Diameter Of Preovulatory Follicle Research Articles

Administration of nerve growth factor-β to heifers with a pre-ovulatory follicle enhanced luteal formation and function and promoted LH release

The objective of this study was to determine the effects of bovine nerve growth factor-β (NGF) on pre-ovulatory follicle vascular area, LH release, ovulation, and luteal function when administered systemically to heifers. Post-pubertal Holstein heifers (n = 12) received an intravaginal progesterone-releasing device (CIDR) and GnRH agonist (100 μg IM). The CIDR was removed 5 d later, and heifers were given dinoprost (25 mg IM) at CIDR removal and 24 h later, followed by a second dose of GnRH agonist 48 h later. Heifers were randomly assigned to treatments using a cross-over design. For example, heifers assigned to NGF (250 μg reconstituted in 12 mL PBS IM) in replicate 1 were assigned to control (12 mL PBS IM) in replicate 2. Transrectal ultrasonography was performed before treatment and repeated every 4 h up to 32 h to determine the pre-ovulatory follicle diameter, vascular area, and ovulation. Serum samples were obtained to assess LH concentrations during the periovulatory period and every 2 d post-ovulation for measuring progesterone concentrations. A subset of heifers had luteal biopsies performed on days 9 (n = 6 per treatment) and 14 (n = 6 per treatment) post-ovulation to count luteal cell numbers and measure relative mRNA abundance for steroidogenic and angiogenic enzymes and LH receptor. Treatment with NGF increased pre-ovulatory follicle diameter (P = 0.02) and serum LH concentrations (P = 0.03) but did not affect time to ovulation (P = 0.42). Heifers treated with NGF had increased serum progesterone concentrations in the subsequent luteal phase (P = 0.03), but no change in vascular area of the follicle (P = 0.16) or CL (P = 0.20). Heifers treated with NGF had a greater number of small luteal cells (P < 0.01) and a tendency for increased LH receptor (LHR) mRNA abundance in the CL (P = 0.10). There was also increased steroidogenic acute regulatory protein (STAR; P = 0.05) and a tendency for increased cytochrome P450 family 11 (CYP11A1; P = 0.10) mRNA abundance in the CL of NGF-treated heifers. There was decreased prostaglandin E2 synthase (PGES; P = 0.03) and its receptor (PGER; P = 0.05) mRNA abundance and a tendency for decreased cytochrome P450 family 17 subfamily A member 1 (CYP17A1; P = 0.08) and hydroxysteroid 17-beta dehydrogenase (HSD17B; P = 0.06) mRNA abundance in the CL of NGF-treated heifers. Administration of NGF improved CL function in heifers potentially as a result of increased LH release.

Presynchronization with CIDR, with or without GnRH, prior to CO-Synch in beef heifers

Objectives were to compare ovarian responses and pregnancy per AI (P/AI) in Angus-cross beef heifers (n = 521; 4 locations) synchronized with CIDR–CO–Synch (CCOS) versus CIDR-GnRH–CO–Synch (CGCOS) protocols. Heifers were assigned a reproductive tract score (RTS: 1, immature, acyclic; 5, mature, cyclic), body condition score (BCS: 1, emaciated; 9, obese) and temperament score (0, calm, 1, excitable). Heifers in the CCOS (n = 261) group received a CIDR on Day −20 (removed on Day −13), 100 μg GnRH on Day −10, 25 mg PGF2α on Day −3 and were timed inseminated 60 h later, with concomitant GnRH (Day 0). Heifers in the CGCOS (n = 260) group received a CIDR on Day −26 (removed on Day −19), 100 μg of GnRH on days −16 and −10, 25 mg of PGF2α on Day −3 and were timed inseminated 60 h later, with concomitant GnRH (Day 0). Ovarian ultrasonography was done in a subset of heifers (n = 60; 30 in each group) to determine number and size of ovarian follicles and presence of corpus luteum (CL). There was increased (P < 0.05) percentage of heifers with CL in CGCOS group compared to heifers in CCOS group on Day −10 (82.3 vs 68.2%) and on Day −3 (88.3 vs 75.1%). Average size of the largest ovarian follicle on Day 0 was greater for heifers in CGCOS group compared to CCOS group (P < 0.05). However, P/AI did not differ between CCOS and CGCOS groups, 55.0% (143/260) and 59.8% (156/261), respectively (P > 0.1). In conclusion, CIDR presynchronization with or without GnRH (CCOS and CGCOS protocols) in beef heifers resulted in similar P/AI. Adding GnRH to presynchronization with CIDR resulted in more heifers with a CL at PGF2α and increased preovulatory follicle diameter at AI. Future studies are needed with bigger sample size and CIDR + CO-Synch treatment as control to determine economic benefit.

Associations among thermal biology, preovulatory follicle diameter, follicular and luteal vascularities, and sex steroid hormone concentrations during preovulatory and postovulatory periods in tropical beef cows

The objectives were to evaluate effects of tropical seasons on thermal biology, preovulatory follicle (POF) diameter, POF and luteal vascularities, and estradiol (E2) and progesterone (P4) concentrations; and to determine the associations among the values for these variables during preovulatory and postovulatory periods in Thai native cows in tropical climates: cold, hot, and rainy seasons. Development and vascularity of the POF and corpora lutea (CL) were evaluated using color Doppler ultrasonography. The temperature-humidity index (THI) was greater when the preovulatory period occurred during the rainy season when compared with the occurrence during the hot and cold seasons of the year. Furthermore, POF diameter was less when the THI was greater. The THI was greater when the postovulatory period occurred during the rainy season when compared to the occurrence of the postovulatory period during the hot and cold seasons of the year. Furthermore, the CL vascularity and P4 concentration were less when the THI was greater. The THI was inversely correlated with CL vascularity and P4 concentrations. When the THI was greatest during the hot and rainy seasons of the year, there were the greatest negative effects on POF size, POF and CL blood flow, and concentrations of E2 and P4 during the preovulatory and postovulatory periods. While native Bos indicus are capable of adapting to tropical conditions, there are still negative effects, such as impaired POF and CL functions, when the THI induces heat stress.

Changes in Ovarian and Uterine Morphology and Estrous Cycle in CD-1 Mice After Vanadium Inhalation.

Vanadium is a metal present in particulate matter and its reprotoxic effects have been demonstrated in males and pregnant females in animal models. However, the effects of this metal on the reproductive organs of nonpregnant females have not been sufficiently studied. In a vanadium inhalation model in nonpregnant female mice, we found anestrous and estrous cycle irregularity, as well as low serum concentrations of 17β-estradiol and progesterone. A decrease in the diameter of secondary and preovulatory follicles, as well as a thickening of the myometrium and endometrial stroma, was observed in the vanadium-treated mice. There was no difference against the control group with respect to the presence of the estrogen receptor α in the uterus of the animals during the estrous stage. Our results indicate that when vanadium is administered by inhalation, effects are observed on the female reproductive organs and the production of female sex hormones.

172 Effect of period of insertion of a progesterone-releasing device and pro-oestrus length on follicular and luteal characteristics and pregnancy rates to fixed-time AI in Bos indicus heifers

An experiment was designed to evaluate the effect of the length of insertion of a progesterone (P4)-releasing device and the length of pro-oestrus on follicular and luteal characteristics and pregnancy rates to AI (P/AI) in Bos indicus heifers treated with oestradiol/P4-based treatments. Bos indicus beef heifers (n=374), 22-26 months of age, with a corpus luteum (CL) or at least one follicle ≥8mm in diameter and with a body condition score between 2.5 and 3.5 (1-to-5 scale) were synchronised using three treatments for fixed-time AI (FTAI). On Day 0, all heifers received 2mg of oestradiol benzoate (Sincrodiol, Ourofino) and an intravaginal device with 1g of P4 (Sincrogest, Ourofino). The P4 device was removed on Day 6 in heifers in the J-Synch 6 group (n=120) and on Day 7 in heifers in the J-Synch 7 group (n=105) and conventional group (n=165). All heifers received 500μg of cloprostenol (Sincrocio, Ourofino) and 300IU of equine chorionic gonadotrophin (SincroeCG 6000UI, Ourofino) at the time of P4 device removal. Furthermore, heifers in the conventional treatment group received 0.5mg of oestradiol cypionate (SincroCP, Ourofino) at the same time. In addition, all heifers were tail-painted for oestrus detection (CeloTest, Biotay). Heifers that had lost ≥50% of the tail paint by 70-74h (J-Synch groups) or 48-52h (conventional group) after device removal were FTAI at that time. Heifers not showing oestrus by 70-74h (J-Synch groups) or 48-52h (conventional group) received 10μg of gonadotrophin-releasing hormone (Sincroforte, Ourofino) at the same time and were FTAI 8h later. All heifers were also examined using ultrasonography (Mindray DP50 Vet) every 12h from the time of device removal to determine the time of ovulation, 6 days after ovulation to determine the diameter of the CL, and 28 days after FTAI for P/AI determination. Data were analysed using the MLGM procedure (InfoStat) for normal data families (follicular dynamics) and binary data family (P/AI). The results are shown in Table 1. The diameter of the dominant preovulatory follicle and the CL did not differ among groups (P&amp;gt;0.12). However, the interval from device removal to ovulation was longer in heifers in the J-Synch groups than in heifers in the conventional group (P&amp;lt;0.05). Furthermore, P/AI was not different among groups. In conclusion, although the J-Synch protocols delayed the interval from P4 device removal to ovulation, the three protocols evaluated in the present study were equally effective in Bos indicus heifers. Table 1.Mean (±s.e.m.) diameter of the preovulatory follicle (P/Foll) and corpus luteum (CL), interval from progesterone (P4) device removal to ovulation, and pregnancy rates to AI (P/AI) in Bos indicus heifers Treatment P/Foll, mm Interval to ovulation, h CL diameter, mm P/AI,% (n) J-Synch 6 10.5±0.7 101.4±2.3a 18.6±1.0 52.0 (62/120) J-Synch 7 10.6±0.7 96.0±2.2a 16.5±0.9 39.0 (41/105) Conventional 9.4±0.7 73.0±1.9b 16.8±0.9 45.0 (74/165) a,bDifferent superscripts denote differences between groups in the interval from P4 device removal to ovulation.

171 Follicular and luteal characteristics and pregnancy rates in Bos indicus suckled cows treated with two oestradiol/progesterone-based protocols with a prolonged pro-oestrus

An experiment was designed to evaluate the effect of the length of insertion of a progesterone (P4)-releasing device on follicular and luteal characteristics and pregnancy rates to fixed-time AI (P/FTAI) in Bos indicus cows treated with oestradiol/P4-based treatments with a prolonged pro-oestrus. Multiparous Bos indicus suckled beef cows (n=659), 60-90 days postpartum, with a corpus luteum (CL) or at least one follicle ≥8mm in diameter and with a body condition score between 2 and 3.5 (1-to-5 scale) were synchronised using three treatments. On Day 0, all cows received 2mg of oestradiol benzoate (Sincrodiol, Ourofino) and an intravaginal device with 1g of P4 (Sincrogest, Ourofino). The P4 device was removed on Day 6 in cows in the J-Synch 6 group (n=195) and on Day 7 in cows in the J-Synch 7 group (n=262) and conventional group (n=202). All cows received 500μg of cloprostenol (Sincrocio, Ourofino) and 400IU of equine chorionic gonadotrophin (SincroeCG 6000UI, Ourofino) at the time of P4 device removal. Cows in the conventional treatment group also received 0.5mg of oestradiol cypionate (SincroCP, Ourofino) at the same time. In addition, all cows were tail-painted for oestrous detection (CeloTest, Biotay). Cows that had lost ≥50% of the tail paint 62-66h (J-Synch groups) or 48-52h (conventional group) after device removal were FTAI at that time. Cows not showing oestrus by 62-66h (J-Synch groups) or 48-52h (conventional group) received 10μg of GnRH (Sincroforte, Ourofino) at the same time and were FTAI 8h later. A subset of 20 cows per group was also examined using ultrasonography (Mindray DP50 Vet) every 12h from the time of device removal to determine the time of ovulation, 6 days after ovulation to determine CL diameter, and 30 days after FTAI to determine P/AI. Data were analysed using the GLM Mixed procedure (InfoStat). The results are shown in Table 1. The diameter of the dominant preovulatory follicle was larger and the interval from device removal to ovulation was longer in cows in the J-Synch groups compared with those in the conventional group (P&amp;lt;0.05). Furthermore, the diameter of the CL was larger and P/FTAI was higher in cows in the J-Synch 7 group compared with those in the J-Synch 6 and conventional groups. In conclusion, delaying the removal of the P4 device by 1 day (i.e. Day 7 vs. Day 6) in the J-Synch groups and prolonging the pro-oestrus period by not giving oestradiol cypionate at P4 device removal (i.e. J-Synch 7 vs. conventional) resulted in a larger preovulatory follicle and CL and higher P/FTAI in Bos indicus suckled cows. Table 1.Mean (±s.e.m.) diameter of the preovulatory follicle (P/Foll) and corpus luteum (CL), interval from progesterone (P4) device removal to ovulation, and pregnancy rates to fixed-time AI (P/FTAI) in Bos indicus suckled cows Treatment P/Foll diameter, mm CL diameter, mm Interval to ovulation, h P/FTAI,% (n) J-Synch 6 15.5±0.3a 18.7±0.8a 83.0±4.0b 41.0 (80/195)a J-Synch 7 16.4±0.2b 21.9±0.8b 86.0±3.0b 52.0 (136/262)b Conventional 13.8±0.2a 16.8±0.9a 64.0±3.0a 37.1 (75/202)a a,bDifferent superscripts denote differences between means or P/FTAI.

164 Gonadotrophin-releasing hormone injection and colour flow Doppler ultrasound of the preovulatory follicle as a tool to increase pregnancy outcome after timed AI in beef cows

The aims of this study were to determine (1) the association between Doppler vascularisation scores (DVS) of the preovulatory follicle (POF) and fertility of beef cows submitted to timed AI (TAI) and (2) whether cows with low DVS benefit from a gonadotrophin-releasing hormone (GnRH) treatment at TAI. Multiparous lactating Nelore cows (Bos indicus; n=69) from a commercial beef farm in the state of Rondônia, Brazil, were enrolled in this study. Cows received 2mg of oestradiol benzoate intramuscularly (Bioestrogen, Biogénesis Bagó) and an intravaginal progesterone-releasing device (1.9g of progesterone; controlled internal drug release, CIDR) to synchronise follicular wave emergence on Day 0. The CIDR device was removed and cows were treated with 150μg of D-cloprostenol intramuscularly (prostaglandin F2α analogue; Croniben), 1mg of oestradiol cypionate intramuscularly, and 300IU of equine chorionic gonadotrophin (Novormon) intramuscularly on Day 8. Cows were then painted with a tail chalk marker to identify those displaying oestrus. All cows were submitted to TAI 48h after CIDR removal. At TAI, occurrence of oestrus was recorded and all cows were examined using transrectal ultrasonography. Blood flow of the POF was evaluated using colour Doppler imaging. Colour Doppler signals present on the follicular wall were subjectively scored using a 1-to-4 scale (1=absence or very low blood flow, and 4=intense blood flow detected on most of the follicular wall surface) adapted from Ginther (2007Ultrasonic Imaging and Animal Reproduction: Color-Doppler Ultrasonography, pages 87-114). Then, cows were divided into three groups according their DVS of the POF: (1) high DVS (DVS ≥3; n=36), (2) low DVS (DVS &amp;lt;3; n=16), and (3) low DVS (DVS &amp;lt;3; n=17) plus a GnRH treatment at TAI. The diameter of the POF was analysed using analysis of variance (PROC GLIMMIX of SAS; SAS Institute Inc.), and the means were compared among groups using Tukey's test. The proportion of cows that displayed oestrus and pregnancy rates was analysed using chi-square test. Cows in the high-DVS group had a larger POF than cows in the low-DVS and low-DVS-GnRH groups (13.2±0.2, 11.7±0.5, and 12.2±0.4, respectively; P&amp;lt;0.05). The proportion of cows that displayed oestrus was greater (P&amp;lt;0.05) in the high-DVS group (72%, 26/36) than in the low-DVS (37.5%, 6/16) or low-DVS-GnRH (53%, 9/17) groups. Finally, greater (P&amp;lt;0.05) pregnancy rates were observed in cows from the high-DVS (47.2%; 17/36) and low-DVS-GnRH (52.9%; 9/17) groups than in cows from the low-DVS group (18.7%; 3/16). The preliminary results from this study demonstrated that diameter of POF is positively associated with DVS. Moreover, cows that presented POF with higher DVS are more likely to become pregnant, and the administration of GnRH to females with low DVS can increase the fertility of beef cows submitted to TAI protocols. This study received funding support from Embrapa (MP1/PC3 project no. 01.03.14.011.00.00) and from CNPq (universal project no. 407307/2016-8).

124 Ovarian follicular development and steroid secretion during oestrous cycle of Lohi sheep

Pakistan has 30.9 million heads of sheep; however, little information is available on their reproductive aspects. The objective of this study was to document ovarian physiology and endocrinology of Lohi ewes during the oestrous cycle. Nine Lohi ewes, synchronized by administering single prostaglandin F2α (PGF2a; Cyclomate, Star Laboratories), were monitored for ovarian follicular dynamics using transrectal ultrasonography (7.5MHz, HS-1500, Honda) for two consecutive oestrous cycles during the breeding season (September to November 2018). Changes in plasma progesterone and oestradiol-17β concentrations of ewes (n=9) were also determined during the oestrous cycle using radioimmunoassay. The interovulatory interval of Lohi ewes averaged 17.0±0.1 days, and the duration of follicular and luteal phases was 4.6±0.2 and 11.3±0.2 days, respectively. Follicles emerged in either 3- or 4-wave patterns, but the frequency of the 3-wave pattern was higher than that of the 4-wave (87 vs. 13%, respectively; P=0.05). Following ovulation (Day 0), follicles (=3mm) in 3-wave cycles (n=14) emerged on Days 0.7, 5.2, and 10.5, whereas in 4-wave cycles (n=2) follicles emerged on Days 0.1, 4, 8.5, and 11.5. The maximum diameter of preovulatory follicles and corpora lutea (CL) were 5.4±0.3 and 10.4±0.3mm, respectively. Regardless of the wave pattern, single ovulation occurred in each cycle. The CL was first detectable on Day 4±0.1, it reached maximum diameter on Day 9±0.1, and luteolysis began on Day 12.2±0.2 of the cycle. The peak plasma oestradiol-17β concentration (42.5±2.6 pgmL−1) was observed 48h before ovulation and correlated with the diameter of the preovulatory follicle during the follicular phase (r=0.84; P&amp;lt;0.05). The peak plasma progesterone concentration (11.8±1.7ngmL−1) was observed on Day 9±0.1 and coincided with the diameter of CL throughout the oestrous cycle (r=0.93; P&amp;lt;0.05). In conclusion, the majority of oestrous cycles in Lohi ewes had a 3-wave pattern and were mono-ovulatory in nature.

170 Embryonic loss and pregnancy rate in response to resynchronisation using oestradiol benzoate or injectable progesterone at 14 days after timed AI in Bos taurus×Bos indicus beef heifers

The effect of the exposure of pregnant cattle to oestradiol benzoate (EB) or injectable long-acting progesterone (iP4), as well as their use for early resynchronisation of non-pregnant (NP) females within the second week after timed AI (TAI), is yet to be assessed. This study aimed to assess the pregnancy rate and embryonic loss of cross-bred Bos taurus×Bos indicus beef heifers in response to an EB- or iP4-based resynchronisation strategy implemented 14 days after the first TAI. A total of 677 cyclic heifers (Braford and Brangus; 24 months old) were subjected to an EB/P4 synchronisation protocol for the first TAI (Day 0=TAI). On Day 14, all heifers received a new P4 intravaginal device (1g, Sincrogest, Ourofino Saúde Animal) and were randomly assigned to two groups: EB (1mg intramuscularly, Sincrodiol; n=339) or iP4 (75 mg intramuscularly, Sincrogest Injetável; n=338). On Day 22, P4 devices were removed and heifers underwent colour Doppler ultrasonography evaluation. Non-pregnant females were identified by the assessment of structural regression of the corpus luteum, as previously reported by our group. On Day 22, NP heifers received 1mg of oestradiol cipionate (SincroCP), 250µg of sodium cloprostenol (Sincrocio), and 200IU of equine chorionic gonadotrophin (SincroeCG), and they were inseminated on Day 24. On Day 22, the diameter of the preovulatory follicle (POF) was also recorded. Pregnancy was diagnosed via B-mode ultrasonography on Days 35-40 and 54. Statistical analyses were performed using SAS software (ver. 9.2, SAS Institute Inc.). The MIXED procedure was used to evaluate the POF diameter, and the GLIMMIX procedure was used to develop a multivariate model including treatment, breed, age, farm (1-7), and possible interactions of treatment with farm, breed, and age to evaluate the binomial data (pregnancy rates and embryonic loss). In NP heifers, the diameter of the POF on Day 22 did not differ (P&amp;gt;0.05) between the EB (9.77±0.25mm) and iP4 (9.92±0.22 mm) groups. Exposure to EB or iP4 on Day 14 post-TAI did not influence pregnancy rates as assessed on Day 22 (56.3% (191/339) vs. 60.1% (203/338)) and on Days 35-40 (49.6% (168/339) vs. 53.3% (180/338)). In addition, embryonic loss between Days 22 and 35-40 was not different between the EB (12%, 23/191) and iP4 (11.3%, 23/203) groups. On the other hand, pregnancy rates in heifers submitted to the second TAI were greater (P=0.01) in the EB group (45.95%, 68/148) than in the iP4 group (31.85%, 43/135). In conclusion, improved pregnancy rates can be achieved using 1mg of EB associated with a P4 intravaginal device on Day 14 of the resynchronisation protocol without affecting the first TAI pregnancy rates and embryonic loss. However, further investigation is warranted to assess the lower pregnancy rates obtained in response to the association of iP4 and P4 device. This research was supported by FAPESP (2015/10606-9; 2019/15514-6), Graduate Studies Support Program (PAPG-UNIPAMPA), and Ourofino Saúde Animal.

Effect of dietary n-3 polyunsaturated rich fish oil supplementation on ovarian function and interferon stimulated genes in the repeat breeding cow

Dietary n-3 polyunsaturated fatty acids (n-3 PUFA) improve utero-ovarian functions and embryonic survival in postpartum dairy cows. Because early embryonic mortality is the major cause of repeat breeding (RB) in cows, there was investigation of the effect of dietary supplementation of n-3 PUFA [eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)] rich fish oil (FO) from −2 to +2 weeks of artificial insemination on the size of preovulatory follicle (POF), serum progesterone (P4) and relative abundance of the mRNA of interferon stimulated genes (ISG) that encode for these proteins in the peripheral blood leukocytes (PBL) in the RB cow (n = 12). The diet of control group was supplemented with palm oil (PO). The results indicated serum concentrations of EPA and DHA were greater by 4.6- and 3.5-fold, respectively at the end of feeding study in the RB cows of the FO group. The diameter of POF was larger by 2.2 mm in FO group; however, serum P4 did not vary from day 14–20 post-artificial insemination. Greater abundance of ISG mRNA transcripts such as ISG15, RTP4, Mx2 and OAS1 in the PBL of pregnant cows of FO group indicates day 20 conceptuses produced more IFN-τ. It is concluded that supplementation of FO during the breeding period increased the size of POF and enhanced the abundance of ISG mRNA transcripts in RB cows that became pregnant.

Effect of manipulating progesterone before timed artificial insemination on reproductive and endocrine outcomes in high-producing multiparous Holstein cows

Our objective was to evaluate the effect of manipulating progesterone (P4) concentrations before timed artificial insemination (TAI) on reproductive and endocrine outcomes in high-producing Holstein cows. Multiparous lactating Holstein cows (n = 80) were synchronized for first TAI using a Double-Ovsynch protocol and were randomly assigned to receive 25 mg of PGF2α 1 d after the first GnRH treatment of the Breeding-Ovsynch protocol that included a once-used P4 insert (low-P4 group) or to receive 2 new P4 inserts during the Breeding-Ovsynch protocol (high-P4 group). Blood samples were collected thrice weekly from -10 to 32 d relative to TAI for all cows and from 32 to 67 d after TAI for pregnant cows and were analyzed for P4 and pregnancy-specific protein B (PSPB) concentrations. Expression of IFNτ-stimulated gene 15 (ISG15) was assessed in blood leukocytes 18 and 20 d after TAI. As expected, P4 concentrations were greater for high-P4 cows than for low-P4 cows from 3 to 8 d before TAI. Incidence of double ovulation was 3-fold greater for low-P4 cows than for high-P4 cows (33 vs. 10%), which resulted in more twin pregnancies 32 d after TAI for low-P4 cows than for high-P4 cows (29 vs. 0%). Low-P4 cows had larger preovulatory follicles at the last GnRH treatment of the Double-Ovsynch protocol and greater P4 concentrations than high-P4 cows after TAI. Relative expression of ISG15 mRNA 18 and 20 d after TAI was greater for low-P4 cows than for high-P4 cows and for pregnant cows than for nonpregnant cows. Overall, PSPB concentrations tended to be greater for low-P4 cows than for high-P4 cows, and pregnant cows had greater P4 concentrations than nonpregnant cows. In summary, cows with low P4 before TAI had increased preovulatory follicle diameter, PSPB concentrations, relative expression of ISG15 mRNA 18 and 20 d after TAI, double ovulations, and twinning compared with cows with high P4 before TAI. Increasing P4 before TAI may effectively decrease double ovulation and twinning in high-producing multiparous Holstein cows.

Effect of eCG on the follicular dynamics and vascularization of crossbred cows with different circulating progesterone concentrations during synchronization of ovulation in an FTAI protocol

ABSTRACT: This study aimed to evaluate the effect of treatment with equine chorionic gonadotrophin (eCG) on the follicular dynamics and function of crossbred cows with different circulating progesterone (P4) concentrations during synchronization of ovulation in a fixed-time artificial insemination (FTAI) protocol. To this end, 30 crossbred cows were submitted to a pre-synchronization protocol to ensure that all of them presented corpus luteum (CL) at the beginning of the protocol, and were evaluated by transrectal ultrasonography (TRUS) to verify the presence of CL. After that, the animals underwent an ovulation synchronization protocol and evaluation of follicular dynamics and vascularization by B-mode and power-Doppler ultrasound (US). High plasma P4 concentrations at the time of ovulation synchronization negatively influenced follicle diameter on day 10 (D10), preovulatory follicle diameter, and preovulatory follicle wall vascularization area (p&lt;0.05). Cows with high P4 concentration at the time of ovulation synchronization that were treated with eCG showed follicle diameter on D10 and preovulatory follicle diameter and wall vascularization area (p&gt;0.05) similar to those of animals with low P4 concentration at the time of ovulation synchronization. Therefore, high P4 concentrations at the time of ovulation synchronization negatively influence follicular diameter and vascularization, and eCG can be used as a strategy to favor better follicular and luteal response in crossbred cows with high P4 concentrations submitted to an FTAI protocol.

Resynchronization of ovulation with new and reused intravaginal progesterone-releasing devices without previous pregnancy diagnosis in Bos taurus indicus cows subjected to timed-artificial insemination.

The study aimed to evaluate pregnancy per artificial insemination (P/AI) of cows subjected to synchronization and resynchronization in ovulation protocols using intravaginal progesterone-releasing insert (P4) before pregnancy diagnosis (PD) and the relationship of PR with the diameter of preovulatory follicles (ØPOF) before TAI. Cows (n=378) were distributed into two groups: a resynchronization group with new devices (GRN; n=185) and resynchronization group with used devices (GRU; n=193). On Day 0, both groups received a new P4 and estradiol benzoate (EB). On D8, P4 removal+D-cloprostenol+eCG+estradiol cypionate (EC) was done. On d10, TAI was conducted. On d32, cows were resynchronized and divided into two groups, GRN (n=185) and GRU (n=193). The GRN group received a new P4+EB, and the GRU group received a used P4+EB. On d40, the P4 was removed+PD. The non-pregnant cows received D-cloprostenol+eCG+EC. US was done again on d42 to determine ØPOF before the second TAI. The P/AI of the GRN and GRU groups after synchronization were 56.2% and 57.0% (p=0.87), respectively, and those after resynchronization were 58.0% and 37.3% (p<0.008), respectively. The P/AI of the GRN and GRU groups observed after TAI (synchronization+resynchronization) were 81.6% and 73.1%, respectively (p=0.047). No difference (p=0.067) in ØPOF between the pregnant and non-pregnant cows in the GRN was found, whereas the GRU group showed a significant difference (p=0.003). Resynchronization protocols optimized the P/AI in both groups. New intravaginal devices resulted in greater P/AI and P/AI accumulation in resynchronization as compared with the GRU; the ØPOF was related with P/AI.

Intensity of estrus following an estradiol-progesterone-based ovulation synchronization protocol influences fertility outcomes

The objective of this study was to examine the association between increased physical activity at the moment of timed artificial insemination (AI), detected by an automated activity monitor (AAM), and fertility outcomes. This paper also investigated factors affecting estrous expression in general. A total of 1,411 AI events from 1,040 lactating Holstein cows were recorded, averaging 1.3 ± 0.6 (±standard deviation) events per cow. Activity (measured as steps/h) was monitored continuously by a leg-mounted AAM located on the rear leg of the cow. Ovulation was synchronized by a timed AI protocol based on estradiol and progesterone. Ovarian ultrasonography was performed in all cows on d -11 (AI = d 0) and in a subset of cows on d 0 (n = 588) and d 7 (n = 819) to determine the presence of a corpus luteum and follicles. The body condition score (1 to 5 scale) was assessed on d 0 and a blood sample was collected for progesterone measurement on d 7. Using the AAM, an estrus event was determined when the relative increase (RI) in physical activity of the cow exceeded 100% of the baseline activity. The physical activity was classified as strong RI (≥300% RI), moderate RI (100-300% RI), or no estrus (<100% RI). Milk production was measured daily and averaged between d -11 and 0. Pregnancy was diagnosed at 32 and 60 d post-AI and pregnancy losses were calculated. The mean RI at estrus was 328.3 ± 132.1%. Cows with strong RI had greater pregnancy per AI than those with moderate RI and those that did not express estrus (35.1 vs. 27.3 vs. 6.2%). When including only cows that successfully ovulated after timed AI, those that displayed strong intensity RI still had greater pregnancy per AI than those with moderate intensity RI or those that did not express estrus (45.1 vs. 34.8 vs. 6.2%). Cows expressing strong RI at timed AI had greater ovulation rates compared with moderate RI and cows that did not express estrus (94.9 vs. 88.2 vs. 49.5%). Furthermore, pregnancy losses were reduced in cows with strong RI compared with cows expressing moderate RI (13.9 vs. 21.7%). Cows with a strong RI at estrus were more likely to have a corpus luteum at the beginning of the protocol and had greater concentration of progesterone 7 d post-AI. Multiparous cows expressed lower RI compared with primiparous cows. Cows with lower body condition score tended to have decreased RI at estrus. No correlation between estrous expression and pre-ovulatory follicle diameter was observed. Also, no correlation was observed between milk production at AI and RI. In conclusion, strong intensity RI of estrus events at timed AI was associated with improved ovulation rates and pregnancy per AI, and reduced pregnancy losses. These results provide further evidence that measurements of estrous expression can be used to predict fertility at the time of AI and possibly be used as a tool to assist decision making strategies of reproduction programs.

Echographie du développement folliculaire et de l’ovulation chez la chèvre du Sahel en œstrus induit

Le développement folliculaire et l’occurrence de l’ovulation ont été étudiés chez la chèvre du Sahel en œstrus induits. Quinze femelles d’âges et de poids moyens respectifs de 37,25 ± 1,5 mois et 32,5 ± 7,3 kg, et de parité comprise entre 1 et 3 ont été traitées à l’aide d’éponges intravaginales (acétate de fluorogestone, 20 mg/éponge) pendant 11 jours. Ce traitement a été associé à l’administration intramusculaire par animal de 400 UI de gonadotrophine chorionique équine (eCG) et de 2 ml de cloprosténol le neuvième jour. La détection de l’œstrus a été réalisée par observation visuelle après l’introduction de deux boucs fertiles dans le troupeau. Le développement folliculaire a été suivi par échographie à intervalles de 12 heures au moyen d’une sonde linéaire de 5 MHz. Le délai moyen d’apparition de l’œstrus a été de 15,4 ± 2,9 h. L’œstrus a été matérialisé par des écoulements vulvaires et des tentatives de chevauchements entre congénères. Sa durée moyenne a été de 32,3 ± 3,8 h. A l’arrêt du traitement, 13 follicules ont été observés sur l’ovaire droit et 13 follicules sur l’ovaire gauche avec des diamètres moyens respectifs de 5,26 ± 1,25 mm et de 4,73 ± 1,45 mm. Le diamètre moyen du follicule préovulatoire a été 7,6 ± 2,1 mm. Ce diamètre a varié selon l’ovaire. Au total 17 ovulations ont été enregistrées dont huit dans l’intervalle entre 45 et 50 heures après la fin du traitement. Ces résultats sont des éléments de repère précieux pour l’application des biotechnologies de la reproduction chez cette race.

Effect of breed and other animal-related factors on conception rate to artificial insemination with frozen semen in mares in Ethiopia.

Equine reproduction is unique by having long behavioral estrus and differences in time of breeding between breeds and individuals of mares. An experimental study was conducted at the Balderas Sport Horses and Recreational Center, Addis Ababa, Ethiopia, from January to June, 2018, to evaluate conception rate to frozen semen in local and exotic crossbreed mares. Mares were teased to characterize estrus behavior and examined by ultrasound in determining imminent ovulation. Inseminations were done post ovulation within an average of 6-9h using frozen-thawed semen. The overall conception rate to frozen semen was 15/21 (71.43%) with 8/11 (72.73%) in crossbreed and 7/10 (70%) in local breed mares. Age and body condition score (BCS) of animals had no significant effect on conception rate to AI with frozen semen. A slightly higher conception rate was obtained when ovulation was from the right ovary than when ovulated from the left ovary. A higher conception rate was obtained when the diameter of the preovulatory follicle was ≤ 45mm than above diameter. The conception rate increased significantly with increased number of services/conception with an overall mean ± (SEM) of 2.2 ± 0.2 services/conception. A more number of services/conception were required for local breed (2.7 ± 0.2) than crossbreed mares (1.8 ± 0.3) and again for lower body condition scores than higher condition scores of mares. In conclusion, the increased number of services improved the conception rate with significant difference between breed of mares, whereas good management of mares for improved body conditions could be required to decrease the number of services per conception.

Environmental effects and repeatability of the follicular diameter in mares

ABSTRACT The objective of this study was to evaluate the effect of environment on follicular diameter and repeatability of the size of preovulatory follicles of mares. Temperature, rainfall, and photoperiod were measured, and their effects were evaluated on follicular dynamics. Data were studied by analysis of variance of follicular size and environment traits. During the five years, 7% double ovulations and 159 anovulatory follicles were recorded. Mean preovulatory follicle diameters of left and right ovaries were 39.3±3.8 and 39.2±3.5 mm [...]

122 Effect of gonadotropin-releasing hormone plus 7-day progesterone (CIDR) with or without eCG on follicular dynamics, estrus response, and pregnancy rate in anestrous Beetal goats during nonbreeding season under subtropical conditions

Oestrus induction is a challenging task during the nonbreeding season in goats. Therefore, our objective was to evaluate the efficacy of gonadotropin-releasing hormone (GnRH) plus a 7-day CIDR-based protocol with or without eCG for the induction of oestrus and fertility in acyclic Beetal goats during the nonbreeding season under subtropical conditions. Based on the absence of corpus luteum via ultrasonography, goats (n=56; mean weight=32.8±0.5 kg; body condition scre=2.5±0.1) were inserted with a CIDR (300mg progesterone) along with GnRH (Day 0), and randomly treated either with eCG (400 IU; n=35) or without eCG (control; n=21) on Day 7. The CIDR was removed on Day 7 and all does were administered single dose of prostaglandin F2α (75µg cloprostenol; Fatro, Italy). Following CIDR insertion, ovarian follicular dynamics was monitored daily through a 7.5-MHz transrectal transducer (Honda HS 1500, Japan) until ovulation in a subset of eCG (n=11) and control goats (n=10). Oestrus was detected at every 12h after CIDR removal with aproned bucks and does were bred naturally. Pregnancy was detected 25 days after breeding by transrectal ultrasound. Data were analysed by t-test, ANOVA, and Chi-squared test using SPSS (version 20.0, SPSS Inc., Chicago, IL, USA). Oestrus occurred more often following CIDR removal in eCG v. control goats (94v. 57%; P&amp;lt;0.05). Similarly, onset of oestrus was earlier (36.0±2.1v. 54.2±4.7 h; P&amp;lt;0.05) and the duration of oestrus was longer in eCG v. control goats (i.e. 30.2±1.3v. 22.1±2.3 h; P&amp;lt;0.05). However, diameter of preovulatory follicles and interval to ovulation after CIDR removal were similar between eCG and control goats (P&amp;gt;0.05). In contrast, growth rate of preovulatory follicle after CIDR removal until ovulation in eCG goats was greater, and ovulation rate was higher than control goats (0.7±0.1v. 0.3±0.1 mm/day; 2.2±0.2v. 1.4±0.2, respectively; P&amp;lt;0.05). In eCG goats, the dominant follicle grew rapidly between Days 8 to 9 (P&amp;lt;0.05), whereas within control goats the dominant follicle grew at consistent rate (P&amp;gt;0.05). After CIDR removal, percentage of ovulating goats in eCG was higher as compared to the control group (100v. 44%; P&amp;lt;0.05). Within control goats, size of the dominant follicle in ovulatory goats was greater at the time of CIDR removal than anovulatory goats (5.7±0.3v. 5.0±0.15 mm; P&amp;lt;0.05), whereas wave emergence tended to be earlier after CIDR insertion in anovulatory goats (4.2±1v. 2.6±0.4 days; P=0.09). Pregnancy rate 25 days postbreeding did not differ between eCG and control goats [i.e. 49% (16/33) v. 25% (3/12); odds ratio 2.9/1; P&amp;gt;0.05]. In conclusion, the eCG protocol resulted in higher oestrus response, ovulation rate, and 3 times higher pregnancy rate in anestrous goats during the nonbreeding season.

118 A retrospective analysis of follicular dynamics, LH, estradiol-17β, and progesterone in prostaglandin F2α-induced estrus of Beetal goats

The objective of the study was to reveal the factors associated with variable interval to ovulation following a single dose of prostaglandin F2α (PGF2) in Beetal goats. In experiment 1, the extent of ovulation time was demonstrated after synchronizing Beetal goats (n=38) by using a single injection of PGF2 based on corpus luteum detection by B-mode transrectal ultrasonography. Ovulations were detected every 6h from PGF2 until ovulation. To explore the changes in follicular dynamics, endocrine profiles and behavioural events due to the variation in the interval to ovulation, another set of goats (n=13; experiment 2) were synchronized by a single dose of PGF2 given at random days of the luteal phase. Follicular dynamics were monitored every 12h from PGF2 until ovulation and oestrus was detected by apronized bucks every 6h from PGF2 treatment. Blood sampling (3mL) for oestradiol-17β and progesterone was done every 12h from PGF2 until ovulation and every 2h for LH assay by radioimmunoassay. Data were analysed by Student’s t-test, general linear models, and Pearson coefficient correlation (SPSS, version 20.0, SPSS Inc., Chicago, IL, USA). Results revealed that ovulation time varied from 60 to 96h after PGF2, and 57% of ovulations occurred by 72h after PGF2 (experiment 1). Based on this observation, goats (n=13) in second experiment were retrospectively divided into early and late ovulators (i.e. 72 and 84h following PGF2, respectively). Onset of oestrus, peak oestradiol-17β concentration, and LH surge after PGF2 were first observed in early than late ovulators (P&amp;lt;0.05). The goats having greater follicle diameter and smaller corpus luteum diameter at the time of PGF2 ovulated earlier than late ovulators (5.4±0.2v. 4.3±0.2mm and 10±0.6v. 11.8±0.3mm, respectively; P&amp;lt;0.05). Likewise, plasma progesterone concentration tended to be lower (P=0.087) in early than late ovulators. However, interval from onset of oestrus to peak LH and ovulation, duration of standing oestrus, interval from LH peak to ovulation, diameter and growth rate of preovulatory follicles, and number of ovulations were similar (P&amp;gt;0.05) between early and late ovulators. In conclusion, the size of dominant follicle and corpus luteum at the time of PGF2 determines the interval to ovulation following a single dose of PGF2 during the luteal phase.

Level of circulating concentrations of progesterone during ovulatory follicle development affects timing of pregnancy loss in lactating dairy cows

The objective of this experiment was to determine the effect of high versus low progesterone (P4) during the pre-dominance or dominance phase (or both) of ovulatory follicle development on follicular dynamics and fertility of lactating dairy cows. Progesterone (P4) was manipulated to reach high (H) or low (L) serum concentrations during the pre-dominance phase (d 0 to 4 of the wave) and dominance phase (d 5 to 7 of the wave) of a second follicular wave ovulatory follicle, creating 4 treatments: H/H, H/L, L/H, and L/L. Luteolysis was induced with PGF2α on d 7 of the wave and ovulation was induced with GnRH 56 h after PGF2α. Cows (n = 558) received artificial insemination (AI) 16 h following GnRH. Pregnancy was determined at 6 intervals during gestation and at calving to quantify pregnancy loss beginning at d 23 post-AI utilizing pregnancy-specific protein B (PSPB) in novel within-cow comparisons. Cows with single ovulations assigned to the L/L treatment had greater pre-ovulatory follicle diameter compared with cows assigned to the L/H or H/L treatments. Cows with single ovulations had greater pre-ovulatory follicle diameter compared with cows with double ovulations. Low P4 in H/L, L/H, and L/L increased double ovulation rate compared with H/H. Cows with double ovulations had greater pregnancies per AI (P/AI) on d 23 post-AI compared with cows with single ovulations but had greater losses if ovulations were unilateral. Cows with low P4 during the entire period of the ovulatory follicle development also had greater P/AI on d 23 post-AI compared with cows with high P4 during both phases. However, full-term P/AI was not different between treatments. This was a result of the greater incidence of pregnancy losses between d 35 and 56 of gestation for cows with unilateral double ovulations compared with bilateral double ovulations and single ovulatory cows. Cows with single ovulation and low circulating P4 during the dominance period of follicle development had increased pregnancy losses between d 35 and 56 of gestation compared with cows with single ovulations and high P4. The PSPB measurements on d 16 and 23 post-AI were highly accurate in the prediction of pregnancy at d 28. The PSPB differed on d 23 and 28 between cows that had versus cows that did not have pregnancy losses between d 28 and 35 of gestation. In summary, circulating concentrations of P4 during ovulatory follicle development affected numbers of follicles ovulated and timing of subsequent pregnancy losses.