Diameter Of Preovulatory Follicle Research Articles

Optimizing timed AI protocols for Angus beef heifers: Comparison of induction of synchronized ovulation with estradiol cypionate or GnRH

This study compared estradiol cypionate (ECP) or GnRH as ovulation inducers at the end of a timed AI (TAI) protocol in Angus heifers. On day 0, heifers (n = 415), between 22 and 24 months of age, were treated with an intravaginal 1 g progesterone (P4) insert and 2 mg of estradiol benzoate. On day 8, heifers had P4 removed, received 500 μg cloprostenol, and were randomized into two groups: ECP [n = 214; 0.5 mg of ECP on day 8] or GnRH [n = 201; 25 μg of GnRH analog licerelin acetate on day 10]. All heifers received TAI on day 10; 48–50 h after P4 insert withdrawal. Estrus was determined by removal of tail paint. Ovaries of heifers were evaluated by ultrasound on day 0 to determine CL presence (with CL = 213, without CL = 202) and on day 10 to measure preovulatory follicle size. Heifers were divided into three categories based on preovulatory follicle diameter: <8.5 mm (smaller than deviation), 8.5–10.9 mm, or ≥11 mm. Pregnancy diagnosis was performed 32 days after TAI. Heifers treated with ECP had greater expression of estrus than GnRH-treated heifers (93.9% vs 67.7%; P < 0.0001), regardless of CL presence at beginning of protocol. Heifers with CL at beginning of protocol had larger preovulatory follicle diameter (10.4 mm vs 9.6 mm; P = 0.0058) and greater pregnancy per AI (P/AI; 61.0% vs 50.5%; P = 0.032) than heifers without CL at day 0. In heifers with CL at day 0, GnRH treatment increased P/AI compared to ECP treatment (68.0% vs 54.9%; P = 0.0498). Expression of estrus was greater in ECP-treated than GnRH-treated heifers that had small (<8.5 mm; 77.1% vs 5.6%; P < 0.001) or medium-sized (8.5–10.9 mm; 98.4% vs. 61.7%) follicles, but not in heifers with large follicles (≥11 mm; 97.9% vs 98.3%). The P/AI was very low in both treatments for heifers with follicles <8.5 mm (ECP-14.3% vs GnRH-16.7%). In heifers with medium-sized follicles (8.5–10.9 mm), ECP treatment tended to increase P/AI compared with GnRH-treated heifers (62.9 vs 46.7%; P = 0.074). In contrast, P/AI was greater for GnRH-treated than ECP-treated heifers with large preovulatory follicles (≥11 mm; 79.7% vs 60.4%; P = 0.032). Thus, the optimal inducer of ovulation in a TAI protocol for beef heifers appears to depend on the presence of a CL at beginning of protocol (GnRH > ECP if CL present) and size of the preovulatory follicle, with ECP increasing expression of estrus and tending to increase fertility in heifers with medium-sized follicles but GnRH increasing fertility in heifers having large preovulatory follicles.

Effects of body condition and supplementation on ovarian function, growth factors and response to estrus synchronization in postpartum beef cows in Tamaulipas, Mexico

ABSTRACTEighteen beef cows with their calves were assigned to 3 groups considering body conditions scores at calving (BCSC): T1 (high BCSC, maintained throughout the study); T2 (low BCSC, increased during the study); and T3 (low BCSC, maintained throughout). An estrus synchronization protocol was implemented, with fixed time artificial insemination on day 70. Follicular or luteal growth, and pregnancy rate were similar between groups. Of all cows, five showed multiple ovulations (2–6); increasing number of ovulations reduced preovulatory follicle diameter and individual luteal volume (P < 0.05), but total luteal volume was greater in cows with 2 or 3 ovulations on day 14 after insemination (P < 0.05). Estradiol and progesterone levels were associated to follicular and luteal growth, respectively. No difference was found in IGF1 or GDF9 levels between groups or associated to fertility. No association was found between BCS, IGF1 or GDF9 levels and the reproductive process.

Oral progesterone supplementation for beef cattle after insemination in TAI programs

Abstract: The objective of this work was to evaluate the effects of supplementation with melengestrol acetate (MGA), after timed artificial insemination (TAI), on the conception rate of Nellore cows, according to cyclicity during the protocol, estrus expression between intravaginal device withdrawal and insemination, and diameter of the largest follicle at the time of insemination, in order to identify predictors of the use of MGA. Multiparous lactating cows (577) were subjected to a hormonal protocol for ovulation synchronization. On day -2 of the protocol, the animals were painted with a marker on the base of their tail. On day 0, they were evaluated for estrus expression; the preovulatory follicle diameter (POFD) was measured by ultrasonography, and the artificial insemination was performed. The animals were assigned to two experimental groups according to the supplementation or not with 0.50 mg MGA, between 13 and 18 days after TAI. Pregnancy diagnosis was performed using an ultrasound examination 35 days after TAI. A positive effect of the treatment with MGA on the conception rate occurred generally in animals in anestrus, irrespective of the estrus expression. For animals treated with MGA, only those with POFD greater than 14 mm showed better conception rates that the control group.

Effect of serum paraoxonase-1 (PON1) activity on follicular development and pregnancy rate in cattle

Paraoxonase-1 (PON1) activity has been associated with improvement in ovarian function in early postpartum dairy cows and improved in vitro embryo development. The aim of the current study was to evaluate the potential association among PON1 activity and follicular growth, diameter of the preovulatory follicle and pregnancy per artificial insemination (AI) service in cattle. In Experiment 1, cows (n=33) were subjected to an estradiol-progesterone based protocol to control time of ovulation. Starting on Day 8 of the protocol, follicular growth and serum PON1 activity were monitored. Cows were separated according to the occurrence of ovulation into two groups: Ovulatory (Ov; n=22) and Anovulatory (Anov; n=11). The serum activity of PON1 was not different between Ov and Anov cows (P=0.94). In addition, using a regression model there was no effect of serum PON1 activity on the diameter of dominant follicle (r2=0.00; P=0.99). In Experiment 2, cows (n=193) were submitted to the same hormonal protocol as in Experiment 1. On the day of the timed artificial insemination (TAI), the diameter of dominant follicle was evaluated and blood samples were collected for analysis of PON1 activity. According to the serum PON1 activity, cows were divided into three groups: Low (<70U/mL), Medium (70–90U/mL) or High (>90U/mL) PON1 activity. The overall pregnancy rate was 62.7% (121/193), with no difference among PON1 activity groups. Additionally, using a regression model there was no effect of serum PON1 activity on the diameter of the preovulatory follicle (r2=0.03; P=0.65) and pregnancy rate (r2=0.005; P=0.94). The results of this study indicate that there is no effect of serum PON1 activity on the diameter of preovulatory follicle or establishment of pregnancy in cows submitted to time of ovulation synchronization protocols.

Effects of preovulatory follicle size on estradiol concentrations, corpus luteum diameter, progesterone concentrations and subsequent pregnancy rate in buffalo cows (Bubalus bubalis)

The present study was undertaken to investigate the effects of preovulatory follicle (POF) size on estradiol concentrations, luteal profile (CL diameter and progesterone concentration) and subsequent pregnancy rate in Murrah buffalo cows. The buffalo cows (n = 49) were synchronized for estrus by two doses of PGF2α given 11 days apart. The buffalo cows were inseminated during standing estrus and again after 24 h. Ovaries were scanned at estrus and 24 h intervals until ovulation, thereafter on days 5, 12 and 16 post-ovulation to examine the POF and CL diameter. Size of POF at estrus was divided into three categories; I: 10 to ≤12; II: >12.0 to ≤14.0; III: >14.0–16.0 mm. Blood samples were collected for estradiol (on day of estrus) and progesterone concentration (on days 5, 12 and 16). The estradiol concentrations were greater (P < 0.05) in category II than category I with the greatest (P < 0.05) concentrations estimated in category III. A positive correlation (P < 0.05) between POF and progesterone concentration, CL diameter and progesterone concentration was observed on all sampling day. Pregnant buffalo cows exhibited greater (P < 0.05) plasma progesterone as compared with their non-pregnant counterpart. Greater pregnancy rates were observed with an increased size of POF (χ2 = 2.9, P > 0.05). It was concluded that the POFs having diameters between 12 and 16 mm are mature enough to be transformed into CL of such optimum diameter and can secrete optimum progesterone concentrations that can sustain the pregnancy in Murrah buffalo cows.

Neurokinin B Regulates Gonadotropin Secretion, Ovarian Follicle Growth, and the Timing of Ovulation in Healthy Women.

Context:Neurokinin B (NKB) is obligate for human puberty, but its role in adult female gonadotropin secretion and ovarian follicle growth is unknown.Objective:To investigate antagonism of NKB on pulsatile gonadotropin-releasing hormone (GnRH) and luteinizing hormone (LH) secretion and ovarian follicle development in healthy women.Design:Open investigation of the effects of a neurokinin-3 receptor (NK3R) antagonist (NK3Ra) vs a no-treatment control cycle.Setting:Clinical research facility.Patients or other participants:Healthy women with regular menses (n = 13).Intervention(s):NK3Ra MLE4901 40 mg taken orally twice daily from cycle day 5 to 6 for 7 days.Main outcome measure(s):LH secretion, ovarian follicle growth, and timing of ovulation.Results:NK3Ra administration reduced basal LH secretion without a change in pulse frequency and delayed the LH surge by 7 days, the duration of treatment [mean cycle day ± standard error of the mean (SEM), 22 ± 1 days vs 15 ± 1 days in control cycles; P = 0.0006]. Follicle growth (mean diameter at the end of administration of NK3Ra administration ± SEM, 9.3 ± 0.4 mm vs 15.1 ± 0.9 mm in control cycles; P < 0.0001) and rising estradiol concentrations (mean ± SEM, 166 ± 29 pmol/L vs 446 ± 86 pmol/L in control cycles; P < 0.0001) were prevented. After treatment, follicle development resumed and normal preovulatory follicle diameter and estradiol concentrations were demonstrated. Postovulatory progesterone rise was similarly delayed (peak cycle day, 30 ± 2 vs 22 ± 1; P = 0.002) and cycle length was prolonged (35 ± 1 days vs 29 ± 1 days in control cycles; P = 0.0003) but luteal progesterone excretion was unaffected by the NK3Ra (LH surge day +7 mean urinary progesterone levels ± SEM, 58 ± 10 pmol/mol vs 48±7 pmol/mol creatinine in control cycles; nonsignificant).Conclusion:These data demonstrate the involvement of NKB-NK3R signaling in the physiological regulation of GnRH/LH secretion, determining normal follicle development in women.

Effect of GnRH on ovulatory response after luteolysis induced by two low doses of PGF2α in lactating dairy cows

This study investigated the effect of gonadotropin-releasing hormone (GnRH) on ovulatory response after complete luteolysis induced by two low doses of prostaglandin (PG)F2α in lactating dairy cows. Cows (n = 18) ranging between 45 and 65 days in milk were recruited for synchronization by a modified Ovsynch-48 protocol (GnRH-7 days-375 μg PGF2α-1 day-250 μg of PGF2α-1 day-GnRH) over a total of 23 estrous cycles. Synchronized cows (n = 16) were randomly assigned to GnRH and Saline groups in stage 1 of the experiment after 9–10 days of ovulation in synchronization. On days 0 and 1 (day 0 = first PGF2α administration), cows were treated with 375 and 250 μg PGF2α, respectively. On day 2, cows in the GnRH and Saline groups were administered 250 μg GnRH or 2.5 mL of 0.9% saline, respectively. Serum progesterone (P4) levels were measured and changes in the corpus luteum (CL) were ultrasonically monitored daily from day 0–3 to assess complete luteolysis. Preovulatory follicle diameter and ovulatory response were evaluated by ultrasonography. In stage 2, cows were treated in a manner converse to that in stage 1. The synchronization rate was 69.6% (16/23). In stages 1 and 2, cows showed complete luteolysis with P4 concentration <1 ng/mL or remaining CL area <50%. Average ovulation time was 29.3 ± 0.5 h, which mostly occurred between 28 and 30 h after GnRH injection. However, all cows in the Saline group ovulated later than 36 h post-injection, with an average time of 52.7 ± 8.6 h. There was no difference in preovulatory follicle diameter between the two groups (16.8 ± 0.5 and 17.3 ± 0.5 mm for GnRH and saline groups, respectively). Although ovulation rate was not correlated with treatment, the rate within 48 h of GnRH injection (93.3%) tended to be higher compared with that in the Saline group (60.0%). Thus, GnRH administration increased ovulation rate following complete luteolysis induced by two low doses of PGF2α. These results indicate that this simple protocol for dairy cows is an effective alternative to timed artificial insemination programs in the field.

Pre-ovulatory follicle affects corpus luteum diameter, blood flow, and progesterone production in mares

Color Doppler ultrasonography was used to study the temporal relationships between pre-ovulatory follicle (POF) and corpus luteum (CL) diameter and blood flow, with systemic progesterone (P4) concentration during two transitional ovulatory seasons in mares. Variables of POF and CL/P4 were evaluated for 6days before and 17days after ovulation, respectively. Evaluations were performed during two consecutive estrous cycles in spring and fall seasons, and during the last estrous cycle of the season. There were significant correlations among POF and CL variables, and P4 concentration that ranged from 0.24 to 0.95, and among the ratios of different variables that ranged from 0.39 to 0.92. There were linear regressions (P<0.01−0.001) for all comparisons among different variables. The POF diameter before the first ovulation of the season was larger (P<0.05), and POF vascularity was less (P<0.05), than in the last estrous cycle during the season. The CL blood flow was less (P<0.01) during the last compared with first pre-ovulatory period of the season. The POF diameters were positively correlated (r=0.67) during the two pre-ovulatory periods of spring and fall. Results provide evidence that the POF affects CL diameter and blood flow, and subsequently P4 production, and that POF diameter is repeatable within the same individual during different seasons.

Effect of insemination site and diameter of the pre-ovulatory follicle on the odds of pregnancy in heifers using sexed or non-sexed semen.

To determine if insemination site or pre-ovulatory follicle diameter at fixed-time artificial insemination (FTAI) affects the odds of pregnancy when heifers are inseminated with sexed semen. The study was conducted in 422 Holstein heifers enrolled into 531 inseminations. Inseminations were randomly allocated to 1 of 16 treatment combinations involving three variables: semen type (sexed vs non-sexed), insemination site (uterine horn vs uterine body) and one of four sires. Ovaries were examined by transrectal ultrasound prior to FTAI to determine the follicle diameter and location. AI technician, times bred, age, weight and temperature-humidity index were also recorded. Pregnancy diagnosis was conducted 29 days post-insemination. Follicle diameter and body weight were categorised according to arbitrary cut-points. Each variable was analysed by logistic regression to determine the effect on pregnancy per AI and compare between sexed and non-sexed inseminations. Insemination site did not affect pregnancy per AI for either sexed (P = 0.528) or non-sexed (P = 0.886) inseminations. Heifers with an 18-22 mm follicle had better odds of pregnancy than heifers that did not (odds ratio (OR) 1.45, 95% confidence interval (CI) 1.004-2.09), although no effect was detected for only sexed or only non-sexed inseminations. Heifers weighing 310-370 kg had a higher pregnancy per AI than heifers weighing > 370 kg for non-sexed inseminations (P = 0.004) and sexed semen from sire 4 caused lower odds of pregnancy than semen from sire 1 (OR 0.40, 95% CI 0.18-0.89). Insemination site did not affect pregnancy per AI, but heifers with an 18-22 mm pre-ovulatory follicle at insemination had better odds of pregnancy for both sexed and non-sexed inseminations.

Effects of leptin administration on development, vascularization and function of Corpus luteum in alpacas submitted to pre-ovulatory fasting

The objective of this study was to determine the effect of leptin administration on the development, vascularization and function of Corpus luteum (CL) in alpacas submitted to pre-ovulatory fasting. Fourteen alpacas were kept in fasting conditions for 72h and received five doses of o-leptin (2μg/kg e.v.; Leptin group) or saline (Control group) every 12h. Ovulation was induced with a GnRH dose (Day 0). The ovaries were examined every other day by trans-rectal ultrasonography (7.5MHz; mode B and power Doppler) from Day 0 to 13 to determine the pre-ovulatory follicle diameter and ovulation, and then to monitor CL diameter and vascularization until the regression phase. Serial blood samples were taken after GnRH treatment to determine plasma LH concentration; and every other day from Days 1 to 13 to determine plasma progesterone and leptin concentrations. The pre-ovulatory follicle and CL diameter, LH, progesterone and leptin plasma concentrations were not affected by treatment (P>0.05). The vascularization area of the CL was, nevertheless, affected by the treatment (P<0.01) with significant differences between groups at Days 3, 7 and 9 (P<0.05). The Leptin group had a larger maximum vascularization area (0.67±0.1 compared with 0.35±0.1cm2; P<0.05). In addition, there was a positive correlation between CL vascularization, CL diameter and plasma progesterone. The exogenous administration of leptin during pre-ovulatory fasting increased the vascularization of the CL in alpacas in vivo.

Preovulatory Follicle Dynamics, and Ovulatory and Endometrial Responses to Different Doses of hCG and Prediction of Ovulation in Mares

A more precise characterization of the effects of human chorionic gonadotropin (hCG) treatment on the preovulatory follicle is needed to improve reproductive efficiency in broodmare management. The objectives of this study were (1) to study the effects of different hCG doses within and among three consecutive estrous cycles on ovulatory response, preovulatory follicle dynamics, and endometrial echotexture; (2) to study the temporal relationship among changes in preovulatory follicle diameter, endometrial echotexture, and estradiol levels; and (3) to analyze a system for prediction of ovulation. In experiment 1, mares were treated with 2,500 IU hCG, 1,500 IU hCG, 500 IU hCG, or saline (controls) and examined by ultrasonography every 6 hours across three consecutive preovulatory periods (n = 135). Similar treatment groups were utilized in experiment 2 (n = 177 preovulatory periods) with examinations performed every six, 12, or 24 hours to test the effectiveness of a prediction of ovulation system. Effective hCG doses (2,500 and 1,500 IU) led to cessation of growth rate and reduction of the preovulatory follicle diameter concurrently with a reduction in systemic estradiol concentration and endometrial echotexture score. A system of prediction of ovulation, based on preovulatory follicle ultrasonographic signs of impending ovulation, was highly accurate for the diagnosis of positive and negative outcomes in both hCG-treated and control groups. However, the effectiveness of this prediction system was reduced due to the false-negative diagnoses that were a reflection of a lack of sensitivity for the detection of nonovulation.

Mastitis outcomes on pre-ovulatory follicle diameter, estradiol concentrations, subsequent luteal profiles and conception rate in Buffaloes

The objectives of this study was to investigate the outcome of mastitis, in its clinical or subclinical forms, on the mean diameter of pre-ovulatory follicle (POF), plasma estradiol concentration on the day of estrus, subsequent luteal profile and subsequent conception rate in buffaloes. Sixty dairy buffalo (Bubalus bubalus) conducted in this study were divided into three groups {healthy (H), n=20; subclinical mastitis (SCM), n=18; and clinical mastitis (CM), n=22}. Ultrasonography of ovaries revealed that mean diameter of POF was larger (P<0.05) in H buffalo (14.35mm) compared to SCM (12.40mm) and CM (10.25mm). Also, plasma estradiol concentration on the day of estrus was higher (P<0.05) in H buffalo compared to SCM and CM counterparts; 34.95 vs. 32.87 and 27.50pg/ml, respectively. Besides, positive correlation was observed between the POF diameter with plasma estradiol concentration in H, SCM and CM buffaloes (r=0.64, 0.74, 0.72 respectively, P<0.05). Moreover, positive correlations (P<0.01) were found on days 9, 12, 16, and 21 post-ovulation between POF diameter and luteal profile. Thus, the conception rate in H buffalo was higher (P<0.05) compared with SCM and CM counterparts; 55% vs. 38.89 and 18.18%, respectively. In conclusion, mastitis in its clinical or subclinical forms disrupts the functioning of the pre-ovulatory follicle on the day of estrus, associated with low follicular estradiol production, resulting in suppression to subsequent luteal profile leading to substantial decrease in pregnancy consequence of buffaloes.

Clinical monitoring of serum sex hormones during normal menstrual follicle growth cycle

Objective To study the changes of six sex hormones corresponding to the follicle growth during the normal menstrual cycle of Chinese women. Methods Thirty Chinese women with regular menstrual period and average age of (28.8±3.2) years were selected for the study by Peking Union Medical College Hospital in September, 2010. Growth of follicles was monitored by using transvaginal sonography. Six sex hormones, including follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), progesterone (P), testosterone (T), and prolactin (PRL) were measured by chemoluminescence immunoassay every day during a menstrual cycle. Nonparametric statistical analysis was used. Results Menstrual cycle of all the patients was in the range of 25 to 39 d, with mean of (29.5 ± 3.1) d. Length of follicular phase and luteal phase was 15.3 and 14.4 d, respectively.Number of days from antral follicle to emergence of dominant follicle, and from the latter to ovulation, was 6.2 and 8.9 d, respectively. Average diameter of preovulatory follicle was 19.33 mm. Both FSH and LH reached peak on the day before ovulation. P started to increase before ovulation and remained at a high plateau from 6th to 9th day after ovulation. Both PRL and T reached peak after ovulation, near the end of a menstrual cycle. Conclusions A small rise of LH and P emerges just 1 to 2 d before ovulation, indicating the maturing of follicle. PRL and T shows cyclic changes as follicle grows. Therefore, PRL and T levels should be measured in the early follicle phases in the clinical practice so that leading the impact of menstrual cycle minimal.(Chin J Lab Med, 2017, 40: 169-173) Key words: Menstrual cycle; Ovarian follicle; Gonadal steroid hormones; Luminescent measurements; Immunoassay

The effect of cooling management on blood flow to the dominant follicle and estrous cycle length at heat stress

The use of ultrasound imaging for the examination of reproductive organs has contributed substantially to the fertility management of dairy cows around the world. This method has many advantages such as noninvasiveness and immediate availability of information. Adding Doppler index to the ultrasound imaging examination, improved the estimation of blood volume and flow rate to the ovaries in general and to the dominant follicle in particular. The aim of this study was to examine changes in the blood flow to the dominant follicle and compare them to the follicular development throughout the cycle. We further set out to examine the effects of different types of cooling management during the summer on the changes in blood flow to the dominant follicle. For this purpose, 24 Israeli-Holstein dairy cows, under heat stress, were randomly assigned one of two groups: one was exposed to five cooling sessions per day (5CS) and the other to eight cooling sessions per day (8CS). Blood flow to the dominant follicle was measured daily using Doppler index throughout the estrous cycle. No differences in the preovulatory dominant follicle diameter were detected between the two cooling management regimens during the cycle. However, the length of the first follicular wave was significantly longer, whereas the second follicular wave was nonsignificantly shorter in the 5CS group as compared to the 8CS group. In addition, no difference in blood flow was found during the first 18 days of the cycle between the two groups. However, from Day 20 until ovulation a higher rate of blood flow was measured in the ovaries of cows cooled 8 times per day as compared to the 5CS group. No differences in progesterone levels were noted. Finally, the estrous cycle length was shorter in the 8CS group as compared to the 5CS group. Our data suggest that blood flow to the dominant follicle and estrous cycle length is affected by heat stress. Using the appropriate cooling management during heat stress can enhance the blood flow to the ovary and may contribute to improved fertility in dairy cows.

Contrasting effects of progesterone on fertility of dairy and beef cows

The role of progesterone in maintaining pregnancy is well known in the bovine. Subtle differences exist between dairy and beef cows because of differing concentrations of progesterone during recrudescence of postpartum estrous cycles, rate of follicular growth and maturation, proportions of 2- and 3-follicular wave cycles, and other effects on pregnancy outcomes per artificial insemination (P/AI). Because proportions of anovulatory cows before the onset of the artificial insemination (AI) period are greater and more variable in beef (usually ranging from 30 to 70%) than dairy (25%) cows, AI programs were developed to accommodate anovulatory and cycling beef cows enrolled therein. Incorporating a progestin as part of an AI program in beef cows improved P/AI by reducing the proportion of cows having premature luteal regression and short post-AI luteal phases. In both genotypes, prolonged dominant follicle growth in a reduced progesterone milieu resulted in increased (1) LH pulses, (2) preovulatory follicle diameter, and (3) concentrations of estradiol and a subsequently larger corpora lutea (CL). In contrast, the progesterone milieu during growth of the ovulatory follicle in an ovulation control program does not seem to affect subsequent P/AI in beef cows, whereas in dairy cows follicle development in an elevated compared with a low progesterone environment increases P/AI. Progesterone status in beef cows at the onset of ovulation synchronization is not related to P/AI in multiparous cows, whereas P/AI was suppressed in primiparous cows that began a timed AI program in a low-progesterone environment. In timed AI programs, elevated concentrations of progesterone just before PGF2α and reduced concentrations at AI are critical to maximizing subsequent P/AI in dairy cows, but seemingly much less important in beef cows. By inducing ancillary CL and increasing concentrations of progesterone, human chorionic gonadotropin may increase P/AI when administered to beef cows 7d after AI or at embryo transfer, and its success seems to depend on induction of ancillary CL, whereas in dairy cows increased fertility was detected in cows with multiple CL, human chorionic gonadotropin-enhanced progesterone from original CL, or both. Pregnancy losses after AI are less frequent in beef cows and are not associated with pre-AI progesterone or cycling status, whereas losses in dairy cows are inversely related to progesterone and adversely affected in anovular dairy cows. Genotype and nutritional management likely influence several physiological differences including circulating concentrations of progesterone and responses to supplemental progesterone.

Synchronization of ovulation in cattle with an aromatase inhibitor–based protocol

A study was designed to determine the effect of stage of the estrous cycle on the proportion of animals that ovulated and the synchrony of ovulation of heifers treated with an aromatase inhibitor–based protocol. Forty-eight heifers were treated intramuscularly with 500 μg of cloprostenol (PGF) followed by 100 μg of GnRH 24 hours later to serve as control data for comparison of the ovulatory response to a subsequent aromatase inhibitor protocol. Daily ultrasound examinations were done to determine the incidence of and interval to ovulation. At the time of ovulation (Day 0), heifers were assigned randomly to five day-groups (n = 8-11/group) and given an intravaginal device containing 3 g of letrozole for 4 days starting on Day 0, 4, 8, 12, or 16. At the time of device removal, heifers were given PGF followed by GnRH 24 hours later. Ultrasound examinations were done daily from 2 days before device insertion to 9 days after the posttreatment ovulation. The preovulatory follicle diameter after letrozole treatment was larger in the Day 4 group compared to the Day 0 and 16 groups and intermediate in the Day 8 and 12 groups (P < 0.001). Compared to control data, the percentage of heifers that ovulated after letrozole treatment was greater (87.1% vs. 69.4%, respectively; P < 0.05) as was the synchrony of ovulation (residuals: 0.24 ± 0.07 vs. 0.68 ± 0.13; P < 0.01). The day on which letrozole treatment was initiated did not affect the proportion of heifers that ovulated or the interval to ovulation. Plasma estradiol concentrations at the time of removal of the letrozole device in the Day 0 and 4 groups was lower (P < 0.05) than in the corresponding controls. Estradiol concentrations in the Day 8 and 12 groups did not differ from already low concentrations in the respective controls. Corpus luteum diameter profiles and progesterone production were not affected by day-group although reduced luteal lifespan after letrozole treatment was observed and requires further investigation. In summary, a protocol involving a letrozole-impregnated intravaginal device for 4 days, PGF treatment at device removal, and GnRH 24 later resulted in a greater ovulation rate and greater synchrony of ovulation than in heifers not given letrozole. Results suggest that the protocol may be initiated effectively at random stages of the estrous cycle and may provide impetus for further studies to assess the efficacy of a letrozole-based synchronization protocol for fixed-time insemination.

Dietary n-3 Polyunsaturated Fatty Acid Affects the Onset of Prostaglandin F2α Induced Oestrus in the Goat (Capra hircus)

The objective of the present study was to examine the effect of dietary n-3 PUFA supplementation on oestrus response in the goat following synchronization with prostaglandin analogue. Parous cycling goats (n=17/group) were fed a concentrate diet supplemented with either fish oil (FO) or palm oil (PO). The FO provided n-3 EPA and DHA @ 156 mg kg−1 body weight while PO was given @ 0.6 ml kg−1 body weight to make the diet isocaloric. Oestrus was synchronized using two dose of prostaglandin F2α (PGF2α) at 11 days apart, with first PG on day 25 of oil supplementation. Goats were observed for oestrus twice a day using a vasectomized teaser buck following second injection of PGF2α. The number and diameter of preovulatory follicle (POF) on the day of oestrus was studied using transrectal ultrasonography. The mean interval from PGF2α administration to the onset of oestrus was significantly (χ2df1=7.003, P=0.008) longer in FO than PO supplemented goats (48.71±3.78 vs 37.41±1.75 h). The proportion of goats showed oestrus within 48 h was 94.11% in the PO group (16/17), while it was 58.82% in the FO group (10/17). However, the oestrus duration was not affected by the FO supplementation. The number of POF was higher in the FO group than the PO (2.23±0.14 vs. 1.82±0.15; P=0.054); however the diameter of POF did not differ among the group (6.90±0.10 vs. 6.77±0.14; P>0.05). In conclusion, the supplementation of goats with n-3 PUFA rich FO delayed the onset of PGF2α induced oestrus and increased the POF number on the day of oestrus.

Improved fertility in suckled beef cows ovulating large follicles or supplemented with long-acting progesterone after timed-AI

We aimed to evaluate the effects and the interaction of size of the preovulatory follicle (POF) and long-acting progesterone (P4) supplementation after timed-AI on CL function and pregnancy success in beef cows. In experiment 1, ovulations of beef cows were synchronized starting on Day −10, and cows were split to receive sodium cloprostenol (large follicle group; LF; n = 31) or nothing (small follicle group; SF; n = 35). Ovulations were induced on Day 0, and cows were inseminated. Ovulated cows were assigned to receive placebo (LF/control group, n = 14; and SF/control group, n = 9) or 150 mg of long-acting P4 on Day 4.5 (LF/P4 group, n = 13; and SF/P4 group, n = 12). Diameter of POF, blood flow in POF wall, ovulation rate, and size and vascularization of CL were greater (P < 0.05) in LF group. In experiments 2 (unknown cyclic status) and 4 (noncycling), ovulations were synchronized, and beef cows received placebo or 150 mg of long-acting P4 on Day 4 after timed-artificial insemination. In experiment 2, pregnancy/AI (P/AI) did not differ (P > 0.1) between P4-treated (53.2%; 209/393) and control cows (56.2%; 219/390), but P/AI was greater in cows with a CL < 0.9 cm2 on Day 4 that were P4-treated (57.9%, 22/38) versus placebo-treated (40.4%, 21/52; P < 0.05). In Experiment 4, P/AI was greater (P < 0.05) in P4-treated cows (55.6%, 105/189 vs. 46.0%, 86/187). In Experiment 3, cyclic–suckled beef cows were treated as described in Experiment 1 to generate animals with small (SF; n = 111) or large POF (LF; n = 109), and subdivided to receive placebo or P4 on Day 4. POF size, ovulation rate, CL area, and P/AI were greater (P < 0.007) in the LF group. Pregnancy/AI in ovulated cows were lower (P = 0.05) in the SF/control group (41.5%, 17/41) compared to LF/control group (62%, 31/50) and were similar for the SF/P4 group (55.6%, 25/45) and LF/P4 group (57%, 28/49) compared to others. In summary, smaller and less vascularized POF results in less functional CL and reduces ovulatory rate and P/AI in cyclic beef cows; the long-acting P4 injection on Day 4 after timed-artificial insemination may attenuate the negative effects of small POF/CL; and postovulatory P4 supplementation improved fertility in anestrous beef cattle.

Expression of estrus modifies the gene expression profile in reproductive tissues on Day 19 of gestation in beef cows

The aim of this study was to test the effect of expression of estrus at artificial insemination (AI) on endometrium, conceptus, and CL gene expression of beef cows. Thirty-six multiparous nonlactating Nelore cows were enrolled on an estradiol- and progesterone (P4)-based timed AI protocol (AI = Day 0) and then slaughtered for the endometrium, CL, and conceptus collection on Day 19. The animals were retrospectively grouped on the basis of cows that (1) showed signs of estrus near AI (n = 19; estrus) and (2) did not show any signs of estrus (n = 17; nonestrus). Body condition score, blood sampling, and ultrasound examination were performed on Days 0, 7, and 18 of the experiment followed by messenger RNA extraction and quantitative reverse transcription polymerase chain reaction analysis of 58 target genes. Data were checked for normality and analyzed by ANOVA for repeated measures using proc GLM, MIXED, and UNIVARIATE of SAS. Only pregnant cows were included in the analyses (n = 12; nonestrus, n = 11). Estrous expression had no correlation with parameters such as body condition score, preovulatory follicle and CL diameter, P4 concentration in plasma on Days 7 and 18 after AI, and interferon-tau concentration in the uterine flushing (P > 0.15); however, a significant increase was observed in conceptus size from cows that expressed estrus (P = 0.02; 38.3 ± 2.8 vs. 28.2 ± 2.9 mm). The majority of transcripts affected by estrous expression in the endometrium belong to the immune system and adhesion molecule family (MX1, MX2, MYL12A, MMP19, CXCL10, IGLL1, and SLPI; P ≤ 0.05), as well as those related with prostaglandin synthesis (OTR and COX-2; P ≤ 0.05). Genes related to apoptosis, P4 synthesis, and prostaglandin receptor were downregulated (CYP11A, BAX, and FPr; P < 0.05) in the CL tissue of cows that expressed estrus. In addition, four genes were identified as differentially expressed in the 19-day-old conceptus from cows that expressed estrus (ISG15, PLAU, BMP15, and EEF1A1; P < 0.05). There was also a significant effect of Day 7 concentration of P4 mainly affecting the immune system, adhesion molecules, and wnt signaling pathway of the endometrium (IGLL1, MX2, SLPI, TRD, APC, WNT2, GLYCAM1, and MYL12A; P < 0.05). A significant interaction between estrous expression and P4 concentration on Day 7 was more pronounced in immune system genes (MX1, MX2, TRD, SLPI, and IGLL1; P < 0.05). This study reported that estrous expression at the time of AI favorably altered the gene expression profile in reproductive tissues during the preimplantation phase toward a more receptive state to the elongating conceptus. These effects seem to be more evident in the endometrium during the time of dynamic remodeling for embryo implantation.

Superovulatory response and embryo quality of Holstein heifers treated with one or two injections of somatotropin

Background: recombinant bovine somatotropin (rbST) is used in bovine embryo donors to improve superovulatory response and embryo quality. Objective: this study evaluated the effect of applying one versus two injections of 500 mg of rbST to donor Holstein heifers on estrus incidence (IE), diameter of the largest preovulatory follicle (dLPF), superovulation response (SR), embryo yield, and pregnancy rate in recipient Holstein cows (PRR). Methods: two superstimulation programs were conducted. Heifers were assigned to one of two treatments: 1) rbST-I, n = 5: heifers received one injection of rbST on day 0 (day of CIDR (controlled internal drug release) removal); 2) rbST-II, n = 5: heifers received the first rbST injection on day -8 and the second one on day 0. Thirty-eight cows were used as recipients and were assigned to receive one embryo from one of the two treatments. Results: there was no effect of treatment (p &gt; 0.05) on PRR, dLPF, IE, and SR. The number of oocytes increased (p &lt; 0.05) in the rbST-I treatment (1.3 ± 0.4 vs 0.2 ± 0.2), but there was no difference in the number (p &gt; 0.05) of degenerated (1.0 ± 0.5 vs 4.5 ± 3.0) or transferable (1.0 ± 0.5 vs 1.4 ± 0.7) embryos between heifers in the rbST-I and rbST-II treatments, respectively. Moreover, no oocytes or embryos were recovered from 36.8% of donor heifers in either treatment. Conclusion: the application of 500 mg of rbST on days -8 and 0 of the follicular wave synchronization program did not increase the superovulatory response but significantly reduced the number of oocytes recovered from superovulated Holstein heifers.