Interruption of the canine estrous cycle with a low and a high dose of the GnRH antagonist, acyline

To test the efficacy and clinical safety of a low and high dose of the GnRH antagonist, acyline, on estrous cycle interruption and anovulation in female dogs, 20 proestrous (<3 d) bitches were randomly assigned to one of the following pharmacological protocols (given sc): acyline 110 μg/kg (ACY-L; n = 6); acyline 330 μg/kg (ACY-H; n = 8); or placebo (PLACE, n = 6). The animals were monitored (clinical and vaginal cytology examinations) daily for 60 d. Blood samples for serum progesterone serum concentrations were collected 14 d after treatment to determine if ovulation had occurred. Appearance of side effects and days to the onset of the first spontaneous estrous cycle after treatment were also recorded. In both ACY groups, but not the PLACE group, estrous cycles were interrupted after treatment (P < 0.05). The interval from treatment to estrus interruption in ACY-L and ACY-H groups was 3.0 ± 0.6 and 3.2 ± 0.2 d, respectively (LSM ± SEM; P > 0.05). In the PLACE bitches, physical, behavioral and cytological proestrus slowly progressed to estrus and diestrus. Ovulation was absent in all ACY, but not in PLACE bitches (P < 0.05). None of the females manifested side effects related to the treatments (P > 0.05). Spontaneous return to a normal estrous cycle during the study period occurred in all ACY (ACY-L 19.5 ± 2.7 d vs ACY-H 24.8 ± 2.0 d; P > 0.05), but in none of the PLACE bitches (P < 0.05). In conclusion, acyline efficiently, safely and reversibly interrupted an early phase of the estrous cycle in bitches by preventing ovulation.     

The GnRH antagonist acyline prevented ovulation, but did not affect ovarian follicular development or gestational corpora lutea in the domestic cat

Two experiments were conducted to investigate the effects of the GnRH antagonist acyline (330 μg/kg, given sc) on ovarian follicular development and ovulation, as well as on pregnancy maintenance in domestic cats. In the first experiment, seven queens in proestrus (total of 24 proestrus periods), were randomly assigned to treatment with either acyline (ACY; n = 17) or a placebo (PLC; n = 7). All queens were mated with a fertile tomcat. In the ACY and PLC groups, cessation of estrus occurred (mean ± SEM) 7.0 ± 1.3 and 7.0 ± 1.7 d after treatment (P > 0.1), ovulation occurred in 2 of 17 and all seven estrus periods (P < 0.05), and pregnancy rates were 1 of 16 and 7 of 7 (P < 0.05), respectively. In the ACY and PLC groups, intervals from treatment to the onset of the ensuing proestrus were 18.4 ± 1.7 and 120 ± 17.2 d. In the second experiment, 14 pregnant queens were randomly allocated, according to their mating date, to treatment with acyline in early pregnancy (from 20 to 25 d, n = 3), mid pregnancy (from 26 to 45 d; n = 4), late pregnancy (> 45 d; n = 3), or injection of a placebo in early (n = 1), mid (n = 2), or late pregnancy (n = 1). Ultrasonographic assessments of the uterus were done every second day for 2 wk post treatment, and serum progesterone (P₄) concentrations were determined before treatment, and at 7 and 14 d after treatment. No pregnancies were prematurely terminated and post-treatment P₄ concentrations did not differ among treatment groups (P > 0.1). In conclusion, in the domestic cat, GnRH withdrawal by acyline prevented ovulation when given in early follicular phase (proestrus), but did not significantly affect luteal function during pregnancy.     

Effects of dietary vitamin supplementation and semen collection frequency on hormonal profile during ejaculation in the boar

To evaluate the effect of dietary and management factors on boar hormonal status during ejaculation, 39 boars were canulated to determine the profiles of luteinizing hormone (LH), follicle-stimulating hormone (FSH), 17β-estradiol (E₂), and testosterone (T) in blood plasma and seminal fluid. Prior to canulation, 18 boars were fed a basal diet (control), whereas the remainder (n = 21) were fed a basal diet supplemented with extra vitamins (supplemented). Within each dietary treatment, two regimens of semen collection were used over the 3 mo preceding the hormonal evaluation: three times per 2 wk (3/2) or three times per wk (3/1). Plasma E₂ was lower (P < 0.01) before ejaculation (232.5 ± 22.6 pg/mL) than at the onset of ejaculation (255.2 ± 27.1 ng/mL). Plasma T increased from 5.14 ± 0.72, before ejaculation to 5.87 ± 0.86 ng/mL at the onset of ejaculation in supplemented boars, whereas it decreased from 5.15 ± 0.65 to 4.87 ± 0.70 ng/mL in controls (diet by time, P < 0.05). At the onset of ejaculation, plasma FSH was higher in 3/2 boars (0.436 ± 0.06 ng/mL) than in 3/1 boars (0.266 ± 0.04 ng/mL; P < 0.05). During ejaculation, plasma LH increased linearly (P < 0.01) from 0.59 ± 0.07 to 0.97 ± 0.10 ng/mL, and plasma E₂ and T concentrations were correlated (r = 0.62, P < 0.01). Plasma FSH before and during ejaculation was negatively correlated with sperm production (r = −0.60, P < 0.01) and testicular weight (r = −0.50, P < 0.01). In conclusion, dietary and management factors had few impacts on hormonal profiles during ejaculation, but homeostasis of some hormones was related to some criteria of reproductive performance in boars.     

The effect of porcine luteinizing hormone in the synchronization of ovulation and corpus luteum development in nonlactating cows

The objective of this study was to determine the effects of different doses of porcine luteinizing hormone (pLH) versus 100 μg gonadotropin-releasing hormone (GnRH) on ovulatory response (during diestrus and proestrus) and corpus luteum (CL) development in nonlactating cows. In Experiment 1, 75 cows received an intravaginal insert containing 1.9 g progesterone (P4) for 10 d to synchronize estrus (Day 0), with prostaglandin F_2α (PGF) at insert removal. On Day 5, all follicles ≥8 mm were ablated, and on Day 12, cows received 8, 12.5, or 25 mg pLH or 100 μg GnRH. Mean (±SEM) plasma P4 concentrations on Day 12 did not differ among treatments (5.6 ± 0.2 ng/mL). Mean plasma LH concentration was greatest (P < 0.01) in cows given 25 mg pLH (4.3 ± 0.4 ng/mL). The ovulatory response to 25 mg pLH (84%) or 100 μg GnRH (72%) was greater (P < 0.05) than that to 8 mg pLH (32%), but not different from that of 12.5 mg pLH (58%). In Experiment 2, 68 cows were given two injections of PGF 10 d apart to synchronize estrus (Day 0). On Day 7, cows received PGF, and, 36 h later, pLH or GnRH (as in Experiment 1). The interval from treatment to ovulation was most variable in cows given 8 mg pLH; only 65% of these cows ovulated during the initial 27 h versus 88% of cows given 25 mg pLH (P < 0.05). Cows given 25 mg pLH or 100 μg GnRH had larger CL area and greater plasma P4 concentrations (P < 0.05) than that of those given 8 mg pLH. In summary, diestrous cows given 25 mg pLH had the greatest plasma luteinizing hormone concentrations, but ovulatory response did not differ from that of those given 100 μg GnRH. Proestrous cows given 25 mg pLH or 100 μg GnRH had greater CL area and P4 concentrations than that of those given 8 mg pLH.     

Follicle suppression of circulating follicle-stimulating hormone and luteinizing hormone before versus after emergence of the ovulatory wave in mares

The effect of the ovarian follicles on plasma concentrations of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) before versus after the expected emergence of the ovulatory follicular wave was studied on Days 0 to 18 (Day 0 = ovulation) in four groups of mares (n = 6/group). In addition to a control group, all follicles ≥6 mm in diameter were ablated on Days 0.5, 6.5, or 12.5 in a herd of mares with reported emergence at 6 mm of the future ovulatory follicle on mean Day 10.5. Concentrations of FSH were not different between the Day-0.5 or Day-6.5 ablation groups and the corresponding controls. However, ablation on Day 12.5 resulted in an immediate FSH increase (group-by-day interaction, P < 0.003). For LH, ablation on Day 0.5 resulted in an interaction (P < 0.02), partially from lower (P < 0.05) concentrations on each of Days 15.5 to 18.0 than that in the controls, whereas ablation on Days 6.5 or 12.5 did not result in a significant group effect or interaction. Testosterone concentration, but not progesterone or estradiol concentration, was lower (P < 0.04) on Day 2 in the Day-0.5 ablation group than that in the controls. We inferred that follicles did not contain adequate FSH suppressors on Days 0.5 and 6.5 and that they were present only in the Day-12.5 ablation group or after the expected emergence of the ovulatory wave. The hypothesis of an association between low postovulatory concentrations of an ovarian steroid and low concentrations of LH after Day 15 was supported.     

Role of LH in luteolysis and growth of the ovulatory follicle and estradiol regulation of LH secretion in heifers

The role of LH in luteolysis and development of the ovulatory follicle and the involvement of GnRH receptors in estradiol (E2) stimulation of LH secretion were studied in heifers. A pulse of PGF_2α, as indicated by a metabolite, was induced by E2 treatment on Day 15 (Day 0 = ovulation) and LH concentration was reduced with a GnRH-receptor antagonist (acyline) on Days 15, 16, and 17. Blood samples were collected every 6 h on Days 14-17 and hourly for 10 h beginning at the Day-15 treatments. Four groups were used (n = 6): control, acyline, E2, and E2/acyline. The number of LH pulses/heifer during the 10 h posttreatment was greater (P < 0.0002) in the E2 group (2.3 ± 0.4, mean ± SEM) than in the acyline group (0.2 ± 0.2) and was intermediate in the E2/acyline group (1.4 ± 0.2). Concentrations of progesterone in samples collected every 6 h on Day 15 showed a group-by-hour interaction (P < 0.02); concentrations decreased in the acyline group but not in the control group. The 12 heifers in the combined acyline and E2/acyline groups had three follicular waves compared to two waves in 10 of 12 heifers in the combined control and E2 groups. Results (1) supported the hypothesis that LH delays the progesterone decrease associated with luteolysis, (2) supported the hypothesis that LH has a positive effect on the continued development and growth of the selected ovulatory follicle, and (3) indicated that E2 stimulates LH production through an intracellular pathway that involves GnRH receptors on the gonadotropes and a pathway that does not involve the receptors.     

Effects of treatment with a prostaglandin analogue on developmental dynamics and functionality of induced corpora lutea in goats

The aim of this study was to compare morphological and functional features of spontaneous and induced corpora lutea (CLs) in goats. Fourteen adult and cycling Anglo Nubian goats (Argentina) were randomly allocated to two groups: Group N (n = 7) included goats with natural spontaneous oestrus and Group PG (n = 7) included does in which oestrus was synchronized by the administration of two i.m. cloprostenol doses, 10 days apart. In both groups, oestrous behaviour was checked twice daily (Day of oestrus = Day 0) and daily transrectal ultrasonographies were performed for evaluating CLs and follicles dynamics through the complete subsequent oestrous cycle; the luteal activity was determined directly, in terms of progesterone (P4) secretion, and indirectly, by assessing effects of CL on follicular dynamics. All goats exhibited oestrous behaviour and ovulation without differences in ovulation rate (N: 1.67 ± 0.2, PG: 2.0 ± 0.1). The total luteal tissue area showed linear growth from Day 4 to Day 15 of oestrous cycle in all goats, but the developmental dynamics differed between groups, treated goats had larger area (P < 0.01). Plasma P4 concentrations also increased from Day 0 to Day 15 in all the does; however, from Day 5 to Day 15, treated does had a lower concentrations than the untreated group (P < 0.001). There were differences in the development of follicular waves between groups; assessment of size-distribution showed that treated group had a higher number of small and larger follicles (P < 0.05). The largest follicles recorded in treated goats had a higher maximum diameter both at the first (PG: 7.6 ± 0.8 mm; N: 4.9 ± 0.7 mm, P < 0.05) and second follicular waves (PG: 6.3 ± 1.4 mm; N: 5.0 ± 0.4 mm, P < 0.05) and a longer growth phase during the second wave (PG: 6.5 ± 1.7 days; N: 4.6 ± 0.7 days, P < 0.05), coincident with the period of maximal luteal secretion. In conclusion, synchronization of oestrus and ovulation by the administration of a prostaglandin analogue causes differences in developmental dynamics and functionality of induced corpora lutea when compared to natural spontaneous ovulation.     

Age-related dynamics of follicles and hormones during an induced ovulatory follicular wave in mares

An ovulatory follicular wave was induced by ablation of follicles ≥6 mm and treatment with prostaglandin F2α (PGF) on Day 10 (ovulation = Day 0). Follicle and hormone dynamics of the induced waves were compared among three age groups: young (5-6 y, n = 14 waves), intermediate (10-14 y, n = 16), and old (≥18 y, n = 15). During the common-growth phase of the induced wave (Days 12-17), diameter of the future ovulatory follicle was not different among ages, but the young group had more (P < 0.05) follicles that reached ≥10 mm. The number was correlated (r = +0.7; P < 0.0001) within mares between consecutive interovulatory intervals, indicating repeatability. Concentrations of LH increased in all age groups during Days 12-17, but were greatest (P < 0.002) in the young group and continued to be greater (P < 0.0001) throughout the ovulatory LH surge. During several days before Day −1, there were no age-related effects on systemic estradiol concentrations, diameter of the preovulatory follicle, or B-mode echo texture or color-Doppler signals of blood flow in the follicle wall. Interpretations were: (1) greater number of follicles in the young group reflected a greater follicle reserve, (2) greater LH concentrations throughout the ovulatory surge in the young group reflected a more positive response to an extraovarian/environmental influence after removal of the negative effect of progesterone, and (3) lower LH concentrations in the older groups were adequate for the preovulatory changes in the follicle.     

Low peripheral progesterone and late embryonic/early fetal loss in suckled beef and lactating dairy cows

Pregnancy failure during placentation in lactating dairy cows was associated with low concentrations of serum progesterone. Beef cows have greater serum progesterone and less pregnancy failure. Experiment 1 determined that reduction of serum progesterone affected late embryonic/early fetal loss in suckled beef cows. Cows (n = 40) received progesterone from two new or used controlled internal drug releasing devices, replaced every 5 d, beginning on Day 28 of gestation (mating = Day 0); CL were enucleated on Day 29. Retention of pregnancy was 77% in treated cows and 97% in 78 control cows (P < 0.05). Experiment 2 determined how pregnant, lactating dairy cows with high or low progesterone concentrations during Days 28-34 differed in luteal function or in serum progesterone during replacement therapy. Luteal tissue from such cows was assayed for progesterone and expression of mRNA for genes of endothelin and prostaglandin (PG) systems. Secretion of progesterone and prostaglandins by dispersed luteal cells was determined during incubation with LH, endothelin-1, or arachidonic acid. Neither luteal progesterone nor mRNAs for endothelin or prostaglandin systems differed. Endothelin-1 inhibited secretion of progesterone more (P < 0.05) in luteal cells from cows with low versus high serum progesterone, when incubated with arachidonic acid. Secretion of prostaglandin F₂α was increased and that of 6-keto-PGF₁α decreased by endothelin-1 in vitro. Serum progesterone during replacement was lower (P < 0.05) for cows with low than high serum progesterone at lutectomy. Thus, clearance, more than luteal production, determined peripheral progesterone in pregnant, lactating dairy cows.     

Normal or induced secretory patterns of luteinising hormone and follicle-stimulating hormone in anoestrous gonadotrophin-releasing hormone-immunised and cyclic control heifers

The objective was to determine the effect of gonadotrophin-releasing hormone (GnRH), GnRH analogue (GnRH-A) or oestradiol administration on luteinising hormone (LH) and follicle-stimulating hormone (FSH) release in GnRH-immunised anoestrous and control cyclic heifers. Thirty-two heifers (477 ± 7.1 kg) were immunised against either human serum albumin (HSA; controls; n = 8), or a HSAGnRH conjugate. On day 70 after primary immunisation, control heifers (n = 4 per treatment; day 3 of cycle) received either (a) 2.5 μg GnRH or (b) 2.5 μg of GnRH-A (Buserelin®) and GnRH-immunised heifers (blocked by GnRH antibody titre; n = 6 per treatment) received either (c) saline, (d) 2.5 μg GnRH, (e) 25 μg GnRH or (f) 2.5 μg GnRH-A, intravenously. On day 105, 1 mg oestradiol was injected (intramuscularly) into control (n = 6) and GnRH-immunised anoestrous heifers with either low (13.4 ± 1.9% binding at 1:640; n = 6) or high GnRH antibody titres (33.4 ± 4.8% binding; n = 6). Data were analysed by ANOVA. Mean plasma LH and FSH concentrations on day 69 were higher (P < 0.05) in control than in GnRH-immunised heifers (3.1 ± 0.16 vs. 2.5 ± 0.12 ng LH ml⁻¹ and 22.5 ± 0.73 vs. 17.1 ± 0.64 ng FSH ml⁻¹, respectively). The number of LH pulses was higher (P < 0.05) in control than in GnRH-immunised heifers on day 69 (3.4 ± 0.45 and 1.0 ± 0.26 pulses per 6 h, respectively). On day 70, 2.5 μg GnRH increased (P < 0.05) LH concentrations in control but not in GnRH-immunised heifers, while both 25 μg GnRH and 2.5 μg GnRH-A increased (P < 0.05) LH concentrations in GnRH-immunised heifers, and 2.5 μg GnRH-A increased LH in controls. FSH was increased (P < 0.05) in GnRH-immunised heifers following 25 μg GnRH and 2.5 μg GnRH-A. Oestradiol challenge increased (P < 0.05) LH concentrations during the 13–24 h period after challenge with a greater (P < 0.05) increase in control than in GnRH-immunised heifers. FSH concentrations were decreased (P < 0.05) for at least 30 h after oestradiol challenge. In conclusion, GnRH immunisation decreased LH pulsatility and mean LH and FSH concentrations. GnRH antibodies neutralised low doses of GnRH (2.5 μg), but not high doses of GnRH (25 μg) and GnRH-A (2.5 μg). GnRH immunisation decreased the rise in LH concentrations following oestradiol challenge.     

Aluminium effects on thyroid gland function: iodide uptake, hormone biosynthesis and secretion

The effects of aluminium (Al) on thyroid function were evaluated in adult Wistar rats intraperitoneally (i.p) injected with 7 mg Al (as lactate)/kg body weight (b.w) per day during a six week period. The time-course kinetics of Na¹²⁵I (3 μCi per 100 g b.w, i.p) was analysed by measuring gamma-radioactivity of thyroid, serum, serum protein precipitate and bile, at times ranging from 2 to 96 h post-dosing. In Al-treated group the ¹²⁵I⁻ thyroid uptake at 24 h (15,840 ± 570 vs. 18,030 ± 630 dpm/mg, P < 0.05) as well as the rate of ¹²⁵I⁻ release from the gland, calculated as the slope of the plot between 24 and 96 h (84 ± 8 vs. 129 ± 11 dpm/mg/h, P < 0.05) were significantly reduced as compared to control. The biliary ¹²⁵I⁻ excretion was not modified at all studied times. The Al content and lipid peroxidation (69.1 ± 8.5 vs. 53.2 ± 7.0 nmol MDA/g wet weight, P < 0.05) of thyroid tissue were increased in Al-treated rats. The serum concentrations of total thyroxine (T4, 3.78 ± 0.14 vs. 4.68 ± 0.12 μg/dL, P < 0.05) and total triiodothyronine (T3, 47 ± 4 vs. 66 ± 5 ng/dL, P < 0.05) were decreased by effect of Al, but free-T4 (1.05 ± 0.05 vs. 1.04 ± 0.04 ng/dL, NS) and thyrotropin (TSH, 2.7 ± 0.4 vs. 2.6 ± 0.5 ng/ml, NS) remain unchanged. In spite of the Al could indirectly affect thyroid iodide uptake and hormones secretion by a mechanism involving the induction of an oxidative stress state, however, these changes could be managed by the hypothalamus-pituitary-thyroid endocrine axis. We can conclude that in adult rats the Al would not act as a thyroid disruptor.     

Characteristics of peaks in serum concentrations of follicle-stimulating hormone and estradiol, and follicular wave dynamics during the interovulatory interval in cyclic ewes

There are three or four ovarian follicular waves in the interovulatory interval of cyclic ewes. Each follicular wave is preceded by a transient peak in serum follicle-stimulating hormone (FSH) concentrations. Serum concentrations of estradiol also increase concurrent with the growth of follicle(s) in each wave. In the current study, we investigated the patterns of follicular wave development and characteristics of FSH and estradiol peaks in all follicular waves of the interovulatory interval and after induction of a supraphysiologic FSH peak in cyclic ewes (Ovis aris). In Experiment 1, 19 ewes underwent daily ovarian ultrasonography and blood sampling for a complete interovulatory interval. In Experiment 2, seven ewes received two administrations of ovine FSH (oFSH), 8 h apart (1 μg/kg; sc), at the expected time of the endogenous FSH peak preceding the second follicular wave of the interovulatory interval. In Experiment 1, the amplitude of the FSH peaks decreased (up to 50%), whereas basal serum FSH concentrations increased across the interovulatory interval (P < 0.05). Maximum follicular diameter was greater (P < 0.05) for Wave 1 and the Ovulatory wave (6.0 ± 0.3 and 6.1 ± 0.2 mm, respectively) than for Waves 2 and 3 (5.3 ± 0.1 and 5.4 ± 0.3 mm, respectively). Life span was greater for follicles in Wave 1 compared with other waves (P < 0.05). Treatment with oFSH increased the amplitude of an FSH peak by 5- to 6-fold. This treatment increased estradiol production (P < 0.05) but had little effect on other characteristics of the subsequent follicular wave. We concluded that changes in the amplitude and duration of the peaks in serum concentrations of FSH that precede follicular waves across the interovulatory interval do not influence the characteristics of the follicular waves that follow.     

Dynamic changes in ovarian follicles measured by ultrasonography during gonadotropin stimulation in rhesus monkeys

The objective was to study dynamic changes of ovaries in rhesus macaques stimulated by gonadotropins to identify an indicator for predicting ovarian response to stimulation. Twenty-one cycling monkeys were given 36 IU/d recombinant human follicle-stimulating hormone (rhFSH) for 8 d. Animals (n = 17) with ≥5 follicles (≥3 mm) in their ovaries on Day 9 of ovarian stimulation were deemed good responders, whereas those with a lesser response were poor responders (n = 4). For these two groups, the mean (±SD) numbers of oocytes retrieved were 44.3 ± 21.4 and 11.0 ± 4.6, respectively. In retrospect, the mean diameters of the ovaries and of the largest follicles, the total number of detectable follicles (diameter >0.5 mm), and serum estradiol concentrations gradually increased during the stimulation period in the good responders but did not increase in the poor responders. Comparing good and poor responders, the number of ovarian follicles >0.5 mm already exhibited a difference (12.9 ± 6.5 vs. 2.9 ± 1.3, respectively, P < 0.05) on Day 1 of stimulation. However, for other end points, differences were not significant until at least Day 5. Moreover, good responders yielded a fivefold higher blastocyst development rate than that of poor responders (P < 0.01). In conclusion, the number of ovarian follicles detected with ultrasonography could be useful to predict the response to FSH stimulation in non-human primates.     

Ovarian and endocrine responses in tropical sheep treated with reduced doses of cloprostenol

The present study aimed to assess the efficacy of reduced doses of cloprostenol for synchronizing estrus and ovulation in hair sheep. With the aim to evaluate the luteolytic activity of reduced cloprostenol doses, a first experiment was performed using a relatively large (group H: 126 μg; n = 8), medium (group M: 68.25 μg; n = 6) and small (group L: 38.5 μg; n = 6) cloprostenol dose. Luteolysis was assessed at Days 3 and 6 after injection (Day 0) by progesterone concentrations (P₄) and transrectal ultrasonography (US). In Experiment 2, sheep were randomly assigned to the same three doses to evaluate a protocol for estrous synchronization using two injections administered 9 days apart. A third trial was performed with ewes treated (9 days apart) with the large dose (H = 126 μg; n = 12) and with a small dose adjusted for facilitating volume management (LA = 43.75 μg; n = 12). Presence of estrous cycling was determined in all the ewes by US and P₄ assay, at Days −9, −6, −2, 0 (Day of second cloprostenol injection), 8 and 11. Bleeding and US were done every 4 h from 16 h of the beginning of the estrus during the third trial to assess the preovulatory LH surge and timing of ovulation. Additionally, blood samples were drawn at Days 0, 1, 2 and 3 to assess estradiol (Experiments 2 and 3) and P₄ (Experiment 2) concentrations during the ovarian follicular phase. In all experiments, percentage of animals showing luteolysis, preovulatory follicular dynamics and function and percentage of ewes showing behavioral estrus in response to treatment was similar among groups. Timing of estrus for group H was earlier than group L (28.6 ± 1.8 h compared with 37.1 ± 2.4 h; P < 0.05). In the third trial, the preovulatory LH peak was higher in the LA group than group H, in terms of maximum mean concentration during the surge (27.7 ± 1.8 ng/mL compared with 21.3 ± 2.2 ng/mL; P < 0.05) and area under the curve (AUC; 183.4 ± 12.7 ng/mL compared with 127.7 ± 10.9 ng/mL; P < 0.01). However, timing of ovulation was similar for H and LA groups. Thereafter, ovulation rate and luteal function at Day 11 were similar. Current results demonstrate that reduced doses of cloprostenol may be applied in a practical manner for reproductive management of sheep, with the additional advantage of reducing treatment costs.     

Plasma follicle stimulating hormone,ovulation rate and fertility in Merino ewes treated with bovine follicular fluid

Charcoal-treated bovine follicular fluid (bFF) given as four 5-ml subcutaneous injections to 13 Merino-Border Leicester ewes around the time of natural luteolysis suppressed (P<0.01) plasma levels of follicle stimulating hormone (FSH) [from 1.08 ± 0.05 to 0.41 ± 0.03, mean ± s.e.m. of log_e (ng+ 1) /mlplasma]. This was followed (P < 0.01) by hypersecretion or a rebound of FSH (to 1.46 ± 0.11) lasting 32 h in 10 of the treated ewes, and then by a further fall (to 0.73 ± 0.03, P < 0.05) before the surge (1.21 ± 0.07, P < 0.05) associated with the preovulatory surge of luteinizing hormone (LH).Plasma FSH at 56–72 h before the LH surge (i.e., at the time of the FSH rebound) was correlated with the subsequent ovulation rate (n=13, r= + 0.73, P < 0.01). Fewer ewes treated with four injections of 2 or 5 ml of bFF than control ewes (injected with bovine plasma) became pregnant (28 of 41 vs. 38 of 41, χ² = 4.05, P < 0.05), although plasma progesterone was similar at Day 11 in treated and control ewes. It is concluded that plasma FSH during such a rebound influences the subsequent ovulation rate in sheep.     

Plasma luteinizing hormone concentrations in cows given repeated treatments or three different doses of gonadotropin releasing hormone

We hypothesized that: (i) repeated GnRH treatments would increase the magnitude and duration of the LH surge and would increase progesterone (P4) concentrations after ovulation; and (ii) the release of pituitary LH would be greater in response to larger doses of GnRH. In Experiment 1, ovary-intact cows were given an intravaginal P4 (1.9 g) insert (CIDR) for 10 d and 500 μg cloprostenol (PGF) at CIDR removal to synchronize estrus. On Days 7 or 8 after estrus, cows received two PGF treatments (12 h apart) and 100 μg GnRH at 36 (Control), 36 and 38 (GnRH38), or 36 and 40 h (GnRH40) after the first PGF. Mean plasma LH concentration (ng/mL) was greater (P < 0.05) in GnRH38 (8.8 ± 1.1) than in Control (5.1 ± 1.3), with that in GnRH40 (5.8 ± 1.3) being intermediate. Although the duration (h) of the LH surge was longer in GnRH40 (8.0 ± 0.4) than in either GnRH38 (P < 0.05; 7.0 ± 0.3) or Control (P < 0.09; 7.1 ± 0.4), mean postovulatory P4 (ng/mL) was greater (P < 0.01) in Control (4.2 ± 0.7) than in GnRH38 (2.9 ± 0.6) or GnRH40 (3.0 ± 0.7) cows. In Experiment 2, ovariectomized cows were given a CIDR for 10 d and 2 mg of estradiol cypionate im at CIDR insertion. Thirty-six hours after CIDR removal, cows received, 50, 100, or 250 μg of GnRH. Cows given 250 μg GnRH released more LH (9.4 ± 1.4 ng/mL) than those given 50 or 100 μg (6.1 ± 1.3 and 5.4 ± 1.4 ng/mL, respectively), and had an LH surge of longer duration than those given 50 μg (6.8 ± 0.4 vs. 5.1 ± 0.3 h). In summary, ovary-intact cows in the GnRH38 group had greater mean and peak LH concentrations, but subsequent plasma P4 concentrations were lower than in Control cows. Ovariectomized cows given 250 μg GnRH had a greater pituitary release of LH.     

Use of deslorelin short-term implants to induce ovulation in cycling mares during three consecutive estrous cycles

Alternatives to human chorionic gonadotropin (hCG) for inducing ovulation in cycling mares over several consecutive cycles were explored. Placebo, one, three or five short-term implants each containing 2.2 mg of gonadotropin-releasing hormone (GnRH) analogue (deslorelin) were administered to cycling mares after identification of a follicle over 30 mm. Mares were treated over three consecutive cycles, and artificially inseminated during the third cycle only. Serum was assayed for concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and progesterone during each cycle. All deslorelin doses decreased the interval to ovulation (4.0 days, 2.6 days, 2.4 days and 2.0 days), increased the proportion of mares ovulating within 48 h (20.0%, 83.3%, 73.3% and 85.7%), and decreased the diameter of the largest follicle at ovulation (45.8 mm, 38.0 mm, 41.0 mm and 37.2 mm) for the placebo, 2.2 mg, 6.6 mg and 11.0 mg groups, respectively (P < 0.05). The interovulatory interval was lengthened in the 11.0 mg group compared with all other groups (21.2 days, 21.8 days, 26.4 days and 32.7 days for the placebo, 2.2 mg, 6.6 mg and 11.0 mg groups, respectively, P < 0.05). No differences (P > 0.05) were detected in serum concentrations of progesterone or pregnancy rate among the groups. Cycle number had no effect (P > 0.05) on the reproductive parameters or serum concentrations of progesterone. Concentrations of LH were higher in Cycle 3 in the 6.6 mg and 11.0 mg groups, and FSH concentrations were decreased in Cycles 2 and 3 in treated mares. Deslorelin was efficacious for inducing ovulation in cycling mares with no diminished activity over three consecutive cycles. However, ovarian suppression and changes in serum concentrations of gonadotropins were noted at higher doses.     

Effects of active immunization against a 13-amino acid receptor-binding epitope of FSHβ on fertility regulation in female mice

Follicle-stimulating hormone (FSH) is crucial for ovarian folliculogenesis and thus essential for female fertility. Here, we developed a novel FSH vaccine based on the tandem of a 13-amino acid receptor-binding epitope of FSHβ (FSHβ13AA-T) and used a mouse model to test its efficacy in female fertility regulation. Compared to placebo-immunized controls, FSHβ13AA-T vaccination: induced a marked (P < 0.05) antibody generation; reduced (P < 0.05) serum concentrations of FSH, inhibin B and 17β-estradiol; disrupted (P < 0.05) normal estrous cyclicity; delayed (P = 0.08) establishment of pregnancy; blocked (P < 0.05) folliculogenesis; and reduced (P < 0.05) litter size. Mechanistically, FSH vaccination reduced (P < 0.05) ovarian estrogen production by decreasing Lhcgr, Cyp19a1 and HSD3β1 expression, and suppressed ovarian follicular development by decreasing ovarian Fshr, Inhα, Foxo3a, Bmp15 and Cdh1 expression. Overall, vaccination of female mice with FSHβ13AA-T substantially disrupted FSH-dependent ovarian steroidogenesis and folliculogenesis, and caused subfertility. Therefore, vaccines based on FSHβ13AA-T have potential as anti-fertility/contraceptive agents in females.     

Administration of sulpiride or domperidone for advancing the first ovulation in deep anestrous mares

Since results with using sulpiride and domperidone are conflicting and since both have not been tested at the same time, the aim of this study was to compare the efficacy of these substances for the induction of ovulation in deep anestrous mares in the same experimental conditions and to determine their fertility after artificial insemination (AI) at the induced estrus. Twenty-six non-pregnant, non-lactating standardbred anestrous mares were randomly assigned to three groups and treated daily for 25 days (from February 3rd to February 28th) with either sulpiride (1 mg/kg of body weight im SID, n = 10), or domperidone (1 mg/kg po SID, n = 10); 6 animals were used as control. The beginning of the transition period and the first ovulation were hastened in sulpiride (16.4 ± 0.8 days) but not in domperidone (46.0 ± 3.3 days) treated mares (P < 0.05). The diameter of the largest follicle was affected by treatment, time and interaction of treatment-by-day (P < 0.05) and significantly increased in the sulpiride group (P < 0.05). Although a main effect of treatment on plasma LH concentration was not observed (P = 0.06), time and interaction of treatment-by-day were statistically significant (P < 0.05). The interval from the beginning of treatment to first ovulation was shorter (P < 0.05) in the sulpiride group (36.9 ± 2.5 days) than in the domperidone (74.7 ± 3.3 days) and control (81.4 ± 3.1) groups. The establishment of pregnancy was significantly (P < 0.05) hastened in sulpiride (61.0 ± 35.2 days) but not in domperidone (83.0 ± 44.0 days) treated mares. Treated mares not pregnant after the first AI, showed normal estrous cycles with regular interovulatory intervals (P > 0.05). It was concluded that sulpiride is effective in advancing the beginning of transition period and the first ovulation whereas domperidone is successful only in some mares.     

Calcium, parathyroid hormone, oxytocin and pH profiles in the whelping bitch

Despite the high prevalence of primary uterine inertia in whelping bitches, the underlying pathogenesis remains unclear. The objectives were to i) determine serum concentrations of total calcium, ionized calcium (iCa), parathyroid hormone (PTH), and blood pH in normally whelping bitches throughout the peri-parturient period; and ii) investigate relationships among iCa, PTH, and acid-base status, and the role that they and oxytocin may have in the underlying pathogenesis of canine uterine inertia. Bitches were randomly selected from a population of German Shepherd Dog bitches with a history of uncomplicated parturition (Group 1; n = 10), and from a population of Labrador bitches with a clinical history of an increased incidence of uterine inertia and stillbirths (Group 2; n = 20). Jugular blood samples were collected daily from -4 d to the onset of whelping (t = 0 h), and then every 4 h until the last pup was born. Overall, bitches from Group 2 had higher mean ± SEM serum concentrations of PTH (4.72 ± 2.45 pmol/L, P < 0.001), lower iCa (1.31 ± 0.08 pmol/L, P < 0.05), and higher venous pH (7.41 ± 0.03, P < 0.005) than bitches from Group 1 (2.9 ± 1.44 pmol/L, 1.38 ± 0.06 mmol/L, and 7.33 ± 0.02, respectively) during the periparturient period. However, there was no significant difference between Groups 1 and 2 for serum oxytocin concentrations during the periparturient period (45.5 ± 40 and 65.5 ± 82 pg/mL). We inferred that low iCa resulting from a rising pH and decreasing PTH during the periparturient period may have contributed to decreased uterine contractility and increased risk of stillbirths. Therefore, manipulating the cationic/anionic difference in diets of pregnant bitches, similar to the bovine model for hypocalcamia, may reduce the incidence of stillbirths in the bitch.