Influence of perioestrous suprabasal progesterone levels on cycle length,oestrous behaviour and ovulation in heifers

In order to induce suprabasal plasma concentrations of progesterone after luteolysis and to determine their effect on oestrous behaviour and ovulation, heifers subcutaneously received silicone implants containing 2.5 (n = 4), 5 (n = 4), 6 (n = 3), 7.5 (n = 3) or 10 (n = 4) g of progesterone, or an empty implant (controls, n = 5) between days 8 and 25 of the cycle (ovulation designated Day 0). Growth of dominant follicles and time of ovulation were determined by ultrasound, and signs of oestrus were recorded and scored. Blood was collected at 2–4 h intervals from Days 15 to 27 and assayed for progesterone concentration. In all heifers, plasma concentrations of progesterone sharply decreased during Days 16–18. Control heifers had their lowest progesterone levels on Days 20.5 and 21, standing oestrus on Day 19.5 ± 0.4 (mean ± SEM), and ovulated on Day 20.7 ± 0.4. A similar pattern was observed in heifers treated with 2.5 and 5 g progesterone. Heifers treated with 6, 7.5 and 10 g of progesterone showed an extended (P < 0.05) interovulatory interval. Onset of prooestrus and time of maximum expression of signs of oestrus were not significantly different from those in controls. However, there was an absence of standing oestrus in most of the cases, signs of oestrus lasted longer (P < 0.05) and were weaker in intensity when doses increased. In these groups, the lowest progesterone concentrations were attained shortly after implant removal. Some heifers treated with 6 and 7.5 g of progesterone had standing oestrus and post oestrous bleeding as seen in the controls but ovulation occurred from Days 24.5 to 27. When plasma progesterone concentrations were over 1 nmol 1⁻¹, disturbed oestrus and delayed ovulation occurred. The extended period of prooestrus and oestrus and delayed ovulation were similar to that described in cases of repeat breeding. It is suggested that suprabasal plasma concentrations of progesterone, after luteolysis, may lead to asynchrony between onset of oestrus and ovulation and consequently be a cause of repeat breeding in cattle.     

Ovulatory responses and plasma luteinizing hormone concentrations in dairy heifers after treatment with exogenous progesterone and estradiol benzoate

The objectives of this experiment were to determine the effects of 0.5 mg estradiol benzoate, administered intramuscularly 24 h after removal of CIDR-B progesterone containing intravaginal devices, on the time to estrus, ovulation and peak LH concentration in dairy heifers. Ovulatory responses and plasma LH concentrations were examined using 14 Friesian dairy heifers in 2 separate treatment periods. All heifers received a CIDR-B progesterone-containing intravaginal device with an attached 10-mg estradiol benzoate capsule for 12 d. Within each period, 24 h after CIDR-B removal, 7 heifers received an intramuscular injection of 0.5 mg estradiol benzoate while the remaining 7 heifers received an intramuscular injection of a placebo. Blood samples for LH assay were collected at 0, 6 and 12 h, and then every 4 h for 60 h after estradiol injection. Detection of estrus was conducted at 4-h intervals, and ultrasonographical examination to detect ovulation was conducted every 8 h for 88 h after removal of the CIDR-B device. Treatment with estradiol benzoate tended to reduce the time from device removal to the LH peak in Period 1 (median time to LH peak 40.1 vs 63.9 h; P = 6.07). In Period 2, treatment with estradiol had no significant effect on the time to the LH peak, standing estrus or ovulation. We hypothesize that the period effect was due to the stage of cycle at the time of treatment. For heifers treated in Period 1, the stage of cycle was random. However, because of the prior synchronization of estrus, which was implicit in the experimental design, heifers in Period 2 tended to be in late diestrus. The administration of estradiol benzoate after treatment with exogenous progesterone appears to overcome the variability in timing of LH peaks typically occurring in a herd of synchronized heifers due to different stages of follicular development.     

Influence of low plasma progesterone concentrations on the release of prostaglandin F2α during luteolysis and oestrus in heifers

A study was conducted to determine the effect of suprabasal plasma concentrations of progesterone on the release of prostaglandin F_2α (PGF_2α) at luteolysis and oestrus. Heifers received silicone implants containing 2.5 (n = 4), 5 (n = 4), 6 (n = 3), 7.5 (n = 3), 10 (n = 4), or 15 (n = 3) g of progesterone, or an empty implant (controls, n = 4) between Days 8 and 25 post ovulation. Blood was collected frequently between Days 14 and 28 and assayed for progesterone and 15-ketodihydroprostaglandin F_2α. Basal progesterone concentrations in control heifers did not differ from those in heifers with 2.5- or 5-g implants and remained around 0.4−0.5 nmol l⁻¹ until ovulation in all three groups. In the heifers treated with 6–15 g of progesterone, basal concentrations were maintained at higher (P < 0.05) levels compared with those in the controls, ranging from 0.8 to 1.6 nmol 1⁻¹. The effect of these elevated progesterone levels was to delay ovulation by prolonging the growth of the ovulatory follicle, which continued growing until the implant was removed. In all experimental groups, the first significant increase of the PGF_2α metabolite occurred between Days 15.3 and 16.3 (P > 0.05) and was associated with the onset of a decrease in progesterone concentrations, which had reached levels below 3 nmol 1⁻¹ by Days 17.4−19.1. PGF_2α metabolite peaks associated with luteolysis were frequent until Day 20. In the period from Day 20 until implant removal, sporadic peaks were observed, ranging in number from 1.0 ± 1.2 (mean ± SEM) in the control group to 3.0 ± 1.4 peaks in the heifers treated with 7.5 g of progesterone (P > 0.05). The number of PGF_2α metabolite peaks during that period was higher (P < 0.05) in heifers treated with 10 and 15 g than in controls. A positive correlation was found between the basal concentration of progesterone and the number of PGF_2α peaks after luteolysis (r = 0.54; P < 0.01). Plasma progesterone concentrations above approximately 1.4 nmol l⁻¹ were able to maintain the release of PGF_2α until the progesterone implants were removed and plasma levels decreased to basal values. These heifers had a preovulatory PGF_2α release pattern resembling that found in repeat breeder heifers.     

Estradiol induces and progesterone inhibits the preovulatory surges of luteinizing hormone and follicle-stimulating hormone in heifers

Objectives were to determine: 1) whether estradiol, given via implants in amounts to stimulate a proestrus increase, induces preovulatory-like luteinizing hormone (LH) and follicle-stimulating hormone (FSH) surges; and 2) whether progesterone, given via infusion in amounts to simulate concentrations found in blood during the luteal phase of the estrous cycle, inhibits gonadotropin surges. All heifers were in the luteal phase of an estrous cycle when ovariectomized. Replacement therapy with estradiol and progesterone was started immediately after ovariectomy to mimic luteal phase concentrations of these steroids. Average estradiol (pg/ml) and progesterone (ng/ml) resulting from this replacement were 2.5 and 6.2 respectively; these values were similar (P greater than 0.05) to those on the day before ovariectomy (2.3 and 7.2, respectively). Nevertheless, basal concentrations of LH and FSH increased from 0.7 and 43 ng/ml before ovariectomy to 2.6 and 96 ng/ml, respectively, 24 h after ovariectomy. This may indicate that other ovarian factors are required to maintain low baselines of LH and FSH. Beginning 24 h after ovariectomy, replacement of steroids were adjusted as follows: 1) progesterone infusion was terminated and 2 additional estradiol implants were given every 12 h for 36 h (n = 5); 2) progesterone infusion was maintained and 2 additional estradiol implants were given every 12 h for 36 h (n = 3); or 3) progesterone infusion was terminated and 2 additional empty implants were given every 12 h for 36 h (n = 6). When estradiol implants were given every 12 h for 36 h, estradiol levels increased in plasma to 5 to 7 pg/ml, which resembles the increase in estradiol that occurs at proestrus. After ending progesterone infusion, levels of progesterone in plasma decreased to less than 1 ng/ml by 8 h. Preovulatory-like LH and FSH surges were induced only when progesterone infusion was stopped and additional estradiol implants were given. These surges were synchronous, occurring 61.8 +/- 0.4 h (mean +/- SE) after ending infusion of progesterone. We conclude that estradiol, at concentrations which simulate those found during proestrus, induces preovulatory-like LH and FSH surges in heifers and that progesterone, at concentrations found during the luteal phase of the estrous cycle, inhibits estradiol-induced gonadotropin surges. Furthermore, ovarian factors other than estradiol and progesterone may be required to maintain basal concentrations of LH and FSH in heifers.     

Plasma progesterone profiles and fertility status of anestrus Zebu cattle treated with norgestomet-estradiol-eCG regimen

Zebu cattle are notorious for poor fertility characterized by late maturity and long intercalving intervals attributed to a variety of factors, including genetic, nutritional and climatic. The aim of the present investigation, therefore, was to induce fertile estrus in acyclic pubertal heifers and postpartum anestrous Zebu cows by hormonal intervention. Pubertal Hariana and Sahiwal anestrous heifers (n=51) and postpartum cows (n=55) were either assigned a placebo (controls, N=6 for each breed and parity) or treated with 10-d norgestomet (3 mg) subcutaneous ear implants, with an initial injection of 3 mg, im norgestomet + 5 mg estradiol valerate, followed by 500 IU eCG at implant withdrawal (NOR-treated groups). Jugular venous plasma samples were obtained from a total of 28 animals (controls : 4 heifers and 4 cows; NOR-treated : 12 heifers and 8 cows) on Days 0 (implant insertion), 3, 7, 9 and Day 10 (implant withdrawal), every 12 h on Days 11 and 12, and then once daily on Days 17, 24 and 31. All the samples were assayed for progesterone. Almost all (97%) heifers and 81% cows were induced to estrus, the majority (92% heifers and 79% cows) within 120 h of implant removal. Synchrony of the induced estrus was better in cows, but interval to estrus and estrus duration were significantly longer in heifers (P<0.05). Post-treatment fertility, based on Day 28 nonretum rate, first service, and overall conception rates, was better in heifers (78.9, 60.5 and 73.7%, respectively) than cows (77.1, 48.6 and 62.9%, respectively), but the differences were significant only for the overall pregnancy rate (71.8% for heifers and 51.2% for cows; P<0.05). Low pre-treatment plasma progesterone values (<0.5ng/mL) were consistent with ovarian inactivity, confirming the true anestrus status of experimental animals. Controls failed to exhibit estrus and maintained low progesterone concentrations throughout the study. In treated animals, high progesterone values from Day 17 onwards suggested ovulatory estrus. These early luteal phase progesterone concentrations in nonpregnant (P=0.06) and nonpregnant, nonretum (P<0.05) animals were low in comparison with those of pregnant animals. Good fertility resulting from breeding according to estrus, inspite of variable intervals to estrus and estrus duration, advocates its advantage over fixed-time insemination in norgestomet-treated anestrous Zebu cattle.     

Luteal function and estrus in peripubertal beef heifers treated with an intravaginal progesterone releasing device with or without a subsequent injection of estradiol

The objectives of this experiment were to determine if treatment of beef heifers with progesterone (P4) using an intravaginal device alone or in combination with estradiol benzoate (EB) would induce estrus and cause development of corpora lutea (CL) with a typical life span. Peripubertal heifers (n = 311) were used when about 40% of the heifers had a functional CL. The heifers were assigned to receive one of the following treatments on Day 0: 1) a sham device for 7 d (C, n = 108); 2) an intravaginal device containing P4 for 7 d (P, n = 102); or 3) an intravaginal device containing P4 for 7 d plus an injection of 1 mg EB 24 to 30 h after device removal (PE, n = 101). Serum concentrations of P4 were determined on Days -7, 0, 8, 15 and 22. Weight and age of the heifers at the start of the trial averaged 292 +/- 45 kg and 365 +/- 38 d, respectively. A greater (P < 0.0001) proportion of the heifers from the PE than P group was in standing estrus (81 vs 37%) and formed normal CL (68 vs 44%) after device removal. Of the heifers exhibiting estrus, a greater (P < 0.05) proportion of PE (94%) than P (80%) heifers was active 1 to 3 d after implant removal. Short-term progesterone treatment increased the proportion of heifers in estrus and those forming normal CL, and adding EB to the progesterone treatment further enhanced these responses.     

Reproductive performance of heifers induced to oestrous asynchrony by suprabasal plasma progesterone levels

Suprabasal progesterone concentrations around oestrus have induced disturbances in oestrous behaviour and ovulation. To determine whether fertility in such an altered oestrus can be maintained at normal levels with additional inseminations (AI) until ovulation, fertility was compared in heifers (n = 11) inseminated in normal oestrous cycles and thereafter in cycles in which the animals were treated with progesterone in order to create suprabasal concentrations after luteolysis. The treatment consisted of silicone implants containing 10.6 mg kg⁻¹ of progesterone inserted subcutaneously on Day 8 of the oestrous cycle (day of ovulation designated Day 0) and removed on Day 25. Both in control oestrous cycles and oestrous cycles under progesterone treatment, growth of the ovulatory follicle and ovulation were determined by frequent ultrasound scanning. Blood was collected frequently for further analysis of progesterone, oestradiol-17β and luteinising hormone (LH). Insemination was performed 12 h after onset of standing oestrus. if ovulation did not occur 24 h after AI, heifers were inseminated again until ovulation. Pregnancy was diagnosed by ultrasound 25 days after ovulation.In control oestrous cycles, plasma progesterone decreased to 0.3 ± 0.3 nmol 1⁻¹. Duration of oestrus was 22.9 ± 2.0 h, the interval from onset of oestrus to ovulation was 32.4 ± 2.3 h and the interval from LH peak to ovulation was 28.6 ± 1.4 h. The interovulatory interval was 20.7 ± 0.6 days. In oestrous cycles in treated heifers, progesterone decreased to 1.0 ± 0.3 nmol l⁻¹ (P > 0.10) and the interovulatory interval was prolonged to 23.5 ± 1.0 days (P < 0.05). Standing oestrus lasted 47.2 ± 12.0 h (P = 0.09, n = 7). The interval from the onset of oestrus to ovulation was 59.4 ± 13.0 h (P = 0.08) and the interval from LH peak to ovulation 25.8 ± 1.3 h (P > 0.10). The prolonged oestrus was associated with increased (P < 0.05) growth of the ovulatory follicle and higher (P < 0.05) release of oestradiol-17β. Conception rates were 90% and 46% (P < 0.05), and the numbers of AI per heifer were 1.1 ± 0.1 and 3.4 ± 0.6 (P < 0.01) for control oestrous cycles and after treatment, respectively.The induction of suprabasal concentrations of progesterone caused asynchronies similar to those observed in cases of repeat breeding. The repeated AI did not maintain fertility at normal levels. It is suggested that the extended growth of the ovulatory follicle may cause impaired oocyte maturation or it may alter the maternal milieu owing to the prolonged release of oestradiol.     

Variable progesterone response and estradiol secretion in prepubertal beef heifers following treatment with norgestomet implants

Two experiments were conducted to determine the effects of norgestomet ear implants on progesterone response and estradiol secretion in prepubertal beef heifers. In the first experiment, 47 beef heifers were treated with norgestomet. The implants were implanted subcutaneously for 9 d. After implant removal, blood samples were taken from heifers 2 to 4 d per week for 40 d. Following progesterone determination in jugular venous plasma, heifers were classified according to their progesterone response: 1) no response (Group 1); no rise in progesterone above 1 ng/ml throughout the sampling period; 2) one cycle (Group 2); one increase in progesterone above 1 ng/ml for at least 2 d followed by no further increase in progesterone during the sampling period; and 3) two cycles (Group 3); a rise in progesterone above 1 ng/ml for at least 2 d followed by another cycle of normal duration. Heifers treated with norgestomet were classified as 23 with no response, 9 with 1 cycle and 15 with 2 cycles. Concentrations of estradiol were measured in jugular venous samples on Day 2 after implant removal. Mean concentrations of estradiol were greater in Group 3 than in Group 1 (P < or = 0.01). In Experiment 2, 29 prepubertal beef heifers were assigned randomly to either a 9-d treatment with norgestomet (n = 14) or to serve as untreated controls (n = 15). Blood plasma samples were collected daily from Days 0 to 44 after implant removal. After progesterone determination, heifers were classified as 8 with no response, 4 with 1 cycle and 3 with 2 cycles in the control group, and 5 with no response, 3 with 1 cycle and 6 with 2 cycles in the norgestomet group (frequencies did not differ; P > 0.1). Jugular venous blood plasma was also collected at 4-h intervals from 0 h to 96 h after implant removal and concentrations of estradiol were measured. Patterns of estradiol secretion differed (P < or = 0.05) and overall mean concentrations of estradiol over the first 96 h following implant removal were greater (P < or = 0.01) in norgestomet-treated heifers versus the controls. We conclude that norgestomet can produce a variable progesterone response with heifers with 2 cycles secreting more estradiol. Implants of norgestomet also causes more acute secretion of estradiol in prepubertal beef heifers.     

Progesterone and estradiol in biodegradable microspheres for control of estrus and ovulation in mares

Progesterone and estradiol 17-beta in poly (DL-lactide) microspheres were used to control estrus and ovulation in mares after luteolysis was induced by prostaglandin F(2)infinity. Mares were given a single intramuscular injection of biodegradable poly (DL-lactide) microspheres, 1 day following prostaglandin treatment, containing no hormones (control), 0.625 g progesterone and 50 mg estradiol (low dose), 1.25 g progesterone and 100 mg estradiol (medium dose), or 1.875 g progesterone and 150 mg estradiol (high dose; n=15 mares per group). Mares treated with the low dose had significantly longer intervals (P<0.05) to estrus and ovulation than the control mares; however, low dose mares had shorter intervals (P<0.05) to estrus than high dose mares and shorter intervals to ovulation than medium and high dose mares. Regression analysis indicated that the medium dose was sufficient for maximizing interval to ovulation while the high dose maximized interval to estrus. All groups of mares exhibited similar (P>0.05) post-treatment estrus lengths. A clinical response scoring system based on synchrony of both estrus and ovulation within a treatment group was also used to measure the effectiveness of treatments on control of estrus and ovulation. Clinical response scores did not differ (P>0.05) among treatment groups. Mares were randomly assigned for insemination at the beginning of the first post-treatment estrus. Rates for embryo recovery performed by uterine lavage 7 days post-ovulation did not differ (P>0.05) among groups. Concentrations of serum progesterone increased in mares receiving progesterone and estradiol microspheres. At 10 to 14 days post-injection of microspheres, progesterone concentrations were higher (P<0.05) and remained above 1 ng/ml in the mares receiving the high dose. Progesterone concentrations were also higher (P<0.05) on Days -3 to -1 (Day 0 = day of post-treatment ovulation) in mares receiving the high dose when compared to control mares. Gonadotropin concentrations were suppressed (P<0.05) in the medium and high dose groups.     

Site of action for endotoxin-induced cortisol release in the suppression of preovulatory luteinizing hormone surges

The study was conducted to identify the mechanisms of endotoxin/cortisol action in the suppression of preovulatory LH surges in heifers infused with Escherichia coli (E. coli ) endotoxin. The hypotheses tested were that 1) endotoxin stimulates the release of progesterone, possibly from the adrenal leading to the LH blockade; 2) cortisol released in response to endotoxin infusion blocks the synthesis of estradiol at the ovarian level, culminating in a failure of the LH surge. Eight Holstein heifers were given two injections of prostaglandin F(2alpha) (PG), 11 d apart, to synchronize estrus. Starting from 25 h after the second injection of PG (PG-2), the uterus of each heifer was infused either with 5 ml of pyrogen-free water (control, n = 3) or with E. coli endotoxin (5 mug/kg of body weight) in 5 ml of pyrogen-free water (treated, n = 5), once every 6 h for 10 treatments. Blood samples were obtained every 15 min for 1 h before infusion and again 2 h after each infusion, then hourly until 1 h before the next infusion. After the tenth infusion, blood was collected daily until estrus. Serum progesterone concentrations remained at baseline values (< 1 ng/ml) in control and treated heifers. The total amount of progesterone measured starting 24 to 84 h after PG-2 injection was not different between control and treated heifers (P 0.05). In the control heifers, serum estradiol concentrations remained basal (< 10 pg/ml) until 4 h before the LH surge. Serum estradiol concentrations increased to 20 +/- 5.6 pg/ml, 4 h before the LH surge in control heifers (LH surge occurred 60 to 66 h after the PG-2 injection). There were no changes in serum estradiol concentrations in treated heifers during the sampling period, and the concentrations remained < 10 pg/ml. The total amount of estradiol measured in control heifers was higher (P < 0.05) than in treated heifers. The results if this study suggest that increases in cortisol concentrations after the infusion of endotoxin might block the synthesis of estradiol at the ovarian level, resulting in the failure of a preovulatory LH surge to occur.     

Controlling interrelationships of progesterone/LH and estradiol/LH in mares

The interrelationships of progesterone, estradiol, and LH were studied in mares (n=9), beginning at the first ovulation (Day 0) of an interovulatory interval. An increase in mean progesterone concentrations began on Day 0 and reached maximum on Day 6, with luteolysis beginning on Day 14. A common progesterone threshold concentration of about 2 ng/ml for a negative effect on LH occurred at the beginning and end of the luteal phase. Progesterone and LH concentrations decreased at a similar rate from Day 6 until the onset of luteolysis on Day 14, consistent with a decreasing positive effect of LH on progesterone. Concentrations of LH during the increase in the ovulatory surge consisted of two linear regression segments involving a rate of 0.4 ng/ml/day for Days 14-22 and 1.8 ng/ml/day for Day 22 to 1 day after the second ovulation. The end of the first segment and beginning of the second segment was 2 days before ovulation and was the day the ovulatory estradiol surge was at a peak.     

Exogenous estradiol reduces inhibition of luteinizing hormone by estradiol in prepubertal heifers

The hypothesis that high levels of exogenous estradiol administered to heifers during the prepubertal period would decrease subsequent negative feedback of estradiol on luteinizing hormone (LH) secretion was tested. Fourteen prepubertal heifers were ovariectomized on Day 0. Ovariectomized heifers received either no further treatment (OVX, n = 4), a single estradiol implant on Day 0 (OVXE, n = 5), or the single implant on Day 0 and two additional implants between Days 16 and 30 (OVXE+ E, n = 5). Ten ovary-intact heifers received either no treatment (INT, n = 5) or were administered the two estradiol implants between Days 16 and 30 (INT+ 5, n = 5). Comparison of LH secretion in OVXE to OVXE+E, and in INT to INT+E resulted in significant time-by-treatment interactions (p less than 0.05 for both). As pubertal age approached, mean concentration of LH (p less than 0.05) and pulse frequency (p less than 0.05) increased more rapidly in OVXE+E and INT+E than in OVXE and INT, respectively. Amplitude of LH pulses was unaffected by treatment. When data were standardized to day of puberty in INT and INT+E heifers, mean LH concentration and LH pulse frequency increased as puberty approached in both groups. These data confirm earlier reports indicating that secretion of LH increases gradually as puberty approaches in heifers. It was concluded that administration of estradiol during the prepubertal period hastened the decline in the subsequent negative feedback of estradiol. Precocious puberty was not induced in ovary-intact females.     

LH and FSH concentration and follicular development in Nellore heifers submitted to fixed-time artificial insemination protocols with different progesterone concentrations

Nine heifers were pre-synchronized (PGF2α, 12 days) and assigned into three groups with 6 repetitions each: (1) CL (～8 days old, n=13); (2) DIB+CL (n=18); (3) DIB+EB (150 μg of PGF2α and 2mg estradiol benzoate, n=18). After progesterone (P4) device removal (8 days) and/or final PGF2α, heifers were injected with either GnRH or EB in a 3×2 factorial totalling 49 observations (5 were excluded). The blood sampling schedule: every 12 h during P4 period; for LH pulse frequency on Days 3-5, every 15 min for 6 h during P4 period; after P4 removal and EB treatment, samples were collected every 3 h for 24 h or after GnRH every 1 h for 10 h. Ovarian follicle number and diameter were evaluated by ultrasonography every 12 h until the last blood sample and then 24 h and 48 h later. After device insertion (12 h), the DIB+CL group had a lesser LH concentration than the DIB+EB group. After 36 h, all DIB+CL-treated heifers had less LH than CL-heifers, and after 60 h, the DIB+EB group had less LH than the CL-group. Considering all P4 groups combined, LH peak amplitude was greater after GnRH compared to EB treatment but total area of LH peak amplitude and time to first peak was less. The CL-group had fewer follicles and a greater largest follicle diameter than DIB+CL and DIB+EB groups. When treated with EB, the DIB+CL group had a lesser ovulation rate at 24 h than the CL- and DIB+EB-groups. Fixed time artificial insemination (FTAI) protocols promoted a pre-ovulatory LH peak, independent of previous exposure to the DIB coupled with a CL or not. The progesterone excess interfered with FSH and LH secretion, follicular development and ovulation within 24 h.     

Effects of immunization against GnRH, melengestrol acetate, and a trenbolene acetate/estradiol implant on growth and carcass characteristics of beef heifers

A 2 x 3 factorial experiment was conducted to determine the effects of an implant (trenbolene acetate/estradiol or no implant) and method of estrus suppression (immunization against GnRH, melengestrol acetate, or no suppression) on growth performance and carcass characteristics of heifers fed for slaughter. At the start of a 21-d feed adaption phase, crossbred beef heifers (n = 144, 390+/-2.8 kg) were given their first dose of an anti-GnRH vaccine or started on melengestrol acetate (MGA). Thereafter, heifers were fed a high-concentrate diet (78% barley grain) for 84 d (Days 0 to 83), received implants on Day 0, a second vaccination on Day 21, and were slaughtered on Days 84 or 85. Implanting increased average daily gain (1.72 vs 1.50 kg/d, P < 0.01), feed efficiency (6.02 vs 6.75 kg dry matter intake/kg gain, P < 0.01), preslaughter weight (532 vs 513 kg, P < 0.01), carcass weight (301 vs 289 kg, P < 0.01), and ribeye area (88.6 vs 85.9 cm2, P < 0.05), but had no affect (P > 0.05) on dry matter intake, grade fat thickness, marbling score, or lean yield. Compared to heifers fed MGA, those immunized against GnRH had a greater ribeye area (90.0 vs 84.6 cm2) and lean yield (63 vs 61%), and had thinner grade fat (7.5 vs 8.6 mm; P < 0.05 for each). Furthermore, immunized heifers had lower (P < 0.001) plasma progesterone concentrations than control heifers on Days 42, 63 and 83. Heifers fed MGA had less estrus mounting activity (P < 0.05) and lower plasma progesterone concentrations (P < 0.001) than the remaining heifers. Method of estrus suppression did not affect (P > 0.05) preslaughter weight, average daily gain, dry matter intake, feed efficiency, carcass weight, or marbling score. In conclusion, implanting significantly increased growth performance and preslaughter and carcass weights. Compared to heifers fed MGA, immunization against GnRH significantly increased ribeye area and lean yield, and reduced grade fat thickness     

Gonadotropin-releasing hormone at estrus: luteinizing hormone, estradiol, and progesterone during the periestrual and postinsemination periods in dairy cattle

Administering gonadotropin-releasing hormone (GnRH) improved conception rates in our previous studies. Our objective was to determine if the effect of GnRH was mediated through serum luteinizing hormone (LH) and/or by altered secretion of serum progesterone (P) and estradiol-17 beta (E) during the periestrual and post-insemination periods. Cattle were given either GnRH (n = 54) or saline (n = 55) at 72 h and inseminated artificially (AI) 80 h after the second of two injections of either prostaglandin F2 alpha or its analog, cloprostenol. Progesterone and E were measured in blood serum collected during 3 wk after AI (estrus) from 60 females. Blood was collected for LH determinations via indwelling jugular cannulae from 14 cows and 11 heifers. Collections were taken every 4 h from 32 to 108 h after the second PGF injection (PGF-2) (periestrual period) and at more frequent intervals during 240 min after administration of GnRH (n = 18) or saline (n = 7). Ten females had a spontaneous preovulatory LH surge before GnRH treatment (GnRH-spontaneous), whereas GnRH induced the preovulatory LH surge in six females. A spontaneous LH surge appeared to be initiated in two heifers at or near the time of GnRH treatment (spontaneous and/or induced). The remaining seven cows had spontaneous LH surges with no subsequent change in LH after saline treatment. Serum P during the 21 days after estrus was lower (p less than 0.05) in both pregnant and nonpregnant (open) cattle treated previously with GnRH compared with saline. Serum P during the first week after estrus was greater (p less than 0.01) and increased (p less than 0.05) more rapidly in saline controls and in GnRH-spontaneous cattle than in those exhibiting GnRH-induced or GnRH-spontaneous and/or-induced surges of LH. Conception rate of cattle receiving GnRH was higher (p = 0.06) than that of saline-treated controls. These data suggest that GnRH treatment at insemination initiated the preovulatory LH surge in some cattle, but serum P in both pregnant and open cows was compromised during the luteal phase after GnRH treatment. Improved fertility may be associated with delayed or slowly rising concentrations of serum progesterone after ovulation.     

Effects of buserelin injection and deslorelin (GnRH-agonist) implants on plasma progesterone, LH, accessory CL formation, follicle and corpus luteum dynamics in Holstein cows

The influence of Buserelin injection and Deslorelin (a GnRH analogue) implants administered on Day 5 of the estrous cycle on plasma concentrations of LH and progesterone (P4), accessory CL formation, and follicle and CL dynamics was examined in nonlactating Holstein cows. On Day 5 (Day 1 = ovulation) following a synchronized estrus, 24 cows were assigned randomly (n = 4 per group) to receive 2 mL saline, i.m. (control), 8 micrograms, i.m. Buserelin or a subcutaneous Deslorelin (DES) implant in concentrations of 75 micrograms, 150 micrograms, 700 micrograms or 2100 micrograms. Blood samples were collected (for LH assay) at 30-min intervals for 2 h before and 12 h after GnRH-treatment from cows assigned to Buserelin, DES-700 micrograms and DES-2100 micrograms treatments and thereafter at 4-h intervals for 48 h. Beginning 24 h after treatment, ovaries were examined by ultrasound at 2-h intervals until ovulation was confirmed. Thereafter, ultrasonography and blood sampling (for P4 assay) was performed daily until a spontaneous ovulation before Day 45. A greater release of LH occurred in response to Deslorelin implants than to Buserelin injection (P < 0.01). Basal levels of LH between 12 and 48 h were higher in DES-700 micrograms group than in DES-2100 micrograms and Buserelin (P < 0.05). The first wave dominant follicle ovulated in all cows following GnRH treatment. Days to CL regression did not differ between treatments, but return to estrus was delayed (44.2 vs 27.2 d; P < 0.01) in cows of DES-2100 micrograms group. All GnRH treatments elevated plasma P4 concentrations, and the highest P4 responses were observed in the DES-700 micrograms and DES-2100 micrograms groups. The second follicular wave emerged earlier in GnRH-treated than in control cows (9.9 vs 12.8 d; P < 0.01). However, emergence of the third dominant follicle was delayed in cows of DES-2100 micrograms treatment (37.0 d) compared with DES-700 micrograms (22.2 d), Buserelin (17.8 d) or control (19.0 d). In conclusion, Deslorelin implants of 700 micrograms increased plasma P4 and LH concentrations and slightly delayed the emergence of the third dominant follicle. On the contrary, Deslorelin implants of 2100 micrograms drastically altered the P4 profiles and follicle dynamics.     

Chronic stress, hormone profiles and estrus intensity in dairy cattle

The objectives of the present study were to determine if lameness, a model for a natural chronic stressor, affects hormone concentrations in milk prior to estrus and/or the subsequent expression of estrus in the postpartum period. Dairy cows>20 days postpartum were scored for lameness and observed for estrus intensity using a weighted scoring system (>100 points=estrus=Day 0). Increasing lameness score was not associated with daily profiles of milk progesterone (throughout Days -18 to 0), estradiol (Days -6 to 2) or cortisol (Days -18 to 2) around estrus, maximum estradiol values or estradiol concentrations on Day 0. However, post hoc pair wise comparisons revealed that prior to estrus, severely lame cows had lower maximum progesterone concentrations compared to nonlame cows (1.3+/-0.1, 1.2+/-0.2, 0.7+/-0.1 ng/ml milk; P=0.042). Furthermore, severely lame cows expressed behavioral estrus with lower intensity (284+/-128 points, n=9) compared to moderately lame (662+/-310 points, n=9) or nonlame animals (583+/-275 points, n=18; P=0.05 and P=0.02, respectively). Resting concentrations of cortisol (Days 20-80 postpartum) did not vary between days postpartum or lameness score. The incidence of behavioral estrus was not affected by increasing lameness score, as 54.2%, 56.2% and 50.0% periods with low progesterone were associated with spontaneous estrus expression, respectively. Concluding, in this biological model of chronic stress, lameness did not affect the incidence of behavioral estrus but did reduce estrus intensity once ovarian cyclicity had resumed after calving. This reduced intensity of estrus was associated with lower maximum progesterone values prior to estrus but not abnormal daily cortisol or estradiol values in milk.     

The variability in the interval between estrus and ovulation in cattle and its determinants

Fertility of Holstein cows has been decreasing for years and, to a lesser extent, the fertility of heifers too but more recently. A hypothesis to explain this phenomenon may be that the chronology of events leading to ovulation is different for those animals bred nowadays when compared to what was reported previously; this would result in an inappropriate time of insemination. Therefore, two experiments were designed to investigate the relationships among estrus behavior, follicular growth, hormonal events and time of ovulation in Holstein cows and heifers. In the first experiment, the onset of estrus, follicular growth, patterns of estradiol-17beta, progesterone and LH, and the time of ovulation were studied in 12 cyclic Holstein heifers that had their estrus synchronized using the Crestar method; this was done twice, 3 weeks apart. The intervals between estrus and ovulation, estrus and the LH peak, and between the LH peak and ovulation were, respectively, 38.5 h +/-3.0, 9.1 +/- 2.0 and 29.4 h +/-1.5 (mean+/- S.E.M). The variation in the interval between estrus and the LH peak explained 80.6% of the variation in the interval between estrus and ovulation. The intervals between estrus and the LH peak, and estrus and ovulation were correlated with estradiol-17beta peak value (r=-0.423, P <0.04 and r=-0.467, P<0.02, respectively). Positive correlation coefficients for the number of follicle larger than 5 mm, and negative correlation coefficients for the size of the preovulatory follicle with the intervals between estrus and LH peak, LH peak and ovulation, and estrus and ovulation suggest an ovarian control of these intervals. In respect to its role to explain the variation in the interval between estrus and ovulation, the variation in the interval between estrus and the LH peak was evaluated further in a second set of experiments utilizing 12 pubertal Holstein heifers and 35 Holstein cows. The duration of the interval between the beginning of estrus and the LH peak was longer in heifers than in cows (4.15 h versus -1.0 h; P <0.002); the variation for this interval was higher in cows than in heifers (S.E.M.= 1.2 h versus 0.8 h; P=0.01). According to the results of these studies it can be proposed that estradiol and other product(s) of ovarian origin regulate not only the duration of intervals between the onset of estrus and the LH surge but also between the LH surge and ovulation. From the results obtained in the first experiment, it may be postulated that differences observed between cows and heifers for the duration of the interval between onset of estrus and the LH surge as well as for the variation of this interval would be observed also for the interval between the onset of estrus and ovulation. Therefore, on a practical point of view, the long interval between the onset of estrus and ovulation and the high variation of this interval, especially in cows, may be a source of low fertility and should be considered when analysing reproductive disorders.     

Neither plasma progesterone concentrations nor exogenous eCG affects rates of ovulation or pregnancy in fixed-time artificial insemination (FTAI) protocols for puberal Nellore heifers

The objective was to evaluate the effects of plasma progesterone (P4) concentrations and exogenous eCG on ovulation and pregnancy rates of pubertal Nellore heifers in fixed-time artificial insemination (FTAI) protocols. In Experiment 1 (Exp. 1), on Day 0 (7 d after ovulation), heifers (n = 15) were given 2 mg of estradiol benzoate (EB) im and randomly allocated to receive: an intravaginal progesterone-releasing device containing 0.558 g of P4 (group 0.5G, n = 4); an intravaginal device containing 1 g of P4 (group 1G, n = 4); 0.558 g of P4 and PGF_2α (PGF; 150 μg d-cloprostenol, group 0.5G/PGF, n = 4); or 1 g of P4 and PGF (group 1G/PGF, n = 3). On Day 8, PGF was given to all heifers and intravaginal devices removed; 24 h later (Day 9), all heifers were given 1 mg EB im. In Exp. 2, pubertal Nellore heifers (n = 292) were treated as in Exp. 1, with FTAI on Day 10 (30 to 36 h after EB). In Exp. 3, pubertal heifers (n = 459) received the treatments described for groups 0.5G/PGF and 1G/PGF and were also given 300 IU of eCG im (groups 0.5G/PGF/eCG and 1G/PGF/eCG) at device removal (Day 8). In Exp. 1, plasma P4 concentrations were significantly higher in heifers that received 1.0 vs 0.588 g P4, and were significantly lower in heifers that received PGF on Day 0. In Exp. 2 and 3, there were no significant differences among groups in rates of ovulation (65-77%) or pregnancy (Exp. 2: 26-33%; Exp. 3: 39-43%). In Exp. 3, diameter of the dominant ovarian follicle on Day 9 was larger in heifers given 0.558 g vs 1.0 g P4 (10.3 ± 0.2 vs 9.3 ± 0.2 mm; P < 0.01). In conclusion, lesser amounts of P4 in the intravaginal device or PGF on Day 0 decreased plasma P4 from Days 1 to 8 and increased diameter of the dominant follicle on Day 9. However, neither of these nor 300 IU of eCG on Day 8 significantly increased rates of ovulation or pregnancy.     

Effects of oxytocin on cloprostenol-induced luteolysis, follicular growth, ovulation and corpus luteum function in heifers

Twenty-five normally cyclic Holstein heifers were used to examine the effects of oxytocin on cloprostenol-induced luteolysis, subsequent ovulation, and early luteal and follicular development. The heifers were randomly assigned to 1 of 4 treatments: Group SC-SC (n=6), Group SC-OT (n=6), Group OT-SC (n=6) and Group OT-OT (n=7). The SC-SC and SC-OT groups received continuous saline infusion, while Groups OT-SC and OT-OT received continuous oxytocin infusion (1:9 mg/d) on Days 14 to 26 after estrus. All animals received 500 microg, i.m. cloprostenol 2 d after initiation of infusion (Day 16) to induce luteolysis. Groups SC-OT and OT-OT received oxytocin twice daily (12 h apart) (0.33 USP units/kg body weight, s.c.) on Days 3 to 6 of the estrous cycle following cloprostenol-induced luteolysis, while Groups SC-SC and OT-SC received an equivalent volume of saline. Daily plasma progesterone (P4) concentrations prior to cloprostenol-induced luteolysis and rates of decline in P4 following the induced luteolysis did not differ between oxytocin-infused (OT-OT and OT-SC) and saline-infused (SC-SC and SC-OT) groups (P >0.1). Duration of the estrous cycle was shortened in saline-infused heifers receiving oxytocin daily during the first week of the estrous cycle. In contrast, oxytocin injections did not result in premature inhibition of luteal function and return to estrus in heifers that received oxytocin infusion (OT-OT). Day of ovulation, size of ovulating follicle and time of peak LH after cloprostenol administration for oxytocin and saline-treated control heifers did not differ (P >0.1). During the first 3 d of the estrous cycle following luteal regression, fewer (P <0.01) follicles of all classes were observed in the oxytocin-infused animals. Day of emergence of the first follicular wave in heifers treated with oxytocin was delayed (P <0.05). The results show that continuous infusion of oxytocin during the mid-luteal stage of the estrous cycle has no effect on cloprostenol-induced luteal regression, timing of preovulatory LH peak or ovulation. Further, the finding support that an episodic rather than continuous administration of oxytocin during the first week of the estrous cycle results in premature loss of luteal function. The data suggest minor inhibitory effects of oxytocin on follicular growth during the first 3 d of the estrous cycle following cloprostenol-induced luteolysis.