Active immunization of beef heifers against luteinizing hormone: II. Evaluation of conjugation technique on antigenicity of LH

This study evaluated the antigenicity of LH-ovalbumin complexes produced using different conjugation techniques. Two homobifunctional cross-linkers, glutaraldehyde (Glut) and carbodiimide (ECDI), were evaluated together with one heterobifunctional reagent, m-maleimido-benzoyl N-hydroxysuccinimide ester (MBS). Polyacrylamide gel electrophoresis and Western transfer techniques were used to confirm conjugation of LH. Forty-four beef heifers were assigned randomly to seven treatment groups. Two groups of heifers were immunized against glutaraldehyde conjugates (Glut-I and Glut-II), two against MBS conjugates (MBS-I and MBS-II) and one against a carbodiimide conjugate (ECDI). Control animals were immunized against nonconjugated LH (LH-only) or ovalbumin alone (Oval). Heifers received one primary injection of antigen followed by two boosters at a 3-wk interval. The Glut conjugates induced the highest (P less than .05) LH antibody activity (Glut-II, 18 +/- 4%; Glut-I, 14 +/- 4%). The ECDI (11 +/- 4%), and MBS-I (11 +/- 2%) conjugates induced greater LH binding than MBS-II (4 +/- 1%), LH-only (4 +/- 1%) or Oval (2 +/- 1%). Glutaraldehyde produced an LH-ovalbumin conjugate of greater LH immunogenicity than either ECDI or the heterobifunctional reagent, MBS.     

Active immunization of beef heifers against luteinizing hormone: III. Evaluation of dose and longevity

Objectives were to evaluate the dose (Exp. 1) and purity of LH preparations (Exp. 2) on the anti-LH antibody response in heifers. Experiment 3 evaluated the longevity of LH immunization on sterility in heifers. In Exp. 1, 115 crossbred heifers were injected every 3 wk for 6 wk with .1, .33, 1.0, 3.0 or 9.0 mg of LH-ovalbumin. Concentrations of anti-LH antibodies generated were quantified by determining the percentage of binding of [125I]LH in serum. Mena LH binding over wk 0 to 12 was greater in heifers immunized with 1.0 mg conjugate than in heifers immunized with other doses (P less than .05). In Exp. 2, LH-ovalbumin conjugates were made from either LH-1, LH-2 or LH-3, which had relative immunological potencies of 2.1, 1.5 and 1.2 x NIH-LH-S1 units/mg, respectively. Forty-eight crossbred beef heifers were immunized against one of these three LH-ovalbumin conjugates, against LH conjugated without ovalbumin (LH-LH), or against ovalbumin alone (Oval). Estrous cycle activity was monitored by measuring serum progesterone concentration. Potency of the LH preparation used in the LH-ovalbumin conjugate was correlated (r = .94) with its ability to produce LH antibodies. In Exp.3, heifers were injected with 1 mg antigen every 2 wk for 10 wk. Five LH-1 heifers and five control heifers were slaughtered for examination of ovaries 10 wk after the last booster injection. The remaining five LH-I and five control animals were placed with a bull 8 wk after the last booster. All five control heifers conceived by 4 +/- 1 wk after placement with the bull whereas the LH-immunized heifers remained acyclic for 42 to 96 wk.     

Active immunization of heifers against luteinizing hormone-releasing hormone, human chorionic gonadotropin and bovine luteinizing hormone

Seventy crossbred heifers were allotted randomly to 10 treatment groups. Treatments consisted of active immunization against ovalbumin (OV) conjugates of luteinizing hormone-releasing hormone (LHRH), human chorionic gonadotropin (hCG) and bovine luteinizing hormone (bLH) with each of three adjuvants. The adjuvants were complete Freund's adjuvant (CFA), M103(6) and 6VR6. Control animals were immunized against OV alone using CFA. Bulls were placed with the heifers following immunization to allow comparison of pregnancy rates between groups. Blood samples were collected weekly for 14 wk to determine antibody concentrations. Significant levels of circulating LH or LHRH antibodies were detected in heifers immunized with each of the hormone conjugates. Complete Freund's adjuvant was the most effective for stimulating antibody response to these antigens; however, M103 was equally effective when used with bLH or hCG conjugates. None of the heifers in the bLH-OV-CFA, bLH-OV-M103 or LHRH-OV-CFA immunization groups was pregnant at slaughter, whereas 71% of the OV-CFA control heifers were pregnant. Fertility suppression may be achieved in the bovine by active immunization against any of these three hormone conjugates. However, the duration of this study (8 wk after immunization) does not allow evaluation of the duration of effectiveness of each of the treatments.     

Hypothalamic deafferentation in prepuberal beef heifers: Effects of gonadotropin-releasing hormone and estradiol benzoate on luteinizing hormone secretion

Hypothalamic control of luteinizing hormone (LH) secretion was investigated in crossbred beef heifer calves by comparing anterior (AHD), posterior (PHD), and complete (CHD) hypothalamic deafferentation with sham operated controls (SOC). Heifers (n = 16) were fitted with an indwelling jugular catheter for 6 days before cranial surgery, and assigned randomly to treatments. Blood for radioimmunoassay of LH was collected sequentially at 15-min intervals during an 8-h period on days ? 1 before and day 6 after hypothalamic deafferentation or sham operation. On the day of surgery, blood samples were collected sequentially at 15-min intervals 2 h before induction of anesthesia and throughout surgery and early recovery. Seven months after hypothalamic deafferentation, all experimental and sham operated heifers were ovariectomized and treated with vegetable oil (i.m.) plus saline (i.v.), vegetable oil plus gonadotropin releasing hormone (GnRH), estradiol benzoate (EB, 1 mg) in vegetable oil. After ovariectomy basal plasma concentrations of LH increased (P < 0.01) compared with the low circulating hormone levels before ovariectomy. The amplitude of LH response to GnRH was greater (P < 0.01) in CHD and PHD when compared with SOC and AHD heifers. Injection of EB failed to induce a LH surge in CHD and PHD 900–1100 min later when compared with the robust response seen in SOC and AHD heifers. Injection of EB plus GnRH elicited LH release in all deafferentated and sham operated heifers. These results indicate a transient change in LH secretion after AHD or CHD in prepuberal heifers with intact ovaries. After OVX, the integrity of the neural connection of the posterior hypothalamus is required for EB-induced LH release in beef heifers.     

Immunization of beef heifers against gonadotropin-releasing hormone prevents luteal activity and pregnancy: Effect of conjugation to different proteins and effectiveness of adjuvants

Three experiments were conducted to evaluate methods of immunization against GnRH on antibody titer, luteal activity, and pregnancy in beef heifers. Experiment 1 evaluated the efficacy of adjuvants with 30 heifers. Control heifers were immunized against human serum albumin (HSA) emulsified in Freund's complete adjuvant (FCA). The other 4 treatments contained GnRH conjugated to HSA (HSA-GnRH) emulsified in FCA, Freund's incomplete adjuvant (FIA), DEAE dextran (DD) + mineral oil (MO), or DD+FIA. Treatment was in the mammary gland for all experiments. Titers against GnRH for heifers immunized against HSA-GnRH with FCA, DD+MO, or DD+FIA were greater than titers for HSA-GnRH with FIA or control heifers (P < 0.01). Body weight was reduced (P < 0.05) in control and FCA heifers compared with FIA, DD+MO, and DD+FIA heifers. Heifers immunized with DD+MO and DD+FIA had fewer granulomas in mammary glands than heifers treated with FCA (P < 0.01). In Exp. 2, 36 heifers were used to determine the effect of the protein conjugated to GnRH on titers against GnRH. Heifers (6/treatment) received a primary immunization against GnRH conjugated to HSA (HSA-GnRH), ovalbumin (OA-GnRH), or keyhole limpet hemocyanin (KL-GnRH), or heifers were immunized against each carrier protein. Antigens were emulsified in DD+FIA. Immunization of heifers against OA-GnRH, KL-GnRH, or HSA-GnRH suppressed luteal activity (P < 0.01) for 23, 16, and 12 wk, respectively, and antibody titers against GnRH were greater (P < 0.01) for 19, 5, and 7 wk, respectively, compared with heifers immunized against the carrier proteins. In Exp. 3, 90 heifers were used to determine the effect of immunization against GnRH on ovarian activity and pregnancy rate. Heifers (30/treatment) received a primary and 2 or 3 booster immunizations against GnRH conjugated to OA, and controls received a primary and 2 booster immunizations against OA. All antigens were emulsified in DD+FIA. At 8 wk after primary immunization, heifers were exposed to fertile bulls for 24 wk. Pregnancy rate was less (P < 0.01) for 3-booster heifers (13%) compared with control (83%) and 2-booster (62%) heifers. We conclude that immunization against GnRH, conjugated to OA and emulsified in DD+FIA, does not influence ADG and produces sufficient titers against GnRH to prevent estrous cycles with few mammary granulomas. Immunization against GnRH with 3 booster immunizations prevented luteal activity and pregnancy in most beef heifers for more than 4 mo.     

Effects of immunization against luteinizing hormone-releasing hormone and treatment with trenbolone acetate on reproductive function of beef bulls and steers

The objectives of this study were 1) to evaluate the ability of trenbolone acetate (TBA) administered in tandem with LHRH immunization to suppress reproductive function in bulls and 2) to examine the effects of LHRH and androgen (TBA) signaling on pituitary gland function. Forty-four Angus × Hereford crossbred calves (BW=225 ± 2 kg; age=187 ± 6 d) received castration, LHRH immunization, or TBA administration in a 2 × 2 × 2 factorial design. Treatment groups receiving LHRH immunization contained 6 animals, whereas other treatment groups contained 5 animals. Animals immunized against LHRH received a primary injection and 2 booster injections of ovalbumin-LHRH-7 fusion protein on d 0, 42, and 196, respectively. Animals treated with TBA were implanted on d 224. Serum LHRH antibodies increased (P<0.05) after each booster for immunized animals, but were negligible in nonimmunized animals throughout the experiment. Serum testosterone concentration (P<0.001) and scrotal circumference (P<0.05) were depressed in LHRH-immunized bulls compared with nonimmunized bulls by d 84 and 168 of the experiment, respectively. Treatment with TBA tended (P=0.08) to decrease serum testosterone concentrations of nonimmunized bulls. Weights of testes at slaughter were decreased (P<0.001) for LHRH-immunized (232 ± 41 g) compared with nonimmunized (752 ± 45 g) bulls, but did not differ (P=0.80) between TBA-implanted (500 ± 49 g) and nonimplanted bulls (484 ± 36 g). Both LHRH immunization and castration decreased pituitary gland stores of LH and FSH (P<0. 001). There was no effect (P>0.10) of TBA on pituitary gland FSH content and only a tendency (P=0.09) to increase pituitary gland LH content. Immunization against LHRH decreased expression of LH β-subunit and common α-subunit genes (P<0.001). Castration increased expression of LH β-subunit and common α-subunit genes (P=0.02). Treatment with TBA further suppressed (P=0.04) α-subunit mRNA expression in LHRH-immunized steers. In summary, LHRH immunization decreased synthesis and storage of LH and decreased storage, but not synthesis of FSH in bulls. The increased synthesis of LH and FSH in nonimmunized, but not LHRH-immunized steers suggests that castration removes the negative feedback on gonadotropin synthesis but that LHRH is still needed for release of these hormones. Androgen replacement with TBA did not restore the negative feedback control of gonadotropin synthesis.     

Induction of precocious puberty in heifers I: enhanced secretion of luteinizing hormone

In beef heifers weaned between 3 and 4 mo of age and fed a high-concentrate diet, approximately 50% reach puberty before 300 d of age (precocious puberty). The objectives of this experiment were 1) to determine whether precocious puberty could be induced experimentally by weaning heifers early and feeding a high-concentrate diet, and 2) to determine the dynamics of secretion of LH associated with precocious puberty. Crossbred Angus and Simmental heifer calves were weaned at 73 +/- 3 d of age and 115 +/- 3 kg of BW and fed a high-concentrate (60% corn; HI, n = 9) or control diet (30% corn; CONT, n = 9). Heifers were fed individually, and target BW gains were 1.50 and 0.75 kg/d for the HI and CONT treatments, respectively. Heifers were weighed every 2 wk. Blood samples were collected weekly and assayed for progesterone concentration to determine age at puberty. Serial blood samples were collected at 20-min intervals for 24 h at mean ages of 102, 130, 158, 172, 190, 203, 217, 231, and 259 d and assayed for LH concentration to evaluate the dynamics of secretion of LH. Heifers fed the HI diet exhibited greater BW gain (P < 0.01) than CONT heifers (1.27 +/- 0.05 vs. 0.85 +/- 0.05 kg/d, respectively). As a result, BW in the HI treatment was greater (P < 0.01) than in the CONT treatment by 188 d of age and remained different through the end of the experiment. Precocious puberty occurred in 8 of 9 heifers fed the HI diet and 0 of 9 heifers fed the CONT diet. Age at puberty was reduced in the HI (P < 0.01) compared with the CONT heifers (262 +/- 10 vs. 368 +/- 10 d of age, respectively). Body weight at puberty was also reduced in the HI (P < 0.05) compared with the CONT treatment (327 +/- 17 vs. 403 +/- 23 kg, respectively). Heifers attaining puberty during the experiment continued with subsequent luteal phases as evidenced by cyclic patterns of progesterone concentrations. Frequency of pulses of LH (pulses/24 h) increased with age (P < 0.01) for both treatments. Heifers in the HI treatment exhibited a greater number of pulses of LH (P < 0.01) than those in the CONT treatment by 190 d of age and in all subsequent collection periods (treatment x age, P < 0.05). Mean LH concentrations also increased with age (P < 0.01) for both treatments but did not differ between treatments. In conclusion, precocious puberty induced by early weaning and feeding of a high-concentrate diet is preceded by increasing frequency of pulses of LH.     

Gonadotropin-releasing hormone-induced ovulation and luteinizing hormone release in beef heifers: effect of day of the cycle

The COSynch protocol has been used to synchronize ovulation and facilitate fixed-time AI in beef cattle. Establishment and maintenance of pregnancy was negatively affected, in previous studies, by GnRH-induced ovulation of small dominant follicles (/=10 mm) and increased ovulatory response after GnRH 2.     

Pulsatile or continuous infusion of luteinizing hormone-releasing hormone and hormonal concentrations in prepubertal beef heifers

An experiment was conducted to determine if exogenous luteinizing hormone-releasing hormone (LHRH) administered iv intermittently as pulses (P) or by continuous sc infusion (I) using osmotic minipumps could sustain pulsatile LH release and induce estrous cyclicity in prepubertal heifers. Prepubertal heifers were assigned randomly to: 1) receive pulses of LHRH (n = 6; 2.5 micrograms LHRH/2 h for 72 h), 2) be infused with LHRH (n = 11; 1.25 micrograms LHRH/h for 72 h), or 3) serve as controls (n = 16). Blood was collected at 20-min intervals for 8 h (0900 to 1700 h) from six heifers in each group on d 1, 2, 3 (during treatment), and on d 4 (during 8 h after terminating LHRH treatments). Heifers given LHRH had higher (P less than .01) LH concentrations than controls. Preovulatory-like LH surges occurred in three I, two P and no control heifers during treatment. Pulse frequencies of LH (no. LH pulses/8 h) were greater (P less than .001) for P heifers than for I and control heifers due to pulsatile LHRH treatment. Serum estradiol was higher (P less than .01) during treatment for LHRH-treated heifers than for controls. Serum follicle-stimulating hormone, cortisol, and progesterone were unchanged during treatment. High levels of cortisol on d 1 declined (P less than .001) to baseline by d 2. Characteristic progesterone rises or short luteal phases occurred within 10 d of treatment initiation in more (P less than .05) LHRH-treated heifers (I = 45%, P = 33%) than controls (6%), although days to first observed estrus and first ovulation were unaffected by treatments. Although both continuous and pulsatile administration of LHRH successfully induced LH and estradiol release as well as preovulatory-like LH surges in some heifers, earlier initiation of estrous cycles was not achieved. Estrous cycles appeared to be delayed by exposure to continuous LHRH infusions during the peripubertal period.     

Influence of insulin and energy intake on ovulation rate, luteinizing hormone and progesterone in beef heifers

Ovarian and gonadotropin responses to insulin and energy restriction were investigated in a 2 X 2 factorial experiment using 2-yr-old Brangus heifers. Thirty heifers were paired by weight and body condition, then assigned to treatment groups receiving 75 (LE) or 180% (HE) of NRC recommendations for dietary energy for maintenance. Diets were adjusted weekly to maintain daily .25 to .5 kg weight loss or 0 to .25 kg weight gain, respectively. On d 10 of the first estrous cycle subsequent to the initial 45 d of feeding, heifers within each dietary group were allocated to receive twice daily infusions of either 40 U insulin (I) or saline (C). Infusions began at 5 and 10 h postprandial and were given in six boluses, 20 min apart. Infusions continued daily until d 20 or estrus, whichever occurred first. On d 11, blood samples were collected at 15-min intervals for 12 h to determine luteinizing hormone (LH) and insulin concentrations. On d 16 to 20, twice daily im injections of 1 mg follicle stimulating hormone (FSH) were administered. Heifers were ovariectomized on d 11 after estrus. Number of corpora lutea (CL) in LE-I heifers was greater (P less than .05) in LE-C, HE-C or HE-I. Total CL weight (g) per heifer was greater (P less than .05) in HE-C and LE-I heifers than in LE-C. Individual CL wt was heavier in HE than in LE heifers (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of active immunization against growth hormone-releasing factor on growth and onset of puberty in beef heifers.

Angus and Charolais heifers (195 +/- 7 kg) were actively immunized against growth hormone-releasing factor (GRF) to evaluate the effect on concentrations of somatotropin (ST), insulin-like growth factor I (IGF-I), insulin (INS), growth, and onset of puberty. Primary immunizations were given at 184 +/- 7 d of age (d 0 of experiment) by injecting (s.c.) 1.5 mg of GRF-(1-29)-Gly-Gly-Cys-NH2 conjugated to 1.5 mg of human serum albumin (GRFi, n = 22) or 1.5 mg of human serum albumin (HSAi, n = 21). Booster immunizations of .5 mg of antigen were given on d 62, 92, 153, and 251. Antibody binding (percentage at 1:2,000 dilution) to [125I]GRF on d 69 was greater (P less than .01) in GRFi (53.7 +/- 4.5) than in HSAi (10.1 +/- .6) heifers. Serum concentration (ng/ml) and frequency (peaks/5 h) of ST release, respectively, on d 78 were lower (P less than .01) in GRFi than in HSAi heifers (3.3 +/- .1 vs 5.6 +/- .2 and .9 +/- .3 vs 2.3 +/- .2). Serum IGF-I (ng/ml) was lower (P less than .01) in GRFi than in HSAi heifers on d 69 (41 +/- 5 vs 112 +/- 4). Serum INS (microU/ml) on d 78 was lower (P less than .05) in GRFi (2.2 +/- .1) than in HSAi (3.8 +/- .2) heifers. Feed intake, ADG, and feed efficiency were lower (P less than .05) in GRFi than in HSAi heifers. Hip height was lower (P less than .01) and fat thickness was greater (P less than .05) in GRFi than in HSAi heifers by d 132 and 167, respectively. Percentage of heifers attaining puberty (progesterone greater than 1 ng/ml for two consecutive weeks) by d 209 and 379 (12.9 and 18.5 mo of age), respectively, was lower (P less than .05) in GRFi (40.9 and 45.5) than in HSAi (81.0 and 100). In conclusion, growing heifers were successively immunized against GRF. Active immunization against GRF resulted in decreased serum concentration of ST, IGF-I, and INS. In addition, GRF immunization led to lowered feed intake, ADG, and feed efficiency, increased fat depth, and delayed onset of puberty in heifers. We propose that ST and IGF-I are important metabolic mediators involved in the initiation of puberty in heifers.     

Effects of 17 beta-estradiol and diets varying in energy on secretion of luteinizing hormone in beef heifers

An experiment was conducted to test the hypothesis that 17 beta-estradiol (E2) would not suppress secretion of luteinizing hormone (LH) in heifers fed a diet limited in energy during the period before the onset of nutritionally induced anestrus. Sixteen of 20 heifers that had been exhibiting normal estrous cycles (20 mo of age, 409 +/- 6 kg body weight) were ovariectomized, and half of them were assigned at random to receive an E2 implant. The ovariectomized heifers were assigned at random to receive diets that contained low (L; 5.8 Mcal X animal-1 X d-1, n = 8) or high levels of energy (H; 20.0 Mcal X animal-1 X d-1, n = 8) for 100 d. The other four heifers remained intact and were fed the L-diet. The intact heifers were utilized to determine the status of reproductive function in animals fed the L-diet. Heifers lost body weight rapidly after initiation of feeding the L-diet. Heifers fed the L-diet then stabilized at a lighter weight until the latter part of the experiment. One of the four intact heifers fed the L-diet became anestrus near the end of the study. Mean concentrations of LH in blood serum increased linearly (P less than .05) in ovariectomized heifers fed the L- and H-diet. Mean concentration of LH in heifers fed the H-diet that were implanted with E2 was similar to ovariectomized heifers fed the H-diet that received no E2. Mean LH in serum of ovariectomized heifers implanted with E2 fed the L-diet was suppressed and remained low throughout the study. Frequency of pulses of LH in ovariectomized heifers fed the L-diet was less (P less than .01) than that in ovariectomized heifers fed the H-diet. Estradiol decreased the number of pulses of LH in heifers fed the L-diet. We conclude that dietary energy restriction in beef heifers has a direct action on the hypothalamo-pituitary axis to lower the number of pulses of LH in the absence of ovarian steroids. However, ovarian E2 appears to suppress further secretion of LH in heifers fed limited levels of dietary energy before the onset of nutritional anestrus occurs, therefore, our working hypothesis is rejected.     

Progesterone concentration,estradiol pretreatment,and dose of gonadotropin-releasing hormone affect gonadotropin-releasing hormone-mediated luteinizing hormone release in beef heifers

We examined whether progesterone (P₄)-induced suppression of LH release in cattle can be overcome by an increased dose of exogenous gonadotropin-releasing hormone (GnRH) or pretreatment with estradiol (E₂). In Experiment 1, postpubertal Angus-cross heifers (N = 32) had their 2 largest ovarian follicles ablated 5 d after ovulation. Concurrently, these heifers were all given a once-used, intravaginal P₄-releasing insert (CIDR), and they were randomly assigned to be given either prostaglandin F_2α (Low-P₄) or no treatment (High-P₄) at follicle ablation, and 12 h later. Six days after emergence of a new follicular wave, half of the heifers in each group (n = 8) were given either 100 or 200 μg of GnRH i.m. Plasma luteinizing hormone (LH) concentrations were higher in the Low- vs High-P₄ groups, and in heifers given 200 vs 100 μg of GnRH (mean ± SEM 15.4 ± 2.2 vs 9.1 ± 1.2, and 14.8 ± 2.1 vs 9.8 ± 1.4 ng/mL, respectively; P ≤ 0.01). Ovulation rate was higher (P = 0.002) in the Low-P₄ group (15/16) than in the High-P₄ group (6/16), but it was not affected by GnRH dose (P = 0.4). In Experiment 2, heifers (n = 22) were treated similarly, except that 5.5 d after wave emergence, half of the heifers in each group were further allocated to be given either 0.25 mg estradiol benzoate i.m. or no treatment, and 8 h later, all heifers were given 100 μg GnRH i.m. Both groups treated with E₂ (Low- and High-P₄) and the Low-P₄ group without E₂ had higher peak plasma LH concentrations compared to the group with high P₄ without E₂ (12.6 ± 1.8, 10.4 ± 1.8, 8.7 ± 1.3, and 3.9 ± 1.2 ng/mL, respectively; (P < 0.04)). However, E₂ pretreatment did not increase ovulation rates in response to GnRH (P = 0.6). In summary, the hypotheses that higher doses of GnRH will be more efficacious in inducing LH release and that exogenous E₂ will increase LH release following treatment with GnRH were supported, but neither significantly increased ovulation rate.     

Active immunization of heifers against prostaglandin F2 alpha: potential as a sterilization vaccine

Four experiments were conducted with 210 heifers in an attempt to develop a sterilization vaccine by active immunization against prostaglandin F2 alpha (PGF2 alpha) and to evaluate feedlot performance following immunization against the combination of PGF2 alpha and estrogens. The objectives were: development of a PGF2 alpha-ovalbumin conjugate that would induce antibody formation when used with complete Freund's adjuvant (CFA); comparison of CFA with other adjuvants in relation to PGF2 alpha antibody binding and maintenance of the corpus luteum; examination of growth performance in immunized heifers against both PGF2 alpha and estradiol-17 beta and evaluation of sterilization in PGF2 alpha-immunized heifers maintained with bulls. A PGF2 alpha-ovalbumin conjugate was developed that resulted in antibody production against PGF2 alpha, although antibody binding was quite low. The antibody response in heifers was higher in Exp. I and II than in Exp. III and IV (P less than .05). Complete Freund's adjuvant was the best adjuvant in inducing antibody formation compared with all other adjuvants tested (P less than .01). Corpus luteum (CL) function was maintained for 2.5 mo and ovulation was apparently blocked in Exp. I. The results of Exp. II confirmed those of Exp. I. Fewer than half of the heifers in Exp. III and IV had prolonged estrous cycles. In Exp. IV, immunized heifers became sterile for at least 4 mo when kept with bulls, although the success rate was only 37%. The low levels of antibody titers to PGF2 alpha in Exp. III and IV may be the reason for the failure to maintain CL function in some heifers.(ABSTRACT TRUNCATED AT 250 WORDS)     

Negative feedback control of luteinizing hormone secretion in prepubertal beef heifers at 60 and 200 days of age

Prepubertal beef heifers at 60 and 200 d of age, born in the fall or spring, were assigned randomly to one of three treatment groups: (1) intact = 1; (2) bilateral ovariectomy (OVX); or (3) OVX plus estradiol-17 beta(E2) administered in silastic implants (OVX + E2). Luteinizing hormone (LH) was measured in serum samples collected at 20-min intervals for 4 h from heifers on -1, +7, +21, +35 and +49 d after OVX. Luteinizing hormone concentrations increased in the serum by 7 d after OVX in heifers at both 60 and 200 d of age (P less than .001; time X treatment). Prior to OVX, the LH patterns were characterized by low levels and infrequent episodic pulses. By 49 d after OVX, the mean LH concentrations increased and the pattern changed to one of rhythmic LH pulses with a periodicity of 1 h (P less than .001; time X treatment). Estradiol-treated OVX heifers did not exhibit a postovariectomy rise in serum LH concentrations. Serum E2 concentration 49 d after OVX in OVX heifers was threefold greater than in 1 or OVX heifers, thus demonstrating that E2 exerted negative feedback on pituitary LH secretion in prepubertal heifers. There was no measurable difference in serum E2 concentrations between I and OVX heifers; however, the contrast in the concentration and pattern of serum LH between the two groups was dramatic and suggested gonadal factors in addition to E2 are involved in controlling LH secretion.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of active immunization against growth hormone releasing factor on concentrations of somatotropin and insulin-like growth factor I in lactating beef cows.

Two experiments were conducted to determine the effects of immunoneutralization of growth hormone-releasing factor [GRF(1-29)-NH2] on concentrations of somatotropin (ST) and insulin-like growth factor I (IGF-I) in lactating beef cows. In Experiment 1, multiparous Hereford cows were immunized against 2 mg GRF(1-29)-(Gly)4-Cys-NH2 conjugated to human serum albumin (GRFi, n = 3) or 2 mg human serum albumin (HSAi, n = 3) at 52 +/- 1 d prior to parturition. Boosters (1 mg) were administered on days 12, 40 and 114 postpartum (pp). Serum samples were collected at 15-min intervals for 5 hr on days 18, 46 and 120 pp, followed by administration (IV) of an opioid agonist (FK33-824; 10 micrograms/kg) and an antagonist (naloxone; .5 mg/kg) at hours 5 and 7, respectively. A GRF-analog ([desamino-Tyr1, D-Ala2, Ala15] GRF (1-29)-NH2; 3.5 micrograms/kg) and arginine (.5 g/kg) were administered at hour 10 on days 47 and 121, respectively. Percentage binding of [125I]GRF (1:100 dilution of serum) 28 d after primary immunization was greater in GRFi (14.3 +/- 4.9) than in HSAi (.7 +/- .3) cows. Binding increased to 29.3 +/- 6.5% after first booster in GRFi cows. Episodic release of ST was abolished by immunization against GRF; concentration and frequency of release of ST were lower (P less than .05) in GRFi than in HSAi cows on all days pp. Concentrations of IGF-I were lower in GRFi than in HSAi cows throughout lactation. Serum ST failed to increase following FK33-824 or arginine in GRFi; however, ST increased after both compounds in HSAi cows. Concentrations of ST following GRF-analog were greater (P less than .05) in HSAi than in GRFi cows. Experiment 2 was conducted to determine if a lower dose of antigen and a single booster would be sufficient to lower ST and IGF-I in lactating cows. Multiparous Hereford and Angus cows were assigned to GRFi (n = 6) or HSAi (n = 6). Primary (1.2 mg) and booster (.5 mg) immunizations were administered -14 and 8 d from calving, respectively. Cows were restricted to 60% of recommended intake of energy during lactation in order to elevate concentrations of ST. Serum samples were collected at 15-min intervals for 6 hr on days 26, 50, 73, 90 and 109 pp. Two of six GRFi cows had binding less than 10% (1:1,000 dilution of serum) and were omitted from further analyses.(ABSTRACT TRUNCATED AT 400 WORDS)     

Increased luteinizing hormone secretion and ovarian function in heifers actively immunized against estrogen and progesterone

This study was designed to test the effects of active immunization against estrogen and progesterone on patterns of luteinizing hormone (LH) and follicle stimulating hormone (FSH) secretion, ovarian characteristics and growth rate of heifers. Heifers were randomly assigned to four treatments: 1) control injection (n = 10); 2) ovariectomy (n = 9); 3) immunization against estrogen (anti-E, n = 10); and 4) immunization against estrogen and progesterone (anti-E+P4, n = 10). Three booster immunizations were administered at 1, 1.5 and 6 mo after primary immunization. Progesterone antibody binding was 40% (34 fmol at 1:600 final dilution) in the anti-E+P4 heifers, and estradiol-17 beta binding was 35% (30 fmol) and 60% (52 fmol at 1:100 final dilution) in the anti-E+P4 and anti-E heifers, respectively, after the final immunization. Anti-E+P4 heifers had more pulses of LH and higher basal concentrations of LH than anti-E or control heifers (P less than .05). Concentrations of LH in anti-E+P4 heifers did not increase to concentrations found in ovariectomized heifers (P less than .05). Immunization against steroids did not alter the secretion of FSH. The number of large follicles (greater than 15 mm diameter) in anti-E+P4 and anti-E heifers was greater than in control heifers (P less than .05). Ovarian weight was increased in anti-E+P4 heifers (P less than .05). Average daily gain was not different among groups (P greater than .05). It was concluded that active immunization against estrogen and progesterone in heifers increased LH secretion and stimulated ovarian function.     

Effects of feeding diets containing endophyte-infected fescue seed on luteinizing hormone secretion in postpartum beef cows and in cyclic heifers and cows.

Two experiments were conducted to investigate the effect of feeding endophyte (Acremonium coenophialum)-infected fescue (Festuca arundinacea Shreb.) seed on LH secretion in postpartum beef cows and in cycling heifers and cows. In Exp. 1, spring-calving primiparous Angus cows (n = 16) were pair-fed for 75 d diets that contained endophyte-free or endophyte-infected (95%) fescue seed that contained 1.3 micrograms/g of ergovaline and 5.2 mg/g of saturated pyrrolizidines. Serial blood samples for basal and GnRH-stimulated serum LH analysis were obtained on d 7, 28, 42, and 56 of the study. The endophyte had no effect on LH secretion (basal, pulse frequency, and amplitude) or milk production. Average daily gain was decreased (P < .05) in cows that consumed infected fescue seed compared with controls (-.20 vs -.01 kg, respectively). Basal serum prolactin concentrations were reduced (P < .01) in treated compared with control cows (8.9 vs 25.4 ng/mL, respectively) on d 70. In Exp. 2, cycling Angus heifers (n = 8; age = 2 yr) and cows (n = 8; age = 4 yr) stratified by age were pair-fed for 40 d diets that contained the noninfected or the highly infected fescue seed. Estrus was synchronized by prostaglandin F2 alpha (d 18 and 28). Serial blood samples for serum LH analysis were obtained on d 28 (luteal phase) and d 30 (follicular phase). The endophyte did not affect LH (P > .28) or prolactin (P > .16) secretion, whereas ADG was decreased (P < .05) in treated compared with control animals (.32 vs .70 kg/d, respectively).