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.     

The effect of vitamin A on trypsin inhibition activity of the blood and cervical mucus after the effects of hormones on the ovaries of sheep in the estrus period

Vitamin A (50,000 I. U.), administered after oestrus synchronization by PGF2 alpha (2 X 125 micrograms; 1st and 11th day) together with PMSG (750 and 1000 I. U.), had a stronger influence on the changes in the trypsin-inhibition activities (TIA) of the blood plasma, as compared with the effect of PMSG. The changes in the average TIA values within 120 hours after the administration of the stimulating dose were also observed more frequently to depend on vitamin A. After administration of 1000 I. U. PMSG, an increase was recorded only in the values of the fraction of low-molecular TIA, whereas the administration of the combinations of PMSG + vitamin A resulted in an increase of all the TIA's under study. This increase was directly correlated with an increased number of non-atretic tertiary follicles, with an increased number of ovulations (at the same dose of PMSG), and with a reduced ratio of changes in the concentration of progesterone (P) and 17-beta oestradiol (E): P/E = 1.1 after the administration of 1000 I. U. PMSG, P/E = 0.81 after I. U. PMSG + vitamin A, and P/E = 0.90 after 1000 I. U. PMSG + vitamin A. The increase in the average TIA of the cervical mucus is due to the increased secretory activity of the cervical glands rather than to the multiplication of these glands after ovary stimulation.     

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.     

Superovulation induction in sheep using PMSG

The effects of application of differentiated PMSG rates (Antex Leo, Denmark) were investigated in 30 ewes of the Slovak Merino breed in the period of May--June in experimental conditions on the induction of superovulation and on the speed of egg transport through the sexual organs. After administration of PMSG at rates of 750, 1000 and 1500 i. u. per head/day, increases were recorded in the average weight of ovaries, number of large follicles and ovulations. Flushing the eggs from different parts of the sexual organs indicated that the higher the PMSG rate, the faster was the egg transport.     

The effect of PMSG on the trypsin inhibitory activity of the blood and cervical mucus and on the reproductive organs in sheep during anestrus

The effect of the doses of 750, 1000, and 1500 I. U. PMSG (Antex Leo) on changes in the trypsin-inhibition activities (TIA) of the blood plasma and cervical mucus of ewes was found to vary: a) in the times (from PMSG administration) of recording the minimum values of total TIA (72, 48, 24 hours); b) in the values of the average total TIA on individual days of the trial (40.3-141.4% of the control values); c) in the fractions of low-molecular TIA after administration of 750 and 1500 I. U. PMSG, the fractions exhibited minimum values after 24 hours, and of 1000 I. U. after 72 hours (53.0 and 60.4, and 69.7% of the control values, respectively); d) in the highest values of the fractions of low-molecular TIA recorded after 96 hours (96.1-183.7% of the control values); e) in the average values for 96 hours of the trial, which ranged from 64.1% (1500 I. U.) to 77.1% (750 I. U.) for total TIA and from 89.6% (750 and 1000 I. U.) to 122.8% (1500 I. U.) for low-molecular TIA; f) in the ratio of atretic and non-atretic (A/N) tertiary follicles, which grew with increasing PMSG doses; this was indirectly correlated with the average values for total TIA; g) in the weights of uteri and ovaria which increased from 176.8 to 322.3% of the control values after the stimulation; h) in the epithelium thickness of cervix and cervical glands increasing from 118% to 316% of the control values; i) in the average TIA values of cervical mucus ranging from 53.3 to 125% of the control values.     

Changes in the biometry of the sex organs of sheep after estrus synchronization and superovulation in the breeding period

In twenty ewes of the Slovak Merino breed coming from a demonstration farm at Zemplínska Teplica, to the age of two-three years biometrical variations of the sex organs and overall follicular response to PMSG and PGF2 alpha administration were investigated in the autumn period (October-November). In the ewes of all groups the heat was synchronized by i. m. administration of PGF2 alpha (Oestrophan Spofa) at a dose of 125 micrograms in the interval of 11 days. On the ninth day the ewes of the second and fourth group were given 1000 i. u. of PMSG (Antex Leo, Denmark), the ewes of the third group 750 i. u. of PMSG. The ewes of the third and fourth group were administered at the same time 50,000 i. u. vitamin A (Axerophtol Spofa) (each group). The weight and dimensions of the sex organs were investigated. The results demonstrate that the administration of PMSG in the mating period increases significantly the weight of oviducts and horns of uterus in the ewes while there are no variations of the weight of the other sex organs and of their length, or it is lower than in the ewes of the control group. The overall follicular response was not influenced by the higher weight of ovaries.     

Serum levels of progesterone and estradiol in relation to tertiary follicles in the sheep ovary in proestrus and after administration of gonadoliberin

Changes in progesterone (P4) and 17 beta-estradiol (E2) concentrations in blood serum as well as changes in counts and quality of tertiary follicles on ovary surfaces of ewes were investigated after administration of different GnRH doses on the 15th and 16th day of the oestral cycle. Progesterone (P4) and 17 beta-estradiol (E2) were determined by radioimmunological assays using RIA-test-Prog and RIA-test-Estra kits. It follows from the results that GnRH in Dirigestran inj. Spofa preparation reduces the E2 concentration and multiplication of the number of selected tertiary follicles of ovaries through stimulation of their growth and ripening in ewes of the Improved Wallachian breed. The greatest number of ovulations was reported in ewes administered three doses of the preparation. A positive correlation (r = 0.803) was found between the E2 concentration and the number of selected tertiary follicles of ewes in proestrus. The results corroborated the data on a pulsatile release of gonadoliberin during pre-ovulation incretion of gonadotropins--mainly lutropin (LH), as well as the inhibiting effect of gonadoliberin on the steroidogenesis of the ovary--mainly on estradiol production.     

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.     

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.     

The Effect of Gonadotrophin Releasing Hormone and Follicle Stimulating Hormone in Conjunction with Pregnant Mare Serum Gonadotrophin on the Superovulatory Response in Crossbred Sheep In India

Thirty-two mature crossbred (Fine Wool Synthetic, FWS) sheep developed for fine wool production in India were treated for superovulation and oestrus synchronization in spring season. The ewes were randomly allocated to four treatment groups in a factorially designed experiment. For induction of superovulation, pregnant mare serum gonadotrophin (PMSG) was administered alone (group 1), in combination with gonadotrophin releasing hormone (GnRH) (group 2) or with follicle stimulating hormone (FSH) (group 3) and with both (group 4). Oestrus was synchronized in all the ewes by two injections of prostaglandin F2 (PGF, 10 mg each) administered at an interval of 10 days. Superovulation treatment started 48 h prior to the second PGF injection. The proportion of ewes in oestrus did not differ significantly (p>0.05) in the four groups. The use of GnRH set the ewes into oestrus earlier than the ewes in the other groups. Treatment with PMSG (800 IU) in conjunction with 4 g of Buserilin (GnRH) increased the ovulation rate (9.1±2.6 corpora lutea (CL)) above that observed when PMSG was used alone (3.0±0.7 CL). The use of FSH (0.5 U ovagen) in conjunction with PMSG was characterized by a decrease in the proportion of ewes with 2 CL (4/8 vs 7/8; p<0.05) and in the number of ovulations, i.e. CL observed (2.4±0.6 vs 9.1±2.6), and a nonsignificant increase in the incidence of large follicles (LF) (4.6±1.28 vs 3.25±0.6; p>0.05). The interaction between treatments of FSH and GnRH was not significant (p>0.05).It is concluded that use of GnRH, in conjunction with either PMSG alone or PMSG plus FSH treatment, advanced the onset of oestrus and increased the ovulation rate in FWS sheep.     

The follicular system of bovine ovaries after estrus synchronization using the luteolytic action of cloprostenol

Oestrus synchronization was studied in samples from six cows of the Black-Pied Lowland breed. Three cows four to five days from oestrus were used as the control; three animals with marked periodic corpora lutea were given an i. m. injection of 0.5 mg cloprostenol. The eighth day from the administration of the preparation, the ovaries of the cows were excised and, after histological processing in a simultaneous series in a 4mm interval, the preparations were subjected to qualitative and quantitative microscopic evaluation. The structure of non-atretic and atretic follicles was described in different stages of the atretic process. The lymphoid cells of atretic follicles were observed to penetrate into the granulosa membrane. A multiplication of non-atretic tertiary follicles was observed after the administration of cloprostenol. This multiplication was more pronounced on the right ovary where the preceding ovulation had taken place (P less than 0.01). The treated animals, compared with the controls, showed a significant multiplication of tertiary follicles at early atresia and at total collapse atresia (P less than 0.001), whereas the number of follicles with contractive atresia showed a significant decrease (P less than 0.001). The results suggest that cloprostenol can influence follicle population mostly through the stimulation of the growth and ripening of tertiary follicles; its modulation effect seems manifest itself in cooperating relation with gonadotrophic hormones, mainly with the follicular secondary hormone (FSH), in the theory of the complex effect of proteohormones .     

Superovulation after administration of PMSG in cows of various breeds

Superovulation response was studied to i. m. administration of 2500 and 3000 i. u. of PMSG (special product Folligon, Intervet Co.) in 67 breeding cows of the Black-Pied Lowland. Slovak Pied and Slovak Pinzgau breeds within the 9th to 12th days of their sexual cycles. The time which had elapsed from these cows' last calving ranged from 50 to 150 days and the number of their prior calvings ranged from one to ten. The best superovulation effect was obtained after the administration of 2500 i. u. PMSG in the Pinzgau breed: these cows had, on the average, 13.20 +/- 2.36 corpora lutea without non-ovulated follicles. At the PMSG dose increased to 3000 i. u., the number of yellow bodies rose to 18.11 +/- 1.12 and that of non-ovulated follicles to 3.46 +/- 0.46. In the Slovak Pied breed the average number of yellow bodies obtained after administration of 2500 i. u. PMSG was 11.74 +/- 1.27 and that of non-ovulated follicles was 0.44 +/- 0.02. At the PMSG dose of 3000 i. u., the number of yellow bodies increased to 15.49 +/- 1.62 and that of non-ovulated follicles increased up to 5.12 +/- 0.81. In the Black-Pied Lowland breed the lowest response was obtained: the i. m. administration of 2500 i. u. PMSG resulted in the formation of 9.5 +/- 0.84 yellow bodies and 1.16 +/- 0.26 non-ovulated follicles, and at the dose of 3000 i. u. the respective numbers were 13.41 +/- 0.89 and 3.07 +/- 0.39. Comparing the superovulation effect in dependence on age, the response of the cows of the Black-Pied Lowland and Slovak Pinzgau breeds to PMSG administration increases until the age of nine years (from 9.79 and 13.6 yellow bodies, respectively, on the average for the first five years to 12.71 and 17.99, respectively, in the ninth year); in the Slovak Pinzgau breed it decreases from 15.83, recorded in the first five years, to 8.28 +/- 1.68 in the ninth year. The number of non-ovulated follicles grows with age in the Slovak Pinzgau breed from 3.46 +/- 0.46 in the first five years to 3.64 between five and nine years of age and up to 4.44 +/- 0.52 after the ninth year, in the Slovak Pied breed from 0.79 to 4.21 +/- 0.38, and in the Black-Pied Lowland breed from 2.69 to 3.20 +/- 0.22 between the fifth and ninth year.     

Ovarian responses to intramuscular or intravenous administration of prostaglandin F2 alpha in control and FSH-treated beef heifers

Sixty Simmental crossbred heifers 18 to 20 mo of age were detected in estrus and assigned at random to a 2 x 2 factorial design study with 30 controls, and 30 given FSH. Half of each group was given prostaglandin F2 alpha (PGF) i.v. and the rest i.m. Injections of follicle-stimulating hormone were started on d 7 to 14 of an estrous cycle and continued for 5 d or until ovariectomy; PGF was administered either i.v. or i.m. at 48 (25 mg) and 60 (15 mg) h after the initial FSH injection. Control females received a similar PGF treatment on a day between d 9 and 15 of the estrous cycle. Blood samples were collected from all animals immediately before PGF administration and every 12 h thereafter until ovariectomy. Within each of the four experimental subgroups, ovariectomies were performed at either 24, 48 or 72 h (five/time group) after initial PGF injection. Ovarian and corpus luteum (CL) weights were recorded as well as number and size of follicles and number of ovulations. Regression of the CL was slower (P less than .05) after administration of PGF i.v. than i.m. (CL weight was 2.6 vs 3.3 +/- .2 g for i.m. and i.v. groups, respectively). Exogenous FSH increased estradiol-17 beta (E2) concentrations, and FSH-treated heifers had more (P less than .05) early ovulations than control heifers did. Ovulations in FSH-treated heifers had begun to occur by 24 h after i.v. and i.m. PGF injection.(ABSTRACT TRUNCATED AT 250 WORDS)     

Follicle stimulating hormone increases serum oestradiol-17 beta concentrations, number of growing follicles and yolk deposition in aging hens (Gallus gallus domesticus) with decreased egg production.

1. The aims of this study were to determine if the number of small yellow follicles (SYF) and large white follicles (LWF) in ovaries of young and old hens differed; and if injection of old hens with follicle stimulating hormone (FSH) changed growth of and yolk deposition into follicles of old hens. 2. Ovaries were removed and follicles were divided according to size and condition and counted. The number of normal SYF and LWF was decreased in old hens compared to young hens, whereas the number of atretic follicles was greater in old hens compared to young hens. 3. Old hens were injected subcutaneously with saline containing 0.1% bovine serum albumin (BSA, vehicle) or 12.5, 50, 200, 400 micrograms porcine (p) FSH or 25 or 50 IU pregnant mare's serum gonadotropin (PMSG) for 5 consecutive days. Blood samples were taken on days 1 and 5 before FSH and PMSG injection and 2 h after FSH and PMSG injection on day 5. Sudan black and Sudan red dyes were injected intravenously on alternative days to monitor yolk deposition into follicles of the hierarchy removed after the fifth day of FSH treatment. 4. Treatment with 200 micrograms pFSH or 25 IU PMSG for 5 d increased serum progesterone (P4) concentrations as compared to controls. Injection of hens with FSH caused a linear dose dependent increase in serum oestradiol-17 beta (E2) concentrations, dose dependent increase in SYF and LWF, and dose related decrease in number of atretic SYF and LWF. The hierarchy of the ovary was disrupted with PMSG, but not FSH. Larger doses of FSH increased the number of small follicles (10 mm diameter) and yolk deposition. 5. We conclude that small follicles which have not entered the rapid growth phase are responsive to FSH. The increased yolk deposition following FSH treatment may have been a direct effect of FSH or may have been caused by the elevation of serum E2 concentrations in response to FSH treatment. It is possible that old chickens may produce inadequate amounts of FSH which result in decreased rate of follicular growth and ultimately decreased egg production.     

An immunohistochemical study of bovine antral follicles,with special attention to non‐atretic follicles with and without atypical granulosa cells

Summary

Inhibin and oxytocin were immunohistochemically demonstrated in all non‐atretic and light‐atretic follicles >2 mm from untreated and pregnant mare's serum gonadotrophin (PMSG)‐treated heifers and cows. Immunostaining for luteinizing hormone (LH) and oestradiol was observed in all non‐atretic follicles >4 mm, but only in follicles from PMSG‐treated cows. Inhibin and oestradiol immunoreactivity was restricted to the granulosa. Oxytocin and LH immunoreactiviity was visualized in both the theca interna and the granulosa. Within the granulosa, LH immunoreactivity was mainly present in cells that were located near the basement membrane. Normal granulosa cells differed from atypical granulosa cells (AGCs) with respect to their ability to bind LH and oestradid It is concluded that immunostaining for α‐inhibin, oxytocin, oestradiol and LH cannot be used as a marker of follicle quality to discriminate between non‐atretic follicles with AGCs and non‐atretic follicles without AGCs in mid‐luteal bovine ovaries.     

The use of bovine anti-PMSG serum in beef cattle after PMSG-superovulation.

Thirteen beef cows were superovulated using 4,000 i.u. of pregnant mare serum gonadotrophin (PMSG) on days 9 to 14 of the estrous cycle, followed by two injections of 500 micrograms prostaglandin F2 alpha analogue (PGF2 alpha) 48 and 55 hrs later. Seven of them were injected intramuscularly with bovine anti-PMSG serum 12 hrs after the first signs of estrus. The remaining 6 cows were served as controls and received no antiserum. Peripheral blood concentrations of progesterone (P) and estradiol-17 beta (E2) were compared in relation to the superovulatory responses. The injection of anti-PMSG serum did not significantly affect the numbers of the corpora lutea (CL), the anovulatory follicles and the transferable embryos at 7 to 8 days after superovulatory estrus, but increased the ratio of embryos classified as excellent or good quality. Although the plasma P concentration showed no significant differences between the anti-PMSG-treated and control cows, the plasma E2 concentration displayed a characteristic difference, suppressing the second E2 peak in the anti-PMSG-treated cows. It is concluded that the use of bovine anti-PMSG serum for PMSG/PGF2 alpha-treated cows at 12 hrs after the beginning of the estrus improves the quality of embryos recovered, probably due to inhibition of high estrogenic environment following ovulation.     

Estrogen receptors alpha and beta and progesterone receptors in normal bovine ovarian follicles and cystic ovarian disease

The purpose of this study was to determine by immunohistochemistry the expression of estrogen and progesterone receptors in ovarian follicular structures from cows with cystic ovarian disease (COD) and to compare these with normal ovarian structures. Secondary, tertiary, atretic, and cystic follicles were evaluated. The follicular cysts of animals with COD presented a significantly higher expression of estrogen receptor alpha in all follicular layers than secondary, tertiary, and atretic follicles in both groups (P < .05). The intensity of estrogen receptor beta in the granulosa cell layer was stronger in tertiary than in secondary and atretic follicles in normal animals (P < .05) and in growing and cystic follicles in animals with COD (P < .05). Theca cells were scarcely stained in the 2 groups. Growing follicles and cysts from COD animals were less stained than tertiary follicles from normal animals (P < .05). Differences did not exist between the 2 groups with regard to the progesterone receptor. Ovaries of animals with COD exhibited altered estrogen receptors expression compared with that in normal animals.     

Superovulation in the cow with pregnant mare serum gonadotrophin: effects of dose and antipregnant mare serum gonadotrophin serum.

The effects of pregnant mare serum gonadotrophin (PMSG) dose and PMSG antiserum on superovulation in crossbred beef cows were studied. In experiment I, three groups were treated with 1200, 2400 or 3600 IU of PMSG and 48 h later with prostaglandin (PGF). The mean numbers of corpora lutea (CL), unovulated follicles, and total ova/embryos collected increased as the PMSG dose increased. The percent of fertilized ova and transferable embryos was lowest in the highest dose group (p < 0.05). In experiment II, all cows received 2500 IU of PMSG; groups 1 and 2 were treated with sheep anti-PMSG serum at 48 h or 60 h after PGF; group 3 cows were PMSG-only controls. The number of CL was lowest and the number of unovulated follicles highest in the PMSG-only group (p < 0.05). The number of CL was higher in group 2 (anti-PMSG at 60 h) than in the control group, with the anti-PMSG at 48 h not different from the other groups. Numbers of total ova/embryos, fertilized ova, and transferable embryos were higher (p < 0.05) in both antiserum-treated groups relative to the PMSG-only group. We conclude that superovulation of beef cows with PMSG and treatment with PMSG antiserum will induce a higher superovulatory response and will result in higher CL numbers and fewer unovulated follicles. Further, the variability in the superovulatory response to PMSG treatment was still evident when PMSG antiserum was administered.     

Changes in neurosecretory activity in the hypothalamus of sheep after administration of PMSG

By means of histological methods of histochemical nature we examined the qualitative alterations in neurosecretory cells of anterior (nucl. paraventricularis - NPV. nucl. preopticus medialis - NPM), medial (nucl. arcuatus s. infundibularis - NARC), and posterior (nucl. tuberomamillaris - NTM) hypothalamus of sheep with a simultaneous caryometric analysis of neurons of the same nuclei after administration of various PMSG doses. The examination was performed on the brain samples of 23 two- to three-year-old sheep, of the Slovak Merino breed, at the live weight of 30 to 40 kg. in the period of physiological anoestrus (June-July). The animals were divided into three experimental groups and were instilled agellin sponges (20 mg). On the 13th day the sponges were removed and the first group was applied 1000 i. u. PMSG, the second group 750 i. u. PMSG. The sheep were killed within 30 to 36 hours after the oestrus determination with a teaser ram. After bleeding of animals the brains were perfused with 2% buffered paraformaldehyde and after taking out the brains from the crania their fixation was finished in buffered picroformol. Paraffin slices were stained with haematoxylin-eosine and crotonaldehyde-fuchsin to neurosecretions. Caryometric analysis was carried out with a 1000 fold magnifying and with a measuring of 200 cells from one sample. We recorded the neurosecretion increase in all hypothalamic nuclei under study. The caryometric analysis has shown a moderate shift to the right, it means the volume increase of nuclei of neurosecretory cells, which demonstrates a stimulation of the function of hypothalamic structures.     

Influence of infusion of prostaglandin F2 alpha (PGF 2 alpha) and weaning on surface and histologic populations of ovarian follicles in early postpartum beef cows

Twenty-four beef cows were infused continuously for 11 d (d 2 to 13 after parturition) into the descending aorta with either prostaglandin F2 alpha-Tham salt (PGF2 alpha; 33.5 mg/d; n = 12) or with .9% NaCl (saline; n = 12) vehicle. Cows from each infusion treatment then were assigned to three slaughter groups (G). Cows in G1 and G2 were suckled until slaughter on d 15 and 35, respectively, whereas calves in G3 were weaned on d 31 and cows were slaughtered on d 35 after parturition. Nonatretic and atretic (greater than 4 pycnosis) antral follicles (greater than .15 mm) on ovaries ipsilateral to the previously gravid uterine horn were evaluated by histological techniques. Compared with saline infusion, PGF2 alpha increased mean diameters of the largest (F1) follicles on the ipsilateral (d 15 and 35; P less than .07) and contralateral (d 15; P less than .07) ovaries (surface evaluation) as well as mean diameters of the largest (F1, d 35; P less than .002), second- and third-largest (F2, F3 d 15 and 35; P less than .01) nonatretic follicles (histologic evaluation) in suckled cows. Compared with suckled cows slaughtered on d 35, weaning on d 31 increased the number of medium follicles (3 to 5 mm), diameters of the three largest nonatretic follicles, percentage of large nonatretic follicles (greater than 3.67 mm) and mitotic index of class 3 (.68 to 1.57 mm) follicles in saline-infused cows. In contrast, in PGF2 alpha-infused cows, weaning decreased (P less than .05 to P less than .001) all of these responses. Surface and histologic evaluations of follicular activity indicated that postpartum infusion of PGF2 alpha stimulated development of large follicles. In response to weaning, PGF2 alpha infusion favored development of those large follicles toward ovulation or atresia on nonovulated follicles.