Comparison of long-term progestin-based protocols to synchronize estrus prior to natural service or fixed-time artificial insemination in Bos indicus-influenced beef heifers.

This experiment was designed to evaluate breeding strategies involving natural service or fixed-time artificial insemination (FTAI) in Bos indicus-influenced beef heifers (n = 1456) when there were field-type management conditions. Body weights and reproductive tract scores (RTS; Scale 1-5) were obtained for heifers before assignment to one of five treatments: 1) Non-synchronized control exposed for natural service (NS), n = 299; 2) melengestrol acetate + natural service (MGA + NS; 0.5 mg/heifer/d), n = 295; 3) 14-d controlled internal drug release insert + natural service (CIDR + NS), n = 289; 4) 14-d MGA-prostaglandin F2α (PG) + FTAI, n = 295; or 5) 14-d CIDR-PG + FTAI, n = 278. Fertile bulls were placed in pastures with heifers of the three NS treatment groups for a 65-day period which began 10 days after progestin treatments (MGA or CIDR) ended. Heifers in FTAI treatment groups were administered PG (25 mg, IM) 16 days after CIDR removal or 19 days following MGA withdrawal, respectively, and FTAI was performed at 66 (CIDR-PG) or 72 h (MGA-PG) after PG. Gonadotropin-releasing hormone (GnRH; 100 µg, i.m.) was administered at FTAI. Pregnancy status was determined at the end of a 65-day breeding period. Pregnancy rates on Days 21 and 65 of the breeding period differed among treatment groups based on pre-treatment pubertal status (P ≤ 0.02) and body weight (P ≤ 0.05) but did not differ by group. These data highlight the need for continued research efforts to improve reproductive management of Bos indicus-influenced females.

Evaluation of the 14-d CIDR-PG and 9-d CIDR-PG protocols for synchronization of estrus in Bos indicus-influenced and Bos taurus beef heifers.

Two long-term, CIDR-based estrus synchronization protocols were evaluated among Bos indicus-influenced and Bos taurus beef heifers. Treatments were evaluated on the basis of estrous response and pregnancy rate resulting from fixed-time artificial insemination (FTAI), and these outcomes were analyzed retrospectively relative to reproductive tract score (RTS; Scale 1-5) at treatment initiation. Estrus was synchronized for 1139 heifers in three locations, and heifers were assigned to one of two treatments within each location based on RTS. Heifers assigned to the 14-d CIDR-PG protocol received a controlled internal drug release (CIDR) insert (1.38 g progesterone) on Day 0, CIDR removal on Day 14, administration of prostaglandin F2α (PG; 25 mg im) on Day 30, and administration of gonadotropin-releasing hormone (GnRH; 100 µg im) concurrent with FTAI on Day 33, 66 h after PG. Heifers assigned to the 9-d CIDR-PG protocol received administration of PG concurrent with CIDR insertion on Day 5, administration of PG concurrent with CIDR removal on Day 14, administration of PG on Day 30, and administration of GnRH concurrent with FTAI on Day 33, 66 h after PG. Estrus detection aids were applied at CIDR removal on Day 14 and at PG on Day 30 to evaluate estrous response rate. Mean RTS differed (P < 0.0001) based on biological type due to higher rates of estrous cyclicity (RTS 4 and 5) among Bos taurus heifers (72%; 416/574) than among Bos indicus-influenced heifers (27%; 150/565). The proportion of heifers expressing estrus following CIDR removal was greater (P = 0.01) among heifers assigned to the 14-d CIDR-PG treatment (88%; 492/559) compared to the 9-d CIDR-PG treatment (83%; 480/580). Estrous response following CIDR removal was also higher (P < 0.0001) among Bos taurus (95%; 547/574) compared to Bos indicus-influenced (75%; 425/565) heifers. Rate of estrous response prior to FTAI did not differ significantly based on treatment but was higher (P < 0.0001) among Bos taurus heifers (60%; 344/574) than among Bos indicus-influenced heifers (45%; 253/565). However, the effect of biological type on estrous response was not significant when RTS was included in the model, as RTS significantly (P < 0.0001) affected the rate of estrous response both at CIDR removal and prior to FTAI. Across treatments and biological types, heifers that expressed estrus prior to AI achieved higher (P < 0.0001) AI pregnancy rates than heifers failing to express estrus. Pregnancy rates to FTAI did not differ significantly based on treatment in either biological type. Higher rates of estrous cyclicity among Bos taurus heifers resulted in higher FTAI pregnancy rates among Bos taurus (51%; 290/574) compared to Bos indicus-influenced heifers (39%; 218/565). However, pregnancy rates of respective RTS did not differ based on biological type. In summary, long-term CIDR-based protocols provide a simple, effective method of estrus synchronization in Bos indicus-influenced and Bos taurus beef heifers. Moreover, these results highlight the importance of management practices that result in high rates of estrous cyclicity prior to protocol initiation, particularly among later maturing breeds and biological types.

Comparison of wet brewers' grains or dried distillers' grains as supplements to conserved bermudagrass forage as winter feeding options for beef cows.

The objective of this study was to examine the use of 2 byproduct supplements and conserved warm-season forage as winter feeding options for primiparous beef cows. Gestating Angus ( = 48) and Brangus ( = 24) 2-yr-old cows were stratified by BW and breed to 1 of 12 pens. Pens were randomly assigned 1 of 2 supplements, wet brewers' grains (WBG) or dried distillers' grains (DDG). Coastal bermudagrass hay or round bale silage (RBS) was fed free choice (6 pens each) and cows received WBG or DDG supplements at a daily rate of 0.05% BW (DM basis) prorated for feeding 3 d/wk. Total BW and BCS changes did not differ ( = 0.65 and = 0.93, respectively) between DDG- and WBG-supplemented cows. Total amount of forage DM offered and mean calculated daily forage DM offered did not differ ( = 0.59 and = 0.20, respectively) between supplement treatments. Estimated daily mean and total supplement DM offered was greater ( < 0.001) for WBG than for DDG treatments. The same 2 supplements and forage sources were used in an unbalanced 6 × 4 design to measure intake, digestibility, and rumen parameters in ruminally fistulated steers. Supplement did not affect forage DMI of hay ( = 0.31) or RBS ( = 0.63). Total DMI was not different ( = 0.37 and = 0.73) for hay-based and RBS-based diets, respectively. Total tract digestibility tended to be greater ( = 0.06) for DDG than for WBG in hay diets but was not different ( = 0.76) for RBS diets. Daily mean ruminal pH was greater ( = 0.03) for WBG than for DDG when supplemented to hay-based diets. In RBS diets, a supplement × hour interaction ( = 0.05) existed for ruminal pH. Daily mean ruminal ammonia N concentration was greater ( < 0.001) for WBG-supplemented diets compared with DDG-supplemented diets. A supplement × hour interaction existed for ruminal ammonia in hay- ( < 0.001) and RBS- ( = 0.09) based diets. Ruminal pH and ammonia N concentrations stayed in ranges adequate to support ruminal metabolism and cattle performance. Marginal differences in ruminal measures between high-moisture and dry byproduct supplements resulted in no differences in cow performance when offered with hay or high-moisture forage. High-moisture forage sources can be coupled with high-moisture byproduct supplements.

Assessment of serum 25-hydroxyvitamin D concentrations of beef cows and calves across seasons and geographical locations.

Vitamin D is critical for the growth and development of calves and positively contributes to immune function of cattle. Serum 25-hydroxyvitamin D (25(OH)D) concentrations above 20 ng/mL have traditionally been considered adequate for growth and development of cattle, but recent evidence has indicated that concentrations below 30 ng/mL are insufficient for immunity. Because little information is available regarding vitamin D status of beef cattle, the objective of this study was to evaluate vitamin D status of beef cow-calf herds on pasture as affected by season and location. Serum samples were collected from 43 cow-calf pairs plus an additional 54 calves in herds located in Florida, Idaho, and Minnesota in the spring calving season. Samples were collected again over the summer months from animals in the Florida and Minnesota herds. Effects of subcutaneous injection of vitamins A, D, and E also were investigated in a subset of calves from the Idaho herd. All cows sampled had serum 25(OH)D concentrations above 30 ng/mL at the time of calving in the spring. The average serum 25(OH)D concentrations of cows rose from near 60 ng/mL in the spring to 75 ng/mL in the summer ( < 0.001). Most calves, on the other hand, had serum 25(OH)D concentrations below 20 ng/mL. The calves in the Florida and Minnesota herds similarly rose from averages of 10 to 15 ng/mL at birth to near 50 ng/mL by the end of summer. Serum 25(OH)D of severely deficient calves increased from 3 ng/mL in nonsupplemented calves to 11 ng/mL at 48 h after birth if given a bolus supplementation of 40,000 IU of vitamin D via subcutaneous injection of a vitamin A, D, and E supplement at birth ( < 0.001). Vitamin D supplementation of cows late in pregnancy has been shown to increase serum 25(OH)D of calves; however, beef cattle generally receive very little supplemental vitamin D, as was the case for the cows studied here. The lower serum 25(OH)D of cows in spring compared with summer and the prevalence of vitamin D deficiency of calves observed here indicate that increased vitamin D supplementation of cows over the winter months or vitamin D supplementation of newborn calves would be beneficial.

Comparison of feed additive technologies for preconditioning of weaned beef calves.

Our objective was to evaluate the response of weaned calves to different supplemental feed additives in a supplement to affect calf performance and mitigate stress response observed during weaning and preconditioning. At weaning in each of 2 yr, 160 Angus and Brangus calves (203 and 227 ± 2.3 and 2.5 kg) were stratified by BW, sex, and breed and were randomly allotted to 1 of 4 treatments ( = 40 calves/treatment): 1) supplement without feed additives (control, CON), 2) supplemented with chlortetracycline, 350 mg/d (CTC), 3) supplemented with monensin, 175 mg/d (RUM), and 4) supplemented with rumen modifier, 5 g/d (ACT). Calves were held by treatment in 1 of 4 drylot pens for 7 d after weaning and were offered ad libitum access to hay and 2.27 kg/d of supplement before placement in one of thirty-two 0.8-ha pastures (5 calves/pasture). On pasture calves were supplemented with 2.27 kg/d (yr 1) or supplemented at 1.0% BW (yr 2). Calf BW and blood samples were collected following weaning (d 0, 1, 4, 7, 11 in yr 1; d 0, 1, 3, 7, 14 in yr 2), at the conclusion of the preconditioning period (d 50, 51 in yr 2), and after transportation (d 52, 55, 59, 65 in yr 2) for analysis of acute phase protein (APP) concentrations. In yr 2, after 44 d on pasture, calves were loaded on 2 semitrucks and transported for 24 h. On return, calves were placed in 4 pastures with hay and fed their respective supplements for 14 d. For each year, data were analyzed with the MIXED procedure of SAS. The model included the main effect of treatment, and pasture was the experimental unit. All variables quantified by day were analyzed using repeated measures. In yr 1, ACT and CTC had greater (P <0.05) 52-d ADG than RUM, whereas CON was intermediate. However, in yr 2, over the 50-d postweaning period there was no difference (P = 0.20; 0.52 kg/d) in ADG response among treatments. After transportation, 7- and 14-d ADG were improved (P < 0.05) for ACT and CTC compared with CON and RUM. In both years, postweaning plasma concentrations of haptoglobin were similar (P > 0.05) among treatments; however an effect of day after transport (P < 0.001) was observed. Feed cost of gain and income over production cost (P ≥ 0.15; mean = $0.51/kg and $73.51, respectively) were not different among treatments. Use of supplemental additives may improve calf performance during a preconditioning period of this duration, but no additive was effective at mitigating stress postweaning. Additives were equally effective in supporting calf growth performance during a posttransportation period.

Subspecies differences in early fetal development and plasma pregnancy-associated glycoprotein concentrations in cattle.

Inclusion of Bos indicus genetics improves production traits of cattle maintained in hot climates. Limited information exists detailing pregnancy-specific events as influenced by variable amounts of Bos indicus genetics. Three experiments were completed to examine the effect of Bos taurus and Bos indicus genotypes on fetal size and plasma pregnancy-associated glycoprotein (PAG) concentrations. In all experiments, cows were bred by AI after synchronization of ovulation. Fetal measurements were completed by transrectal ultrasonography and plasma PAG concentrations were quantified from plasma harvested the day of each fetal measurement. In Exp. 1, fetal size and plasma PAG concentrations were measured at d 53 of pregnancy in cows composed of various fractions of Angus and Brahman (n = 9 to 21 cows/group). Fetus size was greater in cows containing >80% Angus genetics compared with cows containing <80% Angus influence (3.40 ± 0.28 vs. 2.86 ± 0.28 cm crown-rump length; P < 0.01). Plasma PAG concentrations were reduced (P < 0.01) in cows containing >80% Angus genetics when compared with their contemporaries (6.0 ± 1.5 ng/mL vs. 9.4 ± 1.5 ng/mL). In Exp. 2, fetal measurements and plasma PAG concentrations were determined at d 35 and 62 of pregnancy in Angus and Brangus cows. Breed did not affect fetus size at d 35, but Angus cows contained larger fetuses than Brangus cows at d 62 [3.0 ± 0.03 vs. 2.8 ± 0.03 cm crown-nose length (CNL; P > 0.01)]. Plasma PAG concentrations were not different between breed at d 35 and 62 (P > 0.1). In Exp. 3, fetal measurements and plasma samples were collected at d 33/34, 40/41, 47/48, and 54/55 post-AI in Angus and Brangus cows. Fetus size was not different (P > 0.05) between genotypes on d 33/34, 40/41, and 47/48. Angus fetuses were larger than Brangus fetuses at d 54/55 (2.1 ± 0.03 vs. 1.9 ± 0.03 cm CNL; P = 0.001). Plasma PAG concentrations were less in Angus than Brangus cows at each time point (average 4.9 ± 0.9 vs. 8.2 ± 0.9 ng/mL; P = 0.005). In conclusion, these studies determined that the Bos taurus × Bos indicus genotype impacts fetal size and rate of fetal development by 7 wk of gestation. Plasma PAG concentrations were increased in cattle with Bos indicus genetics in 2 of 3 studies, suggesting that genotype is one of several determinants of PAG production and secretion in cattle.

Effect of castration technique on beef calf performance, feed efficiency, and inflammatory response.

The objective of this experiment was to examine the effect of castration technique on daily feed intake (DFI), daily water intake (DWI), growth performance, residual feed intake (RFI), and inflammatory response in weaned beef calves. Seventy-five beef calves (214 ± 3.2 kg; 200 ± 26 d of age) were housed in a GrowSafe 4000 feed intake facility 7 d post weaning (15 calves/pen). Calves were offered a total mixed ration (TDN = 67.3% and CP = 12.2%, DM = 89%) for ad libitum consumption. On d 0, calves were assigned to 1 of 5 treatments (n = 15 calves/treatment): 1) steers castrated surgically pre-weaning (52 d of age; CON); 2) intact bulls (BULL); 3) bulls castrated by the Callicrate Bander on d 0 (No-Bull Enterprises LLC.; BAN); 4) bulls castrated by the Henderson Castrating Tool on d 0 (Stone Mfg & Supply Co.; HEN); and 5) bulls castrated surgically utilizing an emasculator on d 0 (SUR). Average daily gain, DFI, and DWI were recorded over 84 d. Blood was collected from a sub-sample of calves (n = 45) on d 0, 2, 6, 9, 12, and 15 relative to castration. Castration decreased (P = 0.06) ADG for castrates compared with CON from d 0 to 14 but not d 0 to 84. Daily feed intake and DWI were similar (P > 0.10) among treatments during d 0 to 84. Gain:feed was not affected by castration technique; however, RFI tended (P = 0.09) to be negative for CON and BULL compared with castrates on d 0 to 14 but not d 0 to 84. Acute phase protein analyses indicated that surgical castration (SUR or HEN) elicited a short-term inflammatory response in calves, whereas calves castrated with BAN elicited a delayed response. Calves castrated pre-weaning had improved d 0 to 14 ADG, feed intake, and inflammation response compared with calves castrated at weaning. Banding elicited a delayed negative response in ADG, DWI, and inflammation. In weaned calves, castration method did not affect performance, DFI, DWI, or inflammatory response during the 84-d trial.

Evaluation of two progestogen-based estrous synchronization protocols in yearling heifers of Bos indicus × Bos taurus breeding.

Yearling Bos indicus × Bos taurus heifers (n = 410) from three locations, were synchronized with either the Select Synch/CIDR+timed-AI (SSC+TAI) or 7-11+timed-AI (7-11+TAI) treatments. On Day 0 of the experiment, within each location, heifers were equally distributed to treatments by reproductive tract score (RTS; Scale 1-5: 1 = immature, 5 = estrous cycling) and body condition score. The 7-11+TAI treatment consisted of melengestrol acetate (0.5 mg/head/d) from Days 0 to 7, with PGF(2α) (25 mg im) on Day 7, GnRH (100 µg im) on Day 11, and PGF(2α) (25 mg im) on Day 18. The SSC+TAI heifers received the same carrier supplement (without MGA) from Days 0 to 7, and on Day 11 they were given 100 µg GnRH and an intravaginal CIDR (containing 1.38 g progesterone). The CIDR were removed on Day 18, concurrent with 25 mg PGF(2α) im For both treatments, estrus was visually detected for 1 h twice daily (0700 and 1600 h) for 72 h after PGF(2α), with AI done 6 to 12 h after a detected estrus. Non-responders were timed-AI and received GnRH (100 µg im) 72 to 76 h post PGF(2α). The 7-11+TAI heifers had a greater (P < 0.05) estrous response (55.2 vs 41.9%), conception rate (47.0 vs 31.3%), and synchronized pregnancy rate (33.5 vs 24.8%) compared to SSC+TAI heifers, respectively. Heifers exhibiting estrus at 60 h (61.7%) had a greater (P < 0.05) conception rate compared to heifers that exhibited estrus at ≤ 36 (35.3%), 48 (31.6%), and 72 h (36.2%), which were similar (P > 0.05) to each other. As RTS increased from ≤ 2 to ≥ 3, estrous response, conception rate, synchronized pregnancy rate, and 30 d pregnancy rate all increased (P < 0.05), irrespective of synchronization treatment. In conclusion, the 7-11+TAI treatment yielded greater synchronized pregnancy rates compared to SSC+TAI treatment in yearling Bos indicus × Bos taurus heifers.

Effects of acclimation to handling on performance, reproductive, and physiological responses of Brahman-crossbred heifers.

The objective of this study was to evaluate the effects of acclimation to handling on growth, plasma concentrations of progesterone (P4) and cortisol, temperament, and reproductive performance of Brahman-crossbred heifers. Over 2 consecutive years, 37 Braford and 43 Brahman x Angus heifers were initially evaluated, within 30 d after weaning, for BW and puberty status via transrectal ultrasonography and plasma P4 concentrations (d 0 and 10), and for temperament by measurements of chute score, pen score, and exit velocity (d 10 only). On d 11, heifers were stratified by breed, puberty status, temperament score, BW, and age and randomly assigned to receive or not (control) the acclimation treatment. Acclimated heifers were exposed to a handling process 3 times weekly (Mondays, Wednesdays, and Fridays) for 4 wk (d 11 to 39 of the experiment). The acclimation treatment was applied individually to heifers by processing them through a handling facility, whereas control heifers remained undisturbed on pasture. Heifer puberty status, evaluated via plasma P4 concentrations and transrectal ultrasonography, and BW were assessed again on d 40 and 50, d 80 and 90, and d 120 and 130. Blood samples collected before (d 10) and at the end of the acclimation period (d 40) were also analyzed for plasma concentrations of cortisol. Heifer temperament was assessed again on d 40 of the study. No interactions containing the effects of treatment, breed, and year were detected. Acclimated heifers had reduced (P < 0.01) ADG compared with control heifers (0.50 vs. 0.58 kg/d, respectively). Attainment of puberty and pregnancy, however, was hastened (P = 0.02 and 0.04, respectively) in acclimated heifers compared with control. Acclimated heifers had reduced chute score (P < 0.01) and concentrations of cortisol (P < 0.01) and P4 (P = 0.03; prepubertal heifers only) compared with control heifers after the acclimation period (1.37 vs. 1.84 for chute score; 37.8 vs. 50.5 ng/mL of cortisol; 0.52 vs. 0.78 ng/mL of P4). Results from this study indicated that, although acclimation to handling decreased ADG, it resulted in decreased chute score, reduced plasma concentrations of cortisol and prepubertal P4, and enhanced reproductive performance of Brahman-crossbred heifers.

Insulin-like growth factor (IGF) family genes are aberrantly expressed in bovine conceptuses produced in vitro or by nuclear transfer.

Embryos produced through somatic cell nuclear transfer (NT) or in vitro production (IVP) are often associated with increased abortion and abnormalities thought to arise from disruptions in normal gene expression. The insulin-like growth factor (IGF) family has a major influence on embryonic, fetal and placental development; differences in IGF expression in NT- and IVP-derived embryos may account for embryonic losses during placental attachment. In the present study, expression of IGF-I, IGF-II, IGF-I receptor (IGF-IR), and IGF-IIR mRNAs was quantitated in Day 7 and 25 bovine embryos produced in vivo, by NT, IVP, or parthenogenesis, to further understand divergent changes occurring during development. Expression of the IGF-I gene was not detected in Day 7 blastocysts for any treatment. However, there were no differences (P>0.10) among Day 7 treatments in the amounts of IGF-IR, IGF-II, and IGF-IIR mRNA. For Day 25 conceptuses, there was higher expression of IGF-I mRNA for NT and IVP embryonic tissues than for in vivo embryonic tissues (P<0.05). Furthermore, embryonic tissues from NT-derived embryos had higher expression of IGF-II mRNA than IVP embryonic tissues (P<0.05). Placental expression of IGF-IIR mRNA was greater for NT-derived than in vivo-derived embryos (P<0.05). There were no differences in IGF-IR mRNA across all treatments and tissues (P>0.10). In conclusion, these differences in growth factor gene expression during early placental attachment and rapid embryonic growth may directly or indirectly contribute to increased losses and abnormalities in IVP- and NT-derived embryos.

Synchronization of Bos indicus x Bos taurus cows for timed artificial insemination using gonadotropin-releasing hormone plus prostaglandin F2alpha in combination with melengestrol acetate.

Nonlactating Bos indicus x Bos taurus cows were used in three herds to determine the efficacy of different PGF2alpha treatments in combination with GnRH and melengestrol acetate (MGA) for a timed artificial insemination protocol. The start of the experiment was designated as d 0, at which time cows were assigned a body condition score and received 100 microg of GnRH. Cows were fed MGA (0.5 x mg x cow(-1) x d(-1)) on d 1 to 7. On d 7, cows received either a single injection of PGF2alpha (Lutalyse sterile solution; 25 mg; n = 297), a single injection of cloprostenol sodium (Estrumate; 500 microg; n = 297), or half the recommended dose of PGF2alpha (12.5 mg; n = 275) on d 7 and 8. On d 10, all cows were artificially inseminated and received 100 microg of GnRH. Pregnancy rates to the timed artificial insemination (39%) were not affected by treatment, herd, or treatment x herd. There was an effect (P < 0.01) of artificial insemination sire on timed artificial insemination pregnancy rate for one herd, but not the other two herds. Herd influenced (P < 0.05) 30-d pregnancy rates, but there were no treatment or treatment x herd effects as 72.3% of the cows became pregnant during the first 30 d of the breeding season. Results indicate that the type of PGF2alpha treatment administered 7 d after GnRH did not influence timed artificial insemination pregnancy rates in nonlactating Bos indicus x Bos taurus cows.

Efficacy of an intravaginal progesterone insert and an injection of PGF2alpha for synchronizing estrus and shortening the interval to pregnancy in postpartum beef cows, peripubertal beef heifers, and dairy heifers.

The objective was to test the efficacy of an intravaginal progesterone insert and injection of PGF2alpha for synchronizing estrus and shortening the interval to pregnancy in cattle. Cattle were assigned to one of three treatments before a 31-d breeding period that employed artificial insemination. Control cattle were not treated, and treated cattle were administered PGF2alpha or an intravaginal progesterone-releasing insert (CIDR) for 7 d and treated with PGF2alpha on d 6. The treatments were applied in one of three experiments that involved postpartum beef cows (Exp. 1; n = 851; 56+/-0.6 d postpartum), beef heifers (Exp. 2; n = 724; 442.5+/-2.8 d of age), and dairy heifers (Exp. 3; n = 260; 443.2+/-4.5 d of age). Luteal activity before treatment was determined for individual cattle based on blood progesterone concentrations. In Exp. 1, there was a greater incidence of estrus during the first 3 d of the breeding period in CIDR+PGF2alpha-treated cows compared with PGF2alpha-treated or control cows (15, 33, and 59% for control, PGF2alpha, and CIDR+PGF2alpha, respectively; P < 0.001). The improved estrous response led to an increase in pregnancy rate during the 3-d period (7, 22, and 36% for control, PGF2alpha, and CIDR+PGF2alpha, respectively; P < 0.001) and tended to improve pregnancy rate for the 31-d breeding period for cows treated with CIDR+PGF2alpha, (50, 55, and 58% for control, PGF2alpha, and CIDR+PGF2alpha, respectively, P = 0.10). Improvements in rates of estrus and pregnancy after CIDR+PGF2alpha, were also observed in beef heifers. Presence of luteal activity before the treatment period affected synchronization and pregnancy rates because anestrous cows (Exp. 1) or prepubertal heifers (Exp. 2) had lesser synchronization rates and pregnancy rates during the first 3 d of the breeding period as well as during the entire 31-d breeding period. The PGF2alpha, and CIDR+PGF2alpha but not the control treatments were evaluated in dairy heifers (Exp. 3). The CIDR+PGF2alpha-treated heifers had a greater incidence of estrus (84%) during the first 3 d of the breeding period compared with the PGF2alpha-treated heifers (57%), but pregnancy rates during the first 3 d or during the 31-d breeding period were not improved for CIDR+PGF2alpha compared with PGF2alpha-treated heifers. In summary, the concurrent treatment of CIDR and PGF2alpha improved synchronization rates relative to PGF2alpha alone or control. Improved estrus synchrony led to greater pregnancy rates for beef cows and beef heifers but failed to improve pregnancy rates for dairy heifers.

Effectiveness of GnRH plus prostaglandin F2alpha for estrus synchronization in cattle of Bos indicus breeding.

Postpartum and lactating crossbred cows containing a percentage of Bos indicus breeding at three locations were studied to determine the efficacy of GnRH + PGF2alpha combinations for synchronization of estrus and(or) ovulation. Cows were equally distributed to each of three treatments by body condition score at the start of the experiment (d 0). All cows received 100 microg of GnRH on d 0 and 25 mg of PGF2alpha 7 d later. The three insemination protocols included 1) AI 12 h after exhibiting estrus during d 7 to 12 of the experiment (Select-Synch; n = 197); 2) timed-AI + 100 microg of GnRH on d 9 of the experiment (CO-Synch; n = 193); 3) AI 12 h after exhibiting estrus during d 7 to 10 of the experiment. Cows not exhibiting estrus by d 10 were timed-AI and injected with 100 microg of GnRH on d 10 of the experiment (Hybrid-Synch; n = 200). The percentage of cows exhibiting estrus during d 7 to 12 of the experiment was lower (P < 0.05) for CO-Synch (17.6%) cows than for Select-Synch or Hybrid-Synch (45.2 and 33.0%, respectively) cows, which did not differ (P > 0.05). For the Select-Synch and Hybrid-Synch cows that exhibited estrus during d 7 to 10 of the experiment and were artificially inseminated, conception rates were similar across treatments (50.5%). Pregnancy rates were greater (P < 0.01) for CO-Synch and Hybrid-Synch (31.0 and 35.5%, respectively) cows than for Select-Synch (20.8%) cows. A greater (P < 0.01) percentage of cycling cows became pregnant (34.5%) than noncycling cows (25.9%) across all treatments. The CO-Synch and Hybrid-Synch synchronization protocols resulted in greater pregnancy rates compared with the Select-Synch protocol in postpartum and lactating crossbred cows containing a percentage of Bos indicus breeding.

Ovulation and estrus characteristics in crossbred Brahman heifers treated with an intravaginal progesterone-releasing insert in combination with prostaglandin F2alpha and estradiol benzoate.

Crossbred Brahman heifers (n = 60) were studied to determine the effect of a 7-d intravaginal progesterone-releasing insert (INSERT) in combination with PG (Lutalyse; 25 mg i.m.) and estradiol benzoate (EB; .5 mg i.m.) on time of ovulation and estrous behavior. In Phase I, heifers at unknown stages of the estrous cycle were assigned by BW and body condition score to one of the three treatments on d 0: 1) INSERT for 7 d and PG on d 7 (CONTROL; n = 10); 2) INSERT for 7 d, PG on d 7, and EB 24 h after INSERT removal (EB24; n = 10); or 3) INSERT for 7 d, PG on d 7, and EB 48 h after INSERT removal (EB48; n = 10). Blood samples were collected every 8 h after INSERT removal. Also, blood sampling and ultrasonography began 8 h after the onset of estrus, determined with HeatWatch devices, and every 4 h thereafter to detect ovulation. In Phase II, Phase-I treatments (n = 10/treatments) were replicated, but only behavioral estrus data were collected to minimize handling of heifers. Frequent handling of heifers did not influence (P > .1) the interval from INSERT removal to the onset of HeatWatch and visual estrus and duration of estrus, so behavioral estrus data were combined for Phases I and II. Interval from INSERT removal to HeatWatch estrus was decreased (P < .05) in EB24 (45.5 h) vs EB48 (55.9 h) and CONTROL (59.2 h). Interval from INSERT removal to ovulation differed (P < .04) between CONTROL, EB24, and EB48 (93.5, 74.5, and 78.9 h, respectively). Ovulatory follicle size was similar (P > .1) between CONTROL, EB24, and EB48 (14.4, 12.5, and 14.1 mm, respectively). Duration of estrus was similar for CONTROL, EB24, and EB48 (14.0, 15.1, and 17.6 h, respectively). No difference (P > . 1) was observed in number of mounts received between CONTROL, EB24, and EB48 (28.0, 25.7, and 39.4, respectively), but number of mounts received increased in Phase II vs Phase I (40.0 and 22.2, respectively; P < .05). In conclusion, EB hastened the interval from INSERT removal to ovulation without altering duration of estrus or number of mounts received. Frequent handling of heifers did not affect interval to first mount received after INSERT removal or duration of estrus, but it decreased the total number of mounts received.

Endometrial responsiveness to oxytocin during diestrus and early pregnancy in pigs is not controlled solely by changes in oxytocin receptor population density.

These studies were performed to test the hypotheses that: 1) endometrial responsiveness to oxytocin (OT) in pig endometrium is associated with changes in OT receptor (OTr) population density resulting from corresponding regulation of OTr gene transcription, 2) endometrial responsiveness to OT is controlled solely through a mechanism involving changes in OTr population density, and 3) OTr population density and endometrial responsiveness to OT differ between diestrus and early pregnancy in pigs. In experiment 1, OTr population density and dissociation constant (Kd) in cyclic pigs were constant on Days 10-16 but increased (p < 0.05) between Days 10 and 12 of pregnancy before decreasing (p < 0.05) through Day 16. OT induced phosphoinositide (PI) hydrolysis and prostaglandin (PG) F2 alpha secretion in cyclic pigs only on Day 16 (p < 0.05), and during pregnancy only on Day 12 (p < 0.05). Activation of G protein by aluminum fluoride (AIF4-) treatment maximally stimulated (p < 0.05) PI hydrolysis and PGF2 alpha secretion in cyclic pigs on all days, indicating that downstream from the OTr, the PGF2 alpha secretory pathway was fully functional. During pregnancy, PI hydrolysis and PGF2 alpha secretion in response to AIF4- decreased (p < 0.01) on Days 14 compared to Days 10 and 12, and AIF4- did not stimulate PGF2 alpha release on Day 16. In experiment 2, abundance of OTr mRNA in cyclic pigs decreased between Days 0 and 5 before increasing between Days 5 and 12 (p < 0.05), but it was higher (p < 0.05) on Days 10-15 of pregnancy than on equivalent days in cyclic gilts. These results indicate that control of PGF2 alpha secretion in cyclic pigs appeared to occur primarily at the level of OTr coupling to G protein because changes in OTr number were not associated with increased sensitivity to OT or G-protein activation by AIF4-. During pregnancy, control was exerted at multiple levels, which included the OTr, G protein, phospholipase C, and subsequent aspects of the secretory pathway. The present study also indicated that endometrium was responsive to OT during luteolysis in cyclic pigs but not during corpus luteum maintenance in pregnant pigs.

Expression of an inter-alpha-trypsin inhibitor heavy chain-like protein in the pig endometrium during the oestrous cycle and early pregnancy.

In pigs, changes in an unidentified endometrial glycoprotein, pGP30, are temporally associated with rapid trophoblast elongation and initial placental attachment on day 12 of gestation. Identification of endometrial pGP30 was undertaken through protein purification, NH2-terminal amino acid sequencing and cDNA sequencing of products generated through reverse transcription-polymerase chain reaction. Sequencing of 35 amino acids from the NH2-terminal end of pGP30 revealed that the 30 kDa glycoprotein is a cleavage product from the C-terminal region of inter-alpha-trypsin inhibitor heavy chain 4 (I alpha IH4), previously known as inter-alpha-trypsin inhibitor heavy chain-like protein. I alpha IH4 is unique compared with the three other inter-alpha-trypsin inhibitor heavy chains as it does not contain a binding site for bikunin that has serine protease inhibitory activity and is sensitive to cleavage by kallikrein. Endometrial gene expression of I alpha IH4 was detected during the oestrous cycle (days 0-18) and early pregnancy (days 10-18). Gene expression of I alpha IH4 appeared to be enhanced during the midluteal phase (days 12 and 15) of the oestrous cycle and the period of trophoblast attachment (days 12-18). Expression of I alpha IH4 was not detected in day 12 conceptus tissue mRNA. Endometrial expression of I alpha IH4 in pigs may function as an acute phase protein for protection of the uterus from the inflammatory response induced by conceptus attachment to the uterine epithelium.

Ontogeny of elongation and gene expression in the early developing porcine conceptus.

Early porcine conceptus development is characterized by rapid trophoblastic elongation between Days 11 and 12 of pregnancy, a period of embryonic loss in the pig. Growth factors and steroids secreted by the conceptus and uterus, as well as ligand receptors produced by the conceptus, are thought to regulate trophoblastic elongation. Therefore, the objectives of this study were to characterize conceptus gene expression for the steroidogenic enzymes 17alpha-hydroxylase and aromatase and the mesodermal marker brachyury, as well as the expression of receptors for fibronectin (integrin beta-1), progesterone, estrogen, oxytocin, prostaglandin F2alpha, and leukemia inhibitory factor (LIF), prior to and during trophoblastic elongation. Total RNA was extracted from individual conceptuses from Day 10 to Day 12 of pregnancy. Gene expression was determined by reverse transcription polymerase chain reaction on conceptuses having 2- to 4-, 5-, 6-, 7-, 8-, 9-, and 10- to 12-mm spherical, 13- to 25-mm tubular, and > 100-mm filamentous morphologies. There was a stage of development effect on both 17alpha-hydroxylase (p < 0.001) and aromatase (p < 0.003) gene expression. Initial 17alpha-hydroxylase gene expression was detected in early spherical conceptuses (2-4 mm), increasing abruptly through to 7-mm conceptuses. Aromatase gene expression increased dramatically in 6- to 7-mm conceptuses, with increased expression throughout development. Gene expression for LIF receptor (LIFR) (p < 0.02) was similar to that for 17alpha-hydroxylase, while brachyury gene expression began in 6-mm conceptuses and increased (p < 0.001) throughout development. Integrin beta-1 was expressed at all stages of development. Conceptus gene expression was not detected for progesterone, estrogen, oxytocin, and prostaglandin F2alpha receptors. Prior to elongation, dynamic changes in gene expression are occurring that appear to be associated with estrogen production and preparation of the conceptuses for elongation. LIFR expression is highly associated with steroidogenic enzyme production with an initial peak preceding rapid trophoblastic elongation, suggesting that LIF may be involved in early conceptus development in the pig.

Detection of transcripts for retinoic acid receptors, retinol-binding protein, and transforming growth factors during rapid trophoblastic elongation in the porcine conceptus.

Early conceptus development (Days 10-12) in the pig is characterized by rapid remodeling and elongation of the trophoblastic membrane. Endometrial synthesis and secretion of retinol-binding protein (RBP; transporter of retinol) into the uterine lumen parallels the rapid morphological transformation of the conceptus. Hence, retinoic acid, a powerful morphogen, may serve a critical function in trophoblastic remodeling and conceptus elongation through the various isoforms of retinoic acid receptor (RAR). We hypothesize that various transcripts for RAR will be expressed, possibly differentially, in the early developing porcine conceptus on Days 10-12 of gestation. Conceptuses were collected from pregnant gilts and individually frozen in liquid nitrogen after classification for morphological size. A total of 36 conceptuses (4 groups of 9) representing spherical (2-9 mm), early tubular (10-12 mm), late tubular (13-60 mm), and filamentous (> 100 mm) morphologies were assembled for qualitative assessment of mRNA for RAR alpha, RARbeta, RARgamma, RBP, and transforming growth factor beta (TGFbeta)-2 and -3 with reverse transcription polymerase chain reaction (RT-PCR). RBP gene expression was affected by stage of conceptus development (p < 0.05) as RBP expression increased in a linear fashion from 2-mm to 8-mm spherical conceptuses, decreased slightly in the tubular stage, and increased dramatically in filamentous conceptuses. RAR alpha, RARbeta, and RARgamma gene expression was evident at all stages of conceptus development. RAR alpha gene expression increased (p < 0.05) in a linear fashion from 2- to 8-mm spherical conceptuses, and expression plateaued in the 9- to 12-mm conceptuses. Expression of RAR alpha declined slightly at the tubular stage but dramatically increased in filamentous conceptuses. RARbeta gene expression was present across all stages of conceptus development although in diminutive amounts. RARgamma gene expression was evident in all stages of conceptus development but was not affected (p > 0.05) by developmental stage. Gene expression for TGFbeta-2 was not detected by RT-PCR at any stage of conceptus development, whereas TGFbeta-3 expression increased (p < 0.05) from the 2- to 8-mm conceptuses and remained high throughout trophoblastic elongation into the filamentous stage. The presence of mRNA for the RAR alpha, RARbeta, and RARgamma in elongating porcine conceptus provides evidence that RAR, particularly RAR alpha, may function as an important acceptor for retinoic acid and possibly regulate gene expression prior to and during rapid trophoblastic elongation in the porcine conceptus.