Temporal evaluation of follicular dynamics and endocrine patterns of Holstein (Bos taurus), Gir (Bos indicus), and Murrah (Bubalus bubalis) heifers kept under the same nutritional, management and environmental conditions.

The objective of the study was to simultaneously compare ovarian follicular dynamics and endocrine parameters of taurine (Holstein; n = 14), zebuine (Gir; n = 5), and bubaline (Murrah; n = 15) heifers kept under the same environmental, nutritional and management conditions. Heifers were synchronized with two PGF treatments 14 days apart. Ovaries of cyclic heifers were scanned daily during two consecutive ovulations and blood samples were collected every 24 h from each animal. No significant difference was found for length of interovulatory interval, however, zebuine heifers presented a greater number of follicular waves, number of antral follicles on day of ovulation, and higher insulin concentration than the other two breeds. Taurine heifers had highest maximal diameter of first wave dominant and ovulatory follicles and CL volume. Taurine and bubaline heifer's dominant follicle of first wave had longer static and regression phases than zebuine heifers. Bubaline heifers presented overall lowest progesterone concentrations and CL volume, but higher IGF1 levels. No difference was observed between taurine and zebuine heifers regarding IGF1 concentration. Despite higher CL volume found in taurine heifers, no difference in mean progesterone concentration was observed between them and zebuine heifers. Insulin and IGF1 concentrations were greater in follicular phase than in luteal phase when breed was not evaluated. After evaluating the three breeds simultaneously, at the same nutritional and management status it is possible to conclude that each genetic group has a specific follicular development and endocrinology of the estrous cycle.

Synchronization treatments previous to natural breeding anticipate and improve the pregnancy rate of postpartum primiparous beef cows.

Timed AI has become a potential tool to bypass postpartum acyclicity, yet only a small percentage of the world bovine herd is inseminated. Most females are still subjected to bull mating; therefore, the frequent occurrence of postpartum anestrus may compromise their reproductive efficiency. Thus, the aim of this study was to develop an approach that allows the early conception of postpartum primiparous beef cows that are exposed to natural breeding (NB). For this purpose, 350 primiparous Nelore cows 35-65 d postpartum were allocated into three groups: Control (no hormonal treatment), TNB (hormonal protocol for timed-NB without equine chorionic gonadotropin; eCG) and TNB + eCG (hormonal protocol for TNB with eCG) groups. The protocol for TNB consisted of the insertion of a 1-g progesterone device and the intramuscular (IM) administration of 2 mg estradiol benzoate on D-9 (nine days before bull exposure), followed by device removal and the administration of 1 mg estradiol cypionate IM on D0. An additional 300 IU of eCG was given only to TNB + eCG cows. All cows were exposed to bull mating from D0 to D105. Pregnancy was checked by ultrasonography every 50 d, gestational age was estimated, and the number of new gestations every 21-d cycle (P21, P42, P63, P84 and P105) was predicted. Control cows had lower pregnancy at P21 (5.7%c, 7/123) than TNB (30.4%b, 35/115) and TNB + eCG (51.8%a, 58/112; P = 0.001) cows. Pregnancy rate increased across P42, P63 and P84 (P = 0.001) but remained higher for TNB + eCG cows. Regarding time to conception, the TNB + eCG cows achieved a greater than 50% pregnancy rate within the first days after bull exposure, while the TNB and Control cows took more than 40 d and 90 d to achieve 50% pregnancy rates, respectively. At the end of the breeding season (BS), TNB + eCG cows had a 21% higher pregnancy rate than the Control cows and a 16% higher rate than the TNB cows. The probability of conceiving increased 1.5-fold for cows treated with TNB (P = 0.0079) and 2.2-fold for cows additionally treated with eCG (P < 0.0001). The average interval between the onset of the BS and conception was reduced (P < 0.0001) for the TNB + eCG cows (26.5 ± 3.8c) compared to the TNB (35.7 ±â€¯4.1b) and Control (64.7 ±â€¯3.9a days) cows. Thus, the use of TNB, especially when associated with eCG, efficiently improved the early conception of postpartum primiparous beef cows after exposure to NB. The increased number of cows conceiving early in the BS is crucial to improve reproductive efficiency by reducing the interval between parturitions and improving the number of pregnant cows at the end of BS, thus resulting in greater farm income.

Hormonal strategy to reduce suckled beef cow handling for timed artificial insemination with sex-sorted semen.

Two experiments were conducted to assess a hormonal strategy developed to reduce animal handling for timed artificial insemination (TAI) with sex-sorted semen. Four-hundred ninety-one (491) suckled beef cows received a progesterone (P4) intravaginal device and 2 mg intramuscular (im) injection of estradiol benzoate (EB) on a randomly chosen day of the estrus cycle (Day 0) in Experiment 1. Cows were treated with 500 µg of sodic cloprostenol (PGF2α) and with 300 IU of eCG at P4 device removal (Day 8); these cows were also randomly assigned to receive 1 mg of estradiol cypionate (EC) administered at P4 device removal (treatment EC-0h) or 1 mg of EB 24 h after P4 device removal (treatment EB-24h). Both treatments were timed inseminated (TAI) with sex-sorted semen 60 h after P4 device removal. Cows treated with EC-0h presented higher pregnancy rate per AI (P/AI) [45.0% (113/251)] than the ones treated with EB-24h [35.4% (85/240); P = 0.03)]. A subset of cows (n = 26) were subjected to ultrasound examination every 12 h after P4 device removal for 96 h in the row in order to determine the time of ovulation. Similar interval between device removal and ovulation was recorded for EB-24h = 70.0 ±â€¯2.9 h vs. EC-0h = 66.0 ±â€¯2.8 h (P = 0.52). Five-hundred ninety-one (591) cows were subjected to the same synchronization protocols and treatments (EC-0h or EB-24h). In addition, they were randomly assigned to a 2 × 2 factorial arrangement aiming at determining the effects of treatment with estradiol (EC-0h vs. EB-24h) and of semen type (Sex-sorted vs. Non-sex-sorted semen). All animals were timed inseminated 60 h after P4 device removal. There was no interaction (P = 0.07) between the ovulation inducer and semen type. The EC protocol led to greater P/AI than EB (P = 0.03). Greater (P = 0.01) P/AI was achieved through treatments with non-sex-sorted semen rather than with sex-sorted semen [sex-sorted (EB-24h = 49.0%; EC-0h = 51.0%) vs. non-sex-sorted semen (EB-24h = 52.4%; EC-0h = 68.2%)]. Therefore, EC administered at P4 device removal resulted in greater P/AI. Furthermore, the EC-0h protocol allowed reducing suckled beef cow handing for timed artificial insemination with sex-sorted semen.

Review: Using artificial insemination v. natural service in beef herds.

The aim of this review is to compare the performance of different reproductive programs using natural service, estrus synchronization treatment before natural service (timed natural breeding (TNB)), artificial insemination (AI) following estrus detection and timed artificial insemination (TAI) in beef herds. It is well known that after parturition the beef cow undergoes a period of anestrous, when they do not exhibit estrus, eliminating the opportunity to become pregnant in the early postpartum by natural mating or by AI after detection of estrus. Hormonal stimulation is already a consistent and well-proven strategy used to overcome postpartum anestrus in beef herds. Basically, hormones that normally are produced during the estrous cycle of the cow can be administered in physiological doses to induce cyclicity and to precisely synchronize follicular growth, estrus and ovulation. Furthermore, two options of mating may be used after hormonal stimulation: natural service (i.e. utilization of bull service after synchronization, referred to as TNB) and TAI. These strategies improve the reproductive efficiency of the herds compared with natural service without estrus induction or synchronization. After the first synchronized service, the most common strategy adopted to get non-pregnant cows pregnant soon is the introduction of clean-up bulls until the end of the breeding season. However, methods to resynchronize non-pregnant cows after the first service are already well established and offer a potential tool to reduce the time for subsequent inseminations. Thus, the use of these technologies enable to eliminate the use of bulls by using resynchronization programs (i.e. two, three or four sequential TAI procedures). The dissemination of efficient reproductive procedures, such as TNB, TAI and Resynch programs, either isolated or in combination, enables the production of a greater quantity (obtaining increased pregnancy rates early in the breeding season) and quality (maximization of the use of AI with superior genetic sires) of beef calves. These technologies can contribute to improve the production efficiency, and consequently, improve livestock profitability.

Associations of insulin resistance later in lactation on fertility of dairy cows.

The challenge of getting dairy cows pregnant during early lactation is a well-described, worldwide problem. However, specifically in farms with poor reproductive, nutritional, and environmental conditions/management, a low pregnancy rate during early lactation is followed inevitably by an increased number of nonpregnant cows after 150 days in milk, with even more difficulties to achieve pregnancy. Therefore, several studies were designed to understand and develop strategies to mitigate reduced fertility of cows during late lactation. Experiments were performed under tropical regions to determine metabolic status during lactation and association of stage of lactation on oocyte quality and fertility. Lactating cows with extended days not pregnant (e.g.,>150 days in milk) often had systemic metabolic alterations, including development of peripheral insulin resistance and various oocyte alterations, including reduced expression of genes encoding glucose transport proteins, reduced amounts of mtDNA, increased expression of mitochondria-related genes, and increased expression of apoptosis-related genes. Additionally, in vitro embryo production and pregnancy per AI were lower in late- versus early-lactation cows in some but not all studies. Notwithstanding, when a normal embryo was transferred to a cow in late lactation, the pregnancy per transfer was reasonable, reinforcing the assertion that fertility problems in late-lactation cows may be associated with oocyte quality, fertilization, and/or failure of early embryo development. In conclusion, insulin resistance may reduce oocyte competence and consequently fertility in late-lactation dairy cows.

Ovarian follicular growth suppression by long-term treatment with a GnRH agonist and impact on small follicle number, oocyte yield, and in vitro embryo production in Zebu beef cows.

The aim of the present study was to evaluate small follicle number, oocyte yield, and in vitro embryo production (IVEP) in Zebu beef cows treated long term with a GnRH agonist to suppress ovarian follicular growth. Nelore (Bos indicus) cows (n = 20) showing regular estrous cycles were randomly assigned to one of two groups: control (n = 10, placebo ear implant without a GnRH agonist); GnRH agonist (n = 10, GnRH agonist ear implant containing 9.4-mg deslorelin). All cows underwent an ovum pick-up (OPU) session 14 days (Day 14) before the start of treatments (Day 0) followed by seven OPU-IVEP procedures at 30-day intervals (Days 0, 30, 60, 90, 120, 150, and 180). Semen from a single batch of a previously tested bull was used for all the IVEP. Cows treated with agonist reported a decrease over time in the proportion of animals with a (CL; P ≤ 0.05) and large follicles (>10 mm, P ≤ 0.05). These cows had a lesser number of medium + large follicles (>5 mm; 1.74 ± 0.5 vs. 4.13 ± 0.5; P ≤ 0.05), greater number of small follicles (2-5 mm; 44.3 ± 2.8 vs. 30.8 ± 1.8; P ≤ 0.05), greater yield of cumulus-oocyte complexes (COCs; 21.0 ± 2.3 vs. 15.6 ± 1.9; P ≤ 0.05), greater proportion of COCs cultured (79.2 vs. 73.9%; P ≤ 0.05), COCs cleaved (10.6 ± 1.5 vs. 6.8 ± 1.1, P ≤ 0.05), and cleaved rate (52.8 vs. 44.3%; P ≤ 0.05) compared with control cows. The number (3.4 ± 0.7 vs. 3.0 ± 0.6; P > 0.05) and proportion (16.5 vs. 19.1%; P > 0.05) of blastocysts produced were similar between agonist and control cows, respectively. The study has shown that Zebu beef cows treated long term with a GnRH agonist had follicular growth restricted to small follicles. This did not compromise the ability of oocytes to undergo IVF and embryonic development.

Short communication: Field fertility in Holstein bulls: Can type of breeding strategy (artificial insemination following estrus versus timed artificial insemination) alter service sire fertility?

The aim of this study was to compare pregnancy per artificial insemination (P/AI) from service sires used on artificial insemination after estrus detection (EAI) or timed artificial insemination (TAI) breedings. Confirmed artificial insemination outcome records from 3 national data centers were merged and used as a data source. Criteria edits were herd's overall P/AI within 20 and 60%, a minimum of 30 breedings reported per herd-year, service sires that were used in at least 10 different herds with no more than 40% of the breedings performed in a single herd, breeding records from lactating Holstein cows receiving their first to fifth postpartum breedings occurring within 45 to 375 d in milk, and cows with 1 to 5 lactations producing a minimum of 6,804 kg. Initially 1,142,859 breeding records were available for analysis. After editing, a subset of the data (n=857,539) was used to classify breeding codes into either EAI or TAI based on weekly insemination profile in each individual herd. The procedure HPMIXED of SAS was used and took into account effects of state, farm, cow identification, breeding month, year, parity, days in milk at breeding, and service sire. This model was used independently for the 2 types osires f breeding codes (EAI vs. TAI), and service sire P/AI rankings within each breeding code were performed for sires with >700 breedings (94 sires) and for with >1,000 breedings (n=56 sires) following both EAI and TAI. Correlation for service sire fertility rankings following EAI and TAI was performed with the PROC CORR of SAS. Service sire P/AI rankings produced with EAI and TAI were 0.81 (for sires with >700 breedings) and 0.84 (for sires with >1,000 breedings). In addition, important changes occurred in service sire P/AI ranking to EAI and TAI for sires with less than 10,000 recorded artificial inseminations. In conclusion, the type of breeding strategy (EAI or TAI) was associated with some changes in service sire P/AI ranking, but ranking changes declined as number of breedings per service sire increased. Future randomized studies need to explore whether changes in P/AI ranking to EAI versus TAI are due to specific semen characteristics.

Different doses of equine chorionic gonadotropin on ovarian follicular growth and pregnancy rate of suckled Bos taurus beef cows subjected to timed artificial insemination protocol.

This study evaluated the effect of different doses of eCG (control, 300 or 400 IU) administered at progesterone (P4) device removal in suckled Bos taurus beef cows undergoing a timed artificial insemination (TAI) protocol. A total of 966 cows received a P4 insert and 2.0 mg intramuscular estradiol benzoate at the onset of the synchronization. After 9 days, P4 insert was removed, and 12.5 mg of dinoprost tromethamine and 1 mg of estradiol cypionate were administered, followed by TAI 48 hours later. Then, the cows received one of three treatments as follows: control (n = 323), 300 (n = 326), or 400 IU of eCG (n = 317). A subset (n = 435) of cows in anestrus had their ovaries evaluated using ultrasound at the time of P4 removal and at TAI. Data were analyzed by orthogonal contrasts (C): C1 (eCG effect) and C2 (eCG dose effect). Estrous occurrence (control = 53.7%, 300 IU = 70.6%, and 400 IU = 77.0%) and pregnancy per artificial insemination (control = 29.7%, 300 IU = 44.8%, and 400 IU = 47.6%) were improved by eCG treatment (C1; P = 0.0004 and P < 0.0001, respectively). Furthermore, the cows receiving eCG presented larger follicles at TAI (control = 13.5 ± 0.3 mm, 300 IU = 14.0 ± 0.2 mm, and 400 IU = 15.1 ± 0.3 mm; P < 0.0001; C1). However, there was no effect of eCG dose on any response variables studied (C2; P > 0.15). In conclusion, the eCG treatment administered at the time of P4 removal increased the occurrence of estrus, the larger follicles at TAI, and pregnancy per artificial insemination of suckled B taurus beef cows. Despite the greater occurrence of estrus in noncyclic cows receiving 400 IU of eCG, both eCG doses (300 and 400 IU) were equally efficient to improve pregnancy to artificial insemination.

Efficacy of a single intramuscular injection of porcine FSH in hyaluronan prior to ovum pick-up in Holstein cattle.

Plasma FSH profiles, in vitro embryo production (IVP) after ovum pickup (OPU), and establishment of pregnancy with IVP embryos were compared in untreated Holstein oocyte donors and those superstimulated with multiple injections or a single intramuscular (IM) injection of porcine FSH (pFSH) in hyaluronan (HA). Plasma FSH profiles were determined in 23 heifers randomly allocated to one of four groups. Controls received no treatment, whereas the F200 group received 200 mg of pFSH in four doses, 12 hours apart. The F200HA and F300HA groups received 200- or 300-mg pFSH in 5 mL or 7.5 mL, respectively of a 0.5% HA solution by a single IM injection. Plasma FSH levels were determined before the first pFSH treatment and every 6 hours over 96 hours. All data were analyzed by orthogonal contrasts. Circulating FSH area under curve (AUC) in pFSH-treated animals was greater than that in the control group (P = 0.02). Although the AUC did not differ among FSH-treated groups (P = 0.56), the total period with elevated plasma FSH was greater in the F200 group than in the HA groups (P < 0.0001). However, the F300HA group had a greater AUC than the F200HA group (P = 0.006), with a similar total period with elevated plasma FSH (P = 0.17). The IVP was performed in 90 nonlactating Holstein cows randomly allocated to one of the four treatment groups as in the first experiment. A greater proportion of medium-sized (6-10 mm) follicles was observed in cows receiving pFSH, regardless of the treatment group (P < 0.0001). Also, numbers of follicles (P = 0.01), cumulus-oocyte complexes (COCs) retrieved (P = 0.01) and matured (P = 0.02), cleavage rates (P = 0.002), and blastocysts produced per OPU session (P = 0.06) were greater in cows receiving pFSH, regardless of the treatment group. Cows in the F200HA group had a greater recovery rate (P = 0.009), number of COCs cultured (P = 0.04), and blastocysts produced per OPU session (P = 0.06) than cows in the F300HA group. Similar pregnancy rates were observed 50 to 60 days after transferring IVP embryos from donors in the different treatment groups (P > 0.05). In conclusion, a single IM injection of pFSH combined in 0.5% HA resulted in similar plasma FSH profiles as twice-daily pFSH treatments. Treatment of nonlactating donors with pFSH, with or without HA, resulted in increased IVP over untreated controls. A single dose of 200 mg of pFSH in 0.5% HA resulted in greater IVP than 300-mg pFSH in HA. Finally, pregnancy rates with IVP embryos were similar, regardless donor treatment.

Impact of progesterone and estradiol treatment before the onset of the breeding period on reproductive performance of Bos indicus beef heifers.

The present aimed to develop a hormonal therapy based on the insertion of a progesterone (P4) insert (PI) during 10 d plus an estradiol injection (E2) at PI removal before the onset of breeding of beef heifers. In Exp. 1, the plasma P4 profile of prepubertal heifers showed that the insertion of a PI or 24 d previously used P4 insert (UPI) sustained plasma P4 above 1 ng/mL for at least the first 7 d of the treatment. In Exp. 2 there was no positive effect of additional estradiol benzoate (EB) administered at the insertion of UPI on the proportion of heifers with a corpus luteum (CL/Treated) 30 d after UPI removal [UPI + EB = 85.3%(a) (n = 134); EB + UPI + EB = 80.8%(a) (n = 125)]; however, both were greater (P < 0.0001) than the Control group [60.3%(b) (n = 129)]. In Exp. 3, a positive effect (P=0.01) of UPI treatment and both E2 supplementations [EB and estradiol cypionate (EC); P = 0.10] at the UPI removal was observed on CL/Treated [Control = 42.5%(b) (n = 94); UPI = 58.5%(a) (n= 130); UPI + EB = 64.0%(a) (n = 128); UPI + EC = 67.2%(a) (n = 128)]. However, greater pregnancy per treated heifer (P/Treated) following artificial insemination (AI) upon estrus detection was achieved when EC was applied [Control = 20.2%(b); UPI = 29.2%(ab); UPI + EB = 26.6%(b); UPI + EC = 36.7%(a)]. In Exp. 4, the treatment prior to the timed AI (TAI) tended to improve pregnancy per TAI [P/AI; Control 43.6% (n = 298) vs. UPI+EC 51.9% (n = 342); P = 0.08], but increased P/Treated [26.5% vs. 43.3%; P < 0.001]. Thus, UPI + EC treatment were efficient in increasing the CL/Treated, tended to improve the P/AI and consequently enhanced P/Treated of zebu beef heifers.

Impact of puberty status and melengestrol acetate supplementation before the breeding period on reproductive efficiency of Bos indicus beef heifers.

Two experiments were designed to evaluate the impact of puberty status and the administration of melengestrol acetate (MGA) before onset of the breeding period on ovulatory responses (Exp. 1) and conception rate after AI performed on estrus detection during 10 d and the pregnancy rate through 80 d of breeding period (Exp. 2) of pasture-grazed beef heifers. In Exp. 1, heifers (15 pubertal and 15 prepubertal) received 0.5 mg per heifer/d -1 of MGA over 14 d. No differences in the ovulatory responses were found 10 d after the MGA administration (pubertal = 46.7% vs. prepubertal P = 53.3%; P = 0.72). In Exp. 2, 368 heifers were randomly assigned to groups according to pubertal status and the MGA treatment. All heifers were inseminated on estrus detection for up 10 d after MGA administration and following exposure to bulls between 20 and 80 d. The MGA-treated heifers exhibited a greater AI service rate than control heifers (72.1 vs. 41.6%;P < 0.01); however, heifers receiving MGA had lower conception results following AI (51.6 vs. 71.4%; P = 0.01). In addition, MGA-treated heifers were more likely to have a corpus luteum in the middle of the breeding period (95.3 vs. 87.5%;P < 0.01), although the Cox proportional hazard of pregnancy rate was similar (P = 0.29) at the end of the breeding period. At onset of the breeding period, pubertal heifers presented a greater pregnancy rate following AI (pubertal P = 42.2% vs. prepubertal P = 24.9%; P = 0.01). Therefore, pubertal heifers seem to have greater overall reproductive efficiency than prepubertal heifers, particularly at the beginning of the breeding period. Interestingly, administration of MGA before the onset of the breeding period increased AI service rate but did not alter the rate of pregnancy throughout the breeding period of pasture-grazed beef heifers.

Paradoxical effects of bovine somatotropin treatment on the ovarian follicular population and in vitro embryo production of lactating buffalo donors submitted to ovum pick-up.

The aim of the present study was to evaluate the effect of bovine somatotropin (bST; 500mg) administration on lactating buffalo donors submitted to two different ovum pick-up (OPU) and in vitro embryo production schemes with a 7 or 14d intersession OPU interval. A total of 16 lactating buffalo cows were randomly assigned into one of four experimental groups according to the bST treatment (bST or No-bST) and the OPU intersession interval (7 or 14d) in a 2×2 factorial design (16 weeks of OPU sessions). The females submitted to OPU every 14d had a larger (P<0.001) number of ovarian follicles suitable for puncture (15.6±0.7 vs. 12.8±0.4) and an increased (P=0.004) number of cumulus-oocyte complexes (COCs) recovered (10.0±0.5 vs. 8.5±0.3) compared to the 7d interval group. However, a 7 or 14d interval between OPU sessions had no effect (P=0.34) on the number of blastocysts produced per OPU (1.0±0.1 vs. 1.3±0.2, respectively). In addition, bST treatment increased (P<0.001) the number of ovarian follicles suitable for puncture (15.3±0.5 vs. 12.1±0.4) but reduced the percentage (18.9% vs. 10.9%; P=0.009) and the number (1.4±0.2 vs. 0.8±0.1; P=0.003) of blastocysts produced per OPU session compared with the non-bST-treated buffaloes. In conclusion, the 14d interval between OPU sessions and bST treatment efficiently increased the number of ovarian follicles suitable for puncture. However, the OPU session interval had no effect on embryo production, and bST treatment reduced the in vitro blastocyst outcomes in lactating buffalo donors.

A comparison of two different esters of estradiol for the induction of ovulation in an estradiol plus progestin-based timed artificial insemination protocol for suckled Bos indicus beef cows.

The abilities of two different estradiol esters to induce ovulation in a timed AI (TAI) synchronization protocol in suckled Bos indicus cows were evaluated. In Experiment 1 (synchrony of ovulation), 31 cows were submitted to an estradiol/progestin-based synchronization protocol (Day 0) and randomly assigned to one of three treatments at the time of progestin removal on Day 8: 0.5 or 1.0mg of estradiol cypionate (EC) at that time or 1.0mg of estradiol benzoate (EB) 24h later (Day 9). To determine the timing of ovulation, ultrasound examinations were performed every 12h from ear implant removal to 96 h after the removal. Orthogonal comparisons were performed to determine the effects of estradiol ester and the effects of the dose of EC on reproductive parameters. Although neither the E2 ester (P = 0.83) nor the dose of EC (P = 0.55) affected the ovulation rate, the interval from progestin removal to ovulation was longer (P=0.04) in EC-treated cows (1.0mg EC = 71.1 ± 3.6 and 0.5mg EC = 78.0 ± 3.5) than EB-treated cows (EB = 66.0 ± 2.3) was detected. Ovulation in 0.5-mg-EC-treated cows was less synchronous than that in 1.0-mg-EC-treated cows (distribution curves compared using kurtosis). In Experiment 2 (pregnancy per AI; P/AI), 660 cows at two different locations received the same synchronization protocol (n = 361 at Farm A and n = 299 at Farm B) and were treated with estradiol esters as in Experiment 1 [0.5mg EC (n = 220) or 1.0mg EC (n = 219) at the time of progestin removal or 1.0mg EB (n = 221) 24h later]. The cows were inseminated 54 to 56 h after progestin removal. As applied in the Experiment 1, orthogonal comparisons were performed to evaluate the effect of estradiol ester and the dose of EC on P/AI. Although the type of estradiol ester used did not affect the P/AI (P = 0.57; EB - 43.0% vs. EC - 44.6%), the P/AI was higher (P=0.03) in cows treated with 1.0mg EC (55.7%) than in those treated with 0.5mg EC (38.6%). In summary, the administration of 0.5mg EC at the time of progestin removal altered the distribution of ovulation and resulted in a lower P/AI when compared with the use of 1.0mg EC in suckled B. indicus cows. However, the P/AI following the administration of 1.0mg EC at the time of progestin removal did not differ from that after the administration of 1.0mg EB 24h later.

Anti-Mullerian hormone concentration and antral ovarian follicle population in Murrah heifers compared to Holstein and Gyr kept under the same management.

This study was performed to evaluate plasma concentrations of anti-Mullerian hormone (AMH) and the ovarian antral follicle population (AFP) in different genetic groups. Cyclic heifers (13 Bubalus bubalis [Murrah]; 15 Bos taurus [Holstein] and 10 Bos indicus [Gyr]) were maintained under the same management and were synchronized with two doses of 150 µg IM d-cloprostenol administered 14 days apart. After the second d-cloprostenol treatment, heifers had their ovaries scanned daily by ultrasound to define the day of ovulation. On the same day, the AFP was determined and a plasma sample was collected to measure AMH. Murrah heifers had less AFP (25.6 ± 2.1 follicles; p = 0.01) and plasma AMH concentration (0.18 ± 0.03 ng/ml; p < 0.001) than Gyr (60.0 ± 12.2 follicles and 0.60 ± 0.12 ng/ml of AMH); however, data were similar when compared to Holstein (35.9 ± 6.8 follicles and 0.24 ± 0.06 ng/ml of AMH) heifers. Regardless of genetic background, there was a positive relationship between the AFP and plasmatic AMH concentration (Murrah [r = 0.62; p < 0.01], Holstein [r = 0.66; p < 0.001] and Gyr [r = 0.88; p < 0.001]). Also, when heifers were classified according to high- or low-AMH concentration based on the average within each genetic group, high-AMH heifers had greater (p < 0.0001) AFP than low-AMH heifers. In conclusion, both Murrah and Holstein heifers presented lower plasma AMH concentration and AFP when compared to Gyr.

Plasma anti-mullerian hormone: an endocrine marker for in vitro embryo production from Bos taurus and Bos indicus donors.

The aim of this study was to evaluate the association between plasma anti-mullerian hormone (AMH) concentration and in vitro embryo production (IVP) from Bos taurus (Holstein) and Bos indicus (Nelore) donors. A total of 59 Holstein (15 prepubertal heifers aged 8-10 mo, 15 cyclic heifers aged 12-14 mo, 14 lactating cows, and 15 nonlactating cows) and 34 Nelore (12 prepubertal heifers aged 10-11 mo, 10 prepubertal heifers aged 21-23 mo, and 12 cyclic heifers aged 24-26 mo) females were enrolled. All females underwent an ovum pick-up (OPU), without previous synchronization of the follicular wave, and IVP procedure. Immediately before the OPU procedure, blood samples were collected for subsequent AMH determination. A positive correlation was observed between the plasma AMH and number of in vitro embryos produced from Holstein (r = 0.36, P < 0.001) and Nelore (r = 0.50, P = 0.003) donors. For additional analyses, donors within each genotype were classified into 1 of 2 AMH categories (low or high) according to the average AMH concentration for each genotype. The results revealed that females classified as having high AMH presented a greater number of visible aspirated follicles (Holstein: 20.9 ± 1.5 vs 13.6 ± 0.9, P < 0.0001; Nelore: 54.3 ± 6.1 vs 18.6 ± 2.1, P < 0.0001) and a greater number of recovered cumulus-oocyte complexes (Holstein: 17.3 ± 1.5 vs 9.0 ± 0.9, P < 0.0001; Nelore: 45.3 ± 6.4 vs 13.4 ± 1.7, P < 0.0001). However, there was no difference in the blastocyst production rate (Holstein: 20.6% ± 4.0% vs 19.8% ± 4.2%, P = 0.60; Nelore: 33.7% ± 6.5% vs 27.4% ± 5.5%, P = 0.41, high and low AMH, respectively). Moreover, donors classified as having high AMH yielded a greater number of embryos produced per OPU (Holstein: 3.0 ± 0.7; Nelore: 7.0 ± 1.7) compared with those classified as having low AMH (Holstein: 1.2 ± 0.3, P = 0.04; Nelore: 2.2 ± 0.5, P = 0.007). In conclusion, although the plasma AMH concentration did not alter the ability of the cumulus-oocyte complex to reach the blastocyst stage, the AMH concentration in plasma can be an accurate endocrine marker for the in vitro embryo yield from either B. taurus (Holstein) or B. indicus (Nelore) donors. Therefore, AMH is a promising tool to enhance the overall efficiency of OPU-IVP programs in the field as a selective criterion for high embryo producing donors.

Inferences of body energy reserves on conception rate of suckled Zebu beef cows subjected to timed artificial insemination followed by natural mating.

The influence of body condition score (BCS), rump fat thickness (RFAT), and live weight (LW), and the changes in these parameters during the interval from 165 of prepartum (i.e., 125 days of prior gestation) to 112 postpartum on first service conception and pregnancy rates were investigated in suckled Zebu (Bos indicus) beef cows (n = 266) subjected to timed artificial insemination (TAI) followed by natural mating. The aforementioned parameters were recorded at 165 ± 14 days (mean ± standard error) prepartum (concurrent with the weaning of previous calf), at parturition, and at 42 ± 7 days (at the onset of the synchronization of ovulation protocol), 82 ± 7 days (30 days after TAI), and 112 ± 7 days (60 days after TAI) postpartum. At the start of the breeding season (BS), cows were subjected to a synchronization of ovulation program for TAI. Bulls were placed with cows 10 days after TAI and remained until the end of the study (112 days postpartum). Cows with the highest BCS at parturition had an increased probability of first service conception rate at 60 days after TAI (P = 0.02) and a reduced probability of occurrence of pregnancy loss (P = 0.05). Also, cows had a greater likelihood of conceiving postpartum if they had greater RFAT and BCS at 165 ± 14 days prepartum (P = 0.01 and P = 0.03, respectively) and at parturition (P = 0.0007 and P = 0.003, respectively). Cows that had an increase in RFAT and BCS during the dry period (i.e., interval from weaning of the previous calf to parturition) also had a greater likelihood of conceiving (P = 0.03 and P = 0.06, respectively) during the BS. Among the different time points, RFAT and BCS at parturition had the largest impact on risk of conception during the BS. The LW was a poor predictor of conception during the BS (P = 0.11-0.68) except for LW at 165 ± 14 days prepartum (P = 0.01). Collectively, the findings indicated that the likelihood of conception during the BS was highest in cows that had an improvement in RFAT and BCS during the dry period. Therefore, assuring a good nutritional status in the dry period (BCS ≥ 3.0 at 165 ± 14 days prepartum and ≥3.25 at parturition) is an important aim to optimize the postpartum conception rate of suckled Zebu beef cows subjected to TAI followed by natural mating.

Superstimulation prior to the ovum pick-up to improve in vitro embryo production in lactating and non-lactating Holstein cows.

The present study evaluated the efficacy of superstimulation with p-FSH (Folltropin) before the ovum pick-up (OPU) on IVP in lactating and nonlactating Holstein donors. A total of 30 Holstein cows (15 lactating and 15 nonlactating) were blocked by lactation status to one of two groups (control or p-FSH), in a cross-over design. On a random day of the estrous cycle, all cows received an intravaginal progesterone device and 2.0 mg IM of estradiol benzoate (Day 0). Cows in the control group received no further treatment, whereas cows in the p-FSH group received a total dosage of 200 mg of p-FSH on Days 4 and 5 in four decreasing doses 12 hours apart (57, 57, 43, and 43 mg). On Day 7, the progesterone device was removed, and OPU was conducted in both groups (40 hours after the last p-FSH injection in the p-FSH-treated group). There was no difference between groups (P = 0.92) in the numbers of follicles that were aspirated per OPU session (17.2 ± 1.3 vs. 17.1 ± 1.1 in control and p-FSH-treated cows, respectively); however, p-FSH-treated cows had a higher (P < 0.001) percentage of medium-sized follicles (6-10 mm) at the time of the OPU (55.1%; 285/517) than control cows (20.8%; 107/514). Although recovery rate was lower (60.0%, 310/517 vs. 69.8%, 359/514; P = 0.002), p-FSH-treated cows had a higher blastocyst production rate (34.5%, 89/258 vs. 19.8%, 55/278; P < 0.001) and more transferable embryos per OPU session were produced in the p-FSH group (3.0 ± 0.5 vs. 1.8 ± 0.4; P = 0.02). Regardless of treatment, non-lactating cows had a higher blastocyst rate (41.9%, 106/253 vs. 13.4%, 38/283; P = 0.001) and produced more transferable embryos per OPU session (3.5 ± 0.5 vs. 1.3 ± 0.3; P = 0.003) than lactating cows. Thus, superstimulation of Holstein donors with p-FSH before OPU increased the efficiency of IVP. In addition, non-lactating donors had higher percentage of in vitro blastocyst development and produced more embryos per OPU session than lactating cows.

Plasma antimullerian hormone as a predictor of ovarian antral follicular population in Bos indicus (Nelore) and Bos taurus (Holstein) heifers.

In Bos taurus cattle, antimullerian hormone (AMH) has been demonstrated to have a high degree of correlation with ovarian antral follicle count and the number of healthy follicles and oocytes. To document the correlation between the plasma concentration of AMH and follicular number in Bos indicus and Bos taurus heifers, Nelore (Bos indicus, n = 16) and Holstein heifers (Bos taurus, n = 16) had their ovarian follicular waves synchronized. After synchronization, ovarian antral follicular population (AFP) was evaluated three times at 60-day (d) intervals (T-120 d, 120 days before plasma AMH determination; T-60 d, 60 days before; and T0, at the time of plasma AMH determination). The plasma AMH concentration was positively correlated with the number of ovarian follicles on the day of the follicular wave emergence in Bos indicus (Nelore) and Bos taurus (Holstein) heifers at each evaluation time (p < 0.05). The AFP was higher in Bos indicus (Nelore) than in Bos taurus (Holstein) heifers (p < 0.05). Similarly, the AMH concentration was higher in Bos indicus (Nelore) than in Bos taurus (Holstein) heifers (p < 0.0001). When heifers were classified as to present high or low AFP according to the mean of the AFP within each genetic group, high-AFP heifers presented a greater (p < 0.0001) AMH concentration than low-AFP heifers, regardless of the genetic group. In conclusion, the AFP is positively correlated with plasma AMH concentration in both Bos indicus (Nelore) and Bos taurus (Holstein) heifers. Furthermore, Bos indicus (Nelore) heifers presented both greater plasma AMH concentrations and AFP than Bos taurus (Holstein) heifers.

Sperm fertility and viability following 48h of refrigeration: evaluation of different extenders for the preservation of bull semen in liquid state.

Two experiments were conducted to compare the effectiveness of different extenders conventionally used for semen cryopreservation to maintain the viability and fertility of cooled bull semen. In Experiment 1, sperm samples obtained from 20 Nellore bulls were preserved at 5°C for 48h using two extenders containing 20% of egg yolk [Tris (TRIS-R) and Botu-Bov(®) (BB)] and another composed of 1% soy lecithin [Botu-Bov(®)-Lecithin (BB-L)] as substitutes for animal origin products. The samples were evaluated at 6, 24 and 48h for plasma and acrosomal membrane integrity, quantification of thiobarbituric acid reactive substances (ng of TBARS/10(8) cells) and sperm motility parameters by computer-assisted semen analysis (CASA). In Experiment 2, pregnancy rate (P/AI) of 973 fixed-time artificially inseminated Nellore cows were compared when cows were inseminated with conventionally cryopreserved semen in TRIS-egg yolk glycerol (TRIS-C Control, n=253) or semen cooled for 48h in TRIS-R (n=233), BB (n=247) or BB-L (n=240). Although none of the extenders used was effective on maintaining total progressive motility and cellular integrity throughout the 48-h of the refrigeration period (P<0.01), BB-L conferred greater protection against oxidative stress (P<0.05) than egg yolk-based medias. The P/AI for semen samples preserved in TRIS-C, TRIS-R, BB and BB-L were 39.92(a), 25.32(b), 26.32(b) and 33.33(ab), respectively. These results demonstrate that the three conventional extenders used for semen cryopreservation do not provide the protection required to maintain bull semen fertility under refrigeration for a 48-h period, resulting in reduced pregnancy rates. However, the use of lecithin-based medium instead of egg yolk results in greater protection against lipid peroxidation, producing P/AI results comparable to those obtained using frozen semen.

Donor category and seasonal climate associated with embryo production and survival in multiple ovulation and embryo transfer programs in Holstein cattle.

The present study investigated the effect of Holstein donor category (cows vs. heifers) and climate variation (hot vs. cooler season) on the efficiency of in vivo embryo production programs as well as embryo survival after transferred to Holstein recipient cows. A total of 1562 multiple ovulation (MO) procedures (cows: n = 609, and heifers: n = 953) and 4076 embryo transfers (ETs) performed in two dairy herds were evaluated. Donor cows had greater number of CLs (10.6 ± 0.6 vs. 7.5 ± 0.4; P < 0.0001) and ova/embryos recovered (7.6 ± 0.6 vs. 4.6 ± 0.4; P < 0.0001) compared with donor heifers. However, fertilization rate (47.9 vs. 82.4%; P < 0.0001) and proportion of transferable embryos (31.5 vs. 67.4%; P < 0.0001) were lower in donor cows than heifers, respectively. Regardless of donor category, the proportion of freezable embryos was less (P < 0.001) during hot season than in cooler season (21.4 vs. 32.8%). However, greater decline in the proportion of freezable embryos during the hot season was observed in cows (21.7 vs. 10.7%) compared with heifers (46.2 vs. 38.1%; P = 0.01). In contrast, the season on which the embryo was produced (hot or cool) did not affect pregnancy rate on Day 31 (30.5 vs. 31.7%; P = 0.45) and 45 (25.3 vs. 25.1%; P = 0.64) of pregnancy. Regardless of the season in which the embryos were produced, embryonic survival after transferring embryos retrieved from donor cows was greater on Days 31 (36.0 vs. 30.7%; P = 0.001) and 45 (28.3 vs. 23.1%; P = 0.001) of pregnancy when compared with embryos from donor heifers. In conclusion, MO embryo production efficiency decreased during the hot seasons both in cows and heifers; however, the decline was more pronounced in donor cows. Regardless of the embryo source, similar pregnancy rate was observed in the recipient that received embryos produced during the hot and cooler seasons. Curiously, embryos originating from donor cows had higher embryonic survival when transferred to recipient cows than embryos originating from heifers.