of follicular ablation by aspiration to create bilateral twin pregnancies in females with genetic potential for ~3.5 ovulations per cycle (Trio allele carriers). In experiment 1, carriers (n = 30) and noncarriers (n = 10) were synchronized for ovulation and timed artificial insemination (TAI). Follicles (>5 mm) in excess of one per ovary were aspirated ~16 h preceding TAI. Follicle count for females with follicles on only one ovary was reduced to two. Blood was sampled 2 wk post-TAI to assess progesterone (P4) concentrations; embryo count was determined by ultrasound 6 wk post-TAI. Circulating P4 concentration post-TAI was significantly (P < 0.001) associated with both genotype and subsequent pregnancy status (pregnant noncarriers: 7.06 ± 0.68 ng/mL; pregnant carriers: 5.54 ± 0.55 ng/mL; nonpregnant noncarriers: 5.22 ± 1.05 ng/mL; nonpregnant carriers: 3.13 ± 0.42 ng/mL). Experiment 2 was undertaken to offset the negative effects of follicular aspiration on subsequent P4 concentration observed in experiment 1. Carriers (n = 38) and noncarriers (n = 32) were submitted to TAI and follicle ablation as described for experiment 1. Additionally, accessory corpora lutea (CL) were induced in carriers by the administration of human chorionic gonadotropin (carriers) at day 6 post-TAI. Consequently, P4 concentration post-TAI was significantly (P < 0.05) associated with subsequent pregnancy status (pregnant: 8.48 ± 0.61 ng/mL; nonpregnant: 6.70 ± 0.63 ng/mL) but not with genotype (carrier: 8.01 ± 0.59 ng/mL; noncarrier: 7.17 ± 0.64 ng/mL). Embryo number was greater in carriers (exp. 1: 1.64 ± 0.81; exp 2: 1.45 ± 0.09) vs. noncarriers (1.00 ± 0.00, both experiments). Single, twin, and triplet pregnancies occurred in carriers in experiment 1, whereas multiples in experiment 2 were limited to twin pregnancies. Genotype effects on pregnancy rate were not significant (P > 0.10) in either experiment. Results suggest that follicular ablation to create bilateral twin pregnancies in Trio carriers is feasible but requires the induction of accessory CL to offset the negative effects of follicular aspiration on subsequent P4 concentration and associated fertility outcomes.

Changes in circulating progesterone (P4) and estradiol (E2) during proestrus produce dynamic changes in endometrial function and pituitary release of gonadotropins. Independent and combined effects of P4 and E2 on endometrium and pituitary were evaluated. In a preliminary study, an exogenous hormone model of proestrus was created by removal of corpus luteum and follicles ≥5 mm followed by gradual removal of intravaginal P4 implants during 18 h and treatment with increasing doses of estradiol benzoate during 48 h to mimic proestrus using high E2 (n = 9) or low E2 (n = 9). Decreased P4, increased E2, and increased endometrial area (EA) simulated proestrus in high-E2 cows and this was used subsequently. The main experiment used a 2 × 2 factorial design with: high E2 and low P4 (n = 11); high E2 and high P4 (n = 11); low E2 and high P4 (n = 11); low E2 and low P4 (n = 10). At 48 h, gonadotropin-releasing hormone (GnRH)-induced luteinizing hormone (LH) and follicle stimulating hormone (FSH) release was determined. Variables were analyzed using PROCMIXED of Statistical Analysis System. The EA increased dramatically during 48 h only in high-E2 and low-P4 cows. For FSH, high-E2 cows had greater area under the curve (AUC) and FSH peak after GnRH than low E2, with mild negative effects of high P4. For LH, concentration at peak and AUC were 2-fold greater in high E2 compared to low-E2 groups, with low P4 also 2-fold greater than high-P4 groups. Thus, maximal changes in uterus and pituitary during proestrus depend on both low P4 and high E2, but different physiologic responses are regulated differently by E2 and P4. Changes in endometrium depend on low P4 and high E2, whereas GnRH-induced FSH secretion primarily depends on high E2, and GnRH-induced LH secretion is independently increased by high E2 or reduced by high P4.

The aim of these experiments was to study ovarian dynamics and fertility of Bos indicus beef cattle submitted to 7-d progesterone (P4)-based fixed-time AI (FTAI) protocols using different hormonal treatments. In Exp. 1, 2 yr old Nelore heifers (n = 973) were randomly assigned to one of four treatments: EB-0 (estradiol benzoate, EB on D0 and no GnRH at AI), EB-G (EB on D0 and GnRH at AI), G-0 (GnRH on D0 and no GnRH at AI), or G-G (GnRH on D0 and at AI). On D0, heifers received an intravaginal P4 implant (0.5 g) for 7 d and EB (1.5 mg) or GnRH (16.8 μg). On D7, the P4 implant was withdrawn and heifers received cloprostenol (PGF; 0.5 mg) and estradiol cypionate (EC, 0.5 mg). Heifers in G groups also received PGF and eCG (200 IU) on D6, whereas EB heifers received eCG on D7. At FTAI on D9, only EB-G and G-G groups received GnRH (8.4 μg). In Exp. 2, Nelore cows (n = 804) received the same treatments (EB-0, EB-G, G-0, or G-G) using a 1.0 g P4 implant, 2.0 mg EB, and 300 IU eCG. Effects were considered significant when P ≤ 0.05. After treatment on D0, G had more ovulations than EB in heifers (60.3 [287/476] vs. 12.7% [63/497]) and cows (73.7 [83/112] vs. 24.4% [28/113]). Luteolysis after D0 was greater in EB than G in heifers (39.2 [159/406] vs. 20.0% [77/385]) and cows (25.5 [14/55] vs. 1.6% [1/64]). Heifers in G had larger follicles (mm) than EB on D7 (10.3 ± 0.2 vs. 9.2 ± 0.2) and at AI (11.9 ± 0.2 vs. 11.3 ± 0.2). Cows had larger follicles in G than EB on D7 (11.0 ± 0.3 vs. 9.9 ± 0.3) but not at AI. More estrus was observed in G than EB for heifers (80.3 [382/476] vs. 69.6% [346/497]) and cows (67.6 [270/400] vs. 56.2% [227/404]). There was no interaction between D0 and D9 treatments on pregnancy per AI (P/AI) in heifers (EB-0: 56.7 [139/245], EB-G: 53.6 [135/252], G-0: 52.6 [127/241], and G-G: 57.5% [135/235]). However, cows from EB-G had greater P/AI than EB-0 (69.5 [142/204] vs. 60.2% [120/200]), whereas P/AI for G-0 (62.7% [127/203]) was similar to G-G (60.9% [120/197]). In heifers, there was no interaction of GnRH at AI with estrus, however, cows that did not display estrus had greater P/AI if they received GnRH at AI (GnRH = 59.1 [91/154] vs. No GnRH = 48.2% [78/162]). Thus, protocols initiated with EB or GnRH for Bos indicus heifers and cows had differing ovarian dynamics but similar overall fertility, enabling their use in reproductive management programs. Treatment with GnRH at time of AI increased fertility in some instances in Bos indicus cows but not in heifers.

This study aimed to validate a 7 d progesterone (P4)-based fixed-time AI (FTAI) protocol for Bos indicus cattle by comparing to 8 and 9 d-type protocols. The first study compared 7 vs. 8 d protocols in Nelore heifers (Exp. 1.1; n = 742) and cows (Exp. 1.2; n = 2488), and the second study compared 7 vs. 9 d protocols in cows (Exp. 2; n = 1343). On experimental Day −10 and Day −11 the 8 and 9 d groups received an intravaginal P4 implant, 2.0 mg estradiol benzoate (EB) and 0.5 mg cloprostenol sodium (PGF). On Day −9 the 7 d group received the same treatments (P4, EB, and PGF). Then, on Day −2 all groups had the P4 implants removed, and PGF, 0.6 mg estradiol cypionate, and 300 IU equine chorionic gonadotropin (eCG) was administered. Fixed-time AI was performed 48 h later (Day 0) and 8.4 mg buserelin acetate (GnRH) was administered to 7d-G, 8d-G and 9d-G groups, whereas 7d-0, 8d-0 and 9d-0 groups did not receive GnRH at AI. Estrus was detected using tail-chalk between Day −2 and Day 0. Pregnancy per AI (P/AI) was evaluated by ultrasound 30 d after AI. Effects were considered significant when P ≤ 0.05, whereas a tendency was designated when P ≤ 0.10 and P > 0.05. In heifers (Exp. 1.1), incidence of estrus was similar regardless of protocol length (7 or 8 d). There was no independent treatment effect on P/AI or interaction between protocol length and GnRH at AI for P/AI (7d-0: 46.9, 7d-G: 51.4, 8d-0: 47.7, and 8d-G: 43.6%). Heifers in estrus had greater P/AI, and GnRH had no additional effect. More cows (Exp. 1.2) from the 8 d protocol were in estrus than cows submitted to the 7 d protocol. Additionally, despite no interaction between protocol length and GnRH on P/AI (7d-0: 55.9, 7d-G: 60.9, 8d-0: 56.2, and 8d-G: 60.8%), GnRH at AI increased P/AI. There was no interaction between estrus and GnRH, but cows displaying estrus had greater P/AI. Cows not expressing estrus tended (P = 0.06) to have greater P/AI when receiving GnRH. In Exp. 2, more 9 d cows were in estrus than 7 d cows. Protocol length did not affect P/AI but tended (P = 0.08) to interact with GnRH (7d-G had greater P/AI [57.9%] than 7d-0 [47.6%], but 9d-0 [54.6%] and 9d-G [55.4%] were not different from other groups). Moreover, GnRH increased P/AI only for the 7 d protocol. No interaction between estrus and GnRH was detected but estrus improved P/AI, and GnRH tended (P = 0.09) to improve P/AI of cows in estrus. In conclusion, despite longer protocols being more conducive to expression of estrus, there were no detectable effects of protocol length on P/AI. In addition, GnRH at FTAI may improve fertility in cows, particularly when cows are treated with shorter protocols.

Three experiments evaluated ovarian dynamics and circulating progesterone (P4) during P4-based protocols initiated with GnRH, estradiol benzoate (EB), or no additional treatment in Nelore (Bos indicus) cattle. In Exp 1 (n = 59 cows), a 5-d P4-only protocol (P-5d; D0: P4 implant alone (1g); D5: P4 removal, 0.5 mg estradiol cypionate [EC], 0.526 mg cloprostenol [PGF], and 300 IU equine chorionic gonadotropin [eCG]; D7: 8.4 μg buserelin acetate [GnRH]) was compared to a 9d protocol initiated with EB (EB-9d; D0: 2 mg EB + P4; D9: P4 removal + EC + PGF + eCG), and to a 7d GnRH protocol (G-7d; D0: 16.8 μg GnRH + P4; D6: PGF + eCG; D7: P4 removal + PGF; D9: GnRH). Exp 2 (n = 55 cows) compared G-7d and EB-7d protocols (similar to EB-9d, but D9 treatments were done on D7). Exp 3 (n = 64 heifers) compared EB-7d, G-7d, and P-5d protocols. For all experiments, daily ovarian ultrasonography was done from D0 until 4d after implant withdrawal and blood samples were collected at D0 and first PGF. Follicle dynamics were determined for each individual animal, analyzed within individual experiments, and afterwards combined to determine overall effects of treatments. The protocol that began with GnRH, G-7d, had greater ovulation rate after D0 with subsequently greater number of CL and circulating P4 at time of PGF (52.8%, 1.0 ± 0.1 CL, 4.0 ± 0.4 ng/mL) than for EB protocols (12.1%, 0.4 ± 0.05 CL, 2.0 ± 0.2 ng/mL), or P-5d (2.5%, 0.6 ± 0.09 CL, 2.6 ± 0.3 ng/mL). The G-7d and EB protocols had synchronized follicle wave emergence in 92.1% of animals but with distinct patterns. For the G-7d group, wave emergence occurred earlier in ovulating than non-ovulating animals (1.4 ± 0.2 d vs 2.5 ± 0.4 d). By comparison, most animals in EB-7d or EB-9d (80.3%) displayed atresia of the dominant follicle, followed by wave emergence 2–3 d after EB treatment. In contrast, P-5d protocol synchronized wave emergence in only 30.0% of cows. Nevertheless, no differences among treatments were detected for ovulation at end of the protocol (85.7%). In conclusion, the P-5d protocol did not synchronize follicle wave emergence but produced similar final ovulation, whereas, GnRH and EB protocols had follicle dynamics synchronized by distinct mechanisms that produced differences in CL number and P4 at the time of PGF treatment but similar final ovulation. Based on ovarian function, each of these synchronization methods are promising for use in FTAI, although fertility still needs to be evaluated.

© 2014, Bulletin of the Seismological Society of America. All rights reserved. The West Iberia Lithosphere and Asthenosphere Structure (WILAS) project densely covered Portugal with broadband seismic stations for 2 yrs. Here we provide an overview of the deployment, and we characterize the network ambient noise and its sources. After explaining quality control, which includes the assessment of sensor orientation, we characterize the background noise in the short-period (SP), microseismic, and long-period (LP) bands. We observe daily variations of SP noise associated with anthropogenic activity. Temporary and permanent stations present very similar noise levels at all periods, except at horizontal LPs, where temporary stations record higher noise levels. We find that median noise levels are extremely homogeneous across the network in the microseismic band (3–20 s) but vary widely outside this range. The amplitudes of microseismic noise display a strong seasonal variation. The seasonality is dominated by very-long-period double-frequency microseisms (8 s), probably associated with winter storms. Stacks of ambient noise amplitudes show that some microseismic noise peaks are visible across the whole ground-motion spectrum, from 0.3 to 100 s. Periods of increased microseismic amplitudes generally correlate with ocean conditions offshore of Portugal. Some seismic records display an interesting 12 hr cycle of LP (100-s) noise, which might be related to atmospheric tides. Finally, we use plots of power spectral density versus time to monitor changes in LP instrumental response. The method allows the identification of the exact times at which LP response changes occur, which is required to improve the understanding of this instrumental artifact and to eventually correct data., This research was supported by Fundação para a Ciência e Tecnologia through projects WILAS (PTDC/CTE-GIX/097946/2008), QuakeLoc-PT (PTDC/GEO-FIQ/3522/2012), and SCENE (PTDC/CTE-GIX/103032/2008), and by the European Commission FP7 project NERA (Network of European Research Infrastructures for Earthquake Risk Assessment and Mitigation, GA-262330). This is a contribution to PEST-OE/CTE/LA0019/2011 (IDL). The first author acknowledges a Marie Curie International Reintegration Grant from the European Commission, FP7 (PIRG03-GA-2008-230922)