Morphologic and biochemical changes in the hypothalamus in hormonally stimulated sheep

The effect of exogenous hormonal preparations (PMSG, SG, PMSG + PGF2 alpha and FSH) was studied as exerted on neurosecretion and on changes in the nuclear volume of nucleus paraventricularis, nucleus ventromedialis, nucleus infundibularis and nucleus tuberomamillaris, and on histological changes in the third cerebral ventricle. Changes in catecholamine concentrations were determined in the rostral, medial, and caudal hypothalamus of the ewes. The study was performed with 28 ewes of the Slovak Merino breed during the oestrus period. The oestrus was synchronized by the Agelín vaginal tampons (Spofa). The ewes of four experimental groups were stimulated by the administration of 1000 I.U. PMSG (Antex-Leo), 1000 I. U. Serum Gonadotropin (Bioveta Ivanovice na Hané), 1000 I. U. PMSG with 250 micrograms Oestrophan inj. (Léciva Praha), and 450 I. U. Folistiman (VEB Arzneimittelwerk Dresden, GDR). The brain samples were processed by common histological methods. A caryometric analysis was performed at 3000-fold magnification and by measurement of 200 cells of one sample. The data were subjected to mathematical processing after Fischer and Inke (1956). The amount of neurosecretory material was evaluated by light microscopy (Nakahara, 1963). The multiplication of neurosecretion in the hypothalamic nuclei testifies to the fact that the administration of hormonal preparations stimulates the function of hypothalamic structures, which corresponds with the results of the caryometric analysis. It also ensues from the results that the administration of serum gonadotropins (PMSG and SG) at the dose of 1000 I. U. results in a statistically significant decline (P less than 0.001) of the concentrations of norepinephrine in the rostral, medial and caudal hypothalami of the ewes, although the PMSG preparation has a more pronounced effect when compared with SG. The hormonal stimulation with PGF2 alpha and with FSH causes no significant changes in the concentration of norepinephrine in the hypothalamus of the ewes. As norepinephrine is considered as a neurosecretion inhibitor, the multiplication of neurosecretion in the hypothalamus nuclei after stimulation with serum gonadotropins correlates with the decline of hypothalamic norepinephrine concentrations.     

Qualitative and quantitative changes in the hypothalamus and the ependyma of the 3d cerebral ventricle in sheep after irradiation and hormonal stimulation

In the submitted work we concentrated our attention on the study of the changes in neurosecretion. PAS positive mucopolysaccharides in the hypothalamus and ependyma of the IIIrd cerebral ventricle and on studying the changes in the volume of the cell nuclei of the neurons of nucl. supraopticus (NSO), nucl. hypothalamicus centromedialis (NVM) and nucl. tuberomamillaris (NTM) of sheep after irradiation and after hormonal stimulation. We made our observations on 28 sheep of the Slovak Merino breed, of average live weight 35 to 40 kg, two- and three-years old in the period of physiological anoestrum. The first group of four ewes and the second group of four yearling lambs were controls. The third group of six ewes and the fourth group of six yearlings were exposed to irradiation all over with a dosage of 2.46 Gy (250 R) for a period of five days. To synchronize the oestrum of all the sheep we used agelline sponges which we fitted five days before irradiation. After the irradiation and removal of the sponges we gave the test sheep hormonal stimulation with 3 X 500 i. u. of serum gonadotropin. The fifth group of four ewes and the sixth group of four yearlings were only stimulated without irradiation. The fifth day after stimulation we slaughtered the sheep. We processed the brain samples by the usual histological methods. We carried out karyometric analysis with 3000X magnification and the measurement of 200 cells from one sample. We processed the values obtained mathematically according to Fischer and Inke (1956). We evaluated the quantity of neurosecretion material with a light microscope according to Nakahara (1962). We found that the multiplication of neurosecretion and the increased activity of the PAS reaction in the hypothalamic nuclei studied show that the irradiation and hormonal treatment stimulate the function of the hypothalamic structures. The results of the karyometric analysis in the yearlings also confirm this opinion. In the ewes the inhibitive influence of irradiation was probably manifest. The giving of hormones in combination with irradiation causes the multiplication of ependyma cells and the desquamation of surface layers.     

Karyometric changes in the neurons of the hypothalamic and ependymal nuclei of the 3d cerebral ventricle in sheep after administration of Gn-RH and subsequent irradiation

Caryometric analysis was used for the study of changes in the cell nucleus volume of the neurons of nucleus paraventricularis, nucleus arcuatus, and ependyma of the third cerebral ventricle of sheep after the administration of Gn-RH, followed by exposure to X-rays. The test animals were 12 ewes in physiological anoestrus and two rams. The trials were conducted in spring. The first group of four sheep and two rams were left as controls; in the ewes of the second group the hypothalamo-hypophysial region was irradiated by exposure to 516.5 mC/kg (200 R); in the four ewes of the third group, ovaries were directly irradiated at laparotomy by exposure to 64.4 mC/kg (250 R). The ewes of the second and third group were treated with an i. m. administration of 400 micrograms Gn-RH per head before irradiation. The excisions were collected and processed the tenth day from irradiation. The studied material was fixed by injection of 10% formalin and finished by another dose of 10% formalin after the excision of the brains. The paraffin slices were stained with haematoxylin-eosine. The caryometric analysis was performed at 3000-fold magnification, 200 cells being measured in each sample. Changes in neurosecretory cells were described in the regions of nucleus paraventricularis, nucleus arcuatus and in the ependyma of the third cerebral ventricle. The results of the caryometric analysis of nucleus paraventricularis and nucleus arcuatus suggest that the administration of Gn-RH and irradiation of the hypothalamo-hypophysial region, and direct irradiation of ovaries, stimulate the studied cerebral structures. The changes observed in the ependyma of the third cerebral ventricle after the administration of Gn-RH and subsequent irradiation of the hypothalamo-hypophysial region are insignificant; it is only after the direct irradiation of ovaries that these cells are inhibited by an indirect effect through the feedback mechanism.     

Impregno-architectonic study of the medial part of the hypothalamus in sheep

Impregno-architectonics of neurons in the nuclei of the medial part of hypothalamus was studied in two-month and adult female and male sheep using the methods after Ramón-Moliner and Golgi-Cox. Types of neurons in the hypothalamic nuclei are given, morphology of synaptic conjugations and orientation of nerve processes are described. Ventromedial nucleus has the most interesting structure as a part of the medial hypothalamus: it is ring-shaped in cross-section with paucicellular center, contains various types of neurons and all kinds of synapses which can occur in hypothalamus. Only small numbers of neurons in the pars caudalis of tuberomammillary nucleus, similarly like in nucl. supraopticus and nucl. paraventricularis, are impregnated so that they can be supposed to belong to true neurosecretory nuclei.     

Morphologic picture of the ependyma of the 3d cerebral ventricle in ewes after hormonal stimulation

The ependyma in the infundibular region of the third cerebral ventricle was studied. Ageline sponges (20 mg per animal) were applied to nine ewes. On the thirteenth day after the removal of sponges, 750 I. U. PMSG was administered intramuscularly to three ewes, 1000 I. U. PMSG to another three, and the remaining three ewes were left without this treatment. Six ewes were the control. After embedding in paraffin, the material obtained from four control sheep and all the test sheep was stained with haematoxylin-eosine, material from another two control animals was impregnated by the method after Golgi-Cox. The ependyma in the infundibular region of anoestric ewes has a single layer and is cubic to cylindrical; it is only in the recessus infundibuli that it forms two to four layers. After the administration of Ageline, or in combination with PMSG, ependyma can be observed to react within the whole infundibular region of the third cerebral ventricle, but the most expressive reaction is recorded in the caudal part of the middle third of infundibulum where a transient type of cells (between ependymal cells and tanycytes) was found in the control animals after impregnation (they have cilia and one to two short and one long processes). After Ageline administration, ependymal cells in the middle third stretch out like in the anoestric period. The administration of 20 mg Ageline and 750 I. U. PMSG gave rise to low digital excrescences, the ependyma is undulated (pseudo-stratified) or contains small deposits of multiplied cells. After the administration of 20 mg Ageline and 1000 I. U. PMSG, the middle third contained, besides subependymal serous infiltrate, also large digital excrescences, probably filled with a serous fluid, and the surface of the subependyma and ependyma is eroded, containing proliferated deposits of ependymal cells of different thickness. Everywhere proliferation occurs, the surface layer of ependymocytes is desquamated into the cerebrospinal fluid. The histological picture described in the present paper probably suggests an increased secretory activity of ependymal cells after the administration of hormonal preparations. It is confirmed by these results that ependyma is involved in the hypothalamo-hypophyseal control of the sexual activity of sheep.     

Quantitative micromorphological study of the ovaries in sheep after estrus synchronization and superovulation in the autumn mating season

The quantitative micromorphological changes of tertiary follicles and corpora lutea (CL) were studied in ewes in the autumn mating season after oestrus synchronization, induced by administration of PGF2 alpha (Oestrophan Spofa) at a rate of 125 micrograms, and after superovulation, induced by administration of PMSG (Antex Leo, Denmark) at a rate of 1000 I. U., or PMSG at rates of 750 and 1000 I. U. together with 50,000 I. U. vitamin A (Axerophthol Spofa). The highest number of ovulations was obtained in ewes treated with 1000 I. U. together with vitamin A (3.4 +/- 3.0) and after administration of 1000 I. U. PMSG alone (2.6 +/- 2.74). The highest number of tertiary follicles was recorded in ewes after administration of PGF2 alpha. The proportion of tertiary atretic follicles was the highest in ewes after administration of PMSG (64.6%). The occurrence of the luteinizing form of atresia was recorded only in ewes treated with PMSG (4% of the total number of atretic follicles). Using the caryometric analysis of the luteal cells of corpora lutea, the ewes of the experimental groups had two-peak variation curves; this corresponds to the theory of the presence of two luteal types in the tissue of the corpus luteum in ewes. As determined morphometrically, the smallest proportion of connective tissue out of the total volume of ewes' ovaries was found after administration of 1000 I. U. PMSG together with vitamin A. Administration of vitamin A together with PMSG had a favourable influence on the over-all follicular response, on the average number of ovulations, and on the proportion of non-atretic follicles.     

Changes in the hypothalamus in continuously irradiated sheep

Neurosecretion, PAS-positive mucopolysaccharides and the Nissl substance were studied in the neurons of the rostral, medial and caudal hypothalamus of continuously irradiated ewes. The study was performed on 21 ewes of the Slovak Merino breed at the live weight of 34 kg. The animals were in the period of physiological anoestrus and their age was two to three years. The first group of six ewes was the control. The second group included 15 sheep irradiated with a total dose of 6.7 Gy (700 R) for seven days. Co60 was used as the source of irradiation. The animals of this group were killed seven days from treatment. The ewes in the third group were left for the study of mortality. The brains were perfused with 2% buffered paraformaldehyde immediately after the bleeding of the sheep; then the brains were removed from skulls and fixed in buffered picroformol. Paraffin slices were stained with haematoxylin-eosine, aldehyde-fuchsine and alcian blue for neurosecretion, with PAS-reaction for mucopolysaccharides and with cresyl violet for the Nissl substance. As found, the irradiation of the whole body inhibits the activity of neurosecretory cells in rostral and medial hypothalamus, thus reducing neurosecretion. These regions also show a reduced activity of PAS reaction used for the demonstration of mucopolysaccharides. The observed changes also included damage of the endothelium of blood vessels with the occurrence of erythrocyte extravasates and with haemorrhages.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in the hypothalamus and adjacent ependyma after hormonal stimulation of the ovaries in ewes]

Superovulation treatment leaves alternations in the controlling regions of the hypothalamus and in the adjacent ependyme after ovulation. The test ewes were synchronized with Agelin (20 mg chlorsuperlutin in one vaginal sponge) and stimulated (after the removal of the sponges) with 750 IU PMSG + 750 IU HCG and with 1000 IU HCG and 750 IU PMSG + 5 ml Antisergon (goat antiserum against PMSG), administered 68 hours after PMSG (i.e. 40 hours after HCG). The control ewes were in different stages of the ovarial cycle. The experimental ewes were killed 120 to 130 hours after the start of stimulation. Routine histological techniques were used to treat the brain samples; this treatment was followed by assessment under light microscope. The ependyme epithelium of the third cerebral chamber was studied under scanning microscope. Preparations with different FSH:LH ratios had different effects on the nucleus ventromedialis. Antisergon administration influenced the secretion of NPV (prevented persistent stimulation), which was observed after administration of PMSG + HCG. On the surface of the lower part of the third cerebral chamber the administration of Antisergon slowed the formation of the miniblebs. Supraependyme cells disappeared after stimulation for superovulation.     

The effect of the seasons on the ovarian response in sheep after the administration of the serum gonadotropin, PMSG

In the autumn oestrus season, 20 Slovak Merino ewes were exposed to synchronization of oestrus, treated with the PGF2alfa at doses 125 micrograms (Oestrophan, inj. Spofa). followed by an injection of PMSG at doses 1000 IU (Antex Leo Denmark) and 50,000 IU of Vitamin A (Axerophtol Spofa). 23 anoestrus ewes were synchronized with an intravaginal sponges containing 20 mg of chlorsuperlutine (Agelin, Spofa) for 12 days and after sponge withdrawal, the ewes were injected with 750 and 1000 IU of PMSG (Antex Leo Denmark). Ovulatory response was observed and the possibility of ova recovered from the genital organs in ewes after synchronization of oestrus and superovulation in oestrus season. Higher values of the total follicular response (CFO), and the average number of ovulation (PO) after administering equal doses of PMSG were found out both in anoestrus ewes (CFO 6.62 +/- 4.24; PO 4.25 +/- 4.52) and in oestrus ewes (CFO 2.70 +/- 2.10; PO 2.60 +/- 1.74; resp. CFO 2.80 +/- 1.83; PO 3.4 +/- 3.0), if the ewes were treated with PMSG together with vitamin A. The average number of ova flushed was higher in anoestrus ewes (3.0-0.5) than in oestrus ewes (1.67-3.75). In both trials the equal ratio in the number of released ova was gained from ewes of experimental groups (83-88% of the total number). After ova flushing from the genital organs in ewes of the experimental groups most ovas were found in the isthmatal part of the uterine tube (36-60%). On the basis of gained results it was concluded, that synchronized oestrus ewes on receiving PMSG in anoestrus season the ovarial response was more significant than in autumn breeding season.     

The effect of hormonal stimulation on the follicle population in the ovaries of ewes during estrus and anestrus

In the oestral and anoestral periods, 129 ewes of the Slovak Merino breed were treated with different doses of the stimulants PMSG, PGF2 alpha, HCG, SG and FSH after synchronization by means of Ageline tampons. In the anoestral period, PMSG doses of 750 and 1000 i. u. were found to increase the number of tertiary follicles from 12.3 to 17.2 and 18.6, whereas the dose of 1500 i. u. was found to reduce this number from 18.6 to 13.5. PGF2 alpha increases the average number of tertiary follicles from 12.3 to 22.2 and when it is combined with 750 i. u. PMSG, the increase is to 29.3 tertiary follicles. At higher PMSG doses the ratio of atretic (A) to non-atretic (N) tertiary follicles rose and the number of ovulations increased from 2.5 to 4.0. After synchronization with Ageline and administration of PMSG at the doses of 750 and 1000 i. u., the number of tertiary follicles increased from 16.0 to 18.4 and the number of ovulations from 3.1 to 5.7; the number of atretic tertiary follicles showed no change. In the oestral period, the dose of 1000 i. u. PMSG administered after PGF2 alpha reduced the number of tertiary follicles from 18.9 to 14.5 and increased the number of ovulation from 1.6 to 2.6; an increase was also recorded in the number of atretic tertiary follicles.(ABSTRACT TRUNCATED AT 250 WORDS)     

Estrogen receptor alpha expression in the medial preoptic area and the medial basal hypothalamus under different physiological conditions in cattle

The distribution and morphology of immunoreactive estrogen receptor alpha (ERalpha)- containing cells were examined in the cattle brain. The brains were collected from two cows and a male calf. One of the cows was sacrificed one week after parturition. ERalpha expressions in the preoptic area of the rostral forebrain and the medial basal hypothalamic area were evaluated immunohistochemically using a mouse monoclonal antibody. In all animals, the signals that indicate ERalpha were detected in the medial preoptic area, the level of the organum vasculosum of the lamina terminalis and the bed nucleus of the stria terminalis. In the caudal hypothalamic region, they were detected in the arcuate nucleus, the periventricular and ventromedial hypothalamic nuclei. ERalpha-containing cells were characterized by a dense nuclear reaction product. In all the subjects examined, ERalpha signals were clearly detected in the cytoplasm as well. The density of ERalpha expression was different among the three cattle; the number of ERalpha containing cells in the postpartum cow was much lower than that in the other cow and the male calf. The present findings suggest that ERalpha expression in the forebrain is influenced by the reproductive status in the cattle.     

Early Breeding and Pregnancy Diagnosis in Syrian Awassi Sheep Yearlings

Contents
Fifty-nine female yearlings of local Awassi sheep were randomly divided into two groups. Animals in group T (treated) were fitted with intravaginal sponges containing 60 mg of medroxyprogesterone acetate for 14 days followed by 400 IU of pregnant mare's serum gonadotrophin (PMSG) at sponge withdrawal, whereas group C (control) received no treatment. Oestrus rate was 92.7 and 11.2% for groups T and C, respectively. Lambing rate was 78 and 5.6% for groups T and C, respectively. Twinning rate was 31.3% in group T compared to zero in group C. Average birth weight for single born lambs (4.7 ± 0.6 kg) was significantly (p < 0.05) higher than twin born lambs (3.0 ± 0.5 kg) in group T. The average concentration of blood progesterone collected between days 17–19 after mating was 19.30 nmol/l and the accuracy of early pregnancy diagnosis was 100%. It was concluded that, it is possible to induce synchronized oestrus, and to increase the twinning rate in Syrian Awassi sheep yearlings outside the breeding season, using intravaginal sponges and PMSG. In addition, early pregnancy diagnosis could be successfully determined in female Awassi sheep yearlings between days 17–19 after mating.     

Biochemical and histological changes in the hypothalamus in hormone-stimulated sheep]

The effects were investigated of a hormonal preparation of serum gonadotrophin (SC) on neurosecretion, PAS-positive mucopolysaccharides and on histological changes in the third cerebral ventricle; at the same time variations were determined of catecholamine concentrations of ependyma surface in the hypothalamic regions that control directly reproductive functions in ewes of the Merino and Wallachian breeds in the oestrus period. Ewe oestrus was synchronized by insertion of Ageline (Spofa) sponges into the vagina. A dose of 2000 IU of serum gonadotrophin (Bioveta, Ivanovice in Haná) was administered to stimulate the ewes (n = 30). Twelve ewes of the Wallachian breed, six control and six test ones, were used for radioenzymatic detection of catecholamines (Catechola test, Prague). The brains were segmented immediately after bleeding and samples were taken from the eminentia medialis, area preoptica and corpus mamillare pursuant to the stereotactic configuration of the sheep hypothalamus (Welento et al., 1968). The brains of ten ewes of the Merino breed, four control and six test ones, and of eight ewes of the Wallachian breed (four control and four test ones) were used for histological treatment. For the purposes of a histological study, the brains were treated with current histological methods. The amount of neurosecretory material was assessed by light microscopy (Nakahara, 1963). Samples for the study of ependyma of the lower part of the third cerebral ventricle were examined in a scanning electron microscope. The obtained results demonstrate a statistically significant decrease by 36.4% (P less than 0.05) in epinephrin concentrations in the area preoptica of hypothalamus after SG administration (Fig. 3), in comparison with the control group. A decrease in norepinephrin concentration in the corpus mamillare (Fig. 4) was statistically significant (P less than 0.001). The dose of 2000 IU SG influenced most markedly catecholamine concentrations in the eminentia medialis (Fig. 5), where a statistically significant decrease in epinephrin and norepinephrin concentration was observed (P less than 0.01). Dopamine concentrations also decreased significantly (P less than 0.05) in the eminentia medialis (Fig. 5). Following the hormonal stimulation, a significant increase in dopamine and epinephrin concentrations was observed in the hypophysis of sheep (Fig. 6; P less than 0.05) and in the epiphysis there was a significant decrease in epinephrin and dopamine concentration (P less than 0.01).(ABSTRACT TRUNCATED AT 400 WORDS)     

Changes in the follicle system and the population of tertiary follicles in sheep after the administration of PMSG in anestrus

The total quantitative changes of ovaries, proportion of atretic and non atretic follicles and changes of tertiary follicles in sheep after administration of increasing doses of PMSG during the anoestrous period were observed. In experimental groups the statistically significant increase of average weight, volume and dimensions of ovaries in comparison with control group were determined biometrically. The average number of tertiary follicles was greater in experimental groups but at the same time we observed a higher proportion of atretic follicles (64% of the total number in the control group; 71-77% in the experimental groups). In the group of sheep administered a dose of 1500 m.u. PMSG we determined a high proportion of luteinized follicles (as much as 21% of the total number of atretic follicles). The total number of small follicles in the so called transient phase in the comparison of experimental and control groups was not changed significantly. In the experimental group an increased incidence of preovulatory follicles and a reduction of tertiary follicle dimensions in the period of follicle cavity formation was determined.     

The effects of time and dose of pregnant mare serum gonadotropin (PMSG) on reproductive efficiency in hair sheep ewes

The aim of this study was to evaluate the effects of dose and application time of pregnant mare serum gonadotropin (PMSG) on reproductive performance of hair sheep ewes synchronized with fluorogesterone acetate (FGA) under tropical conditions of Northeastern Mexico. Ninety-nine hair ewes (63 Blackbelly and 36 Pelibuey) were treated with intravaginal sponges during 10 days. After insertion of FGA sponges, ewes were divided into four groups, and PMSG was injected intramuscularly at doses of 100, 200, and 400 IU. Relative to FGA sponge removal, PMSG was administrated at −48 h, −24 h, and at sponge removal. PMSG was not administered to the control group. Control ewes had similar (P > 0.05) lambing rate, fertility, and fecundity than those treated with 100 IU of PMSG, but lower (P < 0.05) percentages to these variables than those treated with 200 and 400 IU of PMSG. Time to estrus decreased linearly, and ovulation rate increased quadratically as PMSG dose increased (0 to 400 IU). Administration of PMSG before sponge removal increased (P < 0.01) response to estrus and decreased (P < 0.01) interval to estrus compared with control. Ovulation rate, lambing rate, fertility, and fecundity were not affected (P > 0.05) by administration time of PMSG. Both dose and time of PMSG application did not affect (P > 0.05) pregnancy rate, percentage of single and multiple lambing, and prolificacy. In conclusion, results show that the dose of 400 IU of PMSG administered before sponge withdrawal in an estrus synchronization protocol improved reproductive efficiency of hair sheep ewes.     

Symmetric focal degeneration in the cerebellar and vestibular nuclei in swine caused by ingestion of Aeschynomene indica seeds

A vestibulocerebellar disorder was observed in 2- to 4-month-old swine after consumption of broken rice contaminated with 13% Aeschynomene indica seeds. Affected animals recovered in 2-14 days after removal of the contaminated food. To reproduce the syndrome, 10 pigs were fed 13% A. indica seeds in commercial pig ration or rice. They showed clinical signs similar to those observed in the spontaneous cases, 1 hour to 6 days after ingestion. Three pigs recovered after the withdrawal of the contaminated food. The others were killed. Two pigs that were fed for 25 days with food containing 3% and 6% seeds and two control pigs did not show clinical signs. Histologic lesions were characterized by symmetric focal degeneration in the cerebellar and vestibular nuclei. The initial lesion, observed 24 hours after the first ingestion, was characterized by vacuolation of the neuropil. Subsequently, there was progressive loss of parenchyma, vascular reaction, a few small spheroids, astrocytosis, and accumulation of gitter cells. The brains from four pigs were perfused with a buffered solution of glutaraldehyde-paraformaldehyde 12-48 hours after they started to ingest the seeds. No ultrastructural lesions were observed in the cerebellar nuclei after 12 hours of seed consumption. At 24 hours, endothelial cells and pericytes were separated by empty spaces from astrocyte foot processes, suggesting perivascular edema. The astrocytes were enlarged, consistent with intracellular fluid accumulation. These results suggest that A. indica seeds are toxic, causing functional derangement of the vestibulocerebellar system, followed by alterations in the microvasculature in the cerebellar and vestibular nuclei.     

Comparison of the effect of cronolone sponges and PMSG or cloprostenol on estrous induction in Turkish Saanen goats

The efficiency of cronolone sponges in combination with either pregnant mare serum gonadotrophin (PMSG) or cloprostenol (PGF2alpha) for inducing and synchronizing the estrous cycle in Turkish Saanen does was investigated during the transition from non-breeding to breeding season. All does (n = 80) were treated with 20 mg cronolone sponges for 11 days and divided into 4 equal groups. In addition, each doe received an intramuscular injection of either 1.5 ml sterile saline solution, 0.075 mg PGF2alpha, 500 IU PMSG or 500 IU PMSG and 0.075 mg PGF2alpha, 24 h before the sponge removal. Cervical artificial insemination (AI) with frozen-thawed semen was performed once 16 h after the detection of the first accepted mount. The total estrous response for the first 24 +/- 4 h, total estrous response within 96 h, time to onset of the induced estrus, duration of the induced estrus and pregnancy rate was found to be 75.0%, 97.5%, 31.4 +/- 1.2 h, 29.3 +/- 1.2 h, and 33.3%, respectively. There were significant differences between the first two groups and the last two groups in terms of the onset of induced estrus and estrous response at the first 24 +/- 4 h (P < 0.05). These results indicate that the use of cronolone/PMSG was more effective than cronolone/PGF2alpha in the attainment of early and compact induction of estrus in Turkish Saanen does.     

Impregnoarchitectonic study of the rostral part of the hypothalamus in sheep

The impregnoarchitectonic picture of neurons in the nuclei of the rostral part of hypothalamus was studied by the method after Ramón- Moliner and Golgi-Cox in two months old and adult Merino sheep of both sexes. Neuron types, morphology of synaptic linkages and the spatial orientation of neuron processes are indicated in each hypothalamic structure. The most frequent type of neurons in the regio hypothalamica rostralis is an oval to spindle-shaped neuron, and the cells are exclusively multipolar. As distinct from other hypothalamus nuclei, only a small number of neurons were impregnated in the nuclei of nucleus supraopticus and nucleus paraventricularis because these were true neuroscretory nuclei.     

Ovarian response to PMSG and GnRH in ewes immunised against oestradiol-17 beta

Forty-six adult merino ewes were immunised against oestradiol-17 beta-6 carbomethyloxime:human serum albumin and 48 comparable ewes were used as controls in an experiment to study the effects of gonadotrophin releasing hormone (GnRH) on ovulatory responses after treatment with pregnant mare's serum gonadotrophin (PMSG). All the ewes were treated with progestogen sponges for 14 days and received 1500 iu PMSG on the 12th day. Twenty-four control and 24 immunised ewes received 25 micrograms GnRH 21.5 hours and 23 hours after the sponges were withdrawn. Plasma samples were collected between 17 and 50 hours after the sponges were withdrawn and assayed for luteinising hormone (LH). Immunisation reduced the proportion of ewes which ovulated and their rate of ovulation. Injection of GnRH increased the proportion of immunised ewes ovulating (P less than 0.0005) and their rate of ovulation (P less than 0.0001). More unovulated follicles were observed in immunised ewes regardless of GnRH treatment (P less than 0.0001). The rate of recovery of eggs was reduced after immunisation. Treatment with GnRH produced a surge of LH of equal magnitude in the control and immunised ewes although not as many immunised ewes ovulated.     

Influence of PMSG and time of artificial insemination on fertility of progestogen-treated sheep in confinement

The need for pregnant mates' serum gonadotropin (PMSG) in breeding confined sheep by artificial insemination (AI) at progestogen-synchronized estrus was assessed in 152 adult crossbred ewes brought into season by a controlled light regimen. One-half of the ewes received 500 IU PMSG after intravaginal progestogen treatment; all ewes were inseminated either 54, 57 or 60 h after sponge removal or at 54 and again at 60 h. Based on progesterone determinations 18 d after AI, conception rates with single insemination 54, 57, or 60 h and double insemination at 54 and 60 h were 76, 72, 47 and 72%, respectively, among ewes receiving PMSG, compared to 17, 22, 47 and 43%, respectively, among ewes not give PMSG (P less than .01) Lambing rates were higher (P less than .01) with PMSG (67, 67, 37 and 61%) than without PMSG (11, 11, 26 and 33%). While there was only a small increase (.06 less than P less than .05) in litter size with PMSG, fecundity decreased (P less than .01) from 1.4 to .3 when PMSG was not used. These data indicate that, even with controlled lighting to induce estrous activity, additional stimulation of ovulation by PMSG at progestogen-synchronized estrus is necessary for normal fertility when confined sheep are bred by AI.