Progestin-Based Protocol for Synchronization of Follicular Wave Emergence in Buffaloes During Summer and Winter

The aim of the present study was to evaluate the effects of type of norgestomet auricular implant (new - N or previously used during 5 days - U), season of the year (summer - S and winter - W), and parity (12 heifers - H and 23 cows - C) on synchronization of follicular wave emergence in buffaloes. For this purpose, 35 buffaloes were examined daily by ultrasonography until follicular wave emergence was detected. Data were analysed by ANOVA, using PROC GLIMMIX. No interactions were observed in none variables. Time of follicular wave emergence and number of follicles at emergence were not affected by type of implant or season of the year. Parity also did not influence the number of follicles at emergence. However, follicular wave emergence occurred later in heifers than in cows. In conclusion, the previous use of a norgestomet auricular implant independent of the season of the year does not affect the time or the number of follicles at follicular wave emergence in buffaloes. Nevertheless, although heifers and cows had a similar number of follicles at emergence, the time of follicular wave emergence occurs earlier in cows than in heifers.

Ovarian response to porcine FSH in association with ablation-induced or spontaneous follicular wave development during the estrous cycle in crossbred and Brazilian Warmblood mares

The primary objective of this study was to examine the follicular and ovulatory responses following treatment with pFSH in association with ablation-induced or spontaneous follicular wave emergence or follicle deviation during diestrus in crossbred (Mangalarga × Arabian) and Brazilian Warmblood mares with a propensity for spontaneous multiple ovulations; secondary considerations were given to the collection of embryos In Experiment 1, crossbred mares were administered (im) saline (control, n= 7) or pFSH (25 mg) when the largest follicle of the ablation-induced follicular wave reached ≥13 mm (n= 7) or ≥20 mm (n= 7) or, after pre-treatment ovulation (Day 0) on Day 6 (n= 7) In Experiment 2, crossbred mares were administered (im) saline (control, n= 10) or a larger dose of pFSH (50 mg, n= 7) when the largest follicle of the ablation-induced follicular wave reached ≥13 mm In Experiment 3, Brazilian Warmblood mares were administered (im) saline (control, n= 7), pFSH (25 mg, n= 7 or 50 mg, n= 5) or EPE (12.5 mg, n= 7) as a positive control on Day 6 Ultrasonic technology was used to ablate all follicles ≥8 mm and to monitor follicular development and detect ovulation Treatment with pFSH or EPE was done twice daily until the largest follicle reached ≥32 mm; thereafter, hCG (2500 IU) was administered (iv) when the largest follicle reached ≥35 mm Artificial insemination was done 12 h after hCG and embryo collections were done 8 d after post-treatment ovulations In Experiments 1 and 2, treatment of crossbred mares with pFSH post-ablation in association with the expected time of wave emergence or follicle deviation did not (P> 0.05) enhance the follicular or ovulatory responses or collection of embryos compared to controls In Experiment 3, although the enhanced ovulatory response of mares to EPE at the expected time of spontaneous wave emergence was not different (P> 0.05) from controls, it was greater (P< 0.05) than the response to pFSH In conclusion, the novelty of using follicle ablation prior to pFSH treatment at the time of wave emergence or follicle deviation did not enhance the follicular or ovulatory responses or collection of embryos to treatment in crossbred mares In addition, the hypothesis that Brazilian Warmblood mares with a greater propensity for spontaneous multiple ovulations are as responsive to pFSH compared to EPE was not supported Thus, the combined experimental results of the present study continue to support the general consensus that pFSH is relatively ineffective for follicular superstimulation/superovulation in mares © 2012 Elsevier B.V.

The interval between the emergence of pharmacologically synchronized ovarian follicular waves and ovum pickup does not significantly affect in vitro embryo production in Bos indicus, Bos taurus, and Bubalus bubalis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Superovulation and embryo transfer in Bos indices cattle

Compared to Bos taurus breeds, Bos indices breeds of cattle present several differences in reproductive physiology. Follicular diameter at deviation and at the time of ovulatory capability are smaller in B. indicus breeds. Furthermore, B. indicus breeds have a greater sensitivity to gonadotropins, a shorter duration of estrus, and more often express estrus during the night. These differences must be considered when setting up embryo transfer programs for B. indicus cattle. In recent studies, we evaluated follicular dynamics and superovulatory responses in B. indicus donors with the objective of implementing fixed-time AI protocols in superstimulated donors. Protocols using estradiol and progesterone/progestrogen releasing devices to control follicular wave emergence were as efficacious as in B. taurus cattle, allowing the initiation of superstimulatory treatments (with lower dosages of FSH than in B. taurus donors) at a self-appointed time. Furthermore, results presented herein indicate that delaying the removal of progesterone/progestogen-releasing devices, combined with the administration of GnRH or pLH 12 h after the last FSH injection, results in synchronous ovulations, permitting the application of fixed-time AI of donors without the necessity of estrus detection and without compromising the results. (c) 2005 Elsevier B.V. All rights reserved.

Follicle profile and plasma gonadotropin concentration in pubertal female ponies

Twelve female ponies were examined daily for 30 days and classified as ovulating (OV; N = 6; 197 ± 6 kg) or prepubertal (PP; N = 6; 196 ± 9 kg). Follicles were detected by ultrasound and gonadotropins quantified by radioimmunoassay. The mean diameter of the largest follicles was significantly larger in OV (38 ± 1 mm) than in PP (26 ± 2 mm) but there was no difference between groups in the size of the second largest follicle. There were more small follicles (<24 mm) in the PP than in the OV group, but PP fillies had a smaller number of follicles >29 mm than the OV fillies. Follicle-stimulating hormone (FSH) levels did not differ between groups but PP fillies had lower luteinizing hormone (LH) peak (8 ± 1 ng/ml) and basal (4 ± 0.5 ng/ml) levels, lower peak magnitude (2 ± 0.2 ng/ml) and period average (5 ± 0.6 ng/ml) than OV fillies (32 ± 4.5, 8 ± 1.2, 17.1 ± 6, and 15 ± 2.3 ng/ml, respectively). The PP group, in contrast to the OV group, showed no relationship between FSH surge and follicle wave emergence. We conclude that an LH concentration higher than 8 ng/ml is needed for follicle growth to a preovulatory size. Wave emergence and FSH secretion seem to be independent events, probably due to an inhibitory neural system in these PP animals. PP fillies may provide a physiological model for the study of follicle wave emergence which apparently does not depend on gonadotropin levels.

Morphological characterization of follicle deviation in Nelore (Bos indicus) heifers and cows

Follicle diameter deviation is defined as the beginning of the differential change in growth rates between the largest and next largest follicles subsequent to wave emergence and is considered a key component of follicle selection. Follicle selection has been extensively studied in European breeds of cattle (Bos taurus) but has not been critically studied in Zebu breeds (Bos indicus). The objectives of the present study were to determine and compare the morphological characteristics of deviation associated with the first post-ovulatory wave (Wave 1) of the estrous cycle in Nelore heifers (n = 8) and nonlactating cows (n = 11). Beginning on the day of ovulation (day 0), the three largest follicles (F1-F3, respectively) were individually tracked every 12 h for 6 d using transrectal ultrasonography. In individual animals, deviation was determined graphically using visual inspection of the diameter profiles of F1, F2 and sometimes F3 (observed deviation) and mathematically using segmented regression analysis of the diameter differences between F1 and F2 or sometimes F3 (calculated deviation). Mean day of emergence of Wave 1 when F1 reached >3 rum (approximately 1 d after ovulation) and growth rate of F1 during deviation (approximately 1.4 mm/d) were not significantly different between heifers and cows. The results of determining the beginning of deviation within heifers and cows using the observed and calculated methods were not significantly different. Averaged over both methods, diameter deviation occurred 2.8 d after ovulation when F1 reached 5.7 mm in heifers, and 2.4 d after ovulation when F1 reached 6.1 mm in cows. In conclusion, the emergence of Wave 1 and growth rates and diameters of the future dominant follicles at the beginning of deviation were similar in Nelore heifers and nonlactating cows, regardless of the methods used to determine deviation. Relative to Holstein cattle, emergence of Wave 1 appeared to occur about 1 d later and diameter of the future dominant follicle at the beginning of deviation was about 2 turn smaller in Nelore. (C) 2004 Elsevier B.V. All rights reserved.

Effects of dominant follicle aspiration and treatment with recombinant bovine somatotropin (BST) on ovarian follicular development in Nelore (Bos indicus) heifers

Follicle ablation has been recognized as an efficient method of follicular wave synchronization. Treatment with recombinant bovine somatotropin (BST) has been shown to enhance follicular development in <(Bos taurus)under bar>. This experiment assessed the effects of these treatments in Nelore (<(B. indicus)under bar>) heifers. Eight cycling Nelore heifers were randomly assigned to 3 different treatments. on Day 2 of a synchronized cycle (Day 0 = day of ovulation), heifers assigned to Treatments 1 and 2 received 2 mL of saline, whereas heifers assigned to Treatment 3 received 320 mg of BST. on Day 5, the first-wave dominant follicle was ablated by ultrasoundguided transvaginal aspiration in heifers in Treatments 2 and 3, and all heifers received an injection of prostaglandin on Day 11. Aspiration of the dominant follicle advanced and synchronized (P < 0.05) the day of second-wave emergence (6.9 +/- 0.1 vs. 8.4 +/- 0.4) and the day of the pre-wave FSH peak (6.0 +/- 0.0 vs. 6.9 +/- 0.4), and increased FSH peak concentrations (381 +/- 21 vs. 292 +/- 30; pg/mL; P < 0.01). Recombinant bovine somatotropin treatment caused a two-fold increase in plasma insulin-like growth factor-I (IGF-I) concentrations (P < 0.001) and resulted in a 36% increase in the number of small follicles (<5 mm; P < 0.001) compared with saline-treated heifers. In summary, in agreement with previous reports on <(B. taurus)under bar>, dominant follicle aspiration synchronized ovarian follicular development, and BST treatment increased peripheral concentrations of IGF-I in Nelore heifers. Recombinant bovine somatotropin also increased the number of small follicles, but this response appeared to be inferior to that reported for <(B. taurus)under bar>. (C) 2000 by Elsevier B.V.

Temporal interrelationships among luteolysis, FSH and LH concentrations and follicle deviation in mares

The effect of altered LH concentrations on the deviation in growth rates between the 2 largest follicles was studied in pony mares. The progestational phase was shortened by administration of PGF2α on Day 10 (Day 0=ovulation; n=9) or lengthened by daily administration of 100 mg of progesterone on Days 10 to 30 (n=11; controls, n=10). All follicles ≥5 mm were ablated on Day 10 in all groups to initiate a new follicular wave. The interovulatory interval was not altered by the PGF2α treatment despite a 4-day earlier decrease in progesterone concentrations. Time required for growth of the follicles of the new wave apparently delayed the interval to ovulation after luteolysis. The FSH concentrations of the first post-ablation FSH surge were not different among groups. A second FSH surge with an associated follicular wave began by Day 22 in 7 of 11 mares in the progesterone group and in 0 of 19 mares in the other groups, indicating reduced functional competence of the largest follicle. A prolonged elevation in LH concentrations began on the mean day of wave emergence (Day 11) in the prostaglandin group (19.2 ± 2.2 vs 9.0 ± 0.7 ng/mL in controls; P<0.05), an average of 4 d before an increase in the controls. Concentrations of LH in the progesterone group initially increased until Day 14 and then decreased so that by Day 18 the concentrations were lower (P<0.05) than in the control group (12.9 ± 1.6 vs 20.2 ± 2.6 ng/mL). Neither the early and prolonged increase nor the early decrease in LH concentrations altered the growth profile of the second-largest follicle, suggesting that LH was not involved in the initiation of deviation. However, the early decrease in LH concentrations in the progesterone group was followed by a smaller (P<0.05) diameter of the largest follicle by Day 20 (26.9 ± 1.7 mm) than the controls (30.3 ± 1.7 mm), suggesting that LH was necessary for continued growth of the largest follicle after deviation. (C) 2000 by Elsevier B.V.

Caracterização ultra-sonográfica do desvio folicular ovariano em vacas da raça nelore (Bos taurus indicus)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Eficiência de diferentes doses de acetato de fertirelina na sincronização da emergência da onda folicular e indução da ovulação em vacas leiteiras

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Molecular Evaluation of Developmental Competence of Oocytes Collected In Vivo from Buffalo and Bovine Heifers during Winter and Summer

Buffaloes and bovines are polyestrous and seasonal or annual livestock, respectively, that show reduced fertility during heat stress. To investigate whether reduced fertility is related to oocyte competence in both species, immature oocytes from buffalo and bovine heifers were collected during winter and summer and subjected to molecular analyses. In each season, heifers of both species had their follicular wave emergence synchronized with a standard protocol (Ferreira et al., 2011). Before being subjected to ovum pick up (OPU), cutaneous (CT; degrees C) and rectal (RT; degrees C) temperatures and respiratory rate (RR; breaths/min) were measured. Oocytes' RNA was extracted to evaluate the expression of target genes related to mtDNA replication/transcription (PPARGC1A, TFAM and MT-CO1), apoptosis (BAX and BCL2) and HS (HSP90AA1 and HSPA1AB). ACTB, HIST1H2AG and GAPDH were initially chosen as housekeeping genes. In buffaloes, CT (35.0 +/- 0.4 vs 23.8 +/- 0.5), RT (38.7 +/- 0.1 vs 38.0 +/- 0) and RR (21.3 +/- 1.2 vs 15.4 +/- 1.1) were higher during summer than winter. However, in bovine heifers, RT (38.7 +/- 0.1 vs 38.6 +/- 0.1) and RR (44.8 +/- 1.5 vs 40.6 +/- 1.5) were similar in both seasons, while CT (31.6 +/- 0.3 vs 30.2 +/- 0.3) was increased during summer. Reduced expression of ACTB, HIST1H2AG and GAPDH was evidenced during summer, disqualifying them as housekeeping genes. Similarly, the expression of all target genes was reduced during summer in oocytes of both species. In summary, physiological responses to heat stress seem to be more intense in buffalo than bovine heifers. However, in both species, negative effects of heat stress upon oocyte quality occur at the molecular level and affects genes related to several biological functions.

Relationships between growth of the preovulatory follicle and gestation success in lactating dairy cows

This report summarizes three studies conducted with lactating dairy cows aiming to increase pregnancy rates to fixed time artificial insemination (TAI) protocols. Experiment 1 was designed to determine if changing the timing of PGF2 alpha treatment during an E2/P4-based program would affect fertility to TAI or fixed-time embryo transfer (TET). In experiment 2, pregnancy rates to AI were compared following synchronized ovulation using two protocols that have been developed to reduce the period between follicular wave emergence and TAI. The Ovsynch-type protocol utilizes GnRH to synchronize the follicular wave by inducing ovulation of a dominant follicle at the beginning of the protocol, and to synchronize ovulation at the end of the protocol allowing TAI. In contrast, E2/P4-based protocols utilize E2 products in the presence of P4 to induce atresia of antral follicles and synchronize emergence of a new follicular wave. At the end of E2/P4-based protocol another E2 treatment in the absence of P4 is used to induce LH release and synchronize ovulation and allow TAI. Experiment 3 was designed to determine whether increasing the length time interval with reduced circulating P4 (proestrus) would increase fertility in a TAI program that utilized E2 and P4 to synchronize ovulation of cycling, lactating dairy cows. The overall conclusions are that circulating concentrations of progesterone and estradiol prior to and circulating concentrations of progesterone following ovulation can affect fertility in cattle. In addition, small increases in P4 concentrations near the time of AI, due to lack of complete CL regression, result in reductions in fertility. Earlier treatment with PGF2 alpha should allow greater time for CL regression, an increase in estradiol and subsequent reductions in circulating P4 that could be critical for fertility. Optimization of follicle size in TAI programs is clearly an intricate balance between oocyte quality, adequate circulating E2 near AI, and adequate circulating P4 after AI.

Influence of Parity and Season of the Year on Oocyte Quality and Number in Buffaloes

The aim of the present study was to evaluate the effects of season of the year (summer and winter) and parity (heifers and cows) on oocyte quality and number in buffaloes. For this purpose, 71 buffaloes had follicular wave emergence synchronized before OPU. OPU of all follicles >= 2mm was done 5 days after the beginning of the hormonal protocol, in 4 replicates (two for each season). Data were analyzed by ANOVA using PROC GLIMMIX, in a 2 x 2 factorial arrangement of treatments. No interactions were observed in following variables: number of follicles, number of total and viable oocytes, recovery rate, percentage of viable oocytes, grade I oocytes, grade II oocytes, grade III oocytes, denuded oocytes, expanded cumulus oocytes, and atretic/degenerated oocytes. Number of follicles visualized at OPU and recovery rate were not affected by parity or season. Relative to parity, number of total and viable oocytes were greater in heifers than in cows, respectively. Concerning season of the year, number of viable oocytes and viable oocyte rate were increased in winter. In conclusion, better oocyte quality can be obtained from heifers and during winter in buffaloes. However, the number of total oocytes seems to be more influenced by parity than by season of the year in this species.