Effect of passive immunization against testosterone on reproductive hormone secretion and ovarian function in dairy cows and pubertal beef heifers

Multiparous dairy cows were divided in 3 groups from Day 5 up to Day 56 post partum: high energy level (Group H, n=10), low energy level (Group L, n=10) and low energy level plus anti-testosterone bovine immunoglobulins (Group LI, n= 10). Undernutrition decreased body weight, body condition score, milk yield and energy balance in Groups L and LI compared to Group H (P<0.05), but had no effect on secretory pattern of LH. Passive immunization against testosterone increased LH secretion in Group LI (P<0.05). Follicular score and the presence of follicles >/= 10mm on the ovary were not affected by underfeeding but were higher in Group LI than in Group L after immunization (P<0.01). The duration of the first luteal phase was shorter in Group H than in Groups L and LI and maximum progesterone levels reached were higher in Group LI than in Group H (P<0.01). Reproductive performance was not depressed by underfeeding and immunization. In the pubertal beef heifers maintained in anestrus by undernutrition had very low LH secretion. After passive immunization against testosterone, the increase of LH pulses number became almost significant (P=0.07). Following injection of exogenous LH, the number of follicles >/= 9mm was higher in immunized (Group I, n=8) than in control heifers (Group C, n=7). Group I developed a dominant follicle sooner and of greater size than Group C. Passive immunization against testosterone increased LH secretion and follicular development.     

The effect of photoperiod on serum concentrations of luteinizing and follicle stimulating hormones in prepubertal heifers following ovariectomy and estradiol injection

Eleven heifers, between 63 and 197 days of age, were exposed to 18 hr light/day (L) or natural photoperiods (N), beginning October 19, 1979. They were ovariectomized 8 weeks later. LH concentrations after ovariectomy were not affected by photoperiod, but the rate of increase of FSH after ovariectomy was greater (P<0.10) for group L than for group N. Three weeks after ovariectomy, heiters were injected, IV, with 0.1 mug/kg estradiol-17beta. LH concentrations initially decreased after injection. This was followed by a series of pulses larger than those prior to injection. FSH concentrations declined after injection and remained low throughout the sampling period. The net response of LH concentrations to estradiol (mean post-injection concentration minus mean pre-injection concentration) was greater (P=0.05) for group L (4.7 +/- 0.49 ng/ml) than for group N (2.9 +/- 0.37 ng/ml). Photoperiod did not affect the net response of FSH concentrations to estradiol. We concluded that exposing prepubertal heifers to 18 hr light/day during the winter resulted in a greater rate of increase of FSH after ovariectomy and greater estrogen-induced LH release. Because the response of LH to estradiol-17beta differed from the response of FSH, these hormones may be regulated differently.     

Fertility response of yearling beef heifers after prebreeding energy manipulation, estrous synchronization and timed artificial insemination

The effect of dietary energy and weight class on the fertility of yearling beef heifers (Angus, Hereford, and Angus x Hereford) was investigated over 2 years. In year 1, heifers (n=58) were classed as heavy (HW; > or =340 kg) or light weight (LW; <340kg) and then assigned to receive either a low (LE; 0.23 kg/day) or high energy (HE; 0.68 kg/day) diet. In year 2, heifers (n=60) were also classified as heavy (> or =335kg) or light weight (<335 kg), but the energy content of the diet was raised so that heifers on the LE and HE were targeted to gain 0.46 and 0.79 kg/day, respectively. Heifers in the four groups, LELW (n=14 and 12), LEHW (n=16 and 17), HELW (n=13 and 15), and HEHW (n=15 and 16) received restricted amounts of concentrate (HE > LE) and free choice hay over 47 or 42 days (year 1 and year 2, respectively). To synchronize estrus, heifers were fed capsules containing MGA (0.5 mg/animal each day) beginning 11 days before the end of the feeding trial (day 0), PGF(2alpha) (25mg i.m.) and estradiol benzoate (Ebeta; 400 microg i.m.) was given on days 8 and 10, respectively. Estrous behavior was observed (days 10 and 11) and all heifers were inseminated on day 11. Following AI, heifers were re-grouped and a bull was introduced (days 27-39) for the second service in both years. Pregnancy diagnosis for the first (days 41-42) and second services (days 69-97) was performed by transrectal ultrasonography. Transrectal ultrasonic observations of ovarian follicle number and size were completed for a subset of heifers (n=5-8) from each experimental group at the end of the feeding trial. The effect of year was not significant for any of the reproductive performance variables measured. The mean ADG was (0.72 +/- 0.04 kg/day) and was greater in LW than HW heifers and in heifers in the HE than LE treatment groups (P <0.05). In heifers receiving the LE diet, ADG was lower in HW than LW heifers (weight x diet; P=0.02; 0.54 +/- 0.04 and 0.62 +/- 0.03 kg/day for HW and LW heifers, respectively). The diameter of the largest follicle was greater in heifers receiving the HE diet (P < 0.05; 11.3 +/- 0.4 mm) than those on the LE diet (10.3 +/- 0.3), and in LW (P <0.05) compared to HW heifers. The HE diet increased the size of the largest follicle in LW but not HW heifers (diet x weight, P <0.05). The percentage of pubertal heifers at the end of the feeding period (59.3%), estrous response (56.4%), conception rate (47.7%), ovulation rate (88.9%), and first service pregnancy rate (36.2%) were not significantly affected by initial weight or diet. There was a tendency for first service pregnancy rates to be greater in LW than HW heifers consuming the LE diet (diet x weight, P <0.1; 54.2 +/- 15 and 30.3 +/- 10% for LELW and LEHW heifers, respectively). Pregnancy rate after two services was greater (P=0.01) in LW (82 +/- 10%) than in HW (64.5 +/- 10%). The LE diet achieved moderate rates of gain and allowed high level of reproductive performance in LW but not HW heifers.     

Effect of dietary energy and protein density on body composition,attainment of puberty,and ovarian follicular dynamics in dairy heifers

The objectives were to examine the effects of dietary energy and protein density on age and body composition at puberty, and on ovarian follicular dynamics during the pre- and peripubertal periods in Holstein heifers. In Phase 1, heifers were randomly allotted (n=10 per diet) at 100 kg body weight (BW) to diets with either low (P1L), medium (P1M) or high (P1H) energy and protein formulated for an average daily gain (ADG) of 0.5, 0.8 or 1.1 kg per day, respectively. During Phase 2 (P2), all heifers were fed ad libitum a common diet formulated for an ADG of 0.8 kg per day. Half the animals within the high (n=5) and low groups (n=5) entered P2 either at 12 months of age (P2H-12; P2L-12) or at 330 kg BW (P2H-330; P2L-330). Heifers fed P1H, P1M, P1L, and P2L-12 diets attained puberty at approximately 9, 11, 16, and 14 months of age, respectively (P<0.01). Urea space estimates of body fat and protein percent, and back-fat thickness, were lower in P1L heifers compared to P1H or P1M heifers at similar chronological ages (P<0.05) but did not differ at puberty (P>0.10). Compared to P1L heifers, P1H heifers had high amplitude LH pulses at 8 months, and high frequency low amplitude LH pulses at 10 months of age (P<0.05). The mean diameter (mm) of the dominant follicle was smaller (P<0.05) in P1L heifers (10.6) compared to P1H (12.8) or P1M (12.2) heifers at 8 months. Maximum size and growth rate of the nonovulatory dominant follicle increased with age (P<0.05) but did not differ between P1H and P1M heifers at puberty. The diameter (mm) of the nonovulatory dominant follicle, and the first and second ovulatory follicles were larger in P2L-12 heifers (14.0, 14.7, and 14.9) compared to P1M heifers (13.1, 12.5, and 11.9), while the peak progesterone levels and CL growth were lower (P<0.05) in the first cycle. In conclusion, dairy heifers attained puberty at a constant body weight and body composition independent of dietary manipulation, the size of dominant follicles increased with age in association with increased LH support, and heifers realimented from a low energy diet developed larger first ovulatory follicles and smaller CL with lower peak progesterone concentrations in the first cycle.     

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.     

Pulsatile secretion of gonadotrophins, ovarian steroids and ovarian oxytocin during prostaglandin-induced regression of the corpus luteum in the cow

A luteolytic dose (500 micrograms) of cloprostenol was given on Day 12 of the oestrous cycle to 5 heifers. Blood samples were collected simultaneously from the caudal vena cava and jugular vein at 5-20-min intervals from -6 to 0 (control period), 0 to 12 and 24 to 36 h after PG injection. Pulses of LH were secreted concomitantly with pulses of FSH during all sampling periods. However, during the control period separate FSH pulses were detected resulting in a shorter (P less than 0.01) interpulse interval for FSH than LH (93 versus 248 min). LH and FSH pulse frequencies increased (P less than 0.01) beginning 1-3 h after PG to interpulse intervals of 59 and 63 min, respectively, and continued to be maintained 24-36 h after PG. Concomitantly there was a 2-3-fold increase (P less than 0.01) in basal concentrations and pulse amplitude for LH (but not FSH). FSH basal concentrations and pulse amplitudes decreased (P less than 0.05) in 3 heifers 24-36 h after PG. Pulsatile secretion of oestradiol was observed at frequencies similar to LH during the periods 4-12 h (3 heifers) and 24-36 h (2 heifers) after PG, respectively, resulting in higher (P less than 0.05) mean oestradiol concentrations. Progesterone concentrations in the vena cava increased (P less than 0.01) 5-10 min after PG but decreased (P less than 0.01) 67% by 20 min after PG. This decrease was followed by a rise (P less than 0.05) beginning 2-3 h after PG and lasting for an average of 3.3 h.(ABSTRACT TRUNCATED AT 400 WORDS)     

Effects of progesterone administered to dairy heifers on sensitivity of corpora lutea to PGF(2)alpha and on plasma LH concentration

To determine whether progesterone facilitates PGF(2)alpha-induced luteolysis prior to day 5 of the estrous cycle, 48 Holstein-Friestian heifers were assigned at random to four treatments: 1) 4 ml corn oil/day + 5 ml Tris-HCl buffer (control); 2) 25 mg prostaglandin F(2)alpha (PGF(2)alpha); 3) 100 mg progesterone/day (progesterone); 4) 100 mg progesterone/day + 25 mg PGF(2)alpha (combined treatment). Progesterone was injected subcutaneously daily from estrus (day 0) through day 3. The PGF(2)alpha was injected intramuscularly on day 3. Estrous cycle lengths were decreased by progesterone: 20.2 +/- 0.56, 19.2 +/- 0.31 (control and PGF(2)alpha); 13.2 +/- 1.40, and 11.7 +/- 1.27 (progesterone and combined). The combination of progesterone and PGF(2)alpha did not shorten the cycle any more than did progesterone alone (interaction, P>0.05). PGF(2)alpha treatment reduced progesterone concentrations on day 6 (P<0.05) and both progesterone and PGF(2)alpha reduced plasma progesterone on day 8 (P<0.01 and P<0.05, respectively). LH was measured in blood samples collected at 10- min intervals for 4 hr on day 4 from three heifers selected at random from each of the four treatment groups. Mean LH concentration for control heifers ranged from 0.35 to 0.63 ng/ml (overall mean, 0.49 ng/ml) and for progesterone-treated heifers ranged from 0.12 to 0.30 ng/ml (overall mean, 0.23 ng/ml). LH concentrations were greater in control heifers (P<0.01). The mean LH pulse rate for control heifers was 2.7 pulses/heifers/4 hr, while that for the progesterone-treated heifers was 1.7 pulses/heifer/4 hr. The mean pulse amplitude for control and progesterone treatments was 0.47 ng/ml and 0.36 ng/ml, respectively. Neither pulse amplitude nor frequency were different between treatment groups.     

Embryo yield and plasma progesterone profiles in superovulated dairy cows and heifers.

This study was conducted to compare the superovulatory (SOV) response of dairy cows (n=172) and heifers (n=172), with two SOV treatments started at the mid-luteal-phase of the estrus cycle. Donors were randomly treated either with equine chorionic gonadotrophin (eCG) plus neutra-eCG serum (eCG+N group, n=167) or follicle stimulating gonadotrophin (FSH-P group, n=177).No significant differences were observed among groups in the percentage of superovulatory responsive donors (SR donors; corpora lutea (CL) >/=2), the mean number of total ova, fertilized ova and viable embryos recovered. Cows yielded significantly less total ova and less fertilized ova (P<0.05) and tended to yield less viable embryos (P<0.06) than heifers.Plasma progesterone (P4) concentrations (n=135 donors) on the day of PGF(2alpha) (PGF) injection and on the day of SOV estrus were significantly higher (P<0.01) in eCG+N than in FSH-P donors and, the increase between those 2 days was also significantly higher (P<0.05) in group eCG+N than in group FSH-P, suggesting a higher luteotrophic effect of eCG than FSH-P. SR donors had P4 levels significantly higher (P<0.001) than non-SR donors only on day 5 after the SOV estrus and on the day of embryo recovery. Plasma P4 concentrations at 5 days after the SOV estrus and at embryo recovery correlated significantly (r=0.76, P<0.001).Heifers had significantly higher P4 levels than cows at gonadotrophin injection (P<0.01), PGF injection (P<0.001), 5 days (P<0.01) and 7 days (P<0.001) after the SOV estrus. At day 7 after the SOV estrus, P4 concentrations per ova recovered were significantly higher in heifers than in cows (P<0.01). The increase of plasma P4 per ova recovered, between days 5 and 7 after the SOV estrus, was significantly (P<0.01) higher in heifers than in cows. Also, the increase of plasma P4 between injections of gonadotrophin and PGF was significantly higher (P<0.05) in heifers than in cows.These results suggest that heifers have higher plasma P4 concentrations at diestrus (either before or after the SOV treatment) and this is associated with a higher embryo yield and quality, as compared to lactating cows. These higher plasma P4 concentrations reflect not only differences in ovulation rate as well as the competence of the corpus luteum, which is potentialized by gonadotrophin stimulation.     

Interaction between dietary intake and ovariectomy on concentrations of insulin-like growth factor-I, GH and LH in plasma of heifers

The objective of this study was to determine if alterations in dietary intake and(or) ovariectomy influence plasma concentrations of IGF-I, GH and LH in heifers. Cyclic heifers (n = 23) were individually fed for 10 wk either 1) 1.8% of body weight in dry matter per day (GAIN; n = 7) 2) 1.1% of body weight in dry matter per day (MAINT; n = 8); or 3) 0.7% of body weight in dry matter per day (LOSE; n = 8). After 10 wk of dietary treatment, heifers were ovariectomized 36 to 40 h following the second injection of prostaglandin F2alpha analog (2 injections 11 d apart). Heifers weighed 444 +/- 13, 387 +/- 8, and 349 +/- 9 kg in the GAIN, MAINT and LOSE groups, respectively, at the time of ovariectomy; the average daily weight gains during the 10-wk period were 0.96, 0.17 and -0.31 kg, respectively (P < 0.001), for the 3 groups. Blood plasma was collected for 6 h at 15-min intervals 1 d before and 2 wk after ovariectomy. The MAINT group of heifers had greater IGF-I concentrations than either the LOSE or GAIN groups; IGF-I decreased (P < 0.05) by 23 and 35% after ovariectomy in the MAINT and GAIN groups, respectively, but did not change (P > 0.10) in the LOSE groups. Dietary restriction tended to increase (P < 0.10) GH pulse frequency and mean GH. Ovariectomy had no effect (P > 0.10) on mean GH or GH pulse frequency but increased (P < 0.05) GH pulse amplitude in the GAIN groups. Dietary treatment had no effect (P > 0.10) on mean LH, or LH pulse amplitude and frequency. However, across dietary treatments, ovariectomy increased mean LH and LH pulse frequency but did not affect (P > 0.10) LH pulse amplitude. In summary, dietary restriction increased GH secretion while ovariectomy increased LH secretion. There appears to be a dichotomy of response between GH and IGF-I in the way heifers respond to dietary treatment and(or) ovariectomy.     

Do endogenous opioid peptides mediate the effects of photoperiod on release of luteinizing hormone and prolactin in ovariectomized ewes?

Three experiments were done to determine if endogenous opioid peptides (EOPs) mediate the effects of photoperiod on release of luteinizing hormone (LH) and prolactin (Prl) in ovariectomized (OVX) ewes. Intravenous infusions of 0.5 naloxone X h-1 X kg body weight-1 for 3.5 h increased (P less than 0.01) mean plasma concentrations of LH and decreased (P less than 0.025) mean interpulse interval (period) of LH pulses in OVX ewes exposed to long day lengths (16L:8D). Infusions of either 1.0 or 2.5 mg morphine-SO4 X h-1 X kg-1 for 3 h increased (P less than 0.005) the period of LH pulses and increased (P less than 0.005) concentrations of Prl in OVX ewes during the breeding season. In OVX ewes exposed to long (16L:8D) or short (8L:16D) day lengths infusions of naloxone increased (P less than 0.05) mean concentrations of LH, whereas morphine decreased (P less than 0.01) mean concentrations of LH. These effects were attributed to changes in period of LH pulses (P less than 0.001). The drug X photoperiod interactions were not significant for LH parameters. Naloxone did not affect Prl release in either long- or short-day groups, but morphine increased (P less than 0.001) Prl release during long and short day lengths. The effect of morphine on Prl release was more pronounced in ewes exposed to long day lengths than in those exposed to short day lengths. In conclusion, EOPs inhibit the LH pulse generator in OVX ewes. However, it is doubtful that the EOPs mediate the steroid-independent effects of photoperiod on LH release. The results also suggest that photoperiod may influence Prl release via opiate neurons.     

Difference in luteinizing hormone response to an opioid antagonist in beef heifers and cows

In three experiments, we examined endogenous opioid inhibition of luteinizing hormone (LH) secretion during the bovine estrous cycle. An increase in serum LH in response to the opioid antagonist naloxone (Na; 1 mg/kg i.v.) was the criterion for opioid inhibition. Estrous cycles were synchronized via prostaglandin administration. In Experiment 1, mean serum LH was not different during the luteal phase in yearling heifers (n = 6/group) at Hour 1 after Nal (2.1 ng/ml) compared to controls (1.8 ng/ml). However, LH peak amplitude was increased (p less than 0.05) in the Nal compared to the control group. Serum LH was increased (p less than 0.01) during the follicular phase in heifers at Hour 1 post-Nal compared to controls (4.7 and 3.5 ng/ml, respectively). Again, Nal administration was followed by increased (p less than 0.05) LH pulse amplitude compared to control. In Experiment 2, no effect of Nal upon serum LH was detected in cows (n = 9) during proestrus, metestrus, midluteal and late luteal portions of the estrous cycle. In Experiment 3, the LH response to Nal was examined simultaneously in yearling heifers and cows (n = 5/group) during the luteal and follicular phases. Serum LH increased (p less than 0.001) during Hour 1 post-Nal in heifers compared to cows during the follicular (3.4 vs. 1.7 ng/ml) but not during the luteal phase. LH pulse amplitude also increased (p less than 0.05) during Hour 1 post-Nal in heifers compared to cows during the luteal (2.5 vs. 1.1 ng/nl and follicular (2.5 vs. 1.3 ng/ml) phases.(ABSTRACT TRUNCATED AT 250 WORDS)     

Estradiol regulation of luteinizing hormone secretion in heifers of two breed types that reach puberty at different ages

The working hypothesis was that 17 beta-estradiol (E(2)) negative feedback on the hypothalamic-pituitary axis in regulation of LH secretion decreases during peripuberty in heifers of 2 different genotypes. We investigated whether Bos indicus heifers had a period postpuberty, as compared with prepuberty, of greater E(2) inhibition of LH secretion at a time when heifers of this genotype have been reported to have a period of anestrus. Prepubertal heifers 9 mo of age of 2 genotypes (B. indicus and B. taurus) were assigned to 3 groups (6 animals/group) to either remain intact (control), be ovariectomized, or be ovariectomized and implanted with E(2). Variables evaluated from 10 to 28 mo of age were circulating concentrations of progesterone (P(4)), presence of corpora lutea, and pulsatile pattern of LH release. Results confirmed that B. taurus heifers attained puberty at younger ages (P < 0.001) and at lower live weights (P = 0.015) than did B. indicus heifers (507 +/- 37 days of age vs. 678 +/- 7 days of age; 259 +/- 14 kg vs. 312 +/- 11 kg; respectively). There was cessation of E(2) inhibition of LH pulses coincident with the onset of puberty in heifers of both breed types but at a much younger age in B. taurus heifers. There was no evidence of enhanced negative feedback of E(2) on LH secretion subsequent to puberty in B. indicus heifers nor was there cessation of estrous cycles in control heifers of either breed type after puberty.     

Luteinizing hormone release in intact and castrate rams is altered with immunoneutralization of endogenous estradiol

Changes in the dynamics of luteinizing hormone (LH) release in the adult ram following immunoneutralization of endogenous estradiol were investigated. Castrate rams were actively immunized against estradiol-6-bovine serum albumin for 7 months and then their patterns of episodic LH release and LH response to multiple injections of gonadotropin-releasing hormone (GnRH, two 5-micrograms doses given iv 2 h apart) were assessed (April). In comparison with control rams immunized against rabbit gamma globulin, estradiol-immunized rams (antibody titre approximately 1:5000) exhibited more frequent LH releases (11.7 +/- 0.3 vs. 9.3 +/- 0.8 pulses/8 h, P less than 0.05) and a greater LH response to the first GnRH injection (peak delta value 190 +/- 8 vs. 130 +/- 25 ng/mL, P less than 0.01). Estradiol antiserum collected from the castrate rams was used in the passive immunization of intact rams (antibody titre approximately 1:200) for 1 month (beginning mid-July). Although episodic LH release was always similar for control and immunized rams, testosterone levels in the latter group increased approximately 150%. In contrast with the castrate ram response, GnRH treatment (two 5-micrograms doses given iv 80 min apart) produced a "self-priming" effect on LH release in the intact rams, an effect that was dampened with estradiol immunoneutralization. Consequently, peak 2:peak 1 ratios for delta value and 80-min mean incremental increase were much smaller (P less than 0.01) for the immunized rams (approximately 2:1 vs. 4:1 for the control rams).(ABSTRACT TRUNCATED AT 250 WORDS)     

Steroid secretion rates and plasma binding activity in androstenedione-immune ewes with an autotransplanted ovary

Mature Merino ewes in which the left ovary and its vascular pedicle had been autotransplanted to the neck were divided into control (N = 5) and immunized groups (N = 6). The immunized ewes were treated (2 ml s.c.) with Fecundin 1 and 4 weeks before the start of blood sampling. Ovarian and jugular venous blood was collected every 10 min at two stages of the follicular phase (21-27 h and 38-42 h after i.m. injection of 125 micrograms of a prostaglandin (PG) analogue) and during the mid-luteal phase (8 h at 15-min intervals). The ewes were monitored regularly for luteal function and preovulatory LH surges. Hormone concentrations and anti-androstenedione titres were assayed by RIA and ovarian secretion rates of oestradiol-17 beta, progesterone and androstenedione were determined. After the booster immunization, progesterone increased simultaneously with titre in immunized ewes, reaching 30 ng/ml at the time of PG injection when median titre was 1:10,000. All ewes responded to PG with LH surges 42-72 h later: 2 of the immunized ewes then had a second LH surge within 3-4 days at a time when peripheral progesterone values were 2-3 ng/ml. The frequency of steroid and LH pulses was greater in immunized ewes (P less than 0.05) during the luteal phase but not the follicular phase. The secretion rate of androstenedione was 6-10 times greater (19-37 ng/min; P less than 0.001) in immunized ewes at all sampling stages. Progesterone secretion rates were 3 times greater (16 micrograms/min; P less than 0.001) during the luteal phase in immunized ewes. The amplitude of oestradiol pulses was significantly reduced in immunized ewes (4.8 vs 2.1 ng/min at +24 h and 6.5 vs 2.8 ng/min at +40 h in control and immunized ewes, respectively: P less than 0.05) during the follicular phase. However, the mean secretion rate of oestradiol at each phase of the cycle was not significantly different between treatment groups. Analysis of bound and free steroid using polyethylene glycol showed that greater than 98% of peripheral and ovarian venous androstenedione and 86% of peripheral progesterone was bound in immunized ewes but there was no appreciable binding (less than 0.1%) in control ewes. Similarly, 50% of ovarian venous oestradiol was bound in immunized ewes compared to 15% in control ewes. We conclude that immunization against androstenedione increases the secretion rate of androstenedione and progesterone but not of oestradiol.(ABSTRACT TRUNCATED AT 400 WORDS)     

Gonadotrophin secretion and ovarian responses in prepubertal heifers actively immunized against androstenedione and oestradiol-17 beta

Groups of heifer calves received a primary immunization against androstenedione (Group A; N = 11) or oestradiol-17 beta (Group E; N = 10) at 3 months of age and booster injections on 5 occasions at 2- to 3-month intervals. Controls (Group C, N = 11) were immunized against human serum albumin alone using the same protocol. Immunity was achieved against both steroids as judged by the secondary antisteroid antibody titres in Group A (1126 +/- 261; reciprocal of titre) and Group E (10,357 +/- 4067) heifers. In Groups A and E there was a general decline in the respective peak antibody titres after successive booster injections. From 3 to 9 months of age mean plasma concentrations of LH were higher (P less than 0.05) in Group E heifers (0.89 +/- 0.08 ng/ml) than in Group C (0.46 +/- 0.03 ng/ml) and Group A (0.59 +/- 0.05 ng/ml) heifers which did not differ from one another. There were no differences between groups in plasma FSH concentrations. At 10 months of age the LH response to exogenous LHRH was of higher (P less than 0.05) amplitude for heifers in Group E (2.59 +/- 0.56 ng/ml) than for those in Groups C (0.61 +/- 0.07 ng/ml) and A (1.04 +/- 0.22 ng/ml). Elevated plasma progesterone concentrations at 5 months of age were shown by 2 heifers in Group C, 10 in Group A, and 6 in Group E. From 8 to 14 months of age a consistently higher proportion of Group A heifers exhibited elevated progesterone compared with Group C and Group E heifers. After ovarian synchronization and booster injection at 15 months of age a corpus luteum was present in 2 heifers in Group C, 7 in Group A and none in Group E. The ovaries of Group A heifers were different from those of Groups C and E and were characterized by greater numbers of 2-4 mm follicles. It is concluded that active immunization against gonadal steroids influences both LH secretion and ovarian function in prepubertal heifers. Early increases in ovarian activity in androstenedione-immunized heifers are maintained after puberty and may therefore confer some lifetime reproductive advantages.     

Influence of pregnancy on the onset of oestrus and luteal function after prostaglandin-induced luteolysis in cattle

Luteolysis was induced by an injection of 500 micrograms cloprostenol (a prostaglandin (PG) analogue) in pregnant (P) Holstein heifers on Days 17 or 24 of gestation and in non-pregnant (NP) Holstein heifers on Day 17 of the oestrous cycle (oestrus = Day 0). Heifers in Groups P-17 (N = 8) and P-24 (N = 8) were inseminated twice whereas those in Group NP-17 (N = 8) were not inseminated. Immediately after PG injection, embryos were recovered by uterine flushing (400 ml) to confirm pregnancy in Groups P-17 and P-24. Uterine flushing with an equivalent volume of physiological saline was also done in Group NP-17. The interval from PG injection to oestrus and to the peak of luteinizing hormone (LH) as well as profile of increase in plasma oestradiol concentrations during that period did not differ (P greater than 0.1) among the groups. However, the proportion of heifers exhibiting abnormal luteal phases (primarily of short duration) during the oestrous cycle after PG injection was greater (P less than 0.01) in Group P-24 than in Groups NP-17 + P-17 pooled (6/8 vs 3/16). These results suggest that the previous presence of a conceptus did not have any effect on the onset of oestrus, or on plasma concentrations of oestradiol and LH after PG-induced luteolysis on Days 17 or 24 of gestation. However, luteal function during the subsequent oestrous cycle was impaired if heifers were 24 days pregnant when luteolysis was induced.     

The effect of immunization of heifers against alpha 1-26 inhibin conjugate on the superovulatory response to pFSH

A total of 121 heifers was blocked by time and diet and then randomly assigned, within block, to an inhibin-immunized (I) or a control (C) group. Immunized heifers (n = 61) received a primary immunization (Day 0) with 0.33 mg of an alpha 1-26 bovine inhibin fragment-human serum albumin (HSA) conjugate injected with non-ulcerative Freund's and DEAE-dextran adjuvants. Booster injections were given on Days 28 and 56. Control heifers (n = 60) received HSA and adjuvants. On Days 56 and 83 the ovaries of heifers were examined by ultrasound to determine the ovulation rate, and blood samples were collected for antibody titer determination. On Day 84, 61 heifers (C, n = 30; I, n = 31) received a total of 24 mg of porcine follicle stimulating hormone (pFSH), while 60 heifers (C, n = 30; I, n = 30) received 12 mg im pFSH, which was administered twice daily for 4 d in decreasing doses during the mid-luteal phase of the estrous cycle. Luteolysis was induced with prostaglandin F(2alpha) analog. The heifers were artificially inseminated and were slaughtered 7 d after estrus. Embryos were recovered and morphologically graded on a scale of 1 to 5 (1 = excellent; 5 = degenerated). Antibody titers (percentage binding at 1:125 serum dilution) differed (P < 0.01) between Group C and I heifers at Days 56 (0.1 vs 30%) and 83 (0.2 vs 37%), and 26% of Group I and 1% of Group C heifers (P < 0.01) had twin ovulations on Day 83. The mean number of embryos recovered was reduced (P = 0.02) in Group I heifers (8.9 +/- 1.2) compared with C heifers (12.1 +/- 1.1); however, the mean number of freezable embryos (Grades 1 and 2) was not affected (P = 0.61) by immunization, and there was no interaction with pFSH (P = 0.36). Ovulation rate as well as embryo yield and quality were not different (P > 0.10) between Group C and I heifers when 12 mg pFSH were administered; however, immunization decreased the superovulatory response to 24 mg of pFSH.     

Influence of pre- and post-partum nutrition on LH secretion in suckled postpartum beef heifers

The influence of nutrition during the last trimester of pregnancy and the early postpartum period on postpartum LH secretion was evaluated in two-year-old Hereford and Simmental heifers maintained on a high or low plane of nutrition (experiment 1) or in Hereford heifers fed a high or low energy (150% vs 100% NRC) ration (experiment 2). Amount of LH released with 10 mg estradiol benzoate (IM) at 14 and 28 days postpartum in experiment 1 or at 14,32,50 and 74 days postpartum in experiment 2 was less (P<.01) for heifers fed the low vs high plane or energy ration and less (P<.05) at 14 days postpartum than at subsequent postpartum periods. The interval from estradiol benzoate injection to the LH peak concentration was longer (P<.05) in Simmental than Hereford heifers, longer (P<.05) in heifers fed the low rather than high energy ration, and longer (P<.01) at 14 days postpartum than at subsequent postpartum periods. The amount of LH released was inversely related to the time required for initiation of the release (r = -.62). Tonic LH secretion was higher (P<.05) in heifers fed the high energy ration and was correlated with average daily gain (r = .75), but was unaffected (P.05) by days postapartum or breed of cattle. Results indicate that increased dietary energy intake increases LH secretion and shortens the anovulatory period in suckled postpartum beef heifers.     

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.     

Estradiol influences on pattern of gonadotropin secretion in bovine males during the period of changed responses to estradiol feedback in age-matched females

When ovaries are removed prior to puberty, administration of exogenous 17 beta-estradiol (E2) decreases concentrations of luteinizing hormone (LH) below that of ovariectomized heifers receiving no E2. Subsequent to the time age-matched intact heifers reach puberty, exogenous E2 increases secretion of LH in ovariectomized heifers above that of ovariectomized heifers receiving no E2. The hypothesis that E2 would inhibit gonadotropin secretion in bovine males during the time E2 no longer inhibited gonadotropin secretion in age-matched bovine females was tested. Males (n = 12) and females (n = 12) were gonadectomized at 241 +/- 3 days of age, and half of each sex (6 males and 6 females) were administered a 27-cm E2 implant. An additional group of males (n = 6) and females (n = 6) remained intact and served as controls. Blood samples were collected (to quantify LH and follicle-stimulating hormone [FSH]) from all animals at 15-min intervals for 24 h at 1, 7, 13, 17, 21, 25, 29, 33, 37, and 43 wk after gonadectomy. Additional blood samples were collected twice weekly from control females to monitor progesterone and onset of corpus luteum function (451 days of age). E2 inhibited frequency of pulses of LH (p less than 0.01) and decreased mean concentration of LH and FSH (p less than 0.01) at Week 1 in gonadectomized males treated with E2 compared to gonadectomized males not administered E2.(ABSTRACT TRUNCATED AT 250 WORDS)