Ovulation Synchronization Protocol Research Articles

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.

Comparison between the 5-day cosynch and 7-day estradiol-based protocols for synchronization of ovulation and timed artificial insemination in suckled BOS taurus BEEF cows

The objective was to compare pregnancy per AI and follicular dynamic in suckled Bos taurus beef cows treated with either a 7-day progesterone + estradiol-based protocol or a 5-day progesterone CoSynch protocol for timed artificial insemination (TAI) during four breeding seasons. We hypothesized that estrous cycle status, days postpartum (DPP), fat depth and plasma progesterone concentration differentially modify the effect of treatments. Every year, 9 days before initiation of each breeding season, cows were randomly assigned to one of two groups. Cows in the 7-d P + E group (n = 428) received a progesterone intravaginal device (DIB) and estradiol benzoate on Day −9. On Day −2 the device was removed, and cows received cloprostenol and estradiol cypionate. Forty-eight hours later (Day 0) cows received TAI. Cows in the 5-d P + CoS group (n = 428) received a DIB, and GnRH on Day −8. On Day −3, the device was removed, and cows received cloprostenol. A second dose of cloprostenol was given on Day −2. Cows received GnRH and TAI 72 h after device removal (Day 0). On Day −9, estrous cycle status was determined. In a subset of cows (n = 79) the size of the dominant follicle was determined between Days −2 and 0. In another subset of cows (n = 340), DPP, fat depth (mm) and plasma progesterone concentration (ng/mL) were evaluated on Day −9. Pregnancy per AI was determined 30 d after TAI. Pregnancy per AI was greater for cows in the 5-d P + CoS group than for cows in the 7-d P + E group (50.9% vs. 41.3%, P = 0.01) and was also greater in cyclic than in anestrus cows (54.3% vs. 33.2%, P < 0.0001). There was also a significant effect of breeding season (P = 0.0002) and sire (P = 0.03), and an interaction between treatment group and breeding season (P = 0.03). The dominant follicle was larger (P < 0.0001) in cows in the 5-d P + CoS group than the 7-d P + E group (10.7 ± 0.29 mm vs. 9.0 ± 0.28 mm). Pregnancy per AI was greater in cows with ≥55 DPP (47.0% vs. 29.6%, P = 0.001), fat depth ≥0.50 mm (44.7% vs. 29.7%), and with plasma progesterone concentration ≥1 ng/mL (47.2% vs. 28.7%, P = 0.01). In cows with plasma progesterone ≥1 ng/mL on Day −9, pregnancy per AI was greater in the 5-d P + CoS group (60.5%) than in the 7-d P + E group (34.9%), but there was no difference between treatment groups in cows with plasma progesterone < 1 ng/mL (P = 0.07). In conclusion, the 5-d P + CoS protocol resulted in greater size of the dominant follicle and pregnancy per AI in suckled Bos taurus beef cows subjected to TAI.

Pre-TAI protocol strategies to increase reproductive efficiency in beef and dairy cows.

Ovulation synchronization protocols are well established in beef and dairy cows. However, the protocol response rate is around 70-90%. In beef cows, factors such as inadequate nutrition and calf presence negatively impact the response of progesterone (P4)/estradiol-based ovulation synchronization protocols by interfering with GnRH release and consequently reducing LH pulsatility and final follicular development. In dairy cows, protocols based on GnRH and prostaglandin (Ovsynch) are the most widely used in the world. However, the efficiency of Ovsynch is dependent on the presence of a large follicle at the time of administration of the first GnRH. In these ovulation synchronization protocols, pre-synchronization protocols (Prostaglandins, Double Ovsynch and P4synch) are usually attempted in an effort to increase responses. Thus, the objective of this review was to discuss pre-ovulation synchronization strategies (administration of injectable P4 or energetic/protein supplementation or pre-synchronization with intra-vaginal progesterone devices) aiming to increase the LH pulsatility in beef cows or induce the formation of a GnRH-responsive follicle at the beginning of the Ovsynch protocol in dairy cows.

Effects of short-term intravaginal progestagen treatment on fertility and prolificacy after natural breeding in sheep at different reproductive seasons

ABSTRACT The present study indicates that short-term progestagen-based protocols for synchronization of estrus and ovulation in sheep involving 7 days of progestagen insertion with administration of prostaglandin F2α at either insertion or removal of the progestagen sponge resulted in 80–90% fertility during the breeding season, while a classical long-term protocol of 14 days of progestagen insertion resulted in 77% fertility. During the non-breeding season, fertility was significantly higher for the 7-day protocol with prostaglandin administration at sponge insertion (79.2%) and for the 14-day protocol (80%) than for the 7-day protocol with prostaglandin administration at sponge removal (59.1%; P = 0.018). Prolificacy, in contrast, varied significantly with genotype, being higher in prolific breeds, but it did not vary with progestagen protocol or breeding season. These results suggest that short-term progestagen-based treatments can provide similar reproductive efficiency as long-term treatments, which may help practitioners reduce welfare and health issues while maintaining productivity.

120 Evaluation of ovulation synchronization protocols and alternative pregnancy diagnosis tools in lactating dairy cows.

Our objectives were to appraise three ovulation synchronization protocols and Pregnancy diagnosis(PD) methods in lactating dairy cows. Cows (n=184;4.2 ± 1.2 lactations and average daily milk yield>28 kg with 3X milkings) were randomly assigned to 1 of 3 timed artificial insemination(TAI) protocols: 1)Ovsynch protocol (n=66), GnRH-PGF2α-GnRH; 2)G7G-Ovsynch (n=63) protocol, PGF2α,GnRH 2 days later followed by Ovsynch; 3)PG7G-Ovsynch(n=55) protocol, PGF2α 14d before the start of G7G-Ovsynch protocol.Pregnancy rate(PR) was determined on d30, d60 and d90 post TAI using ultrasonography(US), P4(RIA), milk and Plasma PAGs(ELISA). The PR was compared using chi-square. Milk, plasma PAGs and plasma P4 profiles were analyzed using REG procedures of SAS(P<0.05). Ovarian status was monitored prior to enrollment and at final GnRH of individual protocol. Ovulatory follicle diameter(OFD) was 15.32 ± 0.18(Ovsynch), 15.06 ± 0.16(G7G-Ovsynch) and 15.14 ± 0.13(PG7G-Ovsynch) respectively. The PR on d30 was 44%(29/66), 51%(32/63) and 56%(3/55) in Ovsynch,G7G-Ovsynch and PG7G-Ovsynch(P=0.39) protocols respectively. On d60 post TAI,PR was 39%(26/66),48%(30/63) and 53%(29/55) in all three groups(P=0.33). Similarly, PR was 38%(25/66),48%(30/63) and 51%(28/55) with overall pregnancy loss of 14%,6%(2/32) and 10%(3/31) in Ovsynch,G7G-Ovsynch and PG7G-Ovsynch groups respectively on d90 post TAI. Plasma P4 profile (ng/ml; Mean±SEM) was 7.40 ± 0.53,7.75 ± 0.64 and 7.13 ± 0.42 on d30, d60 and d90 post TAI respectively. Optical density(OD) of milk PAGs was 1.06 ± 0.12,0.48±.04 and 0.96 ± 0.11 on day 30,60 and 90 post TAI respectively. Likewise, plasma OD was 1.82 ± 0.10,0.87 ± 0.05 and 1.66 ± 0.12 on d30,d60 and d90 post TAI. Sensitivity(Se), Specificity(Sp), positive predictive value(PPV) and negative predictive value(NPV) of milk and plasma PAGs based test in present study was 98%(95% CI:91–100),100%(95% CI:94–100), 100% and 98%(95% CI:89–99) respectively in comparison with US on the PD basis. Similarly,Se,Sp,PPV and NPV of plasma P4 test in present study was 100%. In conclusion, introduction of presynch based protocols have been resulted in improved, although non-significant, PR in lactating dairy cows and PAGs are other apposite PD tools.

83 Pregnancy losses after transfer of bovine embryos produced by assisted reproductive technologies

The aim of this study was to evaluate risk factors for pregnancy loss in embryo recipients. We evaluated data from 31,792 embryo transfers (ET), including 859 embryos derived in vivo (IVD), 28,814 produced in vitro (IVP), and 2,119 produced by somatic cell nuclear transfer (SCNT), recorded from 2008 to 2012 on a single commercial farm. Embryos were produced by different in vitro embryo production laboratories, but ET was performed by the same team and within the same herd. Recipients were checked for pregnancy (PR) by ultrasonography 23 days after ET and re-examined on Days 60, 90, and 120 of gestation to identify pregnancy losses (PL). Data were analysed for the main effect of embryo production technique upon PL. For IVD and IVP embryos only, additional risk factors were included in the statistical model, including cryopreservation, type of semen (conventional, sex-sorted, reverse sex-sorted), embryo developmental stage, recipient corpus luteum quality score, use of natural oestrus or synchronization protocol, embryo-recipient synchrony, embryo genetics (taurine v. zebu, dairy v. beef), season (summer v. winter), and fetal sex. Data were analysed using the GLIMMIX procedure of the SAS Software (v 9.3, SAS Institute Inc., Cary, NC, USA). Overall PR was 42.1% (13,395 of 31,792) and PL between days 30 and 60, 60 and 90, and &amp;gt;90 were 8.7, 5.3, and 3.5%, respectively (cumulative PL=17.4%). The SCNT embryos had lower PR compared to IVP and IVD embryos (36.6, 42.5, and 44.2%, respectively; P&amp;lt;0.001) as well as a greater PL in all periods (30-60=45.2, 6.4, and 8.2%; 60-90=3.5, 1.4, and 1.8%; and &amp;gt;90=21.4, 6.3, and 7.1%, respectively; P&amp;lt;0.0001). The PR was similar between fresh and frozen embryos, but lower for vitrified ones (42.9, 43.1, and 39.1%, respectively, P&amp;lt;0.05). Slow freezing resulted in a greater total PL compared to fresh embryos (18.0v. 13.9%, respectively; P&amp;lt;0.05). We observed an interaction between embryo genetics and season. The taurine dairy subgroup had the lowest PR during summer (37.7v. &amp;gt;41.7% in all other groups; P&amp;lt;0.01), but within this group PR increased during winter (37.7v. 44.6%, P&amp;lt;0.05). The PL was greater for taurine than zebu during summer (17.8v. 14.0%; P&amp;lt;0.001), but did not differ during winter (P&amp;gt;0.05). The type of semen, embryo developmental stage, corpus luteum quality score, and embryo recipient synchrony affected pregnancy rates (P&amp;lt;0.05) but not pregnancy losses (P&amp;gt;0.05). Interestingly, the use of ovulation synchronization protocols resulted in similar pregnancy rates but greater pregnancy losses compared to natural oestrus (41.8v. 43.2%, P&amp;gt;0.05; and 15.9v. 22.1%, P&amp;lt;0.05, respectively). Pregnancy loss after 60 days was similar between female and male fetuses (5.0v. 4.9%, respectively; P&amp;gt;0.05). In summary, the assisted technology used to produce and freeze embryos was the main cause of variation in pregnancy losses, which were also affected by the interplay of season by genetic background and by the use of synchronization protocols. Conversely, factors known to affect pregnancy rates, such as embryo developmental stage, corpus luteum quality, and embryo-recipient synchrony, had no effect upon pregnancy loss. Research was supported by Mineirembryo, CAPES, and FAPDF.

Assessment of ovulation synchronization protocols in goats and use of pregnancy specific protein B (PSPB) enzyme linked immunsorbent assay (ELISA) to determine kid number at birth

The efficacy of several protocols for ovulation synchronization and timed artificial insemination (TAI) in goats was examined. In addition, the relationship between levels of pregnancy specific protein B (PSPB) during gestation assessed with a commercially available ELISA and the number of offspring at birth was determined. In Experiment 1, 70 does were randomized into four treatments: (1) breed by estrus [BBE], (2) 6-d treatment with a new [C6N], (3) once-used [C61], or (4) twice-used Controled Internal Drug Release (CIDR) device [C62)]. BBE does received two 15 mg doses of prostaglandin-F2α (PGF) at a 10-d interval and were bred 12 h after estrus onset. CIDR groups received a CIDR for 6 d with 15 mg PGF given at CIDR removal. TAI was performed 48 h after CIDR removal and does were given 50 μg GnRH. All does were inseminated with a single dose of frozen semen using a non-surgical, transcervical technique. Pregnancy rates for the BBE, C6N, C61 and C62 treatment groups were 39% ± 12%, 64% ± 12%, 77% ± 12% and 57% ± 12%, respectively, and did not differ. Reuse of CIDRs, even with reuse extending for a total of 21 d, was as effective as new CIDRs for synchronization of ovulation. In Experiment 2, 68 does were randomized into four treatments: (1) BBE, (2) C6N, (3) NC.Synch [NCS], (4) modified NCS [NCSM]. The BBE and C6N groups were as described for Experiment 1. The NCS and NCSM groups received 15 mg PGF on Day 1, 50 μg GnRH on Day 8 and 15 mg PGF on Day 15 (NCS) or Day 15.5 (NCSM). Does were bred by TAI at 72 h (NCS) or 60 h (NCSM) after the second PGF injection. All does in the NCS and NCSM groups received 50 μg GnRH at TAI. Pregnancy rates were 53% ± 12%, 30% ± 11%, 50% ± 11% and 41% ± 12% for does in the BBE, C6N, NCS and NCSM group, respectively, and did not differ. In Experiment 3, 62 does pregnant to TAI were bled at Days 48 and 85 post-insemination for PSPB. Data on kid numbers and birth weights were subsequently recorded. At Day 48 of gestation, PSPB levels for does birthing singletons were lower than for does birthing twins or triplets (25.0 ± 0.1a, 28.8 ± 0.1b and 30.7 ± 0b ng/mL, respectively, abP<0.05). At Day 85 of gestation, PSPB levels were progressively greater for does birthing singletons versus twins versus triplets (27.0 ± 0.1a, 28.5 ± 0.1b and 31.6 ± 0c ng/mL, abcP<0.05). In conclusion, PSPB concentrations detected using a commercially available ELISA at Day 48 or 85 of gestation could distinguish does carrying single versus multiple fetuses.

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.

Effects of short-term intravaginal progestagens on the onset and features of estrus, preovulatory LH surge and ovulation in sheep

In the present study, there was a comparison among classical long-term progestagen (fluorogestone acetate) protocols for synchronization of estrus and ovulation (14 days; group FGA14, n = 9 ewes) and short-term protocols based on 7 days of progestagen treatment plus a dose of prostaglandin F2α at either insertion (PG-FGA7, n = 11) or removal (FGA7-PG, n = 12). There were no significant differences in the ovulation rate and progesterone secretion among treatments. The FGA7-PG group, however, had a similar percentage of ewes expressing estrous behavior than the group FGA14 (90.9 and 100%, respectively, with a trend for a lesser percentage in the PG-FGA7 group, 63.6%) and about 90% of the ewes in the FGA7-PG group had the preovulatory surge release of LH 8 h after the onset of estrous behavior. These features may be related to a greater number of preovulatory follicles during growing phases (P < 0.05) and a greater plasma estradiol concentration (P < 0.05) in this group than in the classical 14-day group, which suggest these are more functional preovulatory follicles. In conclusion, therefore, the use of the FGA7-PG treatment may favor efficiency of progestagen-based protocols for reproductive management.

Evaluation of buffaloes' follicular dynamics and stress state under different ovulation synchronization protocols.

The aims of the present study were to analyze the effect of different hormonal protocols using an implant containing Norgestomet, at the ovulation synchronization in buffaloes and to verify the effect of the stress caused by manipulation of the herd during the experiment. Twenty-four female Murrah breed buffaloes, lactating, multiparous, aged from three to eight years, with a body condition score of 3.5 or higher (0 to 5) and with more than 45 days post partum, were used. These females were randomly distributed into one of the three groups (group 1, group 2 and group 3) with eight subjects in each. On day 0 (day 0), all animals received an ear progesterone implant with 3.0 mg of Norgestomet and an intramuscular (IM) injection with 2.0 mg of estradiol benzoate (EB). The implants were removed after nine days (day 9) and one single dose of PGF2α (0.15 mg d-cloprostenol, IM) was administered to all animals. On the same day, the group´1 and group 3 buffaloes were treated with 500 UI of eCG (IM). Two days later (day 11), 1000UI of hCG (IM) were administered to the group 1 and group 2 buffaloes. After the implant had been removed, an ultrasound evaluation was performed every 12 h, in order to access the ovarian follicular dynamics, using an Aloka 500 equipment with a 5 MHz transrectal probe. Blood samples were also taken on days 0, 9 and 11 to determine the plasmatic concentrations of cortisol and progesterone. No difference was observed in the ovulation rate (group 1: 62.5%, group 2: 50%, group 3: 75%). However, the size of the preovulatory follicles and the plasmatic concentration of cortisol had (P < 0.05). G3 was the most efficient group in promoting the ovulation synchronization, which suggests that this protocol can be used in Fixed Timed Artificial Insemination programs (FTAI) among postpartum, lactating Murrah breed buffaloes’.

Impact of Different Ovulation Synchronization Protocols on Plasma Profile of Progesterone, Protein and Cholesterol in Cyclic Repeat Breeder Crossbred Cows

Impact of Different Ovulation Synchronization Protocols on Plasma Profile of Progesterone, Protein and Cholesterol in Cyclic Repeat Breeder Crossbred Cows

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.

15 Effects of Timed Artificial Insemination Protocol Using Prostglandin F2α and Estradiol Benzoate on Conception Rate in High-Producing Dairy Cows

For several years, milk production per Holstein cow has been increasing while reproductive performance has declined worldwide. Timed AI (TAI) protocols with synchronization of oestrus and ovulation are tools for the improvement of reproductive performance in high producing dairy herds. In protocols for synchronization of oestrus and ovulation, oestradiol benzoate (EB) has been used to increase the number of animals that show oestrous behaviour, synchronize ovulation, and improve conception rate (CR). The objective of this study was to compare a simple oestrus synchronization and TAI protocol using prostaglandin F2α (PG) and EB with a more complicated TAI protocol or oestrus detection and AI on CR in high-producing Holstein cows. The study was conducted from 2014 to 2016, using 1,036 Holstein cows from 3 commercial dairy herds in central Hokkaido, Japan. The average parity number was 2.6 ± 1.5, and days open averaged 152.2 ± 82.0 days. The average milk yield (305 days) was 11,348 ± 1,619 kg. Oestrus synchronization of cows was achieved using PG followed by EB and TAI (PG was given to cows with a cutoff for CL diameter of &gt;20 mm by ultrasonography; EB (1 mg) was administered 24 h later and AI was carried out 24 to 48 h after EB injection), or by Heatsynch + CIDR [gonadotropin-releasing hormone (GnRH) injection and CIDR insertion at random stages of the oestrous cycle, CIDR removal, and PG injection 7 days later, EB injected 24 h after CIDR removal, and TAI 24 to 32 h after EB injection]. Cows observed in natural oestrus were used as controls. All cows were inseminated after the onset of standing heat or removal of the tail chalk (Paintstik®, LA-CO Industries, Elk Grove Village, IL, USA). Pregnancy diagnoses were performed using ultrasonography between 30 and 45 days after AI. The CR were compared using chi-square test. The CR (no. of pregnant cows/no. of total cows in the treatment group) of the natural oestrus group, PG + EB, and Heatsynch + CIDR cows were 39.2% (302/771), 38.7% (48/124), and 31.2% (44/141), respectively. Overall CR for the 3 groups was 38.0%. There were no significant differences in CR among the 2 groups (P &gt; 0.07). These results show that a simple oestrus synchronization protocol with injection of PG followed by EB can be used effectively for high-producing dairy herds, yielding a satisfactory CR similar to that obtained with the Heatsynch + CIDR protocol or natural oestrus.

7 Ovarian Follicular Dynamics and Conception Rate in Bos indicus Cows with Different Antral Follicle Counts Subjected to Timed Artificial Insemination

Two experiments in Nelore cows subjected to conventional protocol for timed AI (TAI) were designed to compare the influence of antral follicle count (AFC; 1) on the ovarian follicular dynamics and (2) conception rates. First, multiparous cows with high (G-high, ≥45 follicles; n = 43) and low AFC (G-low, ≤15 follicles; n = 32) were selected to receive a conventional oestradiol/progesterone-based TAI protocol for monitoring ovarian follicular dynamics. Second, AFC from 962 multiparous suckling cows (40 ± 15 days postpartum) subjected to a conventional protocol for TAI were classified as G-high (≥45 follicles; n = 194), G-intermediate (≥20 and ≤40 follicles, n = 397), and G-low AFC (≤ 15 follicles; n = 243). Pregnancy status was evaluated 30 to 35 days later. Data were analysed using ANOVA followed by the Tukey test and model of logistic regression (P ≤ 0.05). In the first study, the AFC were greater (P ≤ 0.05) in G-high than in G-low, but remained stable in both groups during the evaluations. The ovarian diameter, perimeter, and area were greater (P ≤ 0.05) in G-high than in G-low (28.3 ± 3.9 v. 20.5 ± 3.2 mm, 100.9 ± 13.2 v. 73.8 ± 13.9 mm, and 67.5 ± 16.4 v. 37.7 ± 11.7 mm2; respectively). Follicular growth rate, time of ovulation, corpus luteum diameter, and progesterone concentration were similar (P &gt; 0.05) for cows with low or high AFC. Dominant follicles showed greater (P ≤ 0.05) diameter in G-low than G-high at D4 (7.3 ± 2.2 v. 6.2 ± 1.4 mm; P = 0.069, tendency), D8 (11.2 ± 1.8v. 9.5 ± 1.8 mm), D9 (12.3 ± 1.7 v. 10.6 ± 1.7 mm), and D10 (13.4 ± 1.3 v. 12.2 ± 1.8 mm), and the estimated diameter of the ovulatory follicle tended (P = 0.08) to be higher in G-low than in G-high (14.4 ± 1.5 v. 13.4 ± 2.1 mm). In the second study, AFC of G-low cows showed higher (P ≤ 0.05) conception rate (61.7%) than those of G-intermediate (52.9%) and G-high (49.5%) cows. In Nelore cows subjected to ovulation synchronization protocol, cows with high AFC showed higher ovarian measurements, but those with low AFC had large follicular diameters. Additionally, the cows with low AFC showed higher conception rate than groups with intermediate and high counts.

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.

Extending the duration of the voluntary waiting period from 60 to 88 days in cows that received timed artificial insemination after the Double-Ovsynch protocol affected the reproductive performance, herd exit dynamics, and lactation performance of dairy cows

This experiment evaluated the reproductive performance, herd exit dynamics, and lactation performance of dairy cows managed with a voluntary waiting period (VWP) of 60 or 88 d. Secondary objectives were evaluating VWP effect on cyclicity status, uterine health, systemic inflammation, and body condition score (BCS) before first service. Lactating Holstein cows from 3 commercial farms in New York State cows were blocked by parity group and total milk yield in their previous lactation and then randomly assigned to VWP of 60 (VWP60; n = 1,352) or 88 (VWP88; n = 1,359) days in milk (DIM). All cows received the Double-Ovsynch protocol (GnRH-7 d-PGF2α-3 d-GnRH-7 d-GnRH-7 d-PGF2α-56 h-GnRH-16 to 20 h-timed artificial insemination; TAI) for synchronization of ovulation and TAI. For second and greater artificial insemination (AI), cows received AI after detection of estrus or the Ovsynch protocol (GnRH-7 d-PGF2α-56 h-GnRH-16 to 20 h-TAI) initiated 32 ± 3 d after AI for cows not re-inseminated at detected estrus. Cyclicity status (progesterone concentration), uterine health (vaginal discharge and uterine cytology), BCS, and systemic inflammation (haptoglobin concentration) were evaluated at baseline (33 ± 3 DIM for both treatments), beginning of the Double-Ovsynch protocol, and 10 d before TAI. Effects of treatments were assessed with multivariable statistical methods relevant for each outcome variable. Extending duration of VWP from 60 to 88 DIM increased pregnancies per AI (P/AI) to first service (VWP60 = 41%; VWP88 = 47%). Nonetheless, the greatest benefit of extending VWP on first-service P/AI was for primiparous cows (VWP60 = 46%; VWP88 = 55%), as P/AI did not differ within the multiparous cow group (VWP60 = 36%; VWP88 = 40%). Physiological status more conducive to pregnancy-characterized by improved uterine health, greater BCS, reduced systemic inflammation, and to a lesser extent more time to resume ovarian cyclicity-explained the increment in P/AI to first service. Our data also indicated that despite having greater P/AI to first service, cows with the longer VWP had delayed time to pregnancy during lactation (hazard ratio = 0.72; 95% confidence interval 0.69-0.98) and greater risk of leaving the herd, particularly for multiparous cows (hazard ratio = 1.34; 95% confidence interval 1.23-1.47). This shift in pregnancy timing led to an overall extension of the lactation length (+13 d), which resulted in greater total milk yield per lactation (+491 kg) but not greater milk yield per day of lactation. In conclusion, data from this experiment highlight the importance of considering the complex interactions between reproductive performance, herd exit dynamics, and lactation performance as well as the effects of parity at the time of defining the duration of the VWP for lactating dairy cows.

Hormonal profile and follicular dynamics concurrent with CIDR and insulin modified Ovsync TAI programs and their impacts on the fertility response in buffaloes

Fifty one cyclic Egyptian buffaloes were used to study the hormonal profile and follicular dynamics concurrent with CIDR and insulin modified Ovsync TAI programs and their impacts on the consequent fertility responses. The buffaloes were randomly assigned into 3 ovulation synchronization protocols: Ovsync-alone (n = 13, control) CIDR-sync (n = 20) and Insulin-sync (n = 18). Ovsync-alone protocol consisted of two im injections of 20 μg bueserlin (GnRHa) on Day 0 (GnRH 1) and on Day 9 (GnRH 2) with an im injection of 500 μg of cloprostenol sodium (PGF2 α) on Day 7. The CIDR-sync protocol consisted of the same treatment protocol as in Ovsync in addition to intra-vaginal insertion of CIDR (contains 1.38 gm of progesterone) on Day 0 followed by removal on Day 7. The Insulin-sync protocol consisted of the same treatment protocol as in Ovsync plus 3 sc injections of insulin at a dose of 0.25 i.u/1 kg, on Days 7, 8, and 9. Buffaloes in all groups were inseminated 16 h after GnRH2 by the same inseminator using frozen semen in straws. Blood samples were collected on Days 0, 3, 5 for serum progesterone assay and on Day 9 to measure serum concentrations of estradiol, insulin and IGF-1. Transrectal ultrasonographic scanning of the ovaries was conducted on Days 7, 8 and 9 to record the diameter of the largest follicle. Pregnancy diagnosis was conducted on Day 30 post-TAI by trans-rectal ultrasonographic scanning of the uterus to calculate conception rate. The serum progesterone concentration showed an increase (p < 0.01) in pregnant compared with non-pregnant buffaloes in both Ovsync-alone and Insulin-sync groups, but not in CIDR-sync group (p > 0.05) on Days 3 and 5. The serum estradiol concentration on Day 9 showed an increase (p < 0.01) in pregnant compared with the non-pregnant buffaloes in all of the treated groups. In Insulin-sync and Ovsync-alone groups, the diameter of the largest follicle (LF) was larger (p < 0.01) in pregnant compared with non-pregnant buffaloes, but in CIDR-sync, the diameter of the (LF) was larger (p < 0.01) in non-pregnant compared with pregnant buffaloes. Also, the results showed that the greatest diameter of LF was observed in pregnant buffaloes in Insulin-sync compared with either pregnant or non-pregnant buffaloes in all groups.It is concluded that modified CIDR-sync and Insulin-sync could improve fertility response through modulating hormonal profile and follicular dynamics in buffaloes during low breeding season.

Impact of days in milk at the initiation of ovulation synchronization protocols on the efficiency of first AI in multiparous Holstein cows

The objective of this study was to evaluate the impact of days in milk (DIM) at the initiation of ovulation synchronization protocols [spontaneous estrus (n=899), Presynch (n=1277), and CIDRsynch (n=978)] on fertility indices of the first AI in multiparous Holstein cows. The days from calving to the initiation of timed-AI (TAI) were categorized into: <50days (D1), 51–65days (D2) and 66–80days (D3). Pregnancy rate (P/AI at day 75) of cows at D2 period was significantly greater than that in the D1 period [OR (odds ratio)=1.26; P=0.050). Additionally, embryonic loss rate was significantly reduced in D2 and D3 groups compared to the reference group (OR=0.68 and 0.63; P=0.012 and 0.001, respectively). Conception (P/AI at day 28) and pregnancy rates in the Presynch group were significantly increased from 26.5 and 19.3% at D1 period to 35.9 and 28.9% at D2 period, respectively (OR=1.59 and 1.72; P=0.009 and 0.001, respectively). In contrast, conception and pregnancy rates using the CIDRsynch protocol did not differ at the different DIM at the initiation of TAI. The results of the Cox regression model reported significant associations for parity and season of calving with the hazard of embryonic loss (P=0.029 and 0.036, respectively). The current results suggest that using the Presynch regimen may achieve satisfactory conception and pregnancy rates at 51–65days in milk; however, the use of CIDRsynch regimen may provide a stable and homogenous conception and pregnancy rates at the different DIM at initiation of TAI.

Injectable progesterone in timed artificial insemination programs in beef cows.

The aims of this study were I) to compare the follicular diameter, corpus luteum diameter and serum progesterone (P4) concentrations in cows treated with conventional protocol vs. injectable P4 protocol; II) to determine the serum P4 profile in ovariectomized heifers; and III) to compare pregnancy rate between protocols. In experiment I, multiparous cows received a protocol for ovulation synchronization with an intravaginal P4 device (n = 38; device + EB day 0; device removal + PGF2α + eCG + EC day 8) or injectable P4 (n = 38; injection + EB day 0; PGF2α + eCG + EC day 8). In experiment II, ovariectomized heifers (n = 8) were treated with injectable P4 and blood samples were collected to determine the serum P4 profile. In experiment III, multiparous cows were timed AI with two different P4 approaches, intravaginal P4 device (n = 48) or injectable P4 (n = 47). In the first experiment, cows treated with P4 device had higher (P < 0.05) diameter of dominant follicle after ovulation induction (11.6 ± 1.8 vs.10.3 ± 1.8 mm) and ovulation rate (97%, 37/38 vs. 47.3%, 18/38) than cows treated with injectable P4. But, the follicular growth daily was higher (P < 0.05) in cows treated with injectable P4 than intravaginal device (1.3 ± 0.4 vs. 1.0 ± 0.3 mm/day, respectively). In experiment II, the P4 concentration peak occurred within 48 hours (6.54 ng/mL) and decreased after 96 hours (P < 0.05) after P4 injection. In experiment III, cows with P4 device had higher (P < 0.05) pregnancy rate than the injectable P4 group (60.4 vs. 34.0%, respectively). These results demonstrate that although the intravaginal P4 devices showed a higher pregnancy rate, a protocol with injectable P4 represents an easier method and a promising alternative for TAI in cattle.

Efeito da prostaglandina F2α no início do protocolo de sincronização da ovulação em novilhas leiteiras Bos indicus, Bos taurus e Bos indicus x Bos taurus

The objective of the present study was to evaluate the effect of treatment with prostaglandin F2α (PGF2α) at the beginning of the protocol for ovulation synchronization on follicular dynamics in Bos indicus (Gyr; n=11), Bos taurus (Holstein Black and White, HBW; n=10), and crossbred animals (Gyr x HBW; n=12). On a random day of the estrous cycles (day 0, D0), the animals received 2.0 mg estradiol benzoate (EB) intramuscularly plus an intravaginal progesterone (P4) device, which was maintained for 8 days (day 8, D8). Half the heifers of each group received a dose of 25 mg PGF2α intramuscularly at the time of insertion of the intravaginal P4 device. When the intravaginal device was removed (D8), all animals received another dose of 25 mg PGF2α intramuscularly, followed by intramuscular injection of 1.0 mg EB 24 h later (day 9, D9). Ultrasonographic evaluations were performed at intervals of 24 hours from D0 to D8 and at intervals of 12 hours from removal of the P4 device to 96 hours thereafter. Samples were collected on days 0, 3, 6, 8, 10 and 22 for the measurement of P4. The mean maximum diameter of the dominant follicle (DF) was smaller (P=0.01) in Gyr heifers (10.0 ± 0.8 mm) than in Gyr x HBW (13.0 ± 0.6 mm) or HBW (12.5 ± 0.8 mm). Furthermore, treatment with PGF2α on D0 increased (P=0.02) the maximum diameter of DF (12.9 ± 0.5 vs. 10.9 ± 0.7 mm) in the three genetic groups evaluated. The growth rate of the DF was lower (P=0.008) in Gyr heifers (0.8 ± 0.1 mm/day) than in crossbred heifers (1.3 ± 0.1 mm/day) or HBW (1.2 ± 0.1 mm/day). Heifers of the three genetic groups treated with PGF2α on D0 had higher (P=0.002) growth rates of DF (1.3 ± 0.1 vs. 0.9 ± 0.1 mm/day). The ovulation rate was lower (P=0.02) in Gyr heifers (36.4%) than in crossbred heifers (83.3%), but did not differ from HBW heifers (70.0%). However, higher (P<0.01) ovulation rates (93.7% vs. 35.3%) were observed in heifers treated with PGF2α on D0. The circulating concentration of P4 on removal of the intravaginal device (D8) was higher (P<0.05) in Gyr heifers (3.8 ± 2.2 ng/ml) than in crossbred heifers (2.2 ± 1.2 ng/ml), but no significant contrast was observed in HBW heifers (2.4 ± 1.6 ng/ml). Throughout the treatment period, the circulating concentrations of P4 were lower (P< 0.05) in heifers treated with PGF2α compared to untreated animals. Gyr heifers exhibited a lower growth rate and smaller maximum diameter of DF and a tendency towards a lower ovulation rate compared to HBW heifers. Treatment with PGF2α on the day of insertion of the intravaginal P4 device reduces circulating P4 concentrations and increases the growth rate and maximum diameter of DF, as well as ovulation rate.