Injectable P4 Research Articles

The efficiency of injectable P4 combined with the male effect on the pregnancy rate of boer goats in the off-season

Goats are seasonally polyestrous, which implies that the kid production is restricted to once a year. However, through estrous cycle manipulation, the production can be distributed throughout the year. Therefore, the present study aimed to estimate the reproductive viability of Boer female goats after administration of injectable P4, either combined with the male effect or independently, during the off-season in the southern region of Paraná. The outcomes evaluated in the study included the pregnancy rate, parturition rate, twin births, number of births, and concentration of births. The study was conducted in a beef goat farm, using 72 goats randomly assigned to three experimental groups. In the G1 group (n=22), females were stimulated by the male effect, in the G2 group (n=25), the animals received 75 mg of P4 intramuscularly at the beginning of the experiment, and in the G3 group (n=25), the goats received 75 mg of P4 intramuscularly at the beginning of the experiment combined with the male effect. The goats were in a breeding season of 60 days. The pregnancy rate, parturition rate, percentage of twin births, and the number of kids born per group were evaluated. The use of injectable P4 showed a similar pregnancy rate to the other groups, with a higher concentration of births in the G1 and G3 groups, both of which were subjected to the male effect (p<0.05). Furthermore, more studies should be carried out using different P4 concentrations.

Treatment alternatives to induce follicular wave emergence for timed-AI in lactating dairy Cows

Two experiments evaluated the effect of different hormonal treatments to synchronize follicle wave emergence on follicle dynamics and pregnancies per AI (P/AI) in estradiol (E2)/progesterone (P4) timed-AI (TAI) protocols in lactating dairy cows. In Experiment 1, lactating, primiparous Holstein cows (n = 36) received a P4 releasing device (Day 0) and were allocated at random to one of the following three treatment groups: Group EB received 2 mg E2 benzoate (EB) intramuscularly (i.m.), Group EB + GnRH received 2 mg EB+20 μg buserelin (GnRH) i.m., or Group EB + P4 received 2 mg EB + 100 mg of injectable P4 (iP4) in oil i.m. All cows received 0.150 mg D-Cloprostenol on Days 7 and 8 followed by P4 device removal, 400 IU eCG and 1 mg ECP on Day 8. Daily ultrasound examinations revealed that although the interval from P4 device removal to ovulation was not affected by treatment, cows that received EB + GnRH had an earlier (P < 0.05) emergence of the new follicular wave (Day 2.6 ± 0.2) than the other two treatment groups (Days 3.5 ± 0.3 and 6.1 ± 0.3, for EB and EB + P4, respectively). In Experiment 2, 808 lactating cows were assigned randomly to the three treatments evaluated in Experiment 1, and all the cows were TAI to determine P/AI. Cows in the EB + GnRH group had greater P/AI (57.4 %, P < 0.01) than those in the EB (44.6 %) or EB + P4 (45.7 %) groups. In conclusion, the administration of GnRH, but not iP4, on the day of insertion of a P4 device improves P/AI in lactating dairy cows synchronized for TAI with an estradiol/P4-based protocol.

Efeito da suplementação com progesterona injetável de longa ação, sete dias após inseminação, sobre o desenvolvimento embrionário e fetal de fêmeas Nelore

ABSTRACT This study evaluated the effects of long-acting injectable progesterone supplementation (P4LA) on embryonic and fetal development and birth weight in Nellore cows seven days after timed artificial insemination (TAI). One hundred and nineteen Nellore female cows became pregnant after the TAI protocol and were divided into two groups: P4LA with cows that received 150mg of P4LA, seven days after insemination, in a single dose, and the control group, with cows who did not receive any hormonal supplementation after TAI. Blood samples were collected on days 17 and 30 after TAI to determine P4 concentration. Embryonic and fetal measurements were performed at 30 and 45 days of gestation, respectively, with the aid of ultrasonography, measuring craniocaudal length and thoracic diameter. No difference was observed in P4 concentration between the groups supplemented with or without injectable P4 on days 17 (P=0.73) and 30 (P=0.62) after TAI. There was no significant difference in embryonic and fetal development or birth weight between the supplemented and non-supplemented groups (P=0.59, P=0.09, and P=0.64, respectively). Supplementation with injectable progesterone seven days after TAI did not interfere with the embryonic and fetal development of Nellore cows, nor did it affect birth weight.

Evolution over the last 40 years of the assisted reproduction technologies in cattle - the Brazilian perspective I - timed artificial insemination.

The reproductive efficiency of the herd is correlated with higher productivity in livestock. Reproduction biotechniques, such as ovulation synchronization protocols, are important to optimize production and accelerate genetic profit in beef and dairy herds. The objective of this review is to describe the evolution over the last 40 years of the artificial insemination (AI) and the timed-AI (TAI) protocols in cattle from a Brazilian perspective. TAI protocols are based on synchronizing emergence of the wave of follicular growth, controlling circulating progesterone (P4) concentrations, stimulating the final growth of the follicle and inducing a synchronized ovulation. Hormonal alternatives that optimize the response at the end of the protocol and strategies to induce final follicle growth and ovulation in categories of females with low expression of estrus are described. Furthermore, the potential positive effect of previous exposure to injectable P4 on fertility of Bos indicus and Bos taurus cows is also discussed.

Effects of injectable and intravaginal progesterone on ewes' reproductive performance at breeding season beginning.

Progesterone (P4) is a steroid hormone that has a regulatory role in the female reproductive system. Studies on the effects of injectable progesterone on ewes are scarce, mainly related to their reproductive responses in the breeding season. This study aimed to compare reproductive performance and serum P4 concentration using injectable or intravaginal P4 in ewes. Two hundred and forty and eight Santa Inês x Dorper ewes (BW; 52.67 ± 11.76 kg; mean ± SE), body condition score (BCS; 2.5 ± 0.8; scale of 1-5), were distributed in four treatments: (i) Control: without administration of P4; (ii) CIDR: intravaginal implantation of 330 mg of P4 for 7 days; (iii) 1P4: 15 mg of P4 intramuscular (IM); and (iv) 2P4I: 30 mg of P4 IM. The first 18 days of breeding season were considered the synchronization period. Except for ewes in the control group, all other ewes received 263 μg IM of cloprostenol sodium for lysis of eventual CL at 24 h before the P4 treatment. After the synchronization period, all ewes were kept together with males for extra 28 days in the breeding season. At the beginning of breeding season, 90% of the ewes had serum P4 concentration less than 1 ng/mL. The estrus rate was greater (P < 0.01) in ewes on the CIDR treatment, with similar estrus rate among the other treatments. The P4 implant was able to keep blood P4 concentration greater than 1 ng/mL in ewes that received an implant of P4 during the 7 days. After implant removal, there was a great increase in the estrus manifestation on ewes in the CIDR treatment, leading to an increase in pregnancy rate at the beginning of breeding season. The current study demonstrated that ewes that received an intramuscular injection of 15 or 30 mg of P4 had similar reproductive performance than ewes that did not receive any P4 intramuscular injection. However, when ewes were implanted with P4 (CIDR), these animals had an increase in estrus manifestation, leading to greater pregnancy earlier during the breeding season.

Effect of increased serum progesterone on luteal morphology and function, pregnancy, and gestational loss in Nelore cows.

The aim of the present study was to determine the impact of different strategies for increasing the level of serum progesterone (P4) on luteal morphology and function in bovine females. The effects of increasing P4 on pregnancy rate and gestational loss (GL) in Nelore cows subjected to timed artificial insemination (TAI) were also evaluated. A total of 939 cows were divided into three groups: P4LA (n=305), 150 mg of long-acting injectable P4 7 days after TAI; GnRH (n=306), 10μg of buserelin acetate 7 days after TAI; and control (n=328), no hormone treatment after TAI. Doppler ultrasound assessments and P4 measurements were performed on days 7 and 16 after TAI. The pregnancy rate and GL as a function of treatment were compared using the SAS GLIMMIX procedure. Corpus luteum (CL) vascular perfusion, volume, and plasma P4 concentration were analysed using the SAS PROC MIXED procedure. No significant difference was found among the treatments in terms of volume, number of pixels, and CL intensity or in the serum P4 concentration at 7 days after ovulation. The CL blood flow at 16 days after ovulation was lower in the P4LA and GnRH groups than that in the control group (p < .01). Serum concentrations of P4 at 16 days after ovulation were higher in the P4LA and GnRH groups than those in the control group (p=.04). A difference in the pregnancy rate (p=.003) and a trend in GL (p=.07) as a function of treatment were found. Overall, long-acting injectable P4 supplementation on day 7 after TAI or GnRH administration affected CL vascularization and increased the serum concentrations of P4 16 days after ovulation, promoting better pregnancy rates than the control.

Induction of puberty in Bos indicus heifers in the western Amazon region

The objective of this study was to evaluate the efficiency and cost of two distinct hormonal protocols for induction of puberty in Bos indicus heifers in the western Amazon region. Nelore prepubertal heifers (n = 127) aged 20 to 24 months (21 ± 0.3 months) and weighing 300 to 340 kg (318 ± 7 kg) were submitted to two distinct protocols for induction of cyclicity. Females were previously submitted to two ovarian ultrasound examinations (12-day interval) to verify the absence of the corpus luteum (CL). Then, the heifers were divided into two experimental groups: G-CIDR/EC (n = 54) and G-P4/EC (n = 73). The G-CIDR/EC group utilized a CIDR® for 12 days and on the day of its withdrawal, 0.6 mg of estradiol cypionate (IM) was administered. The G-P4/EC group received 150 mg of injectable progesterone (IM) and one injection of 0.6 mg EC 12 days after the progesterone injection. Twelve days after the end of the hormonal protocols, the heifers were submitted to another ultrasound evaluation for identification of cyclicity (ovulatory follicle or CL). These females were then submitted to a conventional protocol of synchronization of ovulation for timed artificial insemination (TAI). Only heifers responsive to puberty induction (G-CIDR/EC (n=30) and G-P4/EC (n=51)) were included in the TAI protocol. After seven days of insemination, both groups were exposed to natural breeding. In addition, an economic analysis was performed to evaluate cyclicity induction, the TAI protocol, and pregnancy in both groups. The G-CIDR/EC group showed 81.48% (44/54) of females responsive to induction of cyclicity (presence of CL) while the G-P4/EC group obtained 86.3% (63/73; p=0.463). Regarding the conception rate to TAI and to bull (natural breeding), the G-CIDR/EC group displayed 43.33% (13/30) and 33.33% (10/30), respectively, and the G-P4/EC group exhibited 54.9% (28/51) and 39.22% (20/51), respectively. The overall pregnancy rate (TAI+Bull) was 76.67% (23/30) in the G-CIDR/EC group compared to 94.12% (48/51; P=0.023) in the G-P4/EC group. The cost of the hormonal protocol to cyclicity induction resulted in an economy of 42.8% for injectable P4 compared to the intravaginal P4 group. In addition, the final cost of cyclic induction associated with TAI was 34% lower in the G-P4/EC group compared to the G-CIDR/EC group. We concluded that both cycling induction protocols were efficient in Bos indicus heifers from the western Amazon. However, injectable P4 provided a higher conception rate and lower pregnancy cost at the end of the reproductive season.

Development of a novel 21-day reinsemination program, ReBreed21, in Bos indicus heifers

The aim was to develop a program for resynchronization of ovulation (ReBreed21) that allowed reinsemination of non-pregnant Bos indicus heifers every 21 d using timed AI (TAI) without the need for detection of estrus. The Rebreed21 program begins 12 d after previous TAI (Day 0) by inserting an intravaginal P4 implant (Day 12) that is removed 7 d later (Day 19) combined with treatment with 300 IU of eCG. On Day 21, early pregnancy diagnosis (Doppler PD) is performed based on CL vascularity. Non-pregnant (NP) heifers immediately received AI combined with 100 μg of GnRH. The program is replicated 12 d after second TAI to produce a breeding season (BS) of 42 d with 3 potential TAIs. Two experiments were conducted as a proof of concept for this rapid rebreeding program. In Experiment 1, 76 heifers were enrolled in ReBreed21, as explained above. In Experiment 2, 300 Nellore heifers were synchronized for 1st TAI and randomly assigned to one of two groups: ReBreed21 (n = 147) or another early resynchronization procedure, Resynch14 (n = 153) with P4 implant inserted 14 d after previous TAI plus 50 mg of long-acting injectable P4; 8 d later P4 implant removed (Day 22) and early Doppler PD performed; NP heifers received 150 μg of cloprostenol, 0.5 mg of ECP, and 300 IU of eCG with TAI on Day 24. In both experiments, the largest follicle (LF) was measured at each Resynch TAI. Ultrasound was later used to confirm the early Doppler PD and to determine ovulation (OV) to Resynch at 12 d after TAI in ReBreed21 (Day 33 of pregnancy) and 14 d after TAI in Resynch14 (Day 38 of pregnancy). Final PD was performed 40 d after 3rd TAI. Results for Experiment 1 were: diameter of LF 11.8 ± 0.23 mm; 88.9% OV; 20.5% false positives; 38.1% P/AI at 1st TAI; 44.4% overall P/AI for ReBreed21 TAIs; 72.3% total pregnant at end of BS. In experiment 2, Rebreed21 vs. Resynch14 were different for: diameter of LF (10.9 ± 0.17 vs. 10.0 ± 0.17 mm, P = 0.0003), heifers with LF < 8.5 mm (10.2 vs. 26.4%, P = 0.04), or LF ≥ 11 mm (50.0 vs. 37.2%, P = 0.001), and P/AI at first TAI (29.3% [43/147] vs. 20.3% [31/153], P = 0.074) but similar for OV (overall 86.8% [239/275], P = 0.82), false positives (P = 0.52) overall P/AI for Resynch TAIs (33.6 vs. 28.8%, P = 0.4), and total pregnant at end of BS (58.5% [86/147] vs. 55.6% [85/153], P = 0.64). In addition, median time to pregnancy was 9 d earlier (P = 0.0007) for ReBreed21 than Resynch14. Thus, ReBreed21 is a novel protocol that allows earlier reinseminations than Resynch14 but with similar fertility.

Influence of Injectable Progesterone on the Pregnancy Rate of Heifers Receiving Bovine Embryos

Background: In vitro embryo production (IVEP) allows the spread of superior animal genetics, but pregnancy rates show a high variability with this biotechnique. In the initial stage of pregnancy, progesterone plays a fundamental role in uterine preparation, acting on embryonic growth, implantation, and development. However, on the day of the IVEP transfer to the recipients, progesterone levels may be lower than that expected, influencing the uterine environment and, consequently, the pregnancy rate. Therefore, the objective of this study was to evaluate the pregnancy rate in heifers after the administration of injectable progesterone (P4) in the fixed-time embryo transfer (FTET) protocol.Materials, Methods &amp; Results: The experiment was conducted inside a rural property near the city of Rio Branco, Acre, Brazil. The experimental group consisted of 232 animals, including 78 zebuine (Bos indicus) and 154 mixed (½ blood B. indicus and ½ blood B. taurus) animals, aged between 16 and 24 months, with a mean weight of 300 and 330 kg for zebuine and mixed animals, respectively. The selected animals were previously synchronized using the progesterone-estrogen-prostaglandin-estrogen protocol. Embryo transfer was performed on day 18 of the protocol, which was 9 days after the removal of intravaginal progesterone implant. On day 15 of the protocol, that is, 144 h (6 days) after the device removal, the animals were randomly distributed into two experimental groups: Control Group (CG; 0.5 mL of 0.9% saline solution, intramuscular) and Treated Group (P4G; 0.5 mL of injectable P4, 150 mg, intramuscular). Chi-square test was used for the statistical analysis of the pregnancy rate at a 5% probability. After 23 days of embryo transfer, pregnancy was diagnosed by ultrasonography. The general pregnancy rate, considering all groups (CG and P4G) and breeds included, was 55.17% (128/232). The pregnancy rates of the P4G and CG groups, regardless of breeds, were 55.08% (65/118) and 55.26% (63/114), respectively, with no statistical difference (P = 0.8344). Angus animals presented a similar pregnancy rate in the P4G and CG groups of 54.93% (39/71) and 57.83% (48/83), respectively. Furthermore, similar results were found for the Nellore breed, with no difference in pregnancy rate between the CG (55.81%, 24/43) and P4G (48.57%, 17/35) groups.Discussion: The strategy of using P4 to increase the pregnancy rate is very widespread in fixed-time artificial insemination (FTAI) protocols, although differences still exist. The literature presents zero or negative effects, as in this study, when exogenous P4 was used to increase endogenous P4 concentrations and, consequently, the pregnancy rate. However, some studies demonstrated the beneficial effects of increasing blood P4 concentrations, and that the increased fertility depends on the method and time of P4 supplementation and the animal’s physiological state. In this context, the fact that the injectable progesterone supplementation did not increase the pregnancy rate in this study is justified mainly by factors such as the animal category used (heifers), time of P4 supplementation (day 4 after ovulation), biotechnology used (FTET), and the heterogeneous characteristics related to phases of the estrous cycle of heifers. Supplementation with 150 mg of injectable long-acting progesterone intramuscularly did not interfere in the pregnancy rate of Angus and Nellore heifers receiving bovine embryos.

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.

Supplementation with long-acting progesterone in early diestrus in beef cattle: II. Relationships between follicle growth dynamics and luteolysis

The aims were to characterize follicular dynamics in response to supplemental progesterone (P4) and to investigate the relationships between follicular growth and onset of luteolysis in P4-treated cows, submitted or not to artificial insemination (AI). Nonsuckled beef cows detected in estrus were assigned to receive AI or to remain non-AI. Three days after ovulation (ie, D3), AI cows were injected with 150 mg of long-acting P4 (AI + injectable P4 formulation [iP4]; n = 22), and the non-AI cows were assigned to receive 150 mg iP4 (n = 19) or saline (control, n = 19). Between D3 and D21, growth dynamics of the dominant follicles (DFs) was monitored by ultrasonography. Plasma P4 concentrations were measured every other day from D9 to D19. Pregnancy status (ie, P: pregnant and NP: nonpregnant) was examined by ultrasound on D28 to D32. Injectable P4 formulation supplementation decreased average maximum diameter of first-wave DF (DF1). Neither day of emergence of DF2 or DF3 nor the proportion of two- or three-wave cycles were altered by supplemental P4. Daily mean diameter of DF2 and DF3 was also similar between control and iP4 groups. Consistently, daily mean diameter of DF1 in iP4-treated cows was smaller for cows that underwent luteolysis by D15 compared to a later onset. Progesterone concentrations between D9 and D19 decreased earliest in the iP4 group, latest in the control group and was intermediate for the NP-AI + iP4 group. In addition, three-wave cycles presented a delayed decrease on plasma P4 concentrations than two-wave cycles. Further analysis revealed that on two-wave cycles, P4 concentrations on D15 were lowest in the iP4 and NP-AI + iP4 animals compared to the control and P-AI + iP4 groups. Conversely, for three-wave cycles, on D15, P-AI + iP4, NP-AI + iP4, and controls had greater P4 concentrations than the iP4 group. In summary, our data indicate that impairment of first follicular growth was associated with P4-induced shortened luteal lifespan (D14–D15) and that three-wave cycles after AI can be more supportive for pregnancy maintenance in P4-treated cows. We speculate that such conditions play a critical role in the embryonic ability to inhibit iP4-induced early luteolysis reported in part I of this series.

Supplementation with long-acting progesterone in early diestrus in beef cattle: I. effect of artificial insemination on onset of luteolysis

Progesterone (P4) supplementation in early diestrus advances changes in the endometrial transcriptome, stimulating embryonic development. However, it also induces early onset of luteolysis. Occurrence of luteolysis before D16 postmating can be detrimental to fertility. A potential counteracting role of the elongating conceptus on early luteolysis is understood poorly. The aim of the study was to evaluate the effect of artificial insemination (AI; ie, pregnancy) on the temporal dynamics of luteolysis of cows supplemented with P4. Nonsuckled beef cows were inseminated at 12 h after estrus (D0: ovulation) or were not inseminated (no-AI). On D3, the AI cows were assigned to receive a single dose of 150 mg of injectable long-acting P4 via intramuscular injection (AI + iP4; n = 23), and the no-AI cows were assigned to receive iP4 (iP4; n = 21) or saline (control, n = 22). Corpus luteum (CL) development and regression were determined by ultrasonography (US) between D3 and D21. Plasma P4 concentrations were measured on D3 and every other day from D9 to D21. Pregnancy status was determined by US (D28‒D32). iP4 supplementation reduced luteal development (D5–D10) compared to the control group and increased incidence of luteolysis between D14 and D15. On D15, the proportion of cows that underwent luteolysis and plasma P4 concentrations differed between the iP4 group (47.6; 2.10 ± 0.47) and the control group (13.6; 4.40 ± 0.46) and was intermediate in the AI + iP4 group, respectively (26.1%; 3.70 ± 0.45 ng/mL; P < 0.05). The AI effects were due to the pregnant cows (n = 7). Considering nonpregnant cows only, the proportion of early luteolysis in the AI + iP4 group (37.5%) was similar to the iP4 group. Pregnancy was not established in cows having a shortened luteal lifespan. Indeed, interval to luteolysis in the AI + iP4 group (15.50 ± 0.66 d) was similar to the iP4 group (16.38 ± 0.46 d), but less than the control group (17.38 ± 0.40 d; P = 0.05). In conclusion, the effect of AI on extending luteal lifespan occurred exclusively in cows that maintained pregnancy.

8 Evaluation of puberty induction protocol in peripubertal beef heifers prior to fixed-time AI programs

Beef heifers that conceive early during their first breeding season calve earlier, wean heavier calves, and tend to continue doing so throughout the rest of their reproductive lives. Otherwise, under extensively managed systems in Southern Brazil, only a variable proportion of 2-year-old beef heifers reach puberty before the start of the breeding season. The aim of this study was to evaluate a protocol to induce puberty in peripubertal beef heifers using injectable progesterone (P4) and oestradiol cypionate (EC) to improve pregnancy per AI (P/AI) in fixed-time AI (FTAI) programs. A total of 311 Angus heifers, 24±3 months of age, 3.1±0.3 body condition score (1=emaciated; 5=obese), and 328±41kg of body weight (BW), were examined before FTAI protocol (Day −24) by transrectal ultrasonography to determine reproductive tract score (RTS; 1=prepubertal; 5=pubertal). Prepubertal heifers (RTS 1; 12.8%, 40/311) were excluded from the study. Heifers with RTS=2 (n=271) were assigned to 2 groups: P4 (RTS 2 or 3, n=100) and cyclic (RTS 4 and 5, n=171). Peripubertal heifers were treated on Day −24 with 150mg of injectable P4 IM (Sincrogest® injectable, Ourofino Animal Health, Cravinhos, Brazil) and on Day −12 received 1mg of EC IM (Sincro CP®, Ourofino Animal Health). Cyclic heifers received no hormonal treatment before FTAI protocol. On Day 0, heifers in the P4 group were evaluated again to determine RTS, body condition score, and BW. Induction of puberty was defined by the presence of a follicle &amp;gt;8.5mm (RTS=4) or a corpus luteum (RTS=5). All heifers received an FTAI protocol consisting of 2mg of oestradiol benzoate IM (Sincrodiol®, Ourofino Animal Health) and an intravaginal progesterone implant (1 g; Sincrogest®, Ourofino Animal Health). On Day 8, heifers had implants removed and were injected with 1mg of EC and 500µg of sodium cloprostenol IM (Sincrocio®, Ourofino Animal Health). After 48h, all heifers received an FTAI. Thirty days after AI, the diagnosis of gestation was performed by ultrasonography. Statistics included chi-square and linear regression with coefficients analysed as independent variables, considering a significant P-value of 0.05. On Day 0, percentages of RTS 5, 4, and 3 in the P4 heifers were 37, 27, and 36%, respectively, resulting in 64% of puberty induction. The P/AI was similar (P=0.14) in P4-treated and cyclic heifers (59v. 50.3%, respectively). The regression analysis presented a linear and positive effect of BW on P/AI (P&amp;lt;0.001) in cyclic heifers and a positive influence of BW on RTS (P=0.05) and RTS on P/AI (P=0.008) in P4-treated heifers. The application of RTS system associated with P4 treatment of peripubertal heifers before FTAI protocol improved ovarian activity and might have contributed to higher P/AI, comparable with that achieved in cyclic heifers.

Exacerbated conceptus signaling does not favor establishment of pregnancy in beef cattle

BackgroundInsufficient production of anti-luteolytic signals by the pre-attachment embryo is considered a major cause of pregnancy failure in cattle. We tested the hypothesis that transfer of multiple blastocysts (n = 5/recipient) and progesterone (P4) supplementation amplify anti-luteolytic signaling and reduce embryonic losses in beef cattle. Cows detected in estrus (D0; n = 104) were assigned randomly to receive 150 mg of injectable long-acting P4 (iP4) or vehicle (non-iP4) on D4 and transcervical transfer of none or five, grade 1, not-frozen, in vitro-produced blastocysts, on D7. Luteal development and time of structural luteolysis were monitored by ultrasonography. Plasma P4 concentrations were determined on D4, D5 and D7, and daily between D14 and D20. Conceptus signaling was monitored by transcript abundance of interferon-stimulated gene 15 (ISG15) in peripheral blood mononuclear cells isolated on D14, D16, D18 and D20. Early embryonic mortality (EEM) was defined as the absence of ISG15 mRNA upregulation over time and/or luteal regression up to D20. Late embryonic mortality (LEM) was defined as the absence of a conceptus with a heartbeat on pregnancy diagnosis at D30 (PD30) after observing upregulation of ISG15 mRNA and extension of luteal lifespan. Pregnant cows presented conceptuses with heartbeat at PD30.ResultsOn D5, iP4-treated cows had P4 concentrations 2.07-fold greater than non-iP4 treated (P < 0.001). On D7, P4 concentrations were similar. Pregnant and LEM animals showed a progressive increase in the abundance of ISG15 from D14 to D20. iP4-treated cows detected pregnant at PD30 had 1.53-fold greater abundance of ISG15 mRNA between D14 and D20 than non-iP4 treated cows (P = 0.05). iP4 doubled the frequency of EEM while it did not affect LEM. At PD30, embryonic survival was 37.0% vs. 55.6% for iP4-treated vs. control cows. Majority of pregnant cows (71%) presented only a single viable embryo.ConclusionsA substantial proportion of cows had EEM (31%) and LEM (20%) even after transferring multiple blastocysts. This argues that mortality was due to poor uterine receptivity that could not be reversed by supplemental P4 or overcome by transferring multiple blastocysts. Further, a given uterine environment was not necessarily adequate to all embryos.

Impact of estradiol cypionate prior to TAI and progesterone supplementation at initial diestrus on ovarian and fertility responses in beef cows

In cattle, early diestrus progesterone (P4) supplementation modulates endometrial function to exert pro- and anti-pregnancy establishment effects; specifically, P4 stimulates conceptus growth, but also induces early onset of luteolysis. This paradoxical effect is frequently related to the inconsistent fertility outcomes that result from P4 supplementation experiments. Aim was to investigate the impact of exogenous estradiol (E2) treatment at the end of timed fixed AI (TAI) on frequency of early luteolysis and pregnancy of beef cows supplemented with P4. Ovulations (D0 of study) of suckled multiparous (n = 643) and primiparous (n = 193) Nelore cows (Bos indicus) were synchronized with an E2/P4-based protocol for TAI and assigned to receive 1.0 mg of estradiol cypionate (CP) or nothing (NoCP) on D–2 and 150 mg of injectable long-acting P4 (iP4) or Placebo (NoiP4) on D4 on a 2 × 2 factorial arrangement. On D15, the iP4 supplementation increased (P < 0.05) the frequency of early luteolysis (NoCP + iP4: 26.0%; [13/50] vs. NoCP: 8.0% [4/50]), but CP prevented this effect (CP + iP4: 8.3% [4/48] and CP: 6.4% [3/47]). The CP improved pregnancy/AI (P/AI) of multiparous (CP: 51.6% [165/320] and NoCP: 35.0% [113/323]; P < 0.001) and primiparous cows (CP: 40.4% [40/99] and NoCP: 24.5% [23/94], P < 0.05), regardless of iP4 treatment. The iP4 supplementation affected P/AI of CP and NoCP treated cows according to follicle size at TAI. For the CP treated cows, the iP4 supplementation improved P/AI of sub-populations of cows with follicles <12.35 mm (42.0% [34/81] vs. 53.1% [34/64]), while for NoCP treated cows, the improvements occurred in subpopulations of cows with follicles ≥12.35 mm (46.1% [35/76] vs. 58.7% [37/63]). In conclusion, strategies associating E2 and P4 supplementation decrease the incidence of early onset of luteolysis and improve P/AI of suckled beef cows with smaller follicles.

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.

Evaluation of pregnancy rates of Bos indicus cows subjected to different synchronization ovulation protocols using injectable progesterone or an intravaginal device

This study evaluated the pregnancy rate in Nelore cows (Bos indicus) that were subjected to fixed-time artificial insemination (FTAI) using different protocols consisting of injectable progesterone (P4) or an intravaginal device (impregnated with P4). Multiparous cows 72-84 months in age, 30-45 days postpartum, were selected on the basis of the absence of a corpus luteum (CL) and follicles &lt; 8 mm after transrectal palpation and ultrasound examinations. On a random day of the estrus cycle (D0), the selected animals (n = 135) were randomly assigned to one of three experimental groups (n = 45 each). Group I (injectable P4/FTAI 36 hours) received 250 mg of injectable P4 and 2 mg EB on D0; on D7, they received 500 µg of cloprostenol; on D8, 300 IU of eCG and 1 mg of EB were administered; and finally, FTAI was performed 36 hours after the application of EB. Group II (injectable P4/FTAI 48 hours) received the same protocol as Group I, except that the FTAI was performed 48 hours after ovulation induction. The animals of Group III (Control/CIDR) received a conventional protocol for FTAI using an intravaginal device (D0: P4 and 2 mg EB; D8: device removal, 500 µg cloprostenol, 300 IU eCG, 1 mg EB; and FTAI performed 48 hours after removal of the device). The results showed that cows synchronized with the conventional protocol for FTAI (Control/CIDR) had a higher pregnancy rate (60 %, 27/45) than those synchronized with an injectable P4/FTAI 36 hours (33.33 %; 15/45, P = 0.010). However, the group receiving injectable P4 group/FTAI 48 hours had a similar pregnancy rate (48.9 %; 22/45; P = 0.290) when compared to both the group receiving the conventional protocol and that receiving injectable P4/FTAI 36 hours (P = 0.134). Although the injectable P4 may affect pregnancy rate with the FTAI performed in 36 hours, we found similar pregnancy rates from cows inseminated 48 hours after induction ovulation, considering injectable or intravaginal P4. Therefore, we suggest that injectable P4 represents an alternative source of progesterone for synchronization of cattle for FTAI.

Pregnancy rate evaluation in lactating and non-lactating Nelore cows subjected to fixed-time artificial insemination using injectable progesterone

Most fixed-time artificial insemination (FTAI) protocols utilize progesterone (P4) as a hormonal source to achieve synchronization of estrus in cattle. The use of an injectable P4 source to control estrus would be an interesting pharmacological strategy owing to the practicality of parenteral application. However, the effects of injectable P4 on estrus cycle control in cattle remain poorly studied. In particular, no existing studies have investigated the effect of injectable P4 on the fertility of cows subjected to FTAI. The aim of this study was to evaluate the pregnancy rate of lactating and non-lactating Nelore cows subjected to FTAI with injectable P4. Of the 422 non-lactating cows in this study, 162 (38.3%) became pregnant by 60 days post-FTAI. In the lactating group (n = 516), 166 (32.1%) were pregnant by 60 days after treatment with injectable P4. The proportions of lactating and non-lactating cows becoming pregnant were compared using the chi-square test, adopting a significance level of P &amp;lt; 0.05. It was found that the pregnancy rate of the cows subjected to FTAI with injectable P4 was influenced by lactation status. Lactating cows had lower reproductive performance, possibly because of their higher nutritional requirements. However, the use of injectable P4 shows promising results and may prove to be a useful strategy in large-scale livestock production.

Ovarian follicular dynamics of Nelore (Bos indicus) cows subjected to a fixed-time artificial insemination protocol with injectable progesterone

This study aims to evaluate ovarian follicular dynamics of Nelore cows subjected to fixed-time artificial insemination (FTAI) with injectable progesterone (P4) or intravaginal device. In the device group, 20 cows received an intravaginal device with 1 g P4 and administration of 2 mg estradiol benzoate (EB) intramuscularly (im) on D0. The device was removed on D8, followed by administration of 500 ?g cloprostenol, 300 IU equine chorionic gonadotropin (eCG) and 1 mg estradiol cypionate (EC), im. The cows were inseminated 48h later. In the injectable group, 350 mg P4 (300 mg subcutaneously and 50 mg im) and 2 mg EB im were administered on D0 to 16 cows. On D6, 500 ?g cloprostenol, 300 IU eCG and 1 mg EC were administered im. The FTAI was performed 48h later. Sixty-five percent of cows from the device group (13/20) and 75% from the injectable group (12/16) showed follicles ? 5 mm on D4 (p>0.05). The average rate of follicular growth was 0.92±0.25 mm/day for the device group and 1.06±0.45 mm/day for the injectable group (p>0.05). The average diameter of the dominant follicle (DF) at FTAI was 11.4±1.4 mm for the device group and 10.8±1.8 mm for the injectable group (p>0.05). In the device group, ovulatory follicles (OF) had an average diameter of 12.1±1.4 mm, and in the injectable group 12.3±1.6 mm (p>0.05). Ninety percent of the device group (18/20) and 37.5% of the injectable group (6/16) demonstrated ovulation (p<0.05). The average diameter of the corpus luteum (CL) was 18.7±4.3 mm for the device group and 17.7±3.6 mm for the injectable group. The injectable P4 showed similarity in the presence of follicles ? 5 mm on D4, follicular growth rate, diameters of the DF, OF and CL. However, the ovulation rate was higher in the device group.

180EFFECT OF TIME OF ECG TREATMENT ON PREGNANCY RATES IN BOS INDICUS×BOS TAURUS RECIPIENTS SYNCHRONIZED WITH PROGESTERONE VAGINAL DEVICES AND TRANSFERRED WITHOUT ESTRUS DETECTION

It has been shown recently that treatments with progesterone (P4)-releasing devices combined with estradiol benzoate (EB) plus P4 on Day 0, eCG and PGF on Day 5 and a second application of EB one day after device removal (Day 9) can be used successfully to transfer bovine embryos at a self-appointed time, without the necessity of estrus detection. Although the treatment solved one of the major problems in recipient management, estrus detection, it requires handling the recipients at least five times for treatments and embryo transfer. An experiment was designed to evaluate whether reducing one day of handling, by the administration of eCG and PGF at the time of removal of the P4 device (Day 8), results in comparable pregnancy rates than giving eCG on Day 5. A secondary objective was to determine the effect of injectable P4 at the time of device insertion plus EB treatment. Crossbred Bos taurus×Bos indicus beef heifers (n=301) were randomly assigned to 4 treatment groups in a 2 by 2 factorial design. All Heifers received a P4 device (DIB, Syntex, Argentina) plus 2mg EB i.m. (Syntex) at unknown stages of the estrous cycle (Day 0), with or without 50mg of P4 given i.m. at the same time. Heifers were further subdivided to receive PGF (0.150mg d-cloprostenol, Prolise, Tecnopec, Sao Paulo, Brazil) and 400IU of eCG (Novormon, Syntex) i.m. on Days 5 or 8. In all heifers, DIB devices were removed on Day 8 and 1 mg EB was administered i.m. on Day 9. Day 10 was arbitrarily considered as the day of estrus. On Day 17, heifers were bled for plasma P4 concentrations and examined by ultrasonography to determine the number of CL and their diameter. Heifers that had &amp;gt;1 CL or a single CL with diameter ≥18mm received an in vitro-produced (IVP) embryo by nonsurgical transfer performed by the same veterinarian. Pregnancy rates were determined by ultrasonography 30 days later. The effects of Day of eCG administration (Day 5 or Day 8), P4 of treatment (E2 or E2+P4) and the day-by-P4 treatment interaction on the numbers of CL and plasma P4 were analyzed by ANOVA, and the proportion of recipients selected and pregnant were analyzed using non-parametric tests (NPAR1WAY, SAS). There was no significant effect of P4 treatment or the P4-by-day of eCG interaction in any of the parameters evaluated. However, there was a significant effect of day of eCG administration on plasma P4 concentrations (Day 5=2.4±0.3 v. Day 8=1.7±0.2; P=0.03) and the number of CL (Day 5=1.4±0.1 v. Day 8=1.1±0.0; P=0.02) on Day 17. Furthermore, the proportion of recipients pregnant/treated tended (P=0.1) to be higher in heifers in the Day 5 Group (71/151, 47.0%) than in those in the Day 8 Group (61/150, 40.7%). Although delaying the eCG and PGF administration from Day 5 to Day 8 saves one trip through the chute for treatments, it resulted in lower plasma P4 concentrations and tended to decrease pregnancy rates in bovine embryo recipients synchronized with DIB devices and EB and transferred at a fixed time. Furthermore, the administration of injectable P4 at the time of DIB insertion did not affect pregnancy rates.