Characterisation of pregnancy losses after embryo transfer by measuring plasma progesterone and bovine pregnancy-associated glycoprotein-1 concentrations.

The aim of this analysis was to determine whether pregnancy loss (PL) after embryo transfer (ET) in cattle was related to maternal progesterone (P4) concentrations during and shortly after ET, and maternal bovine pregnancy-associated glycoprotein-1 (bPAG-1) concentrations in plasma at days 25-35 of gestation. Embryos (n=260) were produced either in vivo after superovulation (n=115), or in vitro from oocytes (obtained with ovum pick-up) in co-culture (n=44) or cultured in a synthetic medium (n=101). Overall, PL was 56.9% (148) and no significant differences occurred in calving rate among the three embryo production groups. There was no difference in P4 concentrations on days 7-14 of gestation in the three groups, nor between ongoing and interrupted pregnancies. Between days 25 and 35 of pregnancy, bPAG-1 concentrations were unaffected by embryo production, but in cattle that had PL between days 26 and 120, four bPAG-1 profiles could be detected. Between days 25 and 32, bPAG-1 concentrations were influenced by PL, and concentrations were significantly lower in animals in which PL occurred between days 26 and 120 than in those animals that aborted later or calved at term. Early P4 concentrations suggested that maternal luteal factors were not responsible for PL which appeared to be caused by impaired conceptus development (regardless of embryo type) as reflected by low maternal bPAG-1 concentrations prior to embryonic death.

Effects of superovulation with oFSH and norgestomet/GnRH-controlled release of the LH surge on hormone concentrations, and yield of oocytes and embryos at specific developmental stages.

The objective of this study was to evaluate a new superovulation procedure with oFSH after temporary suppression of the endogenous LH surge by norgestomet followed by administration of GnRH, to collect bovine oocytes and embryos at specific developmental stages. Since 1999, our research group applies this superovulation procedure with controlled release of the endogenous LH surge. The objective of this study is to verify if this procedure is reliable for collection of oocytes and embryos at specific time points of development and if it produces a sufficient number of both oocytes and embryos of good quality. This procedure was validated regarding to hormonal characteristics, superovulatory response and both oocyte and embryo yield at different times of in vivo development. The results demonstrate that the procedure used to control the occurrence of the pre-ovulatory LH surge was effective in 92% of the animals (n = 238) and even in 99% of the animals the oocytes and embryos were collected at the intended stage of development. The superovulatory response and both oocyte, embryo yield and quality were similar to the average yield in Europe reported by Association Européenne de transfert embryonnaire (AETE). In conclusion, this superovulation procedure provides a valid tool to collect oocytes and embryos at specific time points of development.

Concentrations of prolactin, LH, testosterone, TSH and thyroxine in normospermic dogs of different breeds.

Concentrations of prolactin (PRL), LH, testosterone (T), TSH and thyroxine (T(4)) were determined before and at 20, 120 and 180 min after a single iv injection of thyrotropin-releasing hormone (TRH) in eight Beagles, eight Fox Terriers, six Labrador Retrievers and five Great Danes that were normospermic. Mean basal PRL concentrations were lower in the Fox Terriers compared with the Great Danes (p < 0.05). Mean LH concentrations were higher in the Fox Terriers than in the Beagles, and T was lower in the Fox Terriers at some times but not others (p < 0.05). Thyroid Stimulating Hormone (TSH) concentrations did not differ among breeds, while mean basal T(4) values were lower in Fox Terriers compared with Labrador Retrievers and Great Danes (p < 0.05). Stimulation of T(4) secretion 120 and 180 min after iv TRH injection was most pronounced in the Beagles and less in the Fox Terriers (p < 0.05). The results of the present study indicate that potential breed differences in circulating concentrations of PRL, LH, T, TSH and T(4) in male dogs with apparently normal fertility can be encountered, but further studies are needed to determine whether the observed differences are typical features of these breeds, reflect subsets of dogs within breeds, or are in part because of possible uncontrolled parameters such as sample timing, ambient photoperiod, housing conditions or diet.

LH and FSH secretion, follicle development and oestradiol in sows ovulating or failing to ovulate in an intermittent suckling regimen.

The present paper describes LH and FSH secretion, follicle development and ovulation in sows that were subjected to a limited nursing regimen. From Day 14 of lactation, 32 sows were separated from their piglets for 12 h every day (intermittent suckling; IS). Half the sows had boar contact during separation. Nine of 32 sows ovulated spontaneously within 14 days from initiation of IS. The frequency of LH pulses on the first day of IS tended to be higher in anovulatory sows (6.3 v. 4.2 pulses per 12 h; P < 0.10); other characteristics of LH secretion were similar to sows that ovulated. The characteristics of FSH secretion did not differ over the 8-h sampling period. Boar contact did not influence either LH and FSH secretion or the number of sows that ovulated. Up to 58% of anovulatory sows showed an increase in follicle size after initiation of IS and, 4 days after the initiation of IS, one-third still had follicles similar in size to those in ovulatory sows. However, the oestradiol concentration in anovulatory sows did not increase. We conclude that FSH and LH stimulation in anovulatory sows is not limiting for normal follicle development, but that ovarian follicles are not responsive to increased LH secretion.

Intermittent suckling enables estrus and pregnancy during lactation in sows: effects of stage of lactation and lactation during early pregnancy.

Previously we demonstrated that pre-ovulatory LH and post-ovulatory progesterone (P4) concentrations in plasma were low and embryo development was retarded when sows were induced to ovulate during lactation by submitting them to intermittent suckling (IS). The present study investigated whether this was due to: (1) stage of lactation when IS was initiated, and (2) continuation of IS post-ovulation. Multiparous Topigs40 sows were studied under three conditions: conventional weaning at Day 21 of lactation (C21; n=30), intermittent suckling from Day 14 of lactation (IS14; n=32), and intermittent suckling from Day 21 of lactation (IS21; n=33). Sows were separated from piglets for 12h daily during IS. IS sows were either weaned at ovulation or 20d following ovulation. One-third (21/63) of the IS21 and C21 sows had already ovulated or had large pre-ovulatory follicles at Day 21 and were excluded from further study. Initiation of IS at Day 14 instead of Day 21 of lactation tended to reduce P4 at 7d post-ovulation (P=0.07), did not affect pregnancy rate, and tended to reduce embryo survival (P=0.06). Continuation of IS during pregnancy resulted in lower P4 at 7 and 12d post-ovulation, tended to reduce embryo weight and pregnancy rate (P<0.10), whereas embryo survival was not affected. This study presents data for a population of sows in which follicle growth and ovulation are easily triggered under suckling conditions. Further, when these sows are bred during lactation, initiation of IS at 21 rather than 14d of lactation with weaning at ovulation yields the most desirable reproductive performance.

Effects of gonadectomy on prolactin and LH secretion and the pituitary-thyroid axis in male dogs.

The effects of gonadectomy on the secretion of prolactin, LH, TSH, and thyroxine were investigated. Blood serum hormone concentrations were analysed before and at 20, 120, and 180min after a single iv TRH injection in each of eight healthy intact and castrated male beagle dogs before (control) and after 4-week treatment with the dopamine-2 receptor agonist cabergoline. Under control conditions the mean prolactin, TSH, and thyroxine concentrations were similar in intact and gonadectomised dogs, and administration of TRH provoked a significant (p<0.01) increase in concentrations of the three hormones. The overall inhibitory effect of cabergoline treatment on prolactin secretion was more pronounced in the castrated dogs compared with the intact group. Cabergoline significantly suppressed the TRH-induced prolactin increase in each group (p<0.01). Corresponding TRH-stimulated TSH concentrations were not affected by cabergoline. In the gonadectomised dogs, thyroxine concentrations before and at 120 and 180min after TRH injection were significantly lower than under control conditions. LH concentrations were always higher (p<0.01) in gonadectomised dogs compared with the intact dogs, but appeared to be affected neither by TRH nor by cabergoline administration. It can thus be concluded from the results, that gonadectomy does not result in hyperprolactinaemia in male dogs, while LH concentrations are significantly increased due to missing androgen feedback. Thyroid function remains unaffected by gonadectomy. Testicular steroids appear to interact with central dopaminergic and probably other neuroendocrine mechanisms regulating the secretion of prolactin, TSH, and thyroxine. Thus, long-term dopamine-2 receptor agonistic treatment may lead to a hypothyroid condition in castrated male dogs.

Pulsatile plasma profiles of FSH and LH before and during medroxyprogesterone acetate treatment in the bitch.

The aim of this study was to investigate the effect of medroxyprogesterone acetate (MPA) on pulsatile secretion of gonadotropins in the bitch. Five intact Beagle bitches were treated with MPA in a dose of 10mg/kg body weight subcutaneously at intervals of 4 weeks for a total of 13 injections, starting during anestrus. The 6-h plasma profiles of luteinizing hormone (LH) and follicle stimulating hormone (FSH) were determined before, and 3, 6, 9, and 12 months after the start of MPA treatment. After 6 months of MPA treatment basal plasma LH concentration was transiently increased significantly. Basal plasma FSH concentration and the area under the curve above the zero level (AUC0) for FSH were significantly higher after 3 months of MPA treatment than before or after 9 and 12 months of treatment. MPA treatment did not significantly affect pulse frequency, pulse amplitude, or AUC above the baseline for either LH or FSH. During treatment 58 significant LH pulses were identified, and although each LH pulse coincided with an increase in plasma FSH concentration, in 17 cases the amplitude of the increase was too small to be recognized as a significant FSH pulse. In conclusion, MPA treatment did not suppress basal plasma gonadotropin levels in the bitches. On the contrary, it caused a temporary rise in the basal concentration of both FSH and LH, which may have been due to a direct effect of MPA on the ovary. In addition, several LH pulses were not accompanied by a significant FSH pulse, suggesting that MPA treatment attenuated the pulsatile pituitary release of FSH.

Peri-oestrus hormone profiles and follicle growth in lactating sows with oestrus induced by intermittent suckling.

This study describes follicle dynamics, endocrine profiles in multiparous sows with lactational oestrus compared with conventionally weaned sows (C). Lactational oestrus was induced by Intermittent Suckling (IS) with separation of sows and piglets for either 12 consecutive hours per day (IS12, n = 14) or twice per day for 6 h per occasion (IS6, n = 13) from day 14 of lactation onwards. Control sows (n = 23) were weaned at day 21 of lactation. Pre-ovulatory follicles (> or =6 mm) were observed in 100% of IS12, 92% of IS6 and 26% of C sows before day 21 of lactation and in the remaining 74% C sows within 7 days after weaning. All sows with pre-ovulatory follicles showed oestrus, but not all sows showed ovulation. Four IS6 sows and one IS12 sow developed cystic follicles of which two IS6 sows partially ovulated. Follicle growth, ovulation rate and time of ovulation were similar. E(2) levels tended to be higher in IS sows (p = 0.06), the pre-ovulatory LH surge tended to be lower in IS12 (5.1 +/- 1.7 ng/ml) than in C sows (8.4 +/- 5.0 ng/ml; p = 0.08) and P(4) levels were lower in IS12 and IS6 than in C sows (at 75 h after ovulation: 8.8 +/- 2.4 ng/ml vs 7.0 +/- 1.4 ng/ml vs 17.1 +/- 4.4 ng/ml; p < 0.01). In conclusion, sows with lactational oestrus induced by IS are similar to weaned sows in the timing of oestrus, early follicle development and ovulation rates, but the pre-ovulatory LH surge and post-ovulatory P(4) increase are lower.

Hormonal changes in spontaneous and aglépristone-induced parturition in dogs.

To increase our understanding of the endocrine changes associated with parturition in dogs, plasma concentrations of progesterone (P4), 15-ketodihydroprostaglandin F(2alpha) (PGFM), estradiol-17-beta (E2beta), cortisol, ACTH, prolactin (PRL), LH, and FSH were measured in six spontaneously whelping bitches and in six bitches in which parturition was induced with the progesterone-receptor blocker aglépristone on day 58 of pregnancy. Expulsion of pups in the induced group took place in the presence of P4 concentrations that were still elevated. PGFM concentrations increased before parturition in both groups, but levels were lower in the induced bitches. PGFM levels reached a maximum in both groups during parturition and quickly decreased in the spontaneously whelping group after parturition, but remained elevated in the induced group. In both groups, cortisol concentrations reached similar maximum levels during the last 30 h before the onset of expulsion. During the 3 days postpartum, cortisol concentrations were higher in the induced group. The highly variable ACTH concentrations did not differ significantly throughout the study within or between groups. In both groups, E2beta concentrations decreased and PRL concentrations increased between the late gestational period and the 30-h period before parturition. Concentrations of both LH (spontaneously whelping group) and FSH (both groups) decreased between late gestation and the postpartum period. The results of this study illustrate the hormonal changes around parturition in the bitch, and reveal that aglépristone-induced parturition is associated with still incomplete luteolysis, an altered PGFM profile, and elevated postpartum cortisol concentrations as compared with spontaneously whelping dogs.

Feeding level does not affect progesterone levels in intermittently suckled sows with lactational ovulation.

The aim of this study was to examine whether the low post-ovulatory plasma P(4) levels found in intermittently suckled (IS) sows are related to the high feeding level during intermittent lactation. Multiparous sows (n=21) were separated from their piglets for 12h per day starting at day 14 of lactation until 6 days after ovulation. At day 28 of lactation, 9 sows had ovulated (spontaneous) and the remaining 12 sows were treated with PG600 (PG600-induced). At ovulation sows were allocated to either a high (H) feeding level (on average 6.5 kg) or a low (L) feeding level (high feeding level minus 2.5 kg) until 6 days after ovulation. Plasma P(4) levels were not affected by feeding level or type of ovulation (P>0.10), and neither were CL parameters, embryo survival rate and embryo development at day 30 of gestation. In conclusion, low levels of plasma P(4) were found in IS sows with lactational ovulation but these were not affected by feeding level during the first week after ovulation. Further studies are needed to investigate which factors cause the lower plasma P(4) levels in these sows.

Pulsatile release of luteinising hormone during the luteal phase in lactating and weaned sows.

The present paper describes luteal phase luteinising hormone (LH) in sows that ovulated due to a limited nursing regimen (LN). The LN regimen was imposed either at Day 14 or at Day 21 of lactation. At ovulation, lactation was terminated (n = 8) or sows remained lactating throughout early pregnancy (n = 8). Blood samples were collected every 15 min for 8 h during the day, around Day 11 after ovulation. In addition, lactating sows were bled during the night, when piglets were allowed to suckle. The LH pattern was typical for the luteal phase, with one to five pulses per 8 h. The LH characteristics (frequency, base, average, pulse area) did not differ between lactating and weaned sows, except for the amplitude of LH pulses, which was higher in weaned sows compared with lactating sows (1.22 +/- 0.15 v. 0.76 +/- 0.11 ng mL(-1), respectively; P < 0.05). In lactating sows, average LH, basal LH and the frequency of LH pulses were significantly lower during the night, when piglets were allowed to suckle. The sage of lactation at which LN was imposed did not influence LH. In conclusion, it is unlikely that the small difference in LH explains the considerable difference between lactating and weaned sows in progesterone at Day 12 of pregnancy (24.1 +/- 1.3 v. 43.3 +/- 4.0 ng mL(-1), respectively; P < 0.01). Moreover, the difference in progesterone already exists during the early luteal phase (Day 0-10), when secretion of the corpora lutea is still independent of LH.

The acute effect of bull presence on plasma profiles of luteinizing hormone in postpartum, anoestrous dairy cows.

The objective of this study was to investigate whether bull exposure affects LH profiles in postpartum, anoestrous dairy cows. Eight cows between 10 and 17 days after parturition were used. On Day 1, blood samples were taken at 10 min intervals for 8 h. On Day 2, blood sampling continued at 10 min intervals and after 2 h a bull was introduced behind a fence, and blood sampling continued for another 8 h. Time of resumption of luteal activity was between 25 and more than 80 days after parturition for these animals and was not related (P>0.1) with frequency of LH pulses, amplitude of pulses and basal LH concentration on either Day 1 or Day 2. In 6 of the 8 cows, average and basal LH concentration were greater (P<0.001) during the 8 h of bull presence (0.56 +/- 0.33 and 0.39 +/- 0.26 ng/ml, respectively) compared to the 8 h without a bull (0.50 +/- 0.30 and 0.35 +/- 0.24 ng/ml, respectively). Pulse amplitude did not differ (P=0.85) between Day 2 (0.45 +/- 0.24 ng/ml) or Day 1 (0.45 +/- 0.14 ng/ml). LH pulse frequency was greater (P<0.1) on Day 2 (5.3 pulses/8h) compared to the Day 1 (4.6 pulses/8h). In conclusion, fenceline bull exposure early postpartum seems to have an acute effect on LH-release in anoestrous dairy cows. Whether sustained bull exposure can hasten first ovulation after calving through an effect on LH release in dairy cows is an interesting area of research.

A possible role for sperm RNA in early embryo development.

Recently it has been demonstrated that, along with sperm, some of its RNA can be introduced into the oocyte during fertilization, which stays stable until the activation of the embryonic genome. Originally it was thought that RNA present in semen relates to contamination from somatic cells and/or immature sperm both containing substantially higher amounts of RNA than the fertilizing sperm. However, RNA is still found after stringent washing through density gradients resulting in a sperm fraction that is translational silenced and devoid of cytosolic rRNA and thus of potential RNA contamination-which is not transferable to the oocyte. Sperm only delivers a relatively small amount of paternal RNA (5-10 fg) into the fertilized oocyte when compared to the amount of maternal RNA (approximately 1 ng). Pooled human sperm contains about 5000 different mRNA sequences of which half are common between ejaculates. Besides mRNA sperm also contains small sperm RNA molecules that might interfere in gene expression (iRNA). In human sperm already more than 68 putative iRNAs have been identified and 15 of them may specifically inhibit genes that are only active during early embryonic development. The composition and quantity of sperm RNA is considered to be a valuable diagnostic tool for male fertility. However, only a subpopulation of the purified mature sperm fraction (with a yet unknown composition and quantity of RNA) will appropriately respond to capacitation media to become competent to fertilize the oocyte. In this review the origin and function of sperm borne RNA transferred into the oocyte is discussed along with their putative role in early embryogenesis, which still needs to be experimentally proven.

Changes in numbers of large ovarian follicles, plasma luteinizing hormone and estradiol-17beta concentrations and egg production figures in farmed ostriches throughout the year.

In this study we described and analysed changes in the numbers of large ovarian follicles (diameter 6.1-9.0 cm) and in the plasma concentrations of luteinizing hormone (LH) and estradiol-17beta (E(2)beta) in relation to individual egg production figures of farmed ostriches (Struthio camelus spp.) throughout one year. Ultrasound scanning and blood sampling for plasma hormone analysis were performed in 9 hens on a monthly basis during the breeding season and in two periods of the non-breeding season. Our data demonstrated that: (1) large follicles were detected and LH concentrations were elevated already 1 month before first ovipositions of the egg production season took place; (2) E(2)beta concentrations increased as soon as the egg production season started; (3) numbers of large follicles, LH and E(2)beta concentrations were elevated during the entire egg production season; and that (4) numbers of large follicles, LH and E(2)beta concentrations decreased simultaneous with or following the last ovipositions of the egg production season. By comparing these parameters during the egg production season with their pre-and post-seasonal values, significant differences were found in the numbers of large follicles and E(2)beta concentrations between the pre-seasonal, seasonal and post-seasonal period; while LH concentrations were significantly different between the seasonal and post-seasonal period. In conclusion, our data demonstrate that changes in numbers of large follicles and in concentrations of LH and E(2)beta closely parallel individual egg production figures and provide some new cues that egg production in ostriches is confined to a marked reproductive season. Moreover, our data provide indications that mechanism, initiating, maintaining and terminating the egg production season in farmed breeding ostriches are quite similar to those already known for other seasonal breeding bird species.

Basal and GnRH-induced secretion of FSH and LH in anestrous versus ovariectomized bitches.

The basal and gonadotropin releasing hormone (GnRH)-induced plasma concentrations of follicle stimulating hormone (FSH) and luteinizing hormone (LH) were studied in four anestrous and four ovariectomized (OVX) bitches. Blood samples were obtained via jugular venipuncture 40min before and 0, 10, 20, 30, 60, 90, and 120min after the i.v. administration of synthetic GnRH in a dose of 10microg/kg body weight. The basal plasma FSH and LH concentrations were significantly higher in the OVX bitches than in the anestrous bitches. In the anestrous bitches, the plasma FSH concentration was significantly higher than the pretreatment level at 10, 20, and 30min, whereas the plasma LH concentration was significantly elevated at 10 and 20min. The maximal GnRH-induced plasma FSH concentration in the anestrous bitches did not surpass the lowest plasma FSH concentration in the OVX bitches, whereas the GnRH-induced plasma LH concentrations in the anestrous bitches overlapped with the basal plasma LH concentrations in the OVX bitches. In the OVX bitches, GnRH administration did not induce a significant change in the plasma FSH concentration, whereas the plasma LH concentration increased significantly at 10 and 20min. In conclusion, the results of the present study indicate that in anestrous bitches GnRH challenge results in increased plasma levels of both FSH and LH, whereas in the OVX bitches, in which the basal plasma FSH and LH concentrations are higher, only a rise in the plasma LH concentration is present after GnRH stimulation. The results also suggest that a test to measure plasma concentration of FSH in single samples appears to have potential in verification of neuter status in bitches.

LH pulsatile release patterns, follicular growth and function during repetitive periods of suckling and non-suckling in sows.

This paper presents LH, estradiol, and cortisol in 12 sows that were separated from their piglets for 12h a day, beginning around 2w of lactation, until weaning (intermittent suckling, IS). To separate sows from their piglets, the sows either were moved to a different unit (total separation), or were only inhibited from suckling their piglets by a physical barrier (physical separation). Blood samples were frequently collected during 4.5 consecutive days. At the start of IS, four sows showed advanced follicle growth. In the eight remaining sows, total separation resulted in 4/4 sows ovulating, while physical separation resulted in 2/4 sows ovulating. Total and physical separation resulted in different LH secretion patterns. Total separation resulted in a lower amplitude of LH pulses than physical separation throughout the period of sampling (0.26 versus 0.53ng/ml; P<0.01), and seemed to result in an escape from inhibition of LH secretion during suckling. Similarly, sows that ovulated had a lower amplitude of LH pulses (0.30 versus 0.54ng/ml; P<0.05), and also showed a different effect of suckling on LH secretion than anovulatory sows. Total separation, in contrast to physical separation, consistently resulted in increased cortisol after separation (P<0.05). This contrast was not observed between ovulating and non-ovulating sows. We therefore conclude that IS results in an increased LH secretion. Inhibition of all contact between sows and piglets seems to result in a more sustained increase in LH secretion, which increases the chance of ovulation.

The relation between ultrasonographic observations in the oviduct and plasma progesterone, luteinizing hormone and estradiol during the egg laying cycle in ostriches.

In this study we investigated the temporal relationship between ovulation, egg formation, oviposition and the changes in plasma concentrations of progesterone, luteinizing hormone and estradiol-17beta during the egg laying cycle in farmed ostriches. In 10 egg-producing birds, transcutaneous ultrasound scanning was performed at 3h intervals and blood sampling at hourly intervals during a period of at least 48h (one egg laying cycle). In hens (n=8) that ovulated during the observational period, the ovulated egg was first detected 2h after oviposition; thus, ovulation occurred shortly after oviposition in all birds. During the period between two consecutive ovipositions, the developing egg remained for 9h in the proximal part (infundibulum, magnum or isthmus) and for 39h in the distal part of the oviduct (uterus). In ovulating hens, plasma progesterone concentrations showed a characteristic and consistent profile: from basal levels of around 0.1ng/ml concentrations started to increase 12h before oviposition, reached an average maximum of 3.5ng/ml at 3h before oviposition and returned to basal levels 3h and 30min after oviposition. Changes in plasma luteinizing hormone and estradiol-17beta concentrations showed comparable patterns of elevation and decline relative to the timing of oviposition and ovulation. However, variation in their individual basal concentrations was generally larger and peak values were less conspicuous than those of progesterone. In non-ovulating hens (n=2) neither progesterone, nor luteinizing hormone nor estradiol-17beta showed elevations to peak concentrations before oviposition. These data demonstrate that during the egg laying cycle of ostriches, events such as ovulation, egg development and oviposition evolve according to a rather strict time schedule, and that progesterone, luteinizing hormone and estradiol-17beta reach peak concentrations shortly before ovulation. Additionally, our findings also show that on-farm ultrasound scanning is a useful technique to discriminate between ovulating and non-ovulating hens.

Adenohypophyseal function in bitches treated with medroxyprogesterone acetate.

The aim of this study was to investigate the effects of treatment with medroxyprogesterone acetate (MPA) on canine adenohypophyseal function. Five Beagle bitches were treated with MPA (10mg/kg, every 4 weeks) and their adenohypophyseal function was assessed in a combined adenohypophyseal function test. Four hypophysiotropic hormones (CRH, GHRH, GnRH, and TRH) were administered before and 2, 5, 8, and 11 months after the start of MPA treatment, and blood samples for determination of the plasma concentrations of ACTH, cortisol, GH, IGF-1, LH, FSH, prolactin, alpha-MSH, and TSH were collected at -15, 0, 5, 10, 20, 30, and 45 min after suprapituitary stimulation. MPA successfully prevented the occurrence of estrus, ovulation, and a subsequent luteal phase. MPA treatment did not affect basal and GnRH-induced plasma LH concentrations. The basal plasma FSH concentration was significantly higher at 2 months after the start of MPA treatment than before or at 5, 8, and 11 months after the start of treatment. The maximal FSH increment and the AUC for FSH after suprapituitary stimulation were significantly higher before treatment than at 5, 8, and 11 months of MPA treatment. Differences in mean basal plasma GH concentrations before and during treatment were not significant, but MPA treatment resulted in significantly elevated basal plasma IGF-1 concentrations at 8 and 11 months. MPA treatment did not affect basal and stimulated plasma ACTH concentrations, with the exception of a decreased AUC for ACTH at 11 months. In contrast, the maximal cortisol increment and the AUC for cortisol after suprapituitary stimulation were significantly lower during MPA treatment than prior to treatment. MPA treatment did not affect basal plasma concentrations of prolactin, TSH, and alpha-MSH, with the exception of slightly increased basal plasma TSH concentrations at 8 months of treatment. MPA treatment did not affect TRH-induced plasma concentrations of prolactin and TSH. In conclusion, the effects of chronic MPA treatment on adenohypophyseal function included increased FSH secretion, unaffected LH secretion, activation of the mammary GH-induced IGF-I secretion, slightly activated TSH secretion, suppression of the hypothalamic-pituitary-adrenocortical axis, and unaffected secretion of prolactin and alpha-MSH.

Effects of gonadotrophin releasing hormone administration on the pituitary-ovarian axis in anoestrous vs ovariectomized bitches.

The aim of this study was to determine the effects of gonadotrophin releasing hormone (GnRH) administration on the plasma concentrations of reproductive hormones in intact and ovariectomized (OVX) bitches. Therefore, blood samples were collected at multiple times before and after the administration of 10 microg/kg GnRH (Fertagyl)) for the determination of the plasma concentrations of luteinizing hormone (LH), oestradiol, progesterone and testosterone in six anoestrus and in six OVX bitches. The mean plasma LH concentrations before and 60 min after GnRH administration were significantly lower in the anoestrous bitches than in the OVX bitches. In both groups GnRH administration resulted in a significant increase in the plasma LH concentration. The highest plasma LH concentrations were found at 10 min after GnRH administration and these values did not differ significantly between the two groups. Only in the anoestrous bitches a significant increase in plasma oestradiol concentrations was found after GnRH administration and these values were significantly higher than those in the OVX bitches. The plasma concentrations of progesterone and testosterone were low (close to or below the limit of quantitation) both before and after GnRH administration and the differences between anoestrous and OVX bitches were not significant. It can be concluded that (i) basal plasma LH concentration is significantly higher in OVX bitches than in anoestrous bitches, (ii) plasma LH concentration increases after GnRH administration in both anoestrous and OVX bitches, (iii) GnRH administration causes a significant rise in plasma oestradiol concentration only if ovarian tissue is present and (iv) measurement of plasma progesterone and testosterone concentrations before and after GnRH administration does not aid in distinguishing between anoestrous and OVX bitches. The results of this study may provide a basis for the diagnosis of remnant ovarian tissue and verification of neuter status in the bitch.

[Transvaginal aspiration as first treatment of ovarian follicular cysts in dairy cattle under field circumstances]. / Transvaginale aspiratie als primaire behandeling van ovariële follikelcysten bij melkkoeien onder praktijkomstandigheden.

On dairy farms covered by the ambulatory service of the Department of Farm Animal Health, 55 Holstein-Friesian cows with suspected ovarian follicular cysts on the basis of rectal palpation underwent confirmatory transrectal ultrasonography. An ovarian follicular cyst was confirmed in 28 cows (50.9%) and was treated by single transvaginal-guided needle aspiration. Cows with a corpus luteum or a luteinized cyst were not treated. After aspiration, 82.1% (n=23) of the cows showed oestrus behaviour at 13.3 +/- 6.0 days. Artificial insemination was performed during the first heat after cyst aspiration and resulted in a pregnancy rate of 64.2%. In conclusion, single transvaginal-guided needle aspiration of ovarian follicular cysts is an easy and good method for the treatment of follicular cysts. Moreover, it is a safe and good alternative method for the manual, active rupturing of cysts during rectal palpation.