Romney Marsh Ewes Research Articles

Plasma Melatonin and Progesterone Profiles of Suffolk and Romney Marsh Ewes Implanted with Melatonin during Anoestrus Season at Lower Latitudes in Southern Hemisphere

Background: The subcutaneous implants of melatonin are stimulatory and mimic the positive effects of short photoperiod on reproduction in small ruminants. This study investigated the daily plasma melatonin profiles in ewes treated with melatonin implants and kept under natural photoperiod in Southeastern Brazil. The plasma progesterone concentrations were also investigated before and after melatonin implantation.Materials, Methods & Results: Romney Marsh (n = 11) and Suffolk (n = 10) ewes, which had been isolated from rams for at least 2 months prior to the beginning of the trial, were randomly allocated in two groups based on melatonin implant treatment (with or without melatonin implant). For plasma melatonin concentration, 43 days after melatonin implantation and 3 days before the ram introduction blood samples were collected every 2 hours during 24 hours. For plasma progesterone concentrations, blood samples were collected every once to twice a week for 2 different periods: prior to melatonin implantation and 46 days after the melatonin implantation and at the same day of the introduction of rams. The hormonal concentrations were determined by the radioimmunoassay method (RIA). The data were analyzed according to MIXED procedure (SAS) as repeated measurements for random animal effects. The effect of melatonin treatment on plasma melatonin 24-h period varied according to the breed. At the dark-phase, there were no plasma melatonin differences (P > 0.05) between implanted and no-implanted (228.02 ± 58.39 vs. 169.59 ± 48.39) Romney Marsh ewes whereas for Suffolk ewes the plasma melatonin levels were higher in implanted (305.61 ± 68.39 pg/mL) than no-implanted (151.26 ± 38.35 pg/mL) ones. At the light-phase, melatonin treatment effects could be evidenced and these differences (P < 0.01) consisted of higher melatonin values for implanted ewes and basal values for no-implanted ones in both breed groups. Before the melatonin implantation, the plasma progesterone levels were ˂ 1 ng/mL for Romney Marsh (0.41 ± 0.02 ng/mL) and Suffolk (0.47 ± 0.02 ng/mL) ewes. During the ram introduction period, no melatonin treatment effect was observed on plasma progesterone concentrations in both breed groups, but 2 days after ram introduction the plasma progesterone concentrations increased the mean values > than 1 ng/mL in implanted and no-implanted Suffolk ewes. In implanted Romney Marsh ewes the elevation of progesterone mean values was weak whereas in no-implanted Romney Marsh ewes the progesterone levels were maintained ˂ 1 ng/mL during all the blood sample collection times.Discussion: The melatonin treatment also produced a similar model of daily melatonin levels as reported previously by others, which is characterized by high plasma melatonin concentrations during the light phase of the day. The effect of melatonin implants on plasma melatonin profiles interacted with breed confirming an individual response to melatonin implantation which is proportional to genetic individual variation pattern of melatonin secretion. Before the melatonin implantation all Romney Marsh and Suffolk ewes were judged to be in non-ovulatory period (anoestrus) with plasma progesterone mean values lower than 1 ng/mL. The melatonin treatment helped to induce the ovulatory activity in most of the ewes that were in anestrous at the time of melatonin implantation and the efficacy of this treatment depends on the individual variation in ovulatory response to ram introduction. In Southeastern Brazil., melatonin implant altered the daily plasma melatonin profiles of Suffolk and Romney Marsh ewes by increasing the melatonin levels during the light-phase of the day. Melatonin implant also induced an ovulatory response in Suffolk and Romney Marsh after the introduction of the rams. For no-implanted Suffolk ewes, the male effect is sufficient to provoke an ovulatory response.

Progesterona plasmática de ovelhas submetidas ao efeito-macho e mantidas sob diferentes condições nutricionais

Dosou-se a concentração plasmática de progesterona (P4) em ovelhas Santa Inês (SI), Suffolk (SU) e Romney Marsh (RM) em anestro sazonal e submetidas ao efeito-macho, as quais receberam ou não suplementação alimentar. Machos vasectomizados foram introduzidos no grupo de fêmeas após um período prévio de isolamento de 60 dias, e amostras de sangue foram colhidas antes e após a introdução dos machos. Houve efeito (P<0,01) de período, raça, interação período x raça e interação suplementação x período x raça sobre a concentração de P4. Nas ovelhas SI ocorreu aumento (P<0,01) da concentração de P4 após a introdução do macho, indicando que houve aumento na atividade cíclica reprodutiva desse grupo. A suplementação, neste caso, potencializou este aumento. Nas ovelhas SU e RM não ocorreram modificações na concentração de P4 (P>0,01) após a introdução dos machos, nem houve efeito de suplementação. O efeito-macho foi eficaz em induzir a atividade reprodutiva durante o anestro sazonal em ovelhas SI, mas não em ovelhas SU e RM, e a associação dessa prática com a suplementação alimentar é recomendada para ovelhas da raça nativa SI.

Changes to the contractile function of ureter smooth muscle after partial infravesical obstruction in fetal sheep

To measure spontaneous contractile activity, and the responses to agonists using in vitro preparations of sheep ureter after a period of bladder outlet obstruction (BOO) initiated at mid-gestation. Date-mated Romney Marsh ewes, bearing fetuses of 70-75 days of gestation (term in this breed is 145 days) were used. Five fetuses underwent urachal obstruction and partial urethral constriction for 30 days. Six fetuses at the same gestational time underwent a sham operation (control group). Small strips of mid-ureter were cut in the longitudinal axis, and in the cross-sectional (transverse) plane of the ureter. Spontaneous contractile activity and the response to carbachol, high-K(+) (120 mM) solution and alpha,beta-methylene ATP (ABMA) were characterized by measuring the magnitude of evoked responses and the magnitude and frequency of spontaneous activity. The ureters from fetuses with BOO were significantly larger in diameter and had expanded lumens. The proportion of smooth muscle was not significantly different between the BOO and control groups. Spontaneous contractile activity in all preparations was resistant to atropine and the neurotoxin, tetrodotoxin. With transverse sections, the magnitude of spontaneous contractions was smaller in the BOO group, but the frequency was greater. The response to carbachol was also smaller in the BOO group, but the response to high-K(+) solution was similar to that of the control group. ABMA did not generate a response in any preparation. With longitudinal ureteric preparations, spontaneous or agonist-induced activity was negligible in the control group, while preparations from the BOO sheep had spontaneous activity and responded to carbachol and high-K(+) solutions. These results show that in fetuses with BOO and dilated ureters absolute ureteric contractile activity is diminished. However, there is functional reorganization of the muscle layers that generates more force in the longitudinal rather than the transverse axis. This would contribute to a reduced ability of the ureter to propel urine and contribute to the development of raised upper tract pressures.

Annual characteristics of estrous activity in wool and hair ewe lambs under subtropical conditions

The increased emphasis placed on lamb production in Brazil points to the need for a better knowledge of the annual pattern of reproductive activity in ewe lambs. This study evaluates the annual estrous activity pattern of hair and wool ewe lambs raised in southeast Brazil. Estrous characteristics were recorded for 12 months in ten Santa Inês and 21 Romney Marsh and Suffolk ewe lambs. Santa Inês ewe lambs exhibited a greater length of the normal estrous cycle (14 to 19 days) than Romney Marsh and Suffolk ewe lambs (17.0 ± 0.1 days vs. 16.5 ± 0.2 and 15.9 ± 0.2 days). All the ewe lambs presented a greater number of periods of estrus per animal per month for single estrous cycles (up to 26 days) during autumn and winter. During spring and summer there was a reduction of this number but this decline varied according to the breed. For Santa Inês ewe lambs the reduction was only significant in the summer. For wool ewe lambs the reduction was significant in both seasons but was more accentuated for Romney Marsh. The Romney Marsh ewe lambs had a more restricted breeding season than Suffolk and both presented optimal estrus activity in autumn and winter. Santa Inês ewe lambs had a continuous distribution of estrous cycles along the year, although presenting a reduction in the number of estrous periods in the summer. The ewe lambs of three breeds presented characteristics of sexual immaturity, like irregular estrous cycles and silent ovulations.

276. Sex difference in the effect of cortisol on the LH response of the pituitary to exogenous GnRH in hypothalamo-pituitary disconnected gonadectomised sheep

We have used the hypothalamo-pituitary disconnected (HPD) sheep model to investigate direct pituitary actions of cortisol to suppress LH secretion in response to exogenous GnRH. We previously observed that, during the non-breeding season, treatment with cortisol did not suppress the LH response to GnRH in HPD gonadectomised rams or ewes.1 In the present experiment, we tested the effect of cortisol on the LH response to exogenous GnRH in gonadectomised HPD sheep during the breeding season. Using a cross-over design, HPD gonadectomised Romney Marsh rams (n = 6) and ewes (n = 5) received a saline or cortisol (250 μg/kg/h) infusion for 30 h on each of two days, one week apart. All animals were treated with 125 ng i.v. injections of GnRH every 2 h during a 6h control period preceding the infusion and during the infusion. Jugular blood samples were taken during the control period and the first 6 h and last 6 h of the infusion (over 3 LH pulses). Mean plasma concentrations of LH and LH pulse amplitudes, driven by programmed GnRH injections, were similar in gonadectomised rams and ewes and there were no significant effects of saline infusion between the control periods or the saline infusion in either sex. The amplitude of LH pulses was significantly (P < 0.05) reduced in rams during the first 6 h of the cortisol infusion compared to the control period, but there were no effects of the cortisol infusion in ewes. These data show that, in the absence of sex steroids, there is a sex difference in the mechanism by which cortisol acts at the pituitary to reduce LH secretion in response to exogenous GnRH in HPD gonadectomized sheep during the breeding season. We conclude that the effect of cortisol to reduce secretion of LH involves an action on the pituitary, at least in gonadectomised rams. (1)Stackpole CA, Turner AI, Clarke IJ and Tilbrook AJ (2003) Biology of Reproduction 36(Supplement 1), 288.

Sex Differences in the Distribution and Abundance of Androgen Receptor mRNA‐Containing Cells in the Preoptic Area and Hypothalamus of the Ram and Ewe

Rams and ewes show a negative-feedback response to peripheral treatment with testosterone, with both sexes having a similar degree of suppression in luteinizing hormone (LH) secretion during the breeding season. At least part of the action of testosterone to suppress gonadotropin-releasing hormone/LH secretion is exerted via interaction with an androgen receptor. The distribution of androgen receptor-containing cells in the hypothalamus has been described for the ram, but similar studies have not been performed in the ewe. In the present study, we tested the hypothesis that levels of androgen receptor mRNA expression in the preoptic area and hypothalamus would be similar in rams and ewes. Perfusion-fixed brain tissue was obtained from adult Romney Marsh ewes (luteal phase) and rams during the breeding season (n = 4/sex). Androgen receptor mRNA expression was quantified in hypothalamic sections by in situ hybridization using an (35)S-labelled riboprobe and image analysis. Hybridizing cells were found in the medial preoptic area, bed nucleus of the stria terminalis, anterior hypothalamic area, ventromedial nucleus, arcuate nucleus and premamillary nucleus. The level of androgen receptor mRNA expression was higher in rams than ewes in the rostral preoptic area, caudal preoptic area and rostral portion of the bed nucleus of the stria terminalis, with no sex difference in other regions. The preoptic area and bed nucleus of the stria terminalis are important for reproductive behaviour and the sex differences in androgen receptor mRNA expression at these levels may relate to this. The high level of androgen receptor mRNA expression in the basal hypothalamus, with no sex difference, is consistent with the role of this region in the regulation of gonadotropin secretion.

Short term primings with different progestogen intravaginal devices (MAP, FGA and CIDR) for eCG-estrous induction in anestrus ewes

Two experiments with 260 ewes compared the effectiveness of different progestogens and the amount of medroxyprogesterone acetate (MAP) contained in the intravaginal devices with short term primings for equine chorionic gonadotrophin (eCG) estrous induction in anestrus ewes. In the first experiment, 147 Polwarth and Polwarth x Ile de France ewes were allocated to three groups, treated with MAP sponges (MAP group; n=51), fluorogestone sponges (FGA group; n=47) or devices containing progesterone (CIDR group; n=49) for 6 days. At withdrawal of intravaginal devices all animals received 380 IU eCG and were grouped with rams. Ewes in estrus were identified twice daily during a 72 h observation period. Four weeks after estrus, pregnancy was determined by transrectal ultrasonography. There was no significant differences among treatments for ewes in estrus (MAP: 94.1%, FGA: 91.5%, CIDR: 95.9%) or in conception rates (MAP: 62.5%, FGA: 67.4%, CIDR: 59.6%). In the second experiment, 113 Romney Marsh ewes were allocated to four groups in a 2×2 factorial design, based on the dosage of MAP (30 or 60 mg) and priming duration for 14 or 6 days (groups 14D30, n=27; 6D30, n=29; 14D60, n=29; 6D60 n=28). At sponge withdrawal, all ewes received 350 IU eCG and were placed with rams. Ewes that exhibited estrus and pregnancy were recorded. There were no differences in response to estrus (86.2, 82.1, 100 and 89.7%) and conception rates (44.0, 43.5, 44.4 and 34.6%, respectively, for 6D30, 6D60, 14D30 and 14D60). It was concluded that CIDR, FGA or MAP sponges were equally effective with 6 days primings for estrous induction in anestrus ewes. However, 30 mg MAP sponges are as effective as those commercially available.

Suplementación pre y post parto en ovejas. Efecto sobre la pubertad y actividad reproductiva de sus hijas

Fifty mature pregnant Romney Marsh ewes were used to investigate the influence of feed supplementation during the pre and post lambing period and subsequent ...

The Coincidence of Light and Melatonin with a Specific Phase of the Circadian Pacemaker Is Important for the Timing of Seasonal Breeding in the Ewe

The timing of reproductive activity in seasonal breeding sheep relies on daily photoperiodic signals being relayed to provide information on the time of year. Although light and melatonin are involved, the exact mechanism is not understood. In this experiment, three groups of 6 Romney Marsh ewes, a highly seasonal breed, were provided with 8 weeks of short nights (9.6-9.8 h, by artificially advancing dawn) around the winter solstice, near the end of their natural breeding season. One group of animals was infused to a physiological level with melatonin for 5 h during the afternoon prior to the onset of dark, while a second group was identically infused but for5hfromthe time of lights on. A third group received the short-night treatment only. Following the short-night treatment, all groups were exposed to long nights (> 14 h, by delaying dawn) until the summer solstice. Ovarian activity, assessed by progesterone monitoring twice weekly, showed that the noninfused and the morning-infused groups displayed renewed reproductive activity in response to the short-night/long-night treatment. There was no renewed ovarian activity in the afternoon-infused group, indicating that the time of day that melatonin is present, rather than the duration of melatonin exposure, is an important signal in the control of reproductive timing. Measurements of a marker of the endogenous circadian pacemaker, by melatonin measurements under acutely extended darkness, revealed that the short-night treatments phase advanced the onset of the pacemaker in all groups such that the afternoon phase of the pacemaker was coincident with light. The results provide strong support for the model that proposes that an afternoon-located sensitive phase of the pacemaker is responsible for the relay of photoperiodic signals in the timing control of seasonal breeding. The model proposes that the reproductive axis be primed during short nights when the sensitive phase is coincident with light in the afternoon so ovarian activity can be induced when the sensitive phase is located within the longer nights of autumn and coincident with endogenous melatonin.

Alterations of estrous activity in the ewe by circadian-based manipulation of the endogenous pacemaker.

The timing of reproductive activity in the seasonal breeding Romney Marsh ewe depends on the measurement of photoperiodic time. In this experiment, artificial light and dark signals are provided in a measured sequence at an inappropriate time of year to induce breeding out of phase with environmental photoperiod. The endogenous circadian responses and reproductive effects are documented. One group (Group A, control) of 6 Romney Marsh ewes was held in natural photoperiod throughout the experiment. For 8 weeks centered about the winter solstice (Stage 1), an additional 18 animals (Groups B, C, and D) were exposed to an artificial earlier dawn. Measurements of endogenous melatonin performed under acutely extended darkness confirmed a phase advance of the endogenous circadian pacemaker of the suprachiasmatic nucleus compared to control animals. In Stage 2, to the summer solstice (21 December), Group B animals were returned to natural photoperiod, Group C animals were subjected to an earlier artificial dusk, and Group D animals were subjected to an artificial delayed dawn. Melatonin measurements during Stage 2 confirmed that onset and offset times for Group C were earlier and that onset and offset times for Group D were delayed compared to corresponding times for Group B animals. Ovarian activity was monitored throughout. During Stage 2, Groups C and D commenced reproductive activity in mid-spring, and this continued until the experimental conditions changed. Groups A and B commenced reproductive activity at the normal timing in the subsequent autumn. Although not exclusive, these results are consistent with a coincidence model to explain the timing of seasonal breeding in this species with a dusk-located phase of the endogenous pacemaker sensitive to both light and melatonin. The temporal relationship between circadian alterations and the environmental photoperiod warrants further investigation as an explanation for seasonal breeding.

Is a critical interval of the circadian pacemaker at dusk responsive to light and melatonin responsible for the timing of estrus in the Romney Marsh ewe?

Two experiments, using Romney Marsh ewes, tested for the existence and role of a critical interval of the circadian pacemaker located near dusk that may be integrally involved in the precise timing of the breeding season. Groups of Romney Marsh ewes (n = 6) were provided with exogenous melatonin by injection at dusk (Experiment 1) or by infusion at dawn or subjected to extended darkness at dawn (Experiment 2) from the winter to the summer solstice before being exposed to natural photoperiod at latitude 35 degrees S. Other than the experimental protocols, all animals were held in natural photoperiod. The onset of the breeding season (defined as cyclic ovarian activity as indicated by plasma progesterone monitoring) was normal in those animals treated with morning melatonin but was delayed in those animals treated with melatonin at dusk or extended darkness at dawn compared to controls in natural photoperiod (p < .01). Exogenous melatonin at dusk was associated with a phase advance of the onset of the circadian pacemaker (as measured by endogenous melatonin in acutely extended darkness); additional darkness at dawn was associated with a phase delay of both the onset and the offset of the circadian pacemaker. Exogenous morning melatonin did not change the phase of the circadian pacemaker relative to the controls. The results are consistent with an external coincidence model of seasonal breeding in which a critical interval of the circadian pacemaker requires exposure to light during spring/summer to time estrus correctly. The proposed critical interval appears to be located near dusk in this model and is phase locked to the circadian pacemaker. The effect of the exogenous melatonin on the timing of the breeding season is similar to darkness when administered at dusk but is not equivalent to darkness at dawn. The timing of anestrus was not affected by any of the experimental treatments and may reflect a common response to an environmental influence.

Optimum use of subcutaneous melatonin implants to enhance the reproductive performance of seasonal and non-seasonal sheep joined in spring and early summer

This paper reviews data from five research trials and 108 clinical trials conducted in three countries to validate the optimum use of melatonin to advance seasonal breeding patterns of a variety of breeds of sheep. In order to define the optimum time for treatment in breeding flocks, ewes of three different breeds were treated with controlled-release 18-mg melatonin implants (Regulin®), with treatments commencing at various times ranging from 9 to 3 weeks prior to joining with fertile rams. Mating and conception patterns were recorded by regular observation for raddle marks and fecundity was assessed by mid-pregnancy ultrasonography. These research trials were conducted at a latitude of 38°S in Merino ewes of slight seasonality, in Merino-based cross-breeds of intermediate seasonality and in highly seasonal British breed Romney Marsh ewes. A single implant treatment, commencing at 30–40 days prior to spring or early summer joining, resulted in coincidence in timing of the peak of the inductive phase of the ovarian response to melatonin with the peak of oestrus and mating which arose from the pheromonal influence of the rams. Thus, this treatment strategy maximises the potential advantages to be obtained from both the melatonin treatment and the rams. Subsequent clinical trial programmes in Australia, the UK and New Zealand confirmed this treatment strategy and tested the available periods of the year during which effective responses could be obtained under farm conditions. It was shown that, whereas Merino and Merino cross-breeds were responsive to melatonin treatment over a wide part of spring and early summer, the British breed ewes responded only during a relatively short period of the season around the time of the summer solstice. Treatments which commenced too early in British breeds were ineffective or gave negative results, possibly because of a failure of such breeds to be able to fully interpret the pharmacological short-day signal before they have experienced an adequate period of long-day priming. For all breeds, treatments given late in summer at a time of decreasing natural daylight were also ineffective since, by this time, ewes were already experiencing a naturally occurring inductive photoperiodic signal. Overall, the studies reviewed in this paper showed that melatonin pretreatment of spring and early summer joined ewe flocks resulted in both a modest decrease in the number of barren ewes and an increase in the number of multiple births. The effects were seen in flocks of high or low basal performance and in both maiden and mature ewes, but in no case were the responses associated with the induction of supraphysiological litter size. Mean increases in litter size, for different breeds of ewe, ranged from 14.3 to 28.6 additional lambs born per 100 ewes treated, with variation between trials largely accounted for by expected sampling errors associated with the precise measurement of the discontinuous variable of litter size in relatively small sample sizes of 50–200 animals per group. Treatment of mothers with melatonin prior to joining had no effect on the sex ratio, birth weight or appearance of offspring. Melatonin pretreatment of breeding ewes was also associated with an earlier mean conception (lambing) date and a compaction of the conception (lambing) patterns but the magnitude of these measured effects was dependent upon the extent to which untreated control flocks were responsive to the pheromonal influence of rams. Overall the data extends, to a practical field level, the proper use of melatonin to at least partially overcome the seasonal constraint to breeding in sheep, and provides a sound basis for future research and clinical experimentation in this area.

Immunocytochemical localization of oxytocin in corpora lutea and luteinized cysts from anoestrous ewes stimulated with gonadotrophin-releasing hormone

Anoestrous Romney Marsh ewes with or without progesterone pretreatment were injected with multiple low-doses of gonadotrophin-releasing hormone followed by a single, larger bolus. Blood samples were taken at twelve-hourly intervals for progesterone radioimmunoassay. Ewes were slaughtered on day 3 or 5 after the bolus injection, and the ovaries were collected for histology and immunocytochemical examination for oxytocin-immunoreactivity. The corpora lutea of all ewes killed on day 3 had similar weights and morphology. The ovaries of those ewes which were not pretreated with progesterone also contained some luteinized cysts. Ewes slaughtered on day 5 were separated into 2 groups according to plasma progesterone profiles, which were either rising ('normal'), or falling after a transitory rise ('abnormal'). Those ewes pretreated with progesterone all had a 'normal' progesterone profile whereas, of 14 ewes not pretreated with progesterone, 6 were 'normal' and 8 'abnormal'. Corpora lutea were significantly lighter in the 'abnormal' group and the ovaries of most of these ewes also contained luteinized cysts. All corpora lutea and luteinized cysts showed staining for oxytocin-immunoreactivity although the staining intensity was variable. In corpora lutea from 'normal' ewes oxytocin was restricted to large luteal cells. In addition tissues from 'abnormal' ewes also contained many cells with an atypical elongated shape which stained for oxytocin-immunoreactivity. These results show that progesterone pretreatment is needed for both normal morphological and endocrine development of corpora lutea in anoestrous ewes stimulated with gonadotrophin-releasing hormone.

Role of prostaglandin F-2 and oxytocin in the regression of GnRH-induced abnormal corpora lutea in anoestrous ewes

Anoestrous Romney Marsh ewes with (+P) and without (-P) progesterone pretreatment were induced to ovulate by multiple low-dose injection of GnRH followed by a bolus injection of GnRH. Luteal function was assessed by twice daily measurement of plasma progesterone. Animals were slaughtered on Days 3 or 5 after the end of GnRH treatment and CL and endometrium were recovered. In all Day-5 ewes, blood samples were collected at 30-min intervals for 8 h on Days 3 and 5 for measurement of PGFM and oxytocin. At slaughter 92% of the Group +P ewes had ovulated compared with 54% of the Group -P ewes. The ovaries of some of the Group -P ewes only contained luteinized cysts either alone or in association with CL. In the ewes that ovulated, progesterone profiles were normal in all Group +P ewes, whereas Group -P ewes had 'normal' or 'abnormal' profiles in which plasma progesterone was declining prematurely. All of the CL from ewes with abnormal progesterone profiles were associated with follicular cysts, and were significantly smaller and with a lower progesterone content on Day 5. PGFM levels decreased (P less than 0.05) between Days 3 and 5 in ewes in Groups +P and -P with 'normal' CL but increased (P less than 0.01) in Group -P ewes with 'abnormal' CL. Oxytocin levels were lower in Group -P ewes with 'abnormal' CL on Day 5, than in 'normal' ewes in Groups -P (P less than 0.01) or +P (P less than 0.05). In 3/5 Day-5 ewes with 'abnormal' CL there was a clear association between a major peak of oxytocin and a rise in PGFM during the frequent sampling period on Day 3 or Day 5, and endometrial oxytocin binding sites were present at slaughter. This suggests that the premature regression of 'abnormal' CL occurs via the normal luteolytic mechanism. Although ewes in Groups +P and -P with 'normal' CL had similar progesterone profiles, plasma oxytocin was significantly higher (P less than 0.05) in the Group -P ewes and oxytocin binding sites were present only in this group, suggesting that progesterone pretreatment can influence the production of both oxytocin and its receptor.

Function of abnormal corpora lutea in vivo after GnRH-induced ovulation in the anoestrous ewe.

Anoestrous Romney Marsh ewes with and without progesterone treatment (+P, -P) were treated with small-dose (250 ng) multiple injections of GnRH at 2-h intervals for 48 h. Animals were slaughtered on Days 4, 5, 7 and 11 after the end of GnRH treatment and luteal function was assessed by the measurement of daily plasma progesterone concentrations. In all animals which ovulated (29/32, 91%) peripheral progesterone concentrations rose to 0.5-1.0 ng/ml within 3 days of the end of GnRH treatment. In 7/7 (100%) +P animals and 5/22 (23%) -P animals, progesterone concentrations continued to rise and were maintained at levels greater than 1.5 ng/ml until slaughter. In the remaining -P animals, plasma progesterone concentrations declined to reach basal levels by Day 5. Corpora lutea recovered from these animals showed signs of premature regression on Day 5 and were fully regressed by Day 7. Progesterone priming delayed the occurrence of the LH surge which occurred 39.1 +/- 3.6 h after the end of GnRH treatment in the +P animals compared to 20.2 +/- 1.74 h (P less than 0.001) in the -P animals in which luteal function was abnormal and 22.4 +/- 4.35 h in the -P animals in which luteal function was normal. These results show that abnormal luteal function occurs in the majority of GnRH-treated ewes in the absence of progesterone pretreatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of hysterectomy on the short life-cycle corpus luteum produced after GnRH-induced ovulation in the anoestrous ewe

Anoestrous Romney Marsh ewes were treated with small-dose (250 ng) multiple injections of GnRH. Ewes in Groups 1 and 3 were hysterectomized 2 weeks before treatment, while those in Groups 2 and 4 were intact controls. Groups 1 and 2 were primed with progesterone (+P) and treated with 2 h injections of GnRH (250 ng) for 36 h, while Groups 3 and 4 were not pretreated (-P) but were given 2 h injections of GnRH (250 ng) for 18 h. Both treatment regimens were terminated with a bolus injection of GnRH (125 micrograms), given to synchronize the timing of the LH surge and subsequent luteal progesterone production. The plasma progesterone profiles of 5/5 animals in Group 2 (+P controls) and 2/5 animals in Group 4 (-P controls) were indicative of normal luteal function, while the remaining 3/5 animals in Group 4 produced plasma progesterone profiles typical of abnormal luteal function. However, in all the hysterectomized animals (Groups 1 and 3) peripheral plasma progesterone concentrations rose to reach a mean peak value of 1.3 ng/ml plasma on Day 8 which was maintained in all animals irrespective of progesterone pretreatment. The absence of a fall in progesterone concentrations precluded the identification of any animal in Group 4 showing abnormal luteal function. It was also noted that, after hysterectomy, although the corpus luteum was maintained, it was with reduced secretory capacity. The prevention of the expected proportion (70%) of -P animals from displaying a decline in plasma progesterone concentration after hysterectomy provides firm evidence that the uterus is involved in the premature regression of the short-cycle corpus luteum.

Function of abnormal corpora lutea in vitro after GnRH-induced ovulation in the anoestrous ewe.

Normal and abnormal corpora lutea were recovered from anoestrous Romney Marsh ewes on Days 3, 4, 5 and 6 after treatment with small-dose (250 ng) multiple injections of GnRH followed by a bolus injection (125 micrograms) with (+P) and without (-P) progesterone pretreatment and a study made of their characteristics in vitro. Plasma progesterone concentrations initially rose concurrently in all animals but abnormal luteal function occurred in 70% of the -P ewes and was defined on Day 5 when plasma progesterone concentrations declined relative to those in the +P ewes. All corpora lutea recovered on Days 3 and 4 appeared macroscopically similar and there were no significant differences between the +P and -P groups in terms of luteal weight, progesterone content and binding of 125I-labelled hCG on these days. However, corpora lutea from the -P animals only exhibited a decline in progesterone production in vitro on Day 4 (P less than 0.01), and morphological differences became apparent on Days 5 and 6 when the abnormal corpora lutea from the -P animals also decreased in weight (P less than 0.01) and progesterone content (P less than 0.001). Binding of 125I-labelled hCG increased on Day 5 in the normal corpora lutea only. These results show that, although abnormal luteal function induced by GnRH treatment of anoestrous ewes could not be distinguished from normal corpora lutea before Day 5 by measurement of progesterone in peripheral plasma, a significant decline in progesterone production in vitro occurred on Day 4 in the abnormal corpora lutea. This was followed by significant decreases in weight and progesterone content and a failure to increase 125I-labelled hCG binding. Abnormal corpora lutea are therefore capable of some initial growth and progesterone production, before undergoing a rapid and premature regression from Day 4, which has similar characteristics to natural luteolysis.

Treatment with progesterone affects follicular steroidogenesis in anoestrous ewes

Ovaries were recovered from two groups ( n=6/group) of anoestrous Romney Marsh ewes, one group of which had been treated with progesterone implants prior to slaughter. A comparison was made between the maturational characteristics of the follicles > 2 mm diameter recovered from both groups and some significant differences were noted. In particular, the large follicles (> 4 mm diameter) recovered from the progesterone-treated ewes had a significantly ( P<0.01) reduced capacity to secrete oestradiol, but enhanced ( P<0.01) ability to bind hCG when compared to follicles recovered from control ewes. There were also differences in the relationships between follicular characteristics in the two groups of animals including a significant ( P <0.05) correlation between follicular fluid progesterone and hCG binding to theca tissue in large follicles from progesterone-treated animals which did not exist in the control animals. Conversely, in the control animals a significant ( P<0.05) relationship existed between oestradiol production and hCG binding to granulosa cells, but there was no such relationship in the follicles from progesterone-treated ewes. These results demonstrate that the treatment of ewes with progesterone during the anoestrous period clearly affects oestradiol synthesis and hCG binding and thus modifies follicular development.

The effect of season and the introduction of rams on oestrous activity in Somali, Nandi, Merino, Karakul and New Zealand Romney Marsh ewes in Kenya

ABSTRACTA study was made of the levels of oestrous activity of two indigenous breeds of sheep (Somali and Nandi) and three exotic breeds of sheep (Merino, Karakul and New Zealand Romney Marsh) over a period of 3 years, in an equatorial environment. Breed was the only significant source of variation for the length of the oestrous cycle (P&lt; 0·01). The mean lengths of the oestrous cycle were 17·2 (s.d. 3·21), 17·5 (s.d. 2·24), 17·9 (s.d. 2·99), 17·5 (s.d. 2·57) and 16·5 (s.d. 3·41) days for the Somali, Nandi, Merino, Karakul and Romney Marsh breeds, respectively.The mean percentage of ewes of the different breeds showing oestrus in 20-day periods were 69·8 (s.d. 22·57), 49·9 (s.d. 18·67), 63·4 (s.d. 25·70), 79·2 (s.d. 20·30) and 33·2 (s.d. 23·50) % for the Somali, Nandi, Merino, Karakul and Romney Marsh breeds, respectively. Time-series analysis did not detect any evidence of seasonal variation in oestrous activity, although there was an indication that the Merino and Romney Marsh breeds showed a marked increase in oestrous activity following, the introduction of rams. It was concluded that the variation in level of oestrous activity was short term and random.

Progesterone pretreatment has a direct effect on GnRH-induced preovulatory follicles to determine their ability to develop into normal corpora lutea in anoestrous ewes.

In two experiments carried out during seasonal anoestrus, Romney Marsh ewes were treated with small-dose (250 ng) multiple injections of GnRH at 2-h intervals with and without progesterone pretreatment. In Exp. 1, 8/8 progesterone-primed ewes ovulated and produced functionally normal corpora lutea compared with 2/9 non-primed ewes. Follicles were recovered from similarly treated animals 18 or 28 h after the start of GnRH treatment (at least 14 h before the estimated time of the LH peak) and assessed in terms of diameter, granulosa cell number, oestradiol, testosterone and progesterone concentrations in the follicular fluid, oestradiol production in vitro and binding of 125I-labelled hCG to granulosa and theca. There were no significant differences in any of these measures in 'ovulatory' follicles recovered from the progesterone-pretreated compared to non-pretreated animals. In Exp. 2, follicles were removed from similar treatment groups just before and 2 h after the start of the LH surge. Unlike 'ovulatory' follicles recovered from the non-pretreated ewes, those recovered from progesterone-pretreated ewes responded to the LH surge by significantly increasing oestradiol secretion (P less than 0.01) and binding of 125I-labelled hCG (P less than 0.05) to granulosa cells. Overall there was also more (P less than 0.05) hCG binding to granulosa and theca cells from progesterone-pretreated animals. Non-ovulatory follicles recovered from progesterone-primed ewes had more (P less than 0.05) binding of 125I-labelled hCG to theca and a higher testosterone concentration in follicular fluid (P less than 0.05) than did those from non-primed ewes. These results suggest that inadequate luteal function after repeated injections of GnRH may be due to a poor response to the LH surge indicative of a deficiency in the final maturational stages of the follicle.