Effects of different doses of estradiol benzoate used in a cervical relaxation protocol on the success of non-surgical embryo recovery and luteal function in superovulated ewes.

This study investigated the effectiveness of different doses of estradiol benzoate (EB) to promote cervical relaxation and their effects on luteal function and outcomes of non-surgical embryo recovery (NSER) in sheep. Multiparous (MULT) and nulliparous (NULL) crossbred Lacaune X Santa Inês ewes were superovulated and naturally bred. Seven days after progesterone withdrawal, females were randomly assigned to one of three distinct cervical relaxation protocols, consisting of i.m. treatment with 37.5 µg d-cloprostenol and different doses of EB: 0.0 mg (0.0EB group; n = 3 NULL and 14 MULT); 0.5 mg (0.5EB group; n = 4 NULL and 12 MULT) or 1.0 mg (1.0EB group, n = 6 NULL and 11 MULT) 16 h before NSER. All ewes received 50 IU of oxytocin 20 min before NSER (D17). Blood samples were collected and ultrasound exams (B-mode and color Doppler) were performed at two timepoints: immediately before d-cloprostenol and EB treatments and prior to NSER. Estrous behavior, corpora lutea count and NSER success outcomes were not affected by EB treatments nor parity (P > 0.05). Embryo recovery rate was greater for ewes in the 0.5EB group and in the NULL ewes (P < 0.05). Ovarian biometrics differed between the two evaluation timepoints in all groups (P < 0.05). Plasma estradiol increased over time, reaching a significant greater level in 1.0EB ewes compared to controls on D17 (P < 0.05), whereas progesterone concentrations decreased over time in all groups (P > 0.05). In conclusion, treatments did not affect NSER success but they did affect luteal function by altering P4 and E2 concentrations. Therefore, the NSER technique can be successfully performed in ewes with or without prior treatment with EB.

Single dose of 300 IU hCG in the early luteal phase in superovulated ewes: Effects on corpora lutea, progesterone profile, and embryo recovery.

This study aimed to evaluate the effects of hCG treatment during the early luteal phase on ovarian function, progesterone profile, and embryo yield in superovulated ewes. Superovulated sheep were randomly assigned to receive 300 IU hCG i.m. (GhCG, n = 24) or not (GControl, n = 25) at 96 h after the removal of the progesterone (P4) device (D13). Non-surgical embryo recovery (NSER) was performed eight days after P4 withdrawal. Ultrasound evaluations were performed on D13, D14, D16, and D17. Blood samples were collected on D14, D16, and D17. Superovulation scores were recorded based on the number of corpora lutea (CL) as follows: 1 (≤ 2), 2 (3-5), 3 (6-8), and 4 (≥ 9). NSER efficiency, superovulation response, and luteal tissue area were similar in both groups (P > 0.05). Structural luteolysis tended to be higher in GControl (P = 0.07; 47.0 %) while functional luteolysis was similar in both groups (P > 0.05; 0.0 % and 5.9 %). The recovery rate was greater (P < 0.05) in GhCG (89.8 %) compared with GControl (71.0 %), with similar overall ova/embryo numbers observed for both groups (P > 0.05). GhCG showed a higher concentration of animals with a superovulatory response score of 4 (54.5 %; P < 0.05) compared with the lowest scores. Plasma progesterone on D16 was higher (P < 0.05) in GhCG ewes (11.1 ± 1.5 vs 6.9 ± 1.5 ng/mL). In conclusion, the hCG treatment improved circulating P4 and embryo recovery rate, tended to maintain luteal functionality, and thus constitutes an additional tool for improving embryo yield in superovulated ewes.

Differential expression of LHCGR and its isoforms is associated to the variability in superovulation responses of Gir cattle.

The aim of this study was to evaluate the pattern of expression of LHCGR isoforms in Gir heifers characterized as good (10.3 ±â€¯1.2 ova/embryos per flush, n = 5) or poor responders (1.1 ±â€¯0.3 ova/embryos per flush, n = 5) to superovulation protocols. In both groups, an adapted ultrasound-guided follicular aspiration system was used to collect granulosa cells from 8 mm follicles formed either during a synchronized, non-stimulated follicular wave (no stimulation control, NS) or on the fourth day of a superovulation protocol (SOV) induced with 200 IU of pFSH. The recovered follicular fluid was centrifuged and granulosa cells were washed with NaCl 0.9% and kept in RNAlater®. RNA extraction was performed using a commercial RNeasy Micro Kit and eluted samples were quantified and reverse transcribed using the commercial Superscript III kit. cDNA samples were amplified by real-time PCR using a primer to target LH/hCG receptor gene - not selective for LHCGR isoforms (total LHCGR) - and four sets of isoforms selective primers (S1, S10, S10 + 11, and S11). Analyses were performed using the REST software and expression levels are shown as mean ± SEM. Under physiological conditions (NS), poor responders had a higher expression of total LHCGR (4.9 ± 1.7 fold-change, P < 0.01) as well as isoforms S10, S11 and S10 + 11, compared to good responders. In both phenotypes, superovulation down-regulated total LHCGR expression (-0.5 ± 0.2 and -0.9 ± 0.0 for good and poor responders, respectively; P < 0.05). However, in poor responders the exogenous FSH treatment up-regulated the S10 (2.4 ± 2.0; P < 0.05), S10 + 11 (3.8 ± 3.2; P < 0.01), and S1 isoforms (1.8 ± 1.3; P < 0.05), compared to good responders We conclude that down-regulation of total LHCGR, associated to up-regulation of their inactive isoforms, may have compromised follicle development and thus contributed to the low efficiency of superovulation in heifers with a poor responder phenotype.

Viable offspring after successful non-surgical embryo transfer in goats / Nascimento de fetos viáveis após transferência de embriões não-cirúrgica em caprinos

O objetivo do presente estudo foi avaliar a viabilidade da técnica de transferência não cirúrgica em cabras. Quatro cabras não-lactantes pluríparas da raça Toggenburg foram utilizadas como receptoras de embriões, sendo que duas receberam um embriões e duas receberam dois embriões coletados não cirurgicamente cabras doadoras. Os corpos lúteos das receptoras foram detectados um dia antes da transferência de embriões por ultrassonografia transretal. Uma seringa de 5mL contendo 2mL de meio holding foi acoplada em um cateter tomcat, no qual os embriões foram aspirados em uma coluna central a duas outras colunas. Um espéculo Colin número 2 foi inserido na vulva e na vagina, e com o uso de uma fonte de luz, a cerviz foi localizada e imobilizada com uma pinça de Allis. Um cateter uretral número seis acoplado a um mandril e lubrificado com meio PBS foi inserido na cérvix, e assim os aneis cervicais foram gradualmente transpostos. Após perder a resistência, o cateter uretral foi movido lateralmente para o corno uterino desejado. O mandril e a pinça de Allis foram retirados e o conjunto seringa e tomcat foi acoplado ao cateter uretral e o conteúdo injetado no corno uterino ipsilateral ao corpo lúteo com posterior retirada do cateter. Cabras que ovularam em apenas um ovário foram usadas para testar a eficiência da deposição do embrião. O tempo gasto entre a inserção do espéculo e a sua remoção foi inferior a três minutos. O tempo para transpor a cérvix foi inferior a um minuto. A ultrassonografia revelou a deposição de líquido no corno desejado. Receptoras que receberam dois embriões tornaram-se gestantes e pariram três crias. Estes primeiros resultados encorajam a técnica e demonstram que a transferência de embriões em caprinos pode ser feita totalmente por procedimentos não cirúrgicos.(AU)

Viable offspring after successful non-surgical embryo transfer in goats / Nascimento de fetos viáveis após transferência de embriões não-cirúrgica em caprinos

O objetivo do presente estudo foi avaliar a viabilidade da técnica de transferência não cirúrgica em cabras. Quatro cabras não-lactantes pluríparas da raça Toggenburg foram utilizadas como receptoras de embriões, sendo que duas receberam um embriões e duas receberam dois embriões coletados não cirurgicamente cabras doadoras. Os corpos lúteos das receptoras foram detectados um dia antes da transferência de embriões por ultrassonografia transretal. Uma seringa de 5mL contendo 2mL de meio holding foi acoplada em um cateter tomcat, no qual os embriões foram aspirados em uma coluna central a duas outras colunas. Um espéculo Colin número 2 foi inserido na vulva e na vagina, e com o uso de uma fonte de luz, a cerviz foi localizada e imobilizada com uma pinça de Allis. Um cateter uretral número seis acoplado a um mandril e lubrificado com meio PBS foi inserido na cérvix, e assim os aneis cervicais foram gradualmente transpostos. Após perder a resistência, o cateter uretral foi movido lateralmente para o corno uterino desejado. O mandril e a pinça de Allis foram retirados e o conjunto seringa e tomcat foi acoplado ao cateter uretral e o conteúdo injetado no corno uterino ipsilateral ao corpo lúteo com posterior retirada do cateter. Cabras que ovularam em apenas um ovário foram usadas para testar a eficiência da deposição do embrião. O tempo gasto entre a inserção do espéculo e a sua remoção foi inferior a três minutos. O tempo para transpor a cérvix foi inferior a um minuto. A ultrassonografia revelou a deposição de líquido no corno desejado. Receptoras que receberam dois embriões tornaram-se gestantes e pariram três crias. Estes primeiros resultados encorajam a técnica e demonstram que a transferência de embriões em caprinos pode ser feita totalmente por procedimentos não cirúrgicos...

The use of PGF2α as ovulatory stimulus for timed artificial insemination in cattle.

The objective of this study was to evaluate the effect of a PGF2α-analogue (PGF) on ovulation and pregnancy rates after timed artificial insemination (TAI) in cattle. In experiment 1, crossbred dual-purpose heifers, in a crossover design (3 × 3), were given an intravaginal progesterone-releasing insert (controlled internal drug release [CIDR]) plus 1 mg estradiol benzoate (EB) intramuscularly (im) and 250 µg of a PGF-analogue im on Day 0. The CIDR inserts were removed 5 days after follicular wave emergence, and the heifers were randomly divided into three treatment groups to receive the following treatments: (1) 1 mg of EB im (EB group, n = 13); (2) 500 µg of PGF im (PG group, n = 13); or (3) saline (control group, n = 13), 24 hours after CIDR removal. Ovulation occurred earlier in EB (69.81 ± 3.23 hours) and PG groups (73.09 ± 3.23 hours) compared with control (83.07 ± 4.6 hours; P = 0.01) after CIDR removal. In experiment 2, pubertal beef heifers (n = 444), 12 to 14 months of age were used. On Day 0, the heifers were given a CIDR insert plus 2 mg EB im. On Day 9, the CIDR was removed and the heifers were given 500 µg of PGF im. Heifers were randomly assigned into one of three treatment groups: (1) 1 mg of EB (EB group; n = 145); (2) 500 µg of PGF (PG group; n = 149), both 24 hours after CIDR removal; or (3) 600 µg of estradiol cypionate (ECP group; n = 150) at CIDR removal. Timed artificial insemination occurred 48 hours after CIDR removal in the ECP group and 54 hours in the PG and EB groups. The percentage of heifers ovulating was higher in the PG group compared with the other groups (P = 0.08). However, the pregnancy rates did not differ among groups (47.6%, 45%, and 46.6%, for EB, PG, and ECP, respectively; P = 0.9). In experiment 3, 224 lactating beef cows, 40 to 50 days postpartum with 2.5 to 3.5 of body condition score were treated similarly as described in experiment 2, except for the ECP group, which was excluded. The treatments were as follows: 1 mg EB (EB group; n = 117) or 500 µg PGF (PG group; n = 107), 24 hours after CIDR removal. The calves were temporarily separated from their dams from Days 9 to 11. No difference was detected on the pregnancy rate between the EB and PG groups (58.1% vs. 47.6%, respectively; P = 0.11). Taken together, the combined results suggested that PGF2α could be successfully used to induce and synchronize ovulation in cattle undergoing TAI, with similar pregnancy rates when compared with other ovulatory stimuli (ECP and EB).

Occurrence and characteristics of residual follicles formed after transvaginal ultrasound-guided follicle aspiration in cattle.

Ultrasound-guided transvaginal follicle aspiration is used to recover cumulus-oocyte complexes (for IVF) and to synchronize follicular wave emergence (ablation of dominant follicle). Although aspirated follicles are generally supposed to undergo immediate atresia, there are indications that they may remain active. The objective was to evaluate the occurrence and characteristics of residual follicles (RF) after transvaginal follicle aspiration in cattle. Ovarian follicular wave emergence was synchronized in Holstein cows (N = 13) in the presence (groups 1 and 3) or absence (groups 2 and 4) of norgestomet implants. The largest follicle was aspirated at a diameter of 8 mm (groups 1 and 2) or 12 mm (groups 3 and 4). Ovarian follicles were visualized (transrectal ultrasonography) every 12 h after wave emergence. Follicular fluid samples were collected from the largest follicle and from the ensuing RF and concentrations of estradiol and progesterone were determined. After aspiration, 73.2% (52/71) of the follicles refilled with fluid, and a new antrum was detected 12 to 24 h later. Norgestomet did not affect (P > 0.05) RF occurrence or diameter, but in RF from group 4, concentrations of estradiol decreased (-530.7 ± 133.9 ng/mL; P < 0.01) whereas progesterone increased (+429.6 ± 171.7 ng/mL; P < 0.05) relative to preaspiration. In RF, there were three steroidogenesis patterns: (1) high estradiol concentration and high estradiol:progesterone ratio (estradiol-active RF); (2) low estradiol, but high progesterone concentrations (luteinized RF); and (3) low estradiol and low progesterone concentrations (inactive RF). Estradiol-active RF were more likely (P < 0.05) from follicles with high estradiol concentrations (regardless of diameter). In conclusion, fluid-filled structures (RF) with variable steroid production patterns are frequently formed after ultrasound-guided follicle aspiration. The occurrence and features of these RF depended on the diameter and status of these follicles before aspiration.

Doppler ultrasonography as a tool for ovarian management

Doppler ultrasonography allows the characterization and measurement of blood flow, and can be used to indirectly make inference regarding the functionality of different organs, including the ovaries. Several studies highlighted the importance of an adequate blood flow for follicle development and acquisition of ovulatory potential, and for progesterone secretion by the corpus luteum. Due to some particularities of the ovarian vascular anatomy, however, different strategies had to be developed to measure the blood flow detected by Doppler imaging. Some of these approaches were successful to characterize the patterns of blood flow throughout the estrous cycle, pregnancy, and early postpartum, but require post acquisition image processing and do not allow real time decisions to be taken. The subjective evaluation of blood flow has been alternatively used in field conditions aiming to early detect non pregnant animals, select embryo recipients, or predict artificial insemination and in vitro fertilization outcomes. In summary, color Doppler imaging provides important information about the function of corpus luteum and follicles, supporting clinical diagnoses and management decisions. Nonetheless, the adoption of Doppler ultrasonography as a routine exam, instead of a limited use in an individual basis, requires further development of feasible blood flow evaluation procedures and the establishment of reference values.

Three-dimensional modeling of color Doppler images: a new approach to study follicular vascularization in cattle

Ovarian blood supply is directly related to follicle developmental potential and to oocyte quality, and color Doppler ultrasonography might be a valuable tool to predict in vitro fertilization outcomes. In most studies in large domestic animals, however, the evaluation of follicle blood flow is qualitative (presence or absence of color signal) or dependent on the analysis of a single image. The objective of the present study was to first describe the use of a three-dimensional (3D) modeling of color Doppler images for a quantitative assessment of vascularization in bovine ovarian follicles. Follicular wave emergence was synchronized in Holstein and Gir heifers (n = 20), and follicular dynamics were assessed every 12 h using a color Doppler ultrasound device. The recorded cine-loop of the dominant follicle was decomposed into frames and medical image processing software was used to isolate the Doppler signal, generate the 3D model and calculate the volume of vascularization. In experiment 1, the model was validated by comparing the expected and calculated volumes and was used to predict possible variations in the results of the 2D approach. In experiment 2, vascularization was analyzed during follicular development. In both breeds, the volume of vascularization increased after follicle deviation and was positively correlated (P < 0.05) to follicular diameter (r = 0.65 and 0.54 for Holstein and Gir heifers, respectively). Spatial analysis of the three-dimensional model showed an uneven distribution of vascularization in the follicular wall, with a more intense blood flow being detected in the basal (nearest the ovarian hilus) and lateral regions of the dominant follicles. These results demonstrate the potential of this technique as a new tool for in vivo studies of ovarian physiology in large animals.

Three-dimensional modeling of color Doppler images: a new approach to study follicular vascularization in cattle

Ovarian blood supply is directly related to follicle developmental potential and to oocyte quality, and color Doppler ultrasonography might be a valuable tool to predict in vitro fertilization outcomes. In most studies in large domestic animals, however, the evaluation of follicle blood flow is qualitative (presence or absence of color signal) or dependent on the analysis of a single image. The objective of the present study was to first describe the use of a three-dimensional (3D) modeling of color Doppler images for a quantitative assessment of vascularization in bovine ovarian follicles. Follicular wave emergence was synchronized in Holstein and Gir heifers (n = 20), and follicular dynamics were assessed every 12 h using a color Doppler ultrasound device. The recorded cine-loop of the dominant follicle was decomposed into frames and medical image processing software was used to isolate the Doppler signal, generate the 3D model and calculate the volume of vascularization. In experiment 1, the model was validated by comparing the expected and calculated volumes and was used to predict possible variations in the results of the 2D approach. In experiment 2, vascularization was analyzed during follicular development. In both breeds, the volume of vascularization increased after follicle deviation and was positively correlated (P < 0.05) to follicular diameter (r = 0.65 and 0.54 for Holstein and Gir heifers, respectively). Spatial analysis of the three-dimensional model showed an uneven distribution of vascularization in the follicular wall, with a more intense blood flow being detected in the basal (nearest the ovarian hilus) and lateral regions of the dominant follicles. These results demonstrate the potential of this technique as a new tool for in vivo studies of ovarian physiology in large animals.(AU)

Doppler ultrasonography as a tool for ovarian management

Doppler ultrasonography allows the characterization and measurement of blood flow, and can be used to indirectly make inference regarding the functionality of different organs, including the ovaries. Several studies highlighted the importance of an adequate blood flow for follicle development and acquisition of ovulatory potential, and for progesterone secretion by the corpus luteum. Due to some particularities of the ovarian vascular anatomy, however, different strategies had to be developed to measure the blood flow detected by Doppler imaging. Some of these approaches were successful to characterize the patterns of blood flow throughout the estrous cycle, pregnancy, and early postpartum, but require post acquisition image processing and do not allow real time decisions to be taken. The subjective evaluation of blood flow has been alternatively used in field conditions aiming to early detect non pregnant animals, select embryo recipients, or predict artificial insemination and in vitro fertilization outcomes. In summary, color Doppler imaging provides important information about the function of corpus luteum and follicles, supporting clinical diagnoses and management decisions. Nonetheless, the adoption of Doppler ultrasonography as a routine exam, instead of a limited use in an individual basis, requires further development of feasible blood flow evaluation procedures and the establishment of reference values.(AU)

In vivo imaging of cumulus-oocyte-complexes and small ovarian follicles in cattle using ultrasonic biomicroscopy.

The objectives of this study were to: (1) evaluate the feasibility of ultrasonic biomicroscopy (UBM) for imaging ovarian follicles in vivo in cows and heifers, (2) compare the transvaginal to the transrectal approach, (3) compare the number of follicles detected by UBM to conventional ultrasonography (US), and (4) identify cumulus-oocyte-complexes (COC) within follicles by UBM. Mature cows (n=5) and peri-pubertal heifers (n=5) were examined once using conventional B-mode US (Aloka 900) with a 7.5 MHz transrectal, and a 5 MHz transvaginal transducers. A second series of examinations was performed using UBM (Visualsonics Vevo 660) equipped with a 30 MHz transrectal and a 40 MHz transvaginal transducer. A three- to four-fold increase in the number of small follicles (<3 mm) was detected using the transvaginal approach with UBM compared to conventional US in both heifers (32.4 ± 4.24 compared to 7.2 ± 1.4; P<0.0001) and cows (35.0 ± 13.8 compared to 10.7 ± 7.5; P=0.0013). However, fewer follicles (all sizes combined) were detected using the transrectal approach with UBM compared to conventional US in both heifers (8.6 ± 5.6 compared to 17.6 ± 4.9; P=0.02) and cows (5.3 ± 6.1 compared to 20.3 ± 7; P=0.04). In heifers, COC-like structures were identified in 39 of 202 (19.3%) follicles examined. In conclusion, UBM using a transvaginal approach is feasible and may be used for in vivo assessment of early antral follicles as small as 0.4 mm, and COC within follicles.

Features and perspectives of the Brazilian in vitro embryo industry

In the last decade , in vitro fertilization emerged as an alternative to superovulation and has become the technique of choice for bovine embryo production, especially in zebu breeds. The recent growth in the commercial use of in vitro technologies in the Brazilian embryo industry is reviewed here in , highlighting the features and trends during different periods , as well as future challenges and perspectives . The data presented here w ere provided by the Statistics Committee of the Brazilian Embryo Techno logy Society and include reports from breeders associations, commercial IVF companies and ET practitioners . Three different periods were characterized for the use of IVF technologies in the Brazilian embryo industry: 1) the early years (1999 - 2003), when IV F growth was driven by the growing demand from the embryo market, although the technology was still labeled as elit ist ; 2) a period of exponential growth (2003 - 2006), when IVF overc a me conventional ET as the technique of choice for embryo production; and 3 ) a later period , when total numbers tend ed to stabilize but IVF started to increase in importance in dairy breeds. The whole picture shows IVF as an interesting example of innovation, since the development of these new embryo technologies provided new pro ducts, processes and possibilities to satisfy demands and remarkably change the scenario of the Brazilian embryo industry.

Features and perspectives of the Brazilian in vitro embryo industry

In the last decade , in vitro fertilization emerged as an alternative to superovulation and has become the technique of choice for bovine embryo production, especially in zebu breeds. The recent growth in the commercial use of in vitro technologies in the Brazilian embryo industry is reviewed here in , highlighting the features and trends during different periods , as well as future challenges and perspectives . The data presented here w ere provided by the Statistics Committee of the Brazilian Embryo Techno logy Society and include reports from breeders associations, commercial IVF companies and ET practitioners . Three different periods were characterized for the use of IVF technologies in the Brazilian embryo industry: 1) the early years (1999 - 2003), when IV F growth was driven by the growing demand from the embryo market, although the technology was still labeled as elit ist ; 2) a period of exponential growth (2003 - 2006), when IVF overc a me conventional ET as the technique of choice for embryo production; and 3 ) a later period , when total numbers tend ed to stabilize but IVF started to increase in importance in dairy breeds. The whole picture shows IVF as an interesting example of innovation, since the development of these new embryo technologies provided new pro ducts, processes and possibilities to satisfy demands and remarkably change the scenario of the Brazilian embryo industry.(AU)

Ovarian follicular dynamics, follicle deviation, and oocyte yield in Gyr breed (Bos indicus) cows undergoing repeated ovum pick-up.

The objective of this study was to evaluate ovarian follicular dynamics during intervals between successive ovum pick-up (OPU) and determine its effects on the number and quality of recovered cumulus-oocyte complexes (COCs) in Zebu cows (Bos indicus). Pluriparous nonlactating Gyr cows (Bos indicus; n=10) underwent four consecutive OPU sessions at 96-h intervals. The dynamics of ovarian follicular growth between OPU sessions was monitored by twice-daily ultrasonographic examinations. A single dominant follicle (DF) or two codominant (CDF) follicles (>9mm) were present in 63.3% (19 of 30) of intervals studied, with follicle deviation beginning when the future dominant follicle (F1) achieved a diameter of 6.2+/-0.3mm. The phenomenon of codominance was observed in four (13.3%) of the inter-OPU intervals. The remaining intervals (36.6%, 11 of 30) were characterized by a greater follicular population, lower rate of follicular growth, and a smaller diameter F1 (P<0.0001). There was a tendency (P=0.08) toward an increase in the number of recovered COCs when dominant follicles were not present (NDF). The quality of COCs was not affected by the presence of a single dominant follicle, but codominant follicles resulted in recovery of a lower proportion of viable embryos (40.0%, 62.1%, and 63.6%; P<0.05) and higher proportions of degenerate COCs (56.0%, 30.3%, and 28.6%; P<0.05) for CDF, NDF, and DF respectively. We concluded that, in Zebu cows, (a) repeated follicle aspirations altered ovarian follicular dynamics, perhaps by increasing follicular growth rate; (b) follicular dominance could be established in cows undergoing twice-a-week OPU; and (c) the presence of a dominant follicle during short inter-OPU intervals may not affect COC quality, except when a codominant follicle was present.

Pregnancy rates and corpus luteum-related factors affecting pregnancy establishment in bovine recipients synchronized for fixed-time embryo transfer.

The objective was to investigate the influence of corpora lutea physical and functional characteristics on pregnancy rates in bovine recipients synchronized for fixed-time embryo transfer (FTET). Crossbred (Bos taurus taurus x Bos taurus indicus) nonlactating cows and heifers (n=259) were treated with the following protocol: 2mg estradiol benzoate (EB) plus an intravaginal progesterone device (CIDR 1.9g progesterone; Day 0); 400 IU equine chorionic gonadotropin (eCG; Day 5); prostaglandin F(2alpha) (PGF(2alpha)) and CIDR withdrawal (Day 8); and 1mg EB (Day 9). Ovarian ultrasonography and blood sample collections were performed on Day 17. Of the 259 cattle initially treated, 197 (76.1%) were suitable recipients; they received a single, fresh, quality grade 1 or 2 in vivo-derived (n=90) or in vitro-produced (n=87) embryo on Day 17. Pregnancy rates (23 d after embryo transfer) were higher for in vivo-derived embryos than for in vitro-produced embryos (58.8% vs. 31.0%, respectively; P<0.001). Mean (+/-SD) plasma progesterone (P(4)) concentration was higher in cattle that became pregnant than that in nonpregnant cattle (5.2+/-5.0 vs. 3.8+/-2.4 ng/mL; P=0.02). Mean pixel values (71.8+/-1.3 vs. 71.2+/-1.1) and pixel heterogeneity (14.8+/-0.3 vs. 14.5+/-0.5) were similar between pregnant and nonpregnant recipients (P>0.10). No significant relationship was detected between pregnancy outcome and plasma P(4), corpus luteum area, or corpus luteum echotexture. Embryo type, however, affected the odds of pregnancy. In conclusion, corpus luteum-related traits were poor predictors of pregnancy in recipients. The type of embryo, however, was a major factor affecting pregnancy outcome.

Efeito do nível de uréia na dieta sobre o desempenho, a qualidade e o estádio de desenvolvimento embrionário em cabras Alpinas / Effect of the dietary urea level on performance, quality and embryonic development stage on Alpine goats

Utilizaram-se 22 cabras da raça Alpina, distribuídas aleatoriamente em quatro tratamentos (T): as cabras do T1 (n=5) formaram o grupo-controle; as do T2 (n=7) receberam 0,73 por cento de uréia na matéria seca da dieta; as do T3 (n=4) receberam 1,46 por cento de uréia; e as do T4 (n=6), 2,24 por cento de uréia. As cabras foram superovuladas e os embriões, coletados entre sete e oito dias após a primeira monta, foram avaliados quanto à qualidade e ao estádio de desenvolvimento. Amostras de sangue para dosagem dos teores de uréia e glicose foram coletadas nos dias do estro e da coleta de embriões. Houve efeito linear crescente do nível de uréia nas dietas sobre o consumo de MS (kg/dia) e de proteína bruta (kg/dia). O peso das cabras não diferiu (P>0,05) entre os tratamentos nem entre as semanas experimentais. Dezoito cabras (81,8 por cento) manifestaram estro após a sincronização. A duração do estro e o intervalo da remoção da esponja ao início do estro não foram influenciados (P>0,05) pelos tratamentos. Quatorze cabras (77,8 por cento) responderam à superovulação. O número de estruturas e de embriões coletados não diferiu (P>0,05) entre os tratamentos. O número (Y= 10,90 - 11,64NS U + 4,93§U²; R² = 0,67; P<0,10) e a percentagem (Y= 94,08 - 39,59NS U + 18,16ªU²; R² = 0,94; P<0,07) de embriões viáveis, e o número (Y= 10,83- 12,18NS U + 5,02mU²; R² = 0,78; P<0,08) e a percentagem (Y= 94,83- 52,31NS U + 21,56*U²; R² = 0,90; P<0,05) de embriões excelentes e bons apresentaram comportamento quadrático em função do nível de uréia nas dietas. Os teores de uréia e glicose no plasma não foram influenciados (P>0,05) pelos tratamentos. A uréia pode ser fornecida no nível de 2,24 por cento na MS da dieta de cabras não lactantes(AU)

Twenty-two Alpine goats were allocated at random into four treatments: 0.0 percent (T1 - control, n=5); 0.73 percent (T2, n=7); 1.46 percent (T3, n=4) or 2.24 percent (T4, n=6) of urea in the dry matter (DM) of the diet. Embryos collected from 7 to 8 days after mating of superovulated goat were evaluated by quality and development stage. Blood samples for urea and glucose analyses were collected at estrus and at embryos collection day. The DM (kg/day) and crude protein (kg/day) intake increased linearly in function of dietary urea level. Goat body weights were not affected by treatments out experimental weeks (P>0.05). Eighteen goats (81.8 percent) came in estrus after the synchronization. The estrus length and the interval from sponge removal to the beginning of estrus were not affected (P>0.05) by treatments. Fourteen goats (77.8 percent) were responsive to superovulation protocol. The levels of urea (treatments) did not affect structures and embryo numbers (P>0.05). Number (Y= 10.90 - 11.64NS U + 4.93§U²; R²= 0.67; P<0.10) and percentage (Y= 94.08 - 39.59NS U + 18.16ªU²; R²= 0.94; P<0.07) of viable embryos, and number (Y= 10.83- 12.18NS U + 5.02mU²; R²= 0.78; P<0.08) and the percentage (Y= 94.83- 52.31NS U + 21.56*U²; R²= 0.90; P<0.05) of excellent and good embryos were quadraticaly effected by urea dietary level. The treatments did not affect plasma urea and glucose levels (P>0.05). Diets for no nursing goats can be supplied by urea at 2.24 percent of DM(AU)

Efeito do nível de uréia na dieta sobre o desempenho, a qualidade e o estádio de desenvolvimento embrionário em cabras Alpinas / Effect of the dietary urea level on performance, quality and embryonic development stage on Alpine goats

Utilizaram-se 22 cabras da raça Alpina, distribuídas aleatoriamente em quatro tratamentos (T): as cabras do T1 (n=5) formaram o grupo-controle; as do T2 (n=7) receberam 0,73 por cento de uréia na matéria seca da dieta; as do T3 (n=4) receberam 1,46 por cento de uréia; e as do T4 (n=6), 2,24 por cento de uréia. As cabras foram superovuladas e os embriões, coletados entre sete e oito dias após a primeira monta, foram avaliados quanto à qualidade e ao estádio de desenvolvimento. Amostras de sangue para dosagem dos teores de uréia e glicose foram coletadas nos dias do estro e da coleta de embriões. Houve efeito linear crescente do nível de uréia nas dietas sobre o consumo de MS (kg/dia) e de proteína bruta (kg/dia). O peso das cabras não diferiu (P>0,05) entre os tratamentos nem entre as semanas experimentais. Dezoito cabras (81,8 por cento) manifestaram estro após a sincronização. A duração do estro e o intervalo da remoção da esponja ao início do estro não foram influenciados (P>0,05) pelos tratamentos. Quatorze cabras (77,8 por cento) responderam à superovulação. O número de estruturas e de embriões coletados não diferiu (P>0,05) entre os tratamentos. O número (Y= 10,90 - 11,64NS U + 4,93§U²; R² = 0,67; P<0,10) e a percentagem (Y= 94,08 - 39,59NS U + 18,16ªU²; R² = 0,94; P<0,07) de embriões viáveis, e o número (Y= 10,83- 12,18NS U + 5,02mU²; R² = 0,78; P<0,08) e a percentagem (Y= 94,83- 52,31NS U + 21,56*U²; R² = 0,90; P<0,05) de embriões excelentes e bons apresentaram comportamento quadrático em função do nível de uréia nas dietas. Os teores de uréia e glicose no plasma não foram influenciados (P>0,05) pelos tratamentos. A uréia pode ser fornecida no nível de 2,24 por cento na MS da dieta de cabras não lactantes

Twenty-two Alpine goats were allocated at random into four treatments: 0.0 percent (T1 - control, n=5); 0.73 percent (T2, n=7); 1.46 percent (T3, n=4) or 2.24 percent (T4, n=6) of urea in the dry matter (DM) of the diet. Embryos collected from 7 to 8 days after mating of superovulated goat were evaluated by quality and development stage. Blood samples for urea and glucose analyses were collected at estrus and at embryos collection day. The DM (kg/day) and crude protein (kg/day) intake increased linearly in function of dietary urea level. Goat body weights were not affected by treatments out experimental weeks (P>0.05). Eighteen goats (81.8 percent) came in estrus after the synchronization. The estrus length and the interval from sponge removal to the beginning of estrus were not affected (P>0.05) by treatments. Fourteen goats (77.8 percent) were responsive to superovulation protocol. The levels of urea (treatments) did not affect structures and embryo numbers (P>0.05). Number (Y= 10.90 - 11.64NS U + 4.93§U²; R²= 0.67; P<0.10) and percentage (Y= 94.08 - 39.59NS U + 18.16ªU²; R²= 0.94; P<0.07) of viable embryos, and number (Y= 10.83- 12.18NS U + 5.02mU²; R²= 0.78; P<0.08) and the percentage (Y= 94.83- 52.31NS U + 21.56*U²; R²= 0.90; P<0.05) of excellent and good embryos were quadraticaly effected by urea dietary level. The treatments did not affect plasma urea and glucose levels (P>0.05). Diets for no nursing goats can be supplied by urea at 2.24 percent of DM