Effect of recombinant bovine somatotropin on the reproductive efficiency of beef cows subjected to differently timed-artificial insemination protocols.

This study aimed to verify the reproductive efficiency of beef cows treated with recombinant bovine somatotropin (rbST). Study 1, Bos indicus cows were distributed (three groups). The control group (CG) was subjected: on day zero (d0), the animals received a CIDR and oestradiol benzoate (EB); on (d8, CIDR was removed, and PGF2α and oestradiol cypionate (EC) were administered; on d10, timed Artificial Insemination (TAI) was performed; on d45, pregnancy diagnosis was made. The rbST on d0 group (bST0G) was subjected to an identical protocol as CG, except for the addition of 250 mg rbST on d0. The rbST on d8 group (bST8G) was subjected to the same protocol as bST0G, except that the rbST was administered on d8 rather. In study 2, the animals followed the same design which was used in Bos taurus cows. The follicular growth rate (FGR) was calculated between d8 and d10. In study 1, pregnancy/artificial insemination (P/AI) did not differ among the treatments. FGR in bST8G was higher than in other groups. In study 2, bST0G showed higher Pregnancy/Artificial Insemination (P/AI) (p < .05) when compared with other groups. bST0G showed a different FGR (p < .0001) than the other groups. In conclusion, rbST (Bos indicus cows) did not increase P/AI, but it did promote follicular growth when administered on d8; the rbST administered on d0 improved P/AI (p < .05) and the FGR in Bos taurus cows.

Progesterone Device Use Improves Ovum Pick-Up Efficiency in Acyclic Donors.

The aim of this study was to evaluate follicular dynamics and ovum pick-up (OPU) efficacy in untreated mares or mares treated with an intravaginal progesterone (P4) device during seasonal anestrus (acyclic) and during the breeding season (cyclic). Six mares (mean age = 5 years), were recruited into an ovum pick-up scheme that was performed every 14 days with and without the P4 device, during the acyclic and cyclic phases. Aspirations amounted to seven procedures with or without the P4 device during each phase. Five ultrasound assessments were performed at each interval between the OPUs. Data on follicular number and diameter as well as the numbers of recovered and the percentage of recovered oocytes were also collected. The number of follicles from mares in the acyclic phase was higher (P < .005) regardless of the treatment. However, the follicular diameter was smaller for the P4 group (P < .005) from the 2nd to the 5th evaluation post-OPU procedure. The percentage of oocytes recovered during the acyclic phase was higher for mares treated with the P4 device (P < .005). The P4 device resulted in follicles with smaller diameters and facilitated OPU efficacy.

Effects of two protocols of ovulation synchronization on corpus luteum size and blood flow, progesterone concentration, and pregnancy rate in beef heifers.

The objective of this study was to evaluate the effect of 7-day estradiol-progesterone-based [Treat(C)] and 5-day Co-Synch plus progesterone [Treat(Co-Sy)] protocols on ovulation time, pre-ovulatory follicle diameter, corpus luteum (CL) size and blood flow, progesterone (P4) concentration and pregnancy rate (PR) in beef heifers. In Experiment 1, a crossover design was applied (n = 9). For Treat(C), a progesterone intravaginal (PI) device was inserted, plus 2 mg of estradiol benzoate (day 0). On day 7, 500 µg of cloprostenol plus 0.5 mg of estradiol cypionate were administered, and PI was removed. For Treat(Co-Sy), on day 0, a PI was inserted plus 100 µg gonadotropin-releasing hormone (GnRH). On day 5, PI was removed, plus 500 µg of cloprostenol and 100 µg of GnRH were administered at 69-70 h later. From day one to ovulation day, dominant follicle was evaluated by ultrasonography. On days 4 and 8 post-ovulation, CL was evaluated by color Doppler, and P4 concentration was determined by chemiluminescence. In Experiment 2, a split-plot experimental design was used. Protocols followed were the same as in Experiment 1 [Treat(C); n = 310 and Treat(Co-Sy); n = 314]. Heifers were fixed-time artificially inseminated. Pregnancy was determined on day 41. In Experiment 1, the interval between PI removal and ovulation time was different between protocols (P < 0.01). In addition, P4 concentration was related to the CL size (P < 0.001), CL blood flow (P < 0.01) and protocols (P < 0.03). In Experiment 2, PR did not differ between protocols.

Ovarian Follicular Dynamics and Its Functional Significance in Relation with Follicle Deviation, Vaginal Cytology, and Hormone Profiles in Llamas (Lama glama).

The reproductive physiology in camelid species has its particularities. The present study aimed to characterize the ovarian follicular dynamics and its functional significance in relation to follicular deviation, vaginal cytological characteristics, and sexual hormone profiles in llamas as the first report in South American camelids. Non-pregnant, multiparous llamas (Lama glama; n = 10; age: 48−72 mo.; BCS: 2.5−3.0) were enrolled in the study. The ultrasonographic assessment was carried out transvaginally and follicular ablation was performed (day 0) when follicles were larger than 7 mm. The follicle number and diameter were scored daily throughout the process for a proper evaluation of the deviated follicles and to monitor the presence of new follicle pools (1.5 to 2.5 mm diameter). Vaginal cytological evaluation (parabasal, intermediate, and superficial cells) was performed every other day until day 6. Endocrine profiles (17ß estradiol, anti-Mullerian hormone, testosterone, and progesterone) during pre- and post-follicular deviation were determined by using the ELISA assay. Differential follicular dynamics both in the presence of a single dominant follicle (DF) and in codominance during the follicular deviation process were detected in llamas (p < 0.05). The percentage of superficial cells was the most related to the follicular wave phase. However, the percentage of parabasal, intermediate, and superficial cells was not related to the phases of follicular growth, dominance, and regression (p > 0.05). Differential patterns among the different hormone concentration levels regarding the 17ß estradiol, anti-Mullerian hormone, progesterone, and testosterone during follicular deviation were observed, with the latter being significantly different along the deviation process (p < 0.05). In conclusion, the use of vaginal cytology assessment would not be sufficient to determine the follicular phases in llamas. Therefore, complementary analyses, such as ultrasonography and endocrine assessment, are strongly recommended to determine follicular dynamics during the follicular deviation.

Estimation of follicular growth-widely used, seldom studied.

We assessed whether estimation of follicular growth, rather than actual measurement of follicular size on the day of hCG trigger, affected pregnancy rates in intrauterine insemination (IUI) cycles. Patient and cycle characteristics were extracted from an existing database. Comparisons were made between the pregnant (defined as a positive beta hCG) and non-pregnant groups for the following variables: patient's age, number of previous IUI cycles, type of ovarian stimulation, endometrial thickness, number of follicles measuring 14 mm and above, pre and post wash sperm parameters, cycle day when IUI was done and number of days between last ultrasound scan and ovulation trigger. A total of 7302 cycles were included in the final analysis. In 4055 cycles (55.5%) the hCG trigger was on the day of the last ultrasound, in 2285 cycles (31.3%) the hCG trigger was 1 day after the last ultrasound, in 850 (11.6%) it was 2 days after the last ultrasound and in 112 (1.5%) it was 3 or more days after the last ultrasound. Sperm parameters, younger maternal age, and the number of follicles above 14 mm were all associated with pregnancy. No association was found between positive pregnancy test rates and the time from last ultrasound to hCG trigger. Planning IUI based on the estimation of follicular growth 1-4 days before trigger, does not affect pregnancy rates.

Comparing the effect of estradiol benzoate and 17ß-estradiol plus progesterone on follicular turnover and development, and pregnancy outcomes in a timed artificial insemination protocol.

The objective of this study was to compare the efficiency of estradiol benzoate (EB) and 17ß-estradiol (E2) associated with progesterone (P4) in a fixed-time artificial insemination (FTAI) protocol. We hypothesized that E2+P4 induces an earlier emergence of a new follicular wave (NFW), improving pre-ovulatory follicle diameter and pregnancy rates to FTAI (P/FTAI). In Exp.1, on Day 0 (D0), all Bos indicus cows (n = 12/group) received an intravaginal P4 device and a dose of PGF2α analogue. On D0, females were randomly assigned to receive EB or E2+P4. On D8.5, P4 intravaginal devices were removed and a dose of PGF2α and EB were administered in all females followed by fixed-timed AI on D10. Between D0 and D10, the dominant follicular growth was determined by ovary ultrasonography exams. On D8.5 and D10 the percentage of color power-Doppler signals in the dominant follicular wall was evaluated. In Exp. 2, 467 females (2-year-old nulliparous [n = 76], primiparous [n = 92] and pluriparous [n = 299]) were subjected to the similar FTAI and assigned to be treated with EB (n = 243) or E2+P4 (n = 224). Pregnancy diagnosis was performed 30 days after FTAI by ultrasonography. The day to emergence of NFW was similar between treatments (EB: 3.7 ± 0.37 vs. E2+P4: 3.3 ± 0.3, P = 0.76). Females treated with E2+P4 presented greater (P = 0.06) follicular growth between the emergence and D9 (1.18 ± 0.07) than those treated with EB (0.97 ± 0.08). There was also a positive effect (P < 0.05) of E2+P4 on diameter of the dominant follicle on D9 (13.0 ± 0.6 vs. 10.9 ± 0.55) and blood perfusion of the follicle wall on D8.5 (49 vs. 40%). There was a treatment by parity category interaction effect on P/FTAI (P < 0.05). Treatment with E2+P4 was advantageous to P/FTAI of primiparous cows (E2+P4: 58% and EB: 30%). However, for nulliparous and pluriparous cows, P/FTAI was similar between treatments (∼50%). In conclusion, in a E2/P4-based protocol for FTAI, E2+P4 is as efficient as EB in inducing new follicular emergence within a similar day range, but it results similar or greater P/FTAI.

Follicular Dynamics during Estrous Cycle of Pubertal, Mature and Postpartum Crossbred (Nili Ravi × Jianghan) Buffaloes.

The follicular dynamics is used as a reliable indicator for reproductive management in livestock. However, the follicular dynamics (follicle wave emergence, estrus cycle length, diameter of dominant follicle, follicular growth and atretic phases) during the estrous cycle of crossbred (Nili Ravi-Jianghan) buffalo is still unexplored. Therefore, the present study aimed to observe the follicular dynamics in estrous cycle of crossbred buffaloes at different physiological stages (pubertal; n = 28, sexual mature; n = 22 and postpartum; n = 18). In the present study, the follicular dynamics were ultrasonically examined at 12 h intervals throughout an estrous cycle during the breeding season. The results indicate that about 86.76% (59/68) crossbred buffaloes, irrespective of physiological stage, exhibited two follicular waves in estrous cycle with an average estrus cycle length was 20.7 ± 0.4 days. The estrus cycle length was significantly shorter (p < 0.05) in pubertal buffaloes (19.4 ± 0.4 days) compared with sexual mature (21.5 ± 0.3 days) and postpartum (21.9 ± 0.4 days) buffaloes. The first follicular wave emerged on same day during one- (pubertal vs. postpartum), two- (pubertal vs. mature vs. postpartum) or three-wave (mature vs. postpartum) estrous cycle buffaloes. The maximum diameter of dominant follicle (DF) in pubertal, sexually mature and postpartum crossbred buffaloes was 9.6 ± 2.0 mm, 10.6 ± 0.5 mm and 12.6 ± 0.7 mm with growth rate of 1.08 ± 0.04 mm/day, 0.92 ± 0.04 mm/day, and 0.9 ± 0.07 mm/day, respectively. In conclusion, similar to other buffalo breeds, Nili Ravi-Jianghan crossbred buffaloes showed the two-wave follicular pattern dominantly with an average duration of ~20 days estrous cycle. The observed follicular dynamics can be used as a reliable indicator for synchronization and fixed-time artificial insemination (FTAI) programs to improve the fertility of crossbred buffaloes.

Waves of follicle development, growth and degeneration in adult ovary of zebrafish (Danio rerio) on chronic exposure to environmental estrogens in laboratory.

Patterns of quantitative production of follicles, their growth, and degeneration in the adult ovary of zebrafish (Danio rerio) in response to long-term (80 days) exposure to environmental estrogens (EE) in the laboratory, were studied. Experimentally naive female D. rerio procured from fish farm were acclimated to the laboratory (natural temperature, 26 ± 1° C, photoperiod, 11.30 L:12.30 D) for two weeks and divided into 10 groups. Each group (n = 20) was housed in a separate glass aquarium containing 10 L of conditioned water (physico-chemical parameters maintained within the permissible range prescribed for zebrafish) along with either 5 ng or 10 ng/L of 17α-ethynylestradiol (EE2) or diethylstilbestrol (DES) or bisphenol A (BPA) or estradiol 17-ß (positive control) or water with no chemical (negative control). All experimental fish were fed twice daily on commercial pellets (ad libitum) supplemented with Artemia nauplius, the exposure was semi-static and chemical residues in media samples were determined by ultra-performance liquid chromatography (UPLC). Exposure of fish to estrogens increased (p < 0.05) (i) body mass and gonadosomatic indices (GSI) in E2, EE2 and DES groups (ii) previtellogenic and vitellogenic follicles in E2 and EE2 groups (iii) atretic follicles (AF) in DES and BPA groups compared to controls and (iv) decrease in total oocyte volumes (V = 4/3. π. r3) compared to those of E2 group. These results suggest that the chronic exposure of fish to EE (at environmentally relevant concentrations) has a profound influence on ovarian follicular dynamics and the effects of individual EE are discrete on the ovary.

Production, metabolism, and follicular dynamics in multiparous dairy cows fed diets providing different amounts of metabolizable protein prepartum and postpartum.

Our objectives were (1) to determine whether increasing metabolizable protein (MP) supply above requirements in late-gestation cows would benefit health, milk production, and reproduction; (2) to determine whether an increased supply of MP postpartum affects production; and (3) to determine whether supply of MP prepartum interacts with MP supply postpartum. Pregnant nonlactating cows (n = 60) blocked by expected parturition date were assigned to 1 of 3 prepartum diets from 21 d prepartum to parturition: 12% crude protein (CP) soybean meal (SBM) supplement (LSB); 15% CP SBM supplement (HSB); and 15% CP SBM plus animal-marine protein supplement (HMP). Diets were formulated to supply an estimated 924, 988, and 1,111 g/d of MP, respectively, at 11.5 kg of dry matter intake (DMI). After parturition, cows received diets containing 18% CP, either from SBM (SB) or SBM plus animal-marine protein (AMP) supplements, that provided 2,056 (SB) or 2,293 g/d (AMP) of MP at 21 kg of DMI; thus, treatments were in a 3 × 2 factorial arrangement. Milk production and DMI were recorded for 63 d postpartum. Prepartum DMI was lower at wk -3 for cows fed LSB compared with those fed HSB or HMP. Postpartum DMI did not differ significantly between cows fed SB and those fed AMP (20.8 vs. 19.6 kg/d). Milk production did not differ due to prepartum diets or postpartum diets. Milk fat and protein percentages were not affected by prepartum or postpartum diets. Cows fed AMP postpartum tended to produce more milk fat, but 4% fat-corrected milk (FCM) did not differ from SB-supplemented cows (33.6 kg/d vs. 32.2 kg/d). Gross feed efficiency (FCM/DMI) was greater for cows fed AMP postpartum (1.82 vs. 1.68). Prepartum concentrations of urea N in plasma were lower for LSB than for HSB and HMP, and HSB was greater than HMP. Postpartum concentrations of nonesterified fatty acids and ß-hydroxybutyrate were greater for cows fed AMP postpartum than for those fed SB. Postpartum urea N was higher for SB than for AMP (14.4 vs. 12.5 mg/dL). Concentration of total protein in plasma was greater postpartum for cows fed HSB or HMP prepartum than for those fed LSB, and was greater postpartum for cows fed AMP than for those fed SB. Hepatic concentrations of total lipids and triglyceride did not differ among treatments. Hepatic glycogen was greater postpartum for cows fed SB postpartum. Feeding HSB or HMP increased the number of follicles 6 to 9 mm in diameter compared with LSB. The size of the largest follicle was increased by HMP compared with HSB. In conclusion, increasing the amount of MP fed to cows during the last 21 d prepartum did not affect milk production or BCS but increased plasma total protein concentration. Follicular dynamics were improved by increasing the amount of MP prepartum. Feeding HMP prepartum improved follicular dynamics prepartum and increased milk fat yield in wk 1. Feeding AMP postpartum increased efficiency of FCM production and plasma total protein. We found few interactions between prepartum and postpartum MP supply.

Effect of ovarian follicular wave pattern and endocrine characteristics on pregnancy outcome in cows.

The effect of two (2W) versus three (3W) wave patterns of follicular dynamics and concurrent endocrine milieu of follicle-stimulating hormone (FSH), luteinizing hormone (LH), oestradiol 17-ß (E2) and progesterone (P4) were investigated during one interoestrous interval (IEI) before insemination, on ensuing pregnancy, in 70 lactating Jersey crossbred cows. The findings were evaluated for between [included all (overall) 2W-O and 3W-O cows] and within [after separating pregnant (P) and non-pregnant (NP) cows in 2W and 3W] wave patterns. The propensity of two (58.6%, n = 41) and three (41.4%, n = 29) wave patterns was similar (p = .15). The IEI, shorter by 2.6 days for 2W-O versus 3W-O (p < .0009), predicted wave pattern as 100% 2W-O cows had IEI of ≤21 days, present only in 27.6% 3W-O cows (p < .0001). The ovulatory follicle persisted for a significantly shorter duration for 3W-O versus 2W-O cows. The average FSH, LH, E2 and P4 concentrations during the IEI did not differ for between and within the wave patterns. Pregnancy rate (%) of 58.6 versus 41.4 (p = .15) for 2W-O versus 3W-O and 56.1-P versus 43.9-NP (p = .44) for within 2W was similar, but tended to differ for within the 3W pattern (69.0-P versus 31.0-NP, p = .06). The pregnancy outcome was influenced by the age of ovulatory follicle for between the wave patterns and by follicular count as well as FSH surge concentration for within the wave patterns. A shorter luteal phase reduced the pregnancy outcome, a novel finding of the present study.

Exogenous progesterone-dependent modulation in the follicular dynamics of Bos indicus cattle undergoing repeated ovum pick-up sessions.

The objective of the present study was to evaluate the role of serum progesterone (P4) in follicular dynamics, oocytes' recovery and quality and their in vitro developmental competence during consecutive ovum pick-up (OPU) sessions in Bos indicus dairy cows. Wave-synchronized Sahiwal cattle (n = 20) were randomly divided into treatment (n = 10) and control (n = 10) groups. CIDR was used as a source of external progesterone in the treatment group. Four consecutive OPU sessions at 96-hr intervals were conducted and repeated ultrasonography at 12-hr intervals was done to monitor follicular dynamics. The viable oocytes were processed for IVC following IVM and IVF until day 7. The serum P4 concentrations in the P4 and control groups were recorded as 2.31 ± 0.059 versus.0.32 ± 0.065 ng/ml, respectively (p < .05). In the treatment group, the total number of recorded follicles was higher (p < .05; 12.05 ± 0.37 versus. 10.87 ± 0.40), whilst the growth rate (mm/day) of follicles was lower (p > .05). Per session recovered oocytes (5.31 ± 0.19 versus. 3.58 ± 0.21; p < .0001) and recovery rate (54.23 versus. 42.53%; p < .05) were also higher in the treatment group compared to control. Similarly, the viable oocytes (4.54 ± 0.187 versus. 3.06 ± 0.199) and the number of grade I and II oocytes per session (3.37 ± 0.196 versus. 2.06 ± 0.21) were higher (p < .05) in the treatment group compared with the control group. However, the nuclear maturation, cleavage, and blastocyst rate did not differ (p > .05) between the groups. Taken together, during OPU sessions, serum P4 improves oocytes' recovery and quality, whilst does not affect the in vitro developmental competence of recovered oocytes.

Comparison of three doses of estradiol benzoate for synchronization of follicular wave emergence in suckled Bos indicus beef cows.

We aimed to compare the effect of three estradiol benzoate (EB) doses on follicular wave emergence (FWE) and dominant follicle growth of suckled Nelore cows submitted to TAI (D0). On a random day of estrous cycle (D-10), multiparous (MULT; n=36) and primiparous (PRIM; n=20) suckled Nelore cows received an intravaginal progesterone (P4) device and were assigned in three groups. Cows in the EB-1 (n=20), EB-1.5 (n=15) or EB-2 (n=21) groups received, respectively, an im treatment with 1, 1.5 or 2 mg EB. A subgroup (n=10-13 cows/group) were subject to daily ovarian evaluations from D-10 to D0. On D-2, P4 devices were removed, and all cows received the same treatment: 1 mg estradiol cypionate, 0.53 mg sodium cloprostenol, and 300 IU eCG. Statistical analyses were performed considering only the main effects of treatment group and parity order. The proportion of cows with a synchronized FWE and the moment of the FWE did not differ (p>0.05) among the treatment groups (overall: 80% [28/35] and 4.1 ± 0.4 days); however, the FWE occurred earlier (p=0.007) in MULT (3.8 ± 0.2 days) than PRIM (5.1 ± 0.4) cows. The proportion of animals detected in estrus was greater (86% [31/36] vs. 70% [14/20]; p=0.02) and the dominant follicle was larger on D-2 (9.7 ± 0.3 mm vs. 7.8 ± 0.7 mm; p=0.006) and D0 (11.9 ± 0.4 mm vs. 10 ± 0.5 mm; p=0.008) in MULT than PRIM cows. In conclusion, the three EB doses presented similar efficiency to synchronize the FWE in suckled Nelore cows. Moreover, a delayed FWE and smaller dominant follicle is observed in PRIM cows, contributing to the reduced reproductive performance in this parity category when using similar TAI protocols of MULT cows.

Mathematical Modeling and Simulation Provides Evidence for New Strategies of Ovarian Stimulation.

New approaches to ovarian stimulation protocols, such as luteal start, random start or double stimulation, allow for flexibility in ovarian stimulation at different phases of the menstrual cycle. It has been proposed that the success of these methods is based on the continuous growth of multiple cohorts ("waves") of follicles throughout the menstrual cycle which leads to the availability of ovarian follicles for ovarian controlled stimulation at several time points. Though several preliminary studies have been published, their scientific evidence has not been considered as being strong enough to integrate these results into routine clinical practice. This work aims at adding further scientific evidence about the efficiency of variable-start protocols and underpinning the theory of follicular waves by using mathematical modeling and numerical simulations. For this purpose, we have modified and coupled two previously published models, one describing the time course of hormones and one describing competitive follicular growth in a normal menstrual cycle. The coupled model is used to test ovarian stimulation protocols in silico. Simulation results show the occurrence of follicles in a wave-like manner during a normal menstrual cycle and qualitatively predict the outcome of ovarian stimulation initiated at different time points of the menstrual cycle.

Circulating progesterone concentrations and preovulatory follicle diameters affecting ovulatory response in crossbred dairy heifers, following a 7-day progesterone-based synchronization protocol.

The present study was aimed at evaluating the differences of ovarian follicular dynamics and circulating progesterone (P4) concentrations between crossbred Holstein heifers that ovulated and did not ovulate after a P4-based synchronization protocol. Twenty-one crossbred (Holstein × Thai native) heifers with random stages of the oestrous cycle were subjected to the ovulation synchronization protocol, using an intravaginal P4-releasing device (Eazi-Breed CIDR®) for 7 days. Out of 21 CIDR-treated heifers, 14 ovulating heifers were classified as the ovulatory group and 7 non-ovulating heifers were considered the anovulatory group. The heifers having new wave emergence in ovulatory and anovulatory groups were 11/14 (78.6%) and 4/7 (57.1%), respectively. In ovulating heifers, the mean (± SEM) diameter of preovulatory follicle (PF, mm) was significantly larger, compared to non-ovulating heifers (7.21 ± 0.32 versus 4.04 ± 0.44; P = 0.001), while the mean (± SEM) follicular growth rates (mm/d) in non-ovulating heifers tended to be lower, compared to ovulating heifers (0.73 ± 0.17 versus 1.06 ± 0.08; P = 0.07). The mean (± SEM) circulating P4 concentration (ng/ml) throughout the CIDR protocol (0-10 days) in non-ovulating heifers was significantly higher, in comparison with ovulating heifers (2.82 ± 0.27 versus 1.83 ± 0.16; P = 0.02). However, no significant difference in the mean corpus luteum volume between groups was observed. In conclusion, the present results suggested that elevated circulating P4 concentrations and smaller PF diameters could cause ovulation failure in crossbred Holstein heifers, following a 7-day CIDR-based synchronization protocol.

Monitoring follicular dynamics to determine estrus type and timing of ovulation induction in captive brown bears (Ursus arctos).

It is important to understand ovarian physiology when developing an artificial insemination (AI) protocol. Brown bears (Ursus arctos) have a breeding season from May to July, although the type of estrus (polyestrus or monoestrus) is still contested. The present study aimed to define the ovarian dynamics, including follicular waves and ovulatory follicle size, and estrus type in brown bears. Six brown bears were used for ovarian ultrasonography; four were observed between April and October (before the start and after the end of the breeding season) and two in June (breeding season). In addition, we attempted to induce ovulation by administering a gonadotropin releasing hormone (GnRH) agonist. We observed follicular development in April in four bears, but follicles did not develop to greater than 6.0 mm in diameter until May. Thereafter, a group of follicles developed to more than 6.0 mm and grew as dominant follicles, except in one bear. After ovulation and subsequent corpus luteum (CL) formation, the follicular waves disappeared. Furthermore, in three bears treated with GnRH, follicles between 8.2 to 11.2 mm in diameter at the time of treatment ovulated and formed CLs. In two bears, follicles between 5.8 to 8.8 mm ovulated spontaneously within the observation interval. Our results suggest that brown bears may be monoestrous animals. Therefore, AI can only be performed once during the breeding season. Our results also suggest that dominant follicles larger than 8.0 mm are a suitable size for inducing ovulation.

Effect of a 12-h increment in the short-term treatment regimen on ovarian status, estrus synchrony, and pregnancy rate in artificially inseminated dairy goats.

This study was conducted to assess effects of two hormonal treatments on ovarian follicular status, estrous synchrony and fertility in dairy goats during the non-breeding season when duration of progestogen device use varied by 12 h. In both experiments, does were administered 60 mg of medroxyprogesterone acetate via intravaginal devices, respectively, for 6 and 6.5 d (G6 and G6.5). At 24 or 36 h before device removal, 200 IU of eCG im and 30 µg d-cloprostenol im were administered. In Experiment 1 (n = 24), data related to sexual behavior and that were collected using ovarian ultrasonography were recorded, and in Experiment 2 (n = 83) fertility was assessed after Flexible Time Artificial Insemination (FxTAI). The interval from device removal to estrus was shorter (P < 0.05) after imposing the G6.5 treatment regimen. Diameter of largest and second-largest ovarian follicles and interval from device removal to ovulation were similar (P> 0.05) between groups. The does treated with the G6.5 hormonal regimen had greater estrous synchrony, associated with greater development of largest follicles at the time of device removal, which might have led to a lesser fertility rate (P > 0.05). Conversely, treatment with the G6 hormonal regimen resulted in a greater conception rate. In conclusion, increasing time the intravaginal device is inserted from 6 to 6.5 d resulted in greater estrous synchrony, advanced ovarian follicular development, abnormal CL function and lesser pregnancy rates in artificially inseminated dairy goats when there were treatments during the non-breeding season.

Reproductive management in buffalo by artificial insemination.

Artificial insemination (AI) is important for genetic improvement and to control the period of breeding in buffalo and has increased significantly over the past 20 years. AI is more difficult in buffalo compared with cattle due to variable estrous cycles, reduced estrous behavior, and reproductive seasonality. The latter is associated with a higher incidence of anestrus and increased embryonic mortality during the nonbreeding season. Protocols to control the stage of the estrous cycle have undergone recent development in buffalo. These protocols are based on the control of both the luteal phase of the cycle, mainly by prostaglandins and progesterone, and follicle development and ovulation, by prostaglandins, progesterone, GnRH, hCG, eCG and estradiol. Protocols that synchronize the time of ovulation enable fixed timed AI, avoiding estrous detection. Factors to consider when selecting an AI protocol include animal category (heifers, primiparous or pluriparous), reproductive status (cyclic or anestrus), and season. This review looks at the current status of estrus synchronization and AI in buffalo and provides some practical suggestions for application of AI in the field.

Monitoring follicular dynamics using ultrasonography in captive brown bears (Ursus arctos) during the breeding season.

Artificial insemination (AI) may be a useful tool in the reproductive management of endangered animals, including bears. To establish an AI program for bears, we investigated follicular dynamics using weekly transrectal ultrasonography in six captive brown bears. Along with ultrasonography, we monitored plasma progesterone (P4) and estradiol-17ß (E2) concentrations. Furthermore, two bears were administered a gonadotropin releasing hormone (GnRH) agonist to induce ovulation on the first day on which the largest follicle reached more than 10.0 mm in diameter. Brown bears showed two patterns of follicular development in the early and late periods of the breeding season. In the early period (May to mid-June), multiple follicular waves were observed; namely, many follicles developed, and the largest follicles grew to less than 6.0 mm in diameter then regressed. In the late period (mid-June to July), one or two follicles grew to greater than 6.0 mm in diameter and developed as dominant follicles. Moreover, the growth rate of the largest follicle in the late period was faster than that in the early period of the breeding season. One bear with a follicle of 13.1 mm ovulated spontaneously, and one bear ovulated when the follicle was 10.2 mm in diameter after GnRH agonist treatment. Plasma E2 concentrations increased and showed peaks five to seven days before the largest follicles reached their maximum size. Plasma P4 concentrations increased on the day the corpus luteum could be detected using ultrasonography. This is the first study that showed there are two patterns of follicular development in brown bears. Furthermore, the largest follicle reaching greater than 10.0 mm in diameter could be an indicator of the appearance of ovulatory follicles.

Impact of recombinant bovine somatotropin, progesterone, and estradiol benzoate on ovarian follicular dynamics in Bos taurus taurus cows using a protocol for estrus and ovulation synchronization.

This study aimed to evaluate the effect of recombinant bovine somatotropin (bST) in combination with progesterone (P4) and estradiol benzoate (EB) on ovarian follicular dynamics using a protocol for estrus and ovulation synchronization in crossbred Bos taurus taurus cows. Twenty-four non-lactating multiparous cows were randomly assigned to two groups: the recombinant bovine somatotropin group (GbST; n = 11) received an intravaginal P4 device (1.5 g), estradiol benzoate (EB = 1.0 mg IM), bST (500 mg SC), and an ovarian ultrasonography (US) on day zero (d0 = beginning of the study); d-cloprostenol (150 µg, IM), US, and P4 removal on d8; 1.0 mg of EB (IM) on d9; and US on d10 and d15. On the other hand, to the control group (GC; n = 13), the same protocol as the GbST was applied, except for the non-receipt of bST on d0. The follicles were measured and evaluated on d0, d8, and d10, as were the corpora lutea (CL) on d15 (using ultrasonography). The effect of the two treatments (GbST vs. GC) on the follicle size, CL (F-test), and ovulation rate (logistic regression) were evaluated. The GbST showed a greater follicle diameter on d10 (14.5 mm) than the GC (12.1 mm; P < 0.03), as well as a greater diameter of CL on d15 (19.7 vs. 16.9 mm, P < 0.01). In addition, in the former, the ovulation rate (90.9 vs. 69.2%, P = 0.09) was observed to be greater. It was concluded that the combination of bST, P4, and EB in synchronization for estrus and ovulation protocols significantly increased the diameter of the preovulatory follicle, produced a higher follicular growth rate, and a greater diameter of the corpus luteum. Additionally, there was a higher percentage of cows with ovulation compared to the group that did not receive bST.

Impact of short nutrient stimuli with different energy source on follicle dynamics and quality of oocyte from hormonally stimulated goats.

The aim of this study was to verify the effect of the energy source for a short-term diet supplementation on follicular dynamics, ovarian response and oocyte recovery in goats. Thirty Anglo Nubian crossbred does received a diet for 4 weeks to satisfy the nutritional requirements of breeding for adult non-dairy goats. Seven days prior to oocyte recovery (OR), a group of does (n = 10) was supplemented with ground full-fat linseed in the diet (Diet A), whereas a second group of does (n = 10) received crude glycerine in the diet (Diet B). The total mixed ration (TMR) diet was maintained as the Control Diet (n = 10). All animals were oestrous-synchronized by the use of a progesterone insert for 12 days prior to OR. Follicles were stimulated by using pFSH (five 40-mg/ml doses) during the supplementation time. At OR, follicles were counted and recovered oocytes were classified as viable or degenerated. Follicular dynamics was monitored by ultrasonography, and plasma glucose, cholesterol and triglyceride levels were measured during supplementation. Glucose was higher in Diet B and cholesterol in Diet A. Diet B had a lower proportion of small (<3 mm) and large follicles (≥3 mm; p = 0.01). The follicular growth rate was higher in Diet A (p < 0.01), with follicles emerging in the 5th day of supplementation. No differences were observed for follicles counted and oocytes recovered. Thus, the type of energy source supplemented for a short term was capable to alter the follicular dynamics, without affecting the proportion of morphologically viable oocytes upon recovery.