Regulation of uterine function during estrous cycle, anestrus phase and pregnancy by steroids in red deer (Cervus elaphus L.).

Steroid synthesis and production in ruminant uterus is not obvious, especially in seasonally reproduced. We compared steroid production by investigating enzymes involved in red deer uterine steroid metabolism in reproductive seasons. Blood and uteri (endometrium and myometrium) were collected post mortem from hinds on 4th day (N = 8), 13th day of the cycle (N = 8), anestrus (N = 8) and pregnancy (N = 8). The expression of cytochrome P450 aromatase (P450), 3 -beta-hydroxysteroid dehydrogenase (3ß-HSD), 17 -beta-hydroxysteroid dehydrogenase (17ß-HSD), aldo-keto reductase family 1 C1 (AKR1C1), estrogen receptor alpha (ERα), and progesterone receptors (PRs), were analyzed using real-time-PCR and Western Blotting. Plasma samples were assayed for 17-beta-estradiol (E2), progesterone (P4), luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone (T4) concentrations by EIA. Hinds at the beginning of the estrous cycle, mainly in endometrium, were characterized by a high mRNA expression of 3ß-HSD, AKR1C1, PRs and ERα, contrary to the expression in myometrium during pregnancy (P < 0.05). For P4, E2, and FSH, concentration was the highest during the 13th day of the estrous cycle (P < 0.05). Uterine steroid production and output in hinds as a representative seasonally reproduced ruminant occurred mainly during the estrous cycle and sustained in anestrus.

O uso do cloridrato de piridoxina para induzir estro em cadelas / The use of pyridoxine chloridrate to induce estrus in female dogs

Os agonistas dopaminérgicos são utilizados para induzir estro em cadelas, pois atuam na síntese e liberação de prolactina. Objetivou-se avaliar o efeito da piridoxina como indutor de estro em cadelas por agir na neurotransmissão dopaminérgica. Foram selecionadas 40 cadelas em anestro, divididas em quatro grupos experimentais, tratadas com 10mg/kg/dia (G1) e 50mg/kg/dia (G2) de cloridrato de piridoxina, 5µg/kg/dia (G3) de cabergolina e grupo controle/placebo (G4) por até 20 dias. Foram realizadas citologias vaginais a cada 24h para acompanhamento do ciclo estral e análises hormonais (FSH, LH e PRL) no dia zero e 120h do início do tratamento. As cadelas do G3 (100%) manifestaram proestro após 12 dias de tratamento aproximadamente, tempo inferior aos demais grupos (P<0,05). Apenas uma cadela do G1 e uma do G2 ficaram gestantes contra oito fêmeas do G3 e nenhuma do G4 (P<0,05). As concentrações plasmáticas de prolactina foram reduzidas nas fêmeas do G2 e G3 (P<0,05). As demais avaliações hormonais não sofreram influência do tratamento (P>0,05). O cloridrato de piridoxina foi ineficiente para induzir estro em cadelas, mas foi capaz de suprimir a prolactina, de forma semelhante à cabergolina, quando utilizado na dose de 50mg/kg/dia.(AU)

Dopaminergic agonists are used to induce estrus in female dogs as they act in the synthesis and release of prolactin. The objective of this study was to evaluate the effect of pyridoxine as an inducer of estrus by acting on dopaminergic neurotransmission. A total of 40 female dogs in anestrous were divided into four experimental groups treated with 10mg/kg/day (G1) and 50mg/kg/day (G2) of pyridoxine hydrochloride, 5µg/kg/day (G3) of cabergoline and control group/placebo (G4) for up to 20 days. Vaginal cytologies were performed every 24h for follow-up of the estrous cycle and hormonal analyzes (FSH, LH and PRL) on day zero and 120 hours after the start of treatment. The female dogs from G3 (100%) showed proestrus after 12 days of treatment, less time than the other groups (P< 0.05). Only one female from G1 and one from G2 were pregnant against eight from G3 and none from G4 (P< 0.05). Plasma concentrations of prolactin were reduced by treatment in females from G2 and G3 (P< 0.05). The other hormonal evaluations were not influenced by the treatment (P> 0.05). Pyridoxine chloridrate was inefficient to induce estrus in female dogs but was able to suppress prolactin when used at a dose of 50mg/kg/day.(AU)

Systematic review and meta-analysis of the efficacy of reproductive management practices used to induce resumption of ovarian cyclical activity in anestrous does.

Reproductive management practices that use hormones, sociosexual biostimulation, nutritional management, or abiotic factors are used to induce the resumption of reproduction in anestrous does. However, their overall efficacy remains uncertain; therefore, the identification of evidence-based management recommendations to manipulate anestrous in goats is important. Electronic databases were searched to retrieve reports on studies using interventions based on hormonal, sociosexual, nutritional, and abiotic factors. Only experimental studies in which a group of anestrous does was treated and compared against an untreated group were included. Estrus, ovulation, and pregnancy were primary outcomes, whereas the onset of estrus after treatment, the ovulation rate, and the number of anovulatory days were secondary outcomes. Odds ratio (OR) and mean differences were used to synthesize pooled data, and random effects models were used to calculate them. Seventy studies involving 3974 goats met the inclusion criteria. Unclear risk of bias for random sequence generation and allocation concealment predominated across studies. Pooled data for hormonal, sociosexual, and abiotic interventions showed a significant, though variable, increase in estrus (OR range 7.15-144.80), ovulation (OR range 6.08-56.95), and pregnancy (OR range 3.94-30.8). Hormonal treatments significantly reduced the onset of estrus, whereas abiotic interventions failed to reduce the number of anovulatory days. Secondary outcomes were not assessed in trials using sociosexual approaches. Finally, except for pregnancy, no significant efficacy was observed for studies using nutritional management. In conclusion, reproductive management practices using sociosexual approaches showed the highest efficacy for restoring reproductive activity in anestrous does.

Synchronization with controlled internal drug release (CIDR) and prostaglandin F2α (PGF2α) influences oxidant/antioxidant biomarkers and mineral profile in summer-stressed anoestrous buffalo (Bubalus bubalis).

During the summer season, high ambient temperature in tropical and subtropical countries exposes buffaloes to oxidative stress that have antigonadotropic and antisteroidogenic effects. Uses of hormonal therapy can improve the state of ovarian inactivity caused by heat stress and cause anoestrous buffaloes to come into oestrus and successfully achieve pregnancy. Therefore, this study was designed to detect the role of oxidative stress in production of the anoestrous state in summer heat stressed buffaloes and the effects of treatment by Controlled internal drug release (CIDR) in solving of this problem. Also it monitored the changes in Oxidant/antioxidant biomarkers and mineral profile before and after the treatment. Forty buffaloes with no signs of oestrus for more than 120 days after calving were selected. The animals were divided into two groups: the first group (group I, n = 25) was subjected to treatment with CIDR, while the second group (group II) received no treatment and was considered the control group (n = 25). Blood samples were collected before treatment, after treatment and after 45 days of oestrus. The serum level of TAC, MDA, NO, ascorbic acid (vitamin C), P, Cu and Zn were measured. The results showed that 80% of treated buffaloes came into oestrus. The conception rate was 75%. TAC concentrations were significantly higher in group I than in group II. There were significant decreases in the mean values of MDA, NO and ascorbic acid in the buffaloes in oestrus, but these values increased when the buffaloes became pregnant. In contrast there were no significant differences in the mean values of MDA, NO or ascorbic acid in the buffaloes that came into oestrus but failed to conceive. The mean serum P, Cu and Zn values were significantly increased (P < 0.05) in the buffaloes that came into oestrus compared to the control animals. The levels of P and Zn significantly increased when the buffaloes became pregnant and remained unchanged when they failed to conceive. In conclusion, known physiological and pathological variations in the oxidant/antioxidant parameters and mineral profile of summer anoestrous buffaloes may help to understand this problem of infertility.

Effect of the duration of estradiol priming prior to progesterone administration on endometrial gene expression in anestrous mares.

Field data indicate that a longer period of estrus prior to ovulation correlates positively with fertility. To test the hypothesis that the duration of exposure to estrogens prior to progesterone dominance influences endometrial function, we used anestrous mares to simulate varying durations of estrus (3 groups of 5 mares): long (LE), short (SE), and no estrus (NE), as determined by the duration of estradiol priming prior to progesterone treatment: 7, 2 and 0 days for the LE, SE and NE, respectively. Endometrial biopsies were recovered 4 days after progesterone administration in all groups for real time quantitative reverse transcription PCR (RT-qPCR) and immunohistochemical analyses. A total of 17 genes believed to contribute to a "receptive endometrium" for embryo development and viability were evaluated by RT-qPCR. Of the genes evaluated, the expression of FGF-2 (fibroblast growth factor-2) decreased with increased length of preceding estrus, whereas P19 (uterocalin) expression was higher in the LE than in the SE or NE groups. In conclusion, a lower abundance of FGF-2 and higher abundance of uterocalin, a lipocalin protein known to play an important role in providing lipids to the embryo, could contribute to a more receptive endometrium in mares following a long estrus.

Effectiveness of controlled internal drug release device treatment to alleviate reproductive seasonality in anestrus lactating or dry Barki and Rahmani ewes during non-breeding season.

This study aimed to evaluate the effectiveness of hormonal treatments on ovarian activity and reproductive performance in Barki and Rahmani ewes during non-breeding season. Forty-eight multiparous ewes, 24 Barki and 24 Rahmani ewes were divided into two groups, 12 lactating and 12 dry ewes for each breed. Controlled internal drug release (CIDR) device was inserted in all ewes for 14 days in conjunction with intramuscular 500 IU equine chronic gonadotrophin (eCG) at day of CIDR removal. Data were analysed using PROC MIXED of SAS for repeated measures. Breed, physiological status and days were used as fixed effects and individual ewes as random effects. Barki ewes recorded higher (p < .05) total number of follicles, number of large follicles, serum estradiol concentration and estradiol: progesterone (E2 :P4 ) ratio compared to Rahmani ewes. Lactating ewes recorded higher (p < .05) number of small follicles and lower concentration of total antioxidant capacity (TAC) compared to dry ewes. Number and diameter of large follicles recorded the highest (p < .05) values accompanied with disappearance of corpora lutea at day of mating. Serum progesterone concentration recorded lower (p < .05) value at day of mating and the highest (p < .05) value at day 35 after mating. CIDR-eCG protocol induced 100% oestrous behaviour in both breeds, but Rahmani ewes recorded longer (p < .05) oestrous duration compared to Barki. Conception failure was higher (p < .05) in Barki compared to Rahmani ewes. In conclusion, CIDR-eCG protocol was more potent in improving ovarian activity in Barki compared to Rahmani ewes, but this protocol seems to induce hormonal imbalance in Barki ewes that resulted in increasing conception failure compared to Rahmani ewes.

Use of cabergoline for oestrus induction in multiparous anoestrous Beagle bitches.

OBJECTIVE: The purpose of this study was to examine the efficacy of daily cabergoline administration on oestrus induction in bitches. METHODS AND RESULTS: Of 20 multiparous Beagle bitches, 15 were not mated with sires in their last oestrus and 5 bitches that had been mated whelped. All bitches were in early or mid-anoestrus and received 5 µg/kg/day cabergoline PO. Oestrus was induced in the mid-anoestrus bitches at a higher rate than in the early anoestrus bitches. Comparing the different histories of previous pregnancy among the mid-anoestrus bitches, the rate of oestrus induction in the non-pregnant bitches was higher than in the bitches that whelped. CONCLUSION: Cabergoline is effective for oestrus induction in mid-anoestrus bitches.

Effects of GnRH, a progesterone-releasing device, and energy balance on an oestrus synchronisation program in anoestrous dairy cows.

OBJECTIVE: The aim of this research was to study the roles of the day 0 energy balance and gonadotrophin-releasing hormone (GnRH) and progesterone levels on dominant follicle (DF) and corpus luteum (CL) development during the first 7 days of a gonadotrophin-prostaglandin-gonadotrophin (GPG) + progesterone (P4) program in anoestrous dairy cows. METHODS: Cows (n = 81) were allocated to one of the three treatments: (1) GPG + P4 (days 0 and 9, 100 µg GnRH; day 0-7, intravaginal P4 device; day 7, 500 µg PGF2α ); (2) GPG (as for treatment 1 but excluding the P4 device) and (3) prostaglandin + GnRH + P4 (as for treatment 1, but excluding day 0 GnRH). DF and CL size, plasma concentrations of insulin, insulin-like growth factor-I (IGF-I) and non-esterified fatty acid (NEFA) were measured on days 0 and 7. RESULTS: The proportion of cows with a CL on day 7 was significantly different between groups (GPG: 78%, GPG+P4: 69%, PGF2α + GnRH + P4: 42%, P = 0.02). The CL volume on day 7 was significantly associated with treatment, treatment by time postpartum and plasma concentrations of insulin, IGF-I and NEFA. CONCLUSION: In cows without a CL present on day 0 of an oestrus synchronisation program, removal of the day 0 GnRH treatment led to reduced CL development; however, no effect of adding progesterone was found. In contrast, in cows with a CL present on day 0 inclusion of a progesterone device led to a higher CL volume, but removal of the first GnRH injection had no effect. Response to the treatment was affected by plasma concentrations of insulin, IGF-I and NEFA.

Reproductive outcomes of anestrous goats supplemented with spineless Opuntia megacantha Salm-Dyck protein-enriched cladodes and exposed to the male effect.

The possible influence of the "male effect" upon reproductive outcomes of adult anestrous goats under marginal rangeland conditions and supplemented with protein-enriched Opuntia megacantha Salm-Dyck was evaluated. Reproductive variables included: estrus percentage (EST, %), estrus latency (ESL, hours), ovulation percentage (OP, %), ovulation rate (OR, units), average largest follicle at ovulation (LFO, mm), largest corpus luteum (LCL, mm), embryo number (EBN, units), and embryo implantation percentage (EIP, %). During early May, anestrous mix-breed adult goats (Criollo x Alpine-Saanen-Nubian; n = 38, 26° N) were randomly distributed to (1) Control (CC; n = 12), (2), Non-enriched Opuntia (NEO; n = 14), and (3) Protein-enriched Opuntia (PEO; n = 12). Neither LW (P > 0.05) nor BCS (P > 0.05) or any of the evaluated ovarian variables differed (P > 0.05) among treatments; EST = 89.66%, ESL = 53.66 h, OP = 70.33%, OR = 1.07 units, LFO = 4.5 mm, LCL = 9.6 mm, EBN = 0.94 embryos, and EIP = 48.66%. Irrespective of nutritional supplementation regime, all goats denoted an increased response to the male effect just in the middle of the anestrous season and managed under marginal grazing conditions during the dry season (May to June; 26° N). The use of the male effect successfully invoked neurophysiological pathways to re-activate ovarian follicular and luteal pathways during the natural anestrous season in the female goat. Yet, such successful physiological scenario was not equally exerted to promote an increased embryo implantation rate; this issue claims further consideration. Therefore, it is essential to align not only the peri-conceptional but also the peri-implantation stages to the best suited environmental conditions in the rangeland, in order to increase both reproductive and economic efficiency while promoting sustainability in those rangeland-based marginal goat production systems.

Evidence That Endogenous Somatostatin Inhibits Episodic, but Not Surge, Secretion of LH in Female Sheep.

Two modes of gonadotropin-releasing hormone (GnRH) and luteinizing hormone (LH) secretion are necessary for female fertility: surge and episodic secretion. However, the neural systems that regulate these GnRH secretion patterns are still under investigation. The neuropeptide somatostatin (SST) inhibits episodic LH secretion in humans and sheep, and several lines of evidence suggest SST may regulate secretion during the LH surge. In this study, we examined whether SST alters the LH surge in ewes by administering a SST receptor (SSTR) 2 agonist (octreotide) or antagonist [CYN154806 (CYN)] into the third ventricle during an estrogen-induced LH surge and whether endogenous SST alters episodic LH secretion. Neither octreotide nor CYN altered the amplitude or timing of the LH surge. Administration of CYN to intact ewes during the breeding season or anestrus increased LH secretion and increased c-Fos in a subset GnRH and kisspeptin cells during anestrus. To determine if these stimulatory effects are steroid dependent or independent, we administered CYN to ovariectomized ewes. This SSTR2 antagonist increased LH pulse frequency in ovariectomized ewes during anestrus but not during the breeding season. This study provides evidence that endogenous SST contributes to the control of LH secretion. The results demonstrate that SST, acting through SSTR2, inhibits episodic LH secretion, likely acting in the mediobasal hypothalamus, but action at this receptor does not alter surge secretion. Additionally, these data provide evidence that SST contributes to the steroid-independent suppression of LH pulse frequency during anestrus.

Reproductive performance of postpartum ewes treated with insulin or progesterone hormones in association with ram effect.

The reproductive performance of postpartum Santa Inês (SI) and Morada Nova (MN) ewes treated with insulin or progesterone hormones in association with ram effect was evaluated. Ewes from SI (n = 69) and MN (n = 69) breeds were randomly allocated to three groups of each breed (T1-ram effect only; T2-ram effect + insulin; T3-ram effect + progesterone). Progesterone concentrations (ηg/ml; mean ± SD) before and after introduction of rams (n = 6) were 0.51 ± 0.22 and 3.78 ± 0.68 (T1), 0.65 ± 0.21 and 3.77 ± 0.78 (T2) and 0.52 ± 0.21 and 3.84 ± 0.84 (T3) in SI ewes and 0.74 ± 0.19 and 3.71 ± 0.56 (T1), 0.70 ± 0.21 and 3.79 ± 0.75 (T2) and 0.81 ± 0.14 and 3.87 ± 0.80 (T3) in MN ewes, respectively. Thus, lower progesterone concentrations were found before the introduction of rams (p < .05). After the introduction of rams, preovulatory peaks of LH (ηg/ml) occurred at 28 (T1), 44 (T2) and 48 (T3) hr in SI ewes and at 64 (T1), 40 (T2) and 44 (T3) hr in MN ewes. The mean number of ovulations was similar between groups (p > .05), was 1.3 ± 0.51 (T1), 1.5 ± 0.54 (T2) and 1.6 ± 0.51 (T3) in SI ewes and 1.3 ± 0.51 (T1), 1.6 ± 0.51 (T2) and 1.6 ± 0.51 (T3) in MN ewes. In conclusion, the ram effect alone is effective at inducing and synchronizing oestrus in sheep under postpartum anoestrus, irrespective of hormone treatments.

Appetitive and Consummatory Sexual Behaviors of Rams Treated with Exogenous Testosterone and Exposed to Anestrus Dorper Ewes: Efficacy of the Male Effect.

We determine whether Dorper rams treated with testosterone in the sexual resting season trigger both appetitive and consummatory sexual behaviors and sexual and reproductive outcomes in anestrous ewes at 26° North during March-April. Dorper rams (n = 12, 2.5 years) were randomly divided prior to mating in two groups: GT (treated group; n = 6), treated with testosterone propionate (25 mg i.m. every 3 days × 3 weeks) and GC (control group; n = 6), treated with saline. Thereafter, adult anovulatory Dorper ewes (n = 61) were exposed to the GT-rams while the other group (n = 60) was exposed to the GC-rams. Ram's appetitive sexual behavior was similar (50 %) in both groups, yet the GT-rams exerted 100 % of the consummatory sexual behavior. Sexual and reproductive outcomes of the anestrus females exposed to both male groups considered two phases: 0-15 and 16-25 days from exposure to males. Involved variables were: ewes in estrus, ewes ovulating, duration of estrus, pregnant ewes, and lambed ewes. All the average variables favored those ewes exposed to GT-males: ewes in estrus: 85 versus 40 %; ewes ovulating: 80 versus 53 %; duration of estrus: 37 versus 25 h; pregnant ewes: 60 versus 31 %, and lambed ewes: 57 versus 31 %. Results confirm that adult Dorper rams treated with exogenous testosterone had improved consummatory sexual behavior and induced increased ovulation and pregnancy rates when exposed to anestrus ewes. This feasible and inexpensive testosterone-based protocol through the male effect also enhanced both sexual and reproductive outcomes in previously anestrous Dorper ewes during the natural out-of-season. Results are important to speed up reproductive performance of sheep during the natural anestrous season at this latitude, while also they embrace interesting outcomes from a comparative sexual behavior stand point.

FSH stimulation of anestrous cats improves oocyte quality and development of parthenogenetic embryos.

In the domestic cat, the efficiency of in vitro embryo production systems is negatively affected during the nonbreeding season. The objective of this research was to evaluate the effect of FSH stimulation in anestrous cats, on quality of cumulus-oocyte complexes (COCs) and in vitro developmental competence after parthenogenetic activation. To accomplish this purpose, anestrous cats were grouped into: (1) FSH treated (serial doses of 5 mg of porcine FSH each, every 24 hours, for 4 days) and (2) untreated control. The COCs were classified morphologically and a proportion of grade I and II COCs was used for expression analysis of FSHR, LHCGR, EGFR, PTGS2, EGR1, GDF9, and GATM by RT-qPCR. In addition, another proportion of grade I and II COCs was matured in vitro and used for parthenogenetic activation. After 8 days in culture, blastocyst and hatching blastocyst rates were assessed, and the expression of OCT4, SOX2, NANOG, CDX2, and GATA6 was evaluated. The COCs in the FSH group had an enhanced quality, a higher expression of LHCGR and a lower expression of GATM than did COCs from the control group (P < 0.05). Furthermore, embryos in the FSH group had increased blastocyst and hatching blastocyst rates, and those embryos had a higher expression of OCT4 and GATA than their counterparts from the control group (P < 0.05). In conclusion, ovarian stimulation of anestrous cats with FSH improved quality and increased the expression of LHCGR in COCs. The enhanced in vitro developmental competence, after parthenogenetic activation of oocytes from FSH-treated cats, coincided with an increased expression of OCT4 and GATA6 in blastocysts and hatching blastocysts.

Supplementation of Slow-Release Melatonin Improves Recovery of Ovarian Cyclicity and Conception in Summer Anoestrous Buffaloes (Bubalus bubalis).

The role of melatonin as a protective neurohormone against restoring cyclicity in summer anoestrous animals in photoperiod species has gained wider acceptance. This study was designed to uncover the evidence the slow-release melatonin (MLT) has on initiation of ovarian cyclicity and conception rate (CR) in summer anoestrous buffaloes. Thus, buffaloes diagnosed as summer anoestrous (absence of overt signs of oestrus, concurrent rectal examination and radioimmunoassay for serum progesterone at 10 days interval) were grouped as untreated (Group I, sterilized corn oil, n = 8) and treated (Group II, single subcutaneous injection of MLT @18 mg/50 kg bwt in sterilized corn oil, n = 20). Animals treated and detected in oestrus were artificially inseminated (AI) followed by division into Group III (second dose of MLT on 5th day post-AI, n = 8) and Group IV (no melatonin administration, n = 10). Blood samples were collected at 4 days interval for estimation of serum MLT, progesterone and oestrogen using radioimmunoassay kit. Mean oestrous induction rate (OIR), oestrous induction interval (OII), interoestrous interval (IOI) and CR were estimated. Compared to control, concentration of melatonin was significantly (p < 0.05) higher in treated group ranging from 14.34 ± 1.72 to 412.31 ± 14.47 pg/ml whereas other two hormones did not show any concentration difference. Melatonin-administered buffaloes showed significantly (p < 0.05) higher (90%) OIR with OII of 18.06 ± 1.57 days. Results showed improvement in conception rate in buffaloes administered with post-insemination melatonin. It can be concluded from the study that slow-release melatonin supplementation restored cyclicity in summer anoestrous animals resulting in improvement in conception rate in buffaloes.

Effects of progesterone inclusion in a gonadotropin-prostaglandin-gonadotropin programme on follicular dynamics and ovulation synchronisation of pasture-based dairy cows with anoestrous.

The aim of this study was to evaluate how the addition of a progesterone device to a gonadotropin-prostaglandin-gonadotropin (GPG) programme in dairy cows with postpartum anovulatory anoestrus affected ovarian follicular dynamics and the synchronisation of ovulation. Twenty-two dairy cows were randomly assigned to either GPG (Day 0: 100 µg GnRH, Day 7: 500 µg PGF2α, Day 9: 100 µg GnRH, Day 0-7: Day 10) or GPG with a progesterone device from Day 0 to 7 (GPG+P4). Ovarian follicular dynamics and ovulation synchronisation were studied using transrectal ultrasonography. Compared to the GPG+P4, GPG alone resulted in a relatively larger mean dominant follicle size and a higher mean peripheral oestradiol concentration (74.9 pg ∗ day vs. 60.6 pg ∗ day; P=0.002); however, there was much greater variation in follicle diameter in the group treated with GPG (7.8-22 mm vs. 10.8-17.5mm in GPG+P4) and this may, at least partly explain why only 7/11 cows in this group ovulated within 48 h of the Day 9 GnRH injection compared to 10/10 of the cows in the GPG+P4 group. These results suggest that differences in follicular dynamics between GPG and GPG+P4 programmes are not a key driver of the difference in conception rate, but further studies are required to better assess the role of ovulation synchronisation.

Induction of ovulation in anestrus ewes using a dopamine receptor antagonist.

Estradiol decreases the pulse frequency of LH during the nonbreeding season through dopaminergic neurons that results in anestrus in the ewe. Long-term administration of sulpiride, a dopamine antagonist, induced ovulation in seasonally anestrus mares. Accordingly, we tested whether sulpiride would induce ovulatory estrus in seasonally anestrus Malpura ewes. A total of 12 Malpura ewes were divided into sulpiride (at 0.6 mg/kg b.i.d.) or control groups. Anestrus was defined on the basis of the absence of heat signs for 2 months through twice-a-day heat detection during the nonbreeding season (October-November) and progesterone level of less than 1 ng/mL. Rates of estrus induction, ovulation, multiple ovulations, and lambing in the sulpiride-treated ewes were 83.3%, 100%, 16.6%, and 66.7%, respectively. The mean interval from treatment to estrus was 5.25 ± 1.49 days. Progesterone levels were elevated after ovulation significantly on Days 5 and 7 after estrus as compared to Day 0 after sulpiride treatment (P < 0.05). In contrast, none of the control group ewes showed either estrus or ovulation. There was a significant association between sulpiride treatment and estrus induction rate as well as ovulation rate (P < 0.05). It is concluded that the result provides proof of concept that the dopamine antagonist can induce ovulation in seasonally anestrus ewes.

Effect of stimulation with a gonadotropin mixture on reproductive outcome in nulliparous ewes bred during seasonal anestrus and early breeding season.

To evaluate the effect of gonadotropic stimulation on the reproductive response of nulliparous acyclic female sheep during mid-anestrous and early breeding season, fall-born ewe lambs of mixed breeding (N=191; ∼7 months old; study 1) and yearlings (Dorset X Texel (DT), N=120, study 2) were assigned randomly during May to July to be either pre-treated with progesterone using CIDR-g (0.3g of progesterone) devices for 5 days or to also receive gonadotropins (240IU eCG and 120IU hCG, 3mL i.m. injection of P.G. 600(®)) at CIDR-g removal. Study 3 was conducted with nulliparous acyclic DT yearling ewes (N=104) in late August and was similar to study 2 except that a group of control ewes that were not treated was included. Gonadotropic stimulation increased estrous response (P=0.0002), pregnancy rate to the first service period (P=0.0007), proportion of ewes lambing (P=0.03) and the lambing rate (P=0.01) in fall-born ewe lambs (study 1), but did not modify reproductive outcome in yearling females (study 2) during mid-anestrus. During the transition into the breeding season (study 3), progesterone pre-treatment increased the pregnancy rate and the proportion of ewes lambing to first service after ram introduction (P=0.003 and P=0.02, respectively). However, gonadotropic stimulation of progesterone pre-treated yearlings did not modify reproductive performance variables. In conclusion, gonadotropic stimulation improved reproductive outcome in fall-born ewe lambs bred during the mid-anestrous period, but did not affect reproductive outcome in yearling females.

Plasma and ovarian oestradiol and the variability in the LH surge induced in ewes by the ram effect.

The proportion of anoestrous ewes ovulating after exposure to a sexually active ram is variable mainly due to whether an LH surge is induced. The aim of this study was to determine the role of oestradiol (E2) in the ram-induced LH surge. In one study, we measured the plasma concentrations of E2 in ewes of different breeds before and after the 'ram effect' and related these patterns to the presence and latency of the LH surge, while another compared ovarian responses with the 'ram effect' following exposure to rams for 2 or 12 h. In all ewes, the concentration of E2 increased 2-4 h after rams were introduced and remained elevated for 14.5 ± 0.86 h. The quantity of E2 secreted before the LH surge varied among breeds as did the mean concentration of E2. The granulosa cells of IF ewes collected after 12 h exposure to rams secreted more E2 and progesterone and had higher levels of StAR than the 2 h group but in MV ewes there was no differences between these groups for any of these parameters. These results demonstrate that the LH surge induced by the rams is a result of increased E2 secretion associated with increased levels of STAR in granulosa cells and that these responses varied among breeds. The results suggest that the variable occurrence of a LH surge and ovulation may be the result of variable ovarian responses to the 'ram effect' and insensitivity of the hypothalamus to the E2-positive feedback signal.

Effects of senktide, a neurokinin 3 receptor agonist, on luteinizing hormone secretion and follicular development in anestrous Shiba goats: a pilot study.

BACKGROUND: Recent studies suggest that neurokinin B and its receptor, neurokinin 3 receptor, have an essential role in the regulation of gonadotropin-releasing hormone and luteinizing hormone (LH) release in several mammalian species. As the first trial, this pilot study reports the effect of intravenous treatment with senktide, a selective agonist of neurokinin 3 receptor, on LH secretion, follicular development in female goats that were clinically diagnosed with anestrus. FINDINGS: Anestrous goats were intravenously administered 200 nmol senktide at 4-h intervals for 24 h. Most of them examined (5/6 cases) showed a pulsatile increase in LH secretion after each injection of senktide, whereas the remaining one case showed a surge-like increase of LH secretion. Ovulation was confirmed in 5/6 cases at the range of 48-96 h after the beginning of treatment. CONCLUSIONS: This pilot study demonstrated that intravenous treatment with senktide has therapeutic action in goats with anestrus by inducing LH release, which could promote follicular development and ovulation.

Progesterone improves the maturation of male-induced preovulatory follicles in anoestrous ewes.

The first ovulation induced by male effect in sheep during seasonal anoestrus usually results in the development of a short cycle that can be avoided by progesterone priming before ram introduction. In elucidating the involvement of the hypothalamic-pituitary-gonadal axis in the occurrence of short cycles, the effects of progesterone and the time of anoestrus on the development of male-induced preovulatory follicles were investigated in anoestrous ewes using morphological, endocrine and molecular approaches. Ewes were primed with progesterone for 2 (CIDR2) or 12 days (CIDR12) and untreated ewes used as controls during early (April) and late (June) anoestrus. The duration of follicular growth and the lifespan of the male-induced preovulatory follicles were prolonged by ∼1.6 days in CIDR12 ewes compared with the controls. These changes were accompanied by a delay in the preovulatory LH and FSH surges and ovulation. Intra-follicular oestradiol concentration and mRNA levels of LHCGR and STAR in the granulosa and theca cells of the preovulatory follicles were higher in CIDR12 ewes than the control ewes. The expression of mRNA levels of CYP11A1 and CYP17A1 also increased in theca cells of CIDR12 ewes. CIDR2 ewes gave intermediate results. Moreover, ewes ovulated earlier in June than in April, without changes in the duration of follicular growth, but these effects were unrelated to the lifespan of corpus luteum. Our results give the first evidence supporting the positive effect of progesterone priming on the completion of growth and maturation of preovulatory follicles induced by male effect in seasonal anoestrous ewes, thereby preventing short cycles.