[The effect of hormonal preparations on superovulation and egg transport in ewes]. / Ucinok niektorých hormonálnych prípravkov na superovuláciu a pasáz vajícok u bahníc.

The relation between the prevalence of contagious mastitis with the finding of Streptococcus agalactiae and Staphylococcus aureus and the number of somatic cells in the tank samples of milk was studied on the basis of a clinical and bacteriological examination of 11 129 dairy cows on four large farms, with a capacity of 600 to 1240 animals. A statistically significant correlation was demonstrated between the analyzed characteristics (r = 0.621). The relation was demonstrated at a prevalence up to 25.3%. The number of somatic cells provides a marked indication of prevalence only from 10%. We failed to demonstrate a statistically significant correlation between the prevalence of mastitis with a cultivation finding significant correlation between the prevalence of mastitis with a cultivation finding of Staphylococcus aureus and the number of somatic cells in tank samples of milk (r = 0.054). The results suggest that on large farms it will not be possible to use only the number of somatic cells in tank milk samples as a signal of diagnostic improvement and preventive mastitis-control measures, since a 10% prevalence of these diseases usually excludes the possibility of herd sanitation.

[Changes in the concentration of estrone, estradiol and estradiol in the blood of pregnant ewes]. / Zmeny koncentrácie estrónu, estradiolu a estriolu v krvi kotných oviec

A chemical method was used for the study of the concentration of the three main groups of oestrogens in the blood of five ewes after mating in natural heat. The concentration values of all the three groups of oestrogens was found to increase in proportion with the length of gravidity. From the 30th to the 140th day of gravidity the average concentration values of oestrone increased from 309 to 1380 ng per 100 ml blood, those of oestradiol from 48 to 192 ng per 100 ml blood. Significant changes were recorded in oestrone between the 50th and 60th day and between the 130th and 140th day of gravidity; in oestradiol and oestriol such changes occurred on the 140th day of gravidity.

[Concentration of estrone, estradiol and estriol in the blood of pregnant sheep after induction of estrus with chlormadinone acetate and medroxyprogesterone acetate]. / Chlormadinonacetátom a medroxyprogesterón-acetátom

The radioisotopic method of 131J-labelled albumin was employed to determine the distribution of acidic proteinase activity in some organs and tissues of chickens. The highest enzymatic activities were found in intestine wall, in pancreas, and in liver. Considerably lower activities were ascertained in kidneys, brain, lungs, and heart. The different proportions of these enzymes in homogenates and supernatant fractions (106 000 g) testify to a lack of uniformity in the solubility of cathepsins in the organs tested. The tested organs, with the exception of pancreas, did not show any enzymatic activity of neutral proteinases.

PIP: Estrus was artificially induced in 10 Merino sheep by a vaginal swab administration of 30 mg of chlormadionone acetate (in 5 animals for 13 days) or of 60 mg of medroxyprogesterone acetate (in 5 animals for 17 days), both on a background administration of 750 IU pregnant mare's serum gonadotropin. After impregnation in this condition, the levels of esterone, estradiol, and estriol were determined. A progressive increase was found in the levels of all 3 of these hormones over the course of the pregnancy, in both groups. Comparison of estrogen concentrations showed that the use of the 2 progesterone derivatives did not affect the endocrine ciruclation of the mother and the fetus during pregnancy.

[Concentration of free 17-beta estradiol in the plasm of ewes following treatment with chlormadione acetate (CAP) in 2 synchronized cycles]. / Koncentrácia vol'ného 17-beta estradiolu v krvnej plazme oviec po osetreni chlormadinonacetátom (CAP) v dvoch synchronizovaných cykloch.

Six ewes of the Merino breed were subjected to radioimmunological assay (RIA) for the study of free 17-beta estradiol during the instillation of vaginal tampons soaked with 30 mg chlormadinone acetate (CAP), and 17 and 34 days after removal of the tampons. On the day before the administration of the preparation, the concentrations of free 17-beta estradiol were comparatively low, ranging within the limits of 38.6 +/- 5.6 pg ml-1 of plasma. These values showed no significant changes during the instillation of the tampons. When the tampons were removed, three periods of increased 17-beta estradiol levels were recorded: 1. on the 1st to 3rd day after removal of the tampons with a peak of 93.2 +/- 26.7 pg ml-1, 2. on the 13th to 16th day with the highest value of 59.7 +/- 23.6 pg ml-1, 3. on the 30th to 33rd day after removal of the tampons 76.2 +/- 23.5 pg ml-1. The results showed that the endocrine activity of the ovaries, expressed by a change in the concentration of plasma 17-beta estradiol after removal of the tampons (CAP), repeated in similar time intervals as under natural conditions and was probably associated with repeated growth and ripening of the follicles of ovaries and their ovulation.

[The effect of prolonged exposure to gamma rays and hormonal stimulation with serum gonadotropin on the levels of catecholamines in the hypothalamus, epiphysis and adrenal glands in sheep]. / Vplyv protrahovanej expozície gama-ziarenia a hormonálnej stimulácie sérovým gonadotropínom na hladiny katecholamínov v hypotalame, epifýze a v nadoblickách oviec.

The effect of the exposure of the whole body to continuous radiation and of the administration of serum gonadotropin (SG) was studied as exerted on the concentration of catecholamines (epinephrine and norepinephrine) in the hypothalamus, epiphysis and adrenal glands of ewes during the anoestric period with synchronized oestrus. The first group (young barren ewes) and second group (older ewes) were exposed to continuous radiation of 60Co for five days. The radiation was provided at the rate of 0.020 Gy per hour. After the termination of irradiation the ewes were subjected to hormonal stimulation by fractionated administration of 1500 I. U. SG. The third and fourth experimental groups of ewes were stimulated with 1500 I. U. SG without irradiation. Catecholamines were separated from the tissue supernatants by the adsorption chromatographic method and the catecholamine contents in the eluates were determined spectrofluorometrically. Protracted exposure to gamma radiation and hormonal stimulation with SG reduces the concentration of norepinephrine in the whole hypothalamus of the sheep. A statistically significant decrease (P less than 0.001) was recorded in the medial and caudal hypothalamus of the adult ewes and in the rostral and caudal hypothalamus regions of the young ewes. A decrease of norepinephrine concentration, statistically significant in the caudal (P less than 0.01) and medial hypothalamus, was recorded in the group of adult ewes after hormonal stimulation with SG without irradiation. The experimental group of young ewes responds to hormonal stimulation by a greater reduction of norepinephrine contents, as compared with combined exposure to radiation and hormonal stimulation. It is assumed that the decrease in catecholamine concentration after hormonal stimulation with SG is associated with the increase in the content of oestrogens, which act on the adrenergic receptors of the hypothalamus.

[Changes in the trypsin-inhibiting activity of blood and cervical mucus in sheep after the administration of PGF2 alpha and PMSG in anestrus]. / Zmeny trypsín inhibicných aktivít krvnej plazmy a cervikálneho hlienu oviec po podaní PGF2alfa a PMSG v anestri.

The changes in the concentrations of proteolytic enzyme inhibitors after the administration of hormones influencing the activity of the ovaries can cause an imbalance in the development of follicles. Important roles are attributed to serine proteases during the development of follicle, ovulation, and activity of the corpus luteum. In order to find out the changes in the trypsin inhibiting activities (TIA) of the blood plasma and cervical mucus after the administration of PGF2 alpha (100 + 100 micrograms in January and 250 + 250 micrograms in May on the 0th and 11th day) and after the single administration of 750 I. U. PMSG on the 10th day, a 14-day trial was performed to investigate the circadian changes. TIA was determined by means of slowing down the hydrolysis of the synthetic substrate N-alpha-tosyl-p-nitro-anilide (TAPA) by bovine trypsin. When the blood plasma TIA is expressed as the percent of the control values, a large drop is observed on the 11th and 12th days of the January trial (total as well as low-molecular TIA). In the May experiment this drop was recorded only on the 11th day in total TIA (c TIA); low-molecular TIA (n TIA), on the other hand, increased, particularly after the administration of PMSG. The average values of plasma TIA's, recorded in the course of the 14-day trial, increased after the administration of PGF2 alpha (c TIA increased to 109.8 +/- 22.1%, n TIA to 135.42 +/- 54.0% in January; c TIA to 112.59 +/- 50.5%, n TIA to 109.48 +/- 55.9% in May). After the administration of PMSG the TIA values remained higher than the controls (c TIA 107.13 +/- 24.0%, n TIA 133.40 +/- 52.1% in January; c TIA 113.87 +/- 51.3%, n TIA 118.33 +/- 55.6% in May). The average TIA values of the cervical mucus increased after PGF2 alpha administration to 3.06 +/- 1.7% in comparison with the control value of 2.59 +/- 1.1%, and also after the administration of PMSG to 2.82 +/- 1.9% (January, TIA per 1 mg of mucus).

[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]. / Vplyv vitamínu A na trypsín inhibicné aktivity (TIA) krvnej plazmy a cervikálneho hlienu po hormonálnom ovplyvnení ovárií oviec v estrickom období.

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 neurosecretory activity in the hypothalamus of sheep after administration of PMSG]. / Zmeny neurosekrecnej aktivity u hypotalame oviec po podaní 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.

[Morphologic picture of the ependyma of the 3d cerebral ventricle in ewes after hormonal stimulation]. / Morfologický obraz ependýmu tretej mozgovej komory u bahníc po hormonálnej stimulácii.

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.

[Morphologic and biochemical changes in the hypothalamus in hormonally stimulated sheep]. / Morfologické a biochemické zmeny v hypotalame u hormonálne stimulovaných oviec.

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.

[The effect of prolonged whole-body gamma irradiation of 6,7 Gy and 4,8 Gy (700 and 500 R) on the trypsin inhibiting activity (TIA) of blood, the cervical mucus and the morphological structure of the cervix in ewes]. / Vplyv protrahovaného celotelového gama ziarenia 6,7 Gy a 4,8 Gy (700 a 500 R) na trypsín inhibicnú aktivitu (TIA) krvi, cervikálneho hlienu a morfologickú struktúru cervixu bahníc.

The pattern of changes in the trypsin-inhibition activities (TIA) of blood plasma, cervical mucus and in the morphological structure of cervix was studied in ewes exposed to 60Co radiation for seven and five days, the radiation doses being 6.7 Gy and 4.8 Gy (700 and 500 R). During exposure, the group of ewes irradiated with 4.8 Gy was given the Roboran vitamin premix in addition to standard feed. After termination of irradiation the animals in this trial were given the ampicillin antibiotic (5250 mg). TIA was determined from the retardation of the hydrolysis of the synthetic substrate N-alpha-tosyl-p-nitroanilide (TAPA) by bovine trypsin; the TIA was expressed as the percentage of inhibited trypsin. As found on the day of the termination of irradiation, almost all the studied TIA values of blood plasma and cervical mucus were increased in the irradiated animals, the range being from 103.1 to 155.0% of the levels for non-irradiated ewes. A reduction was recorded only in total TIA of blood plasma in the group irradiated with the dose of 6.7 Gy (83.1% of the values for non-irradiated animals). In the group of animals irradiated with 4.8 Gy and fed no Roboran, the TIA of cervical mucus was observed to decrease to 92.4%. When evaluating the average TIA of different test groups throughout the experiment it was found that all the studied TIA values of the irradiated animals increased within a range from 104.1 to 184.0% of the values for untreated ewes. It was only in the average values of total TIA in blood plasma that, after irradiation with 6.7 Gy, a decrease to 92.7% of the level without irradiation was recorded. It was found during the study of changes in the proportions of glands in the stroma and changes in epithelium thickness in the mucous membrane of cervix uteri that the irradiated ewes had the epithelium thickness reduced to 95.3%-65.5% and that their stromal gland number decreased to 75.4%-79.7% of that recorded in non-irradiated animals. It was only in the group given a Roboran supplement that an increase to 123.7% of the gland number for untreated ewes was recorded on the tenth day after the termination of irradiation.

[Distribution of catecholamines and monoamine oxidase activity in the system controlling sexual function in sheep]. / Distribúcia katecholových amínov a aktivita monoamínoxidázy v riadiacom systéme pohlavných funkcií oviec.

A study was performed on the distribution of catecholamines (noradrenaline, dopamine and adrenaline) and L-DOPA in the hypothalamo-hypophysial region, as well as in the region of n. caudatus, which participate in the control of sexual activities in sheep. After isolation in the activated aluminium oxide, the catecholamine concentration was determined spectrophotometrically. Hypothalamus was divided into three regions: rostral, medial and caudal. In the same regions of the cerebroneural system (CNS), the activity of the degradation enzyme monoaminooxidase (MAO) was determined by radiochemical method. 14C-tryptamine was as a substrate and the results were measured by means of scintillation spectrophotometer Packard. The distribution of catecholamines and L-DOPA in the hypothalamus of sheep is different. The largest proportion is that of noradrenaline in caudal hypothalamus (1.84 +/- +/- 0.36). The dopamine levels in hypothalamus are quite balanced though substantially lower than those of noradrenaline (from 0.22 to 0.31 micrograms/g). The concentrations of adrenaline and L-DOPA in the sheep hypothalamus are low. The sheep hypophysis contains more noradrenaline (1.70 +/- 0.38), adrenaline content amounts to 1.30 +/- 0.28. L-DOPA and dopamine occur in this region at low concentrations. In comparison with the other regions of sheep brain, high dopamine concentrations (2.0 +/- 0.58 micrograms/g) and higher L-DOPA levels were determined in n. caudatus. The activity of the degradation enzyme monoaminooxidase in the cerebroneural system of sheep is different. The highest MAO activity was determined in the rostral hypothalamus (1100 pmol X mg-1 X min-1), lower in its caudal and medial region.(ABSTRACT TRUNCATED AT 250 WORDS)

[Levels of catecholamines in the hypothalamus, hypophysis and in the adrenal in sheep after whole-body exposure to gamma rays]. / Hladiny katecholových amínov v hypotalame, hypofýze a v nadoblickách oviec po celotelovej expozícii gama-ziarenia.

The changes were studied in the levels of catecholamines and L-DOPA in the control system of reproduction cycle (hypothalamus, hypophysis) and in adrenal glands of sheep after all-body irradiation with 60Co at the total dose of 6.7 Gy for seven days. The power input per hour of irradiation source was 0.039 Gy. The catecholamines (noradrenaline, dopamine and adrenaline) and L-DOPA were determined, after having been isolated from the tissues, by the method of spectral fluorimetry. After all-body exposition to gamma-radiation noradrenaline dropped in hypothalamus in comparison with the control group of sheep, most significantly in the rostral (by 74.2%) and caudal part (by 40%). A similar drop like in the case of noradrenaline was also observed in dopamine, the concentrations of which decreased in rostral hypothalamus by 72%, in medial hypothalamus by 94% and in caudal hypothalamus by 60%. Adrenaline shows a drop in hypothalamus, most significant in caudal region (by 62%). In relation with that the level of a precursor for the synthesis of catecholamines and L-DOPA has been changed which achieved, in the studied regions of hypothalamus in sheep, significantly lower levels than were determined in the control group of sheep. As regards the hypophysis, after irradiation no significant changes in the levels of noradrenaline and adrenaline were recorded, however, dopamine and L-DOPA dropped significantly (P less than 0.01). The exposition to gamma-radiation also causes a decrease in the concentrations of catecholamines and L-DOPA in the adrenal glands of sheep, most significant in noradrenaline (by 61%).(ABSTRACT TRUNCATED AT 250 WORDS)

[Changes in antiproteolytic activity in the blood and cervical mucus after superovulation stimulation]. / Zmeny antiproteolytickej aktivity krvnej plazmy a cervikálneho hlienu po superovulacných stimuláciách.

The function of proteins, peptides, proteases and inhibitors of proteases in modulations of regulation mechanisms of gonadotrophins during the development of ovarian folicles has not been fully explained up to now. We can see difference reactions of ewes to superovulation stimulations in oestrous and anoestrous periods as shown by the variation of the antiproteolytic activity of blood plasma and cervical mucus. Trypsin is used as a model for serine protease, and trypsin inhibitory activity (TIA) was measured from the reduced rate of trypsin hydrolysis of the chromogenic substrate N-alpha-tosyl-L-arginine-4-nitroanilide (TAPA, Bartík et al., 1974). Full hydrolytic activity was determined as a change in absorbency at 405 nm = 1.0 after ten-minute incubation at 25 degrees C and pH = 8.1, and inhibition was expressed in percentage of full activity. Statistical analyses were performed by Student's t-test. Twenty-three ewes in anoestrus and twenty-eight ewes in oestrus were included in this experiment. They were of the Slovak Merino breed, two to three years old, with the mean live weight of thirty to forty kg. The ewes were treated with Ageline vaginal sponges (20 mg chlorsuperlutin/sponge) to provide for synchronization of ovarian activity, in the interval of eleven to twelve days. After removal of sponges, the ewes were stimulated for superovulation as shown in Tab. I. Heparinized blood plasma samples were stored at -25 degrees C. Low molecular TIA activities were determined in HClO4--treated blood plasma. Samples of cervical mucus were taken on cotton-wool tampons which were evaluated in 0.2 M Tris-HCl buffer, pH = 8.1. Figs. 1-6 shows TIA changes in blood plasma and cervical mucus on particular days. Fig. 7 shows the mean values of controls (I-initial) after synchronization (A) and after stimulation (S). TIA changes were different in anoestrous and oestrous periods. Differences in TIA changes in blood plasma and cervical mucus in oestrous period suggested certain local regulation mechanisms of synthesis and/or secretion of this activity in the cervix. Some values of TIA were maximum at the time of expected heat or ovulation and may be some of the factor(s) which influenced fertility of females after superovulation stimulations.

[Antiproteolytic activity of blood and cervical mucus in sheep after hormonal ovarian stimulation and gamma ray irradiation (2.45 Gy) during anestrus]. / Antiproteolytické aktivity krvnej plazmy a cervikálneho hlienu oviec po hormonálnom ovplyvnení ovárií a ozarovaní gama lúcmi (2.45 Gy) v anestri.

The antiproteolytic activities of the blood plasma (BP) and cervical mucus (CM) determined as trypsin inhibiting activities (TIA) where trypsin served as a model serine protease, were variable after superovulatory stimulations of ewes and after their gamma irradiation (2.45 Gy). TIA's were determined from the reduction in the bovine trypsin hydrolysis of the low-molecular chromogenic substrate N-alpha-tosyl-L-arginyl-p-nitro anilide (TAPA) (Bartík et al., 1974). The inhibition was expressed in per cent when delta A405 = 1.0 for 10-minute incubation at 25 degrees C, pH = 8.05, 0.2 mol/l tris-HCl buffer, was taken as 100%. Student's t-test was used for the statistical evaluation. A hundred ewes of the Merino breed were applied Agelin vaginal tampons (20 mg chlor-superlutin per head) on day 1 for 10 days in the anoestric period (May). A part of these ewes were subjected to whole-body gamma irradiation on days 6 to 11. When the irradiation was completed and the tampons were taken out, the ewes (three to four years old lambing ewes, yearling ewes) were stimulated to superovulations by an administration of 1500 IU serum gonadotropin (SG) or 450 IU follicle stimulating hormone (FSH). These parameters were followed in the subsequent 5-6 days: BP TIA (Figs. 1 and 4), fraction of low-molecular BP (n) TIA (Figs. 2 and 5), CM TIA (Figs. 3 and 6). Fig. 7 shows the average values of the results. The lambing ewes and yearling ewes had various responses to irradiation and superovulatory stimulation. Gamma irradiation eliminated the increase in BP TIA (P < 0.001) in the ewes after stimulations (Fig. 1a, b). The yearling ewes showed nonsignificant changes (ns) as their responses to SG stimulations and gamma irradiation, but the responses on the particular days of the trial were different (Fig. 1c, d--FSH stimulation). Similar changes were observed in the BP nTIA fraction. The changes in the average values of CM TIA were nonsignificant in the lambing ewes, the gamma irradiation reduced CM TIA only after FSH stimulation (P < 0.002). The amount of cervical mucus decreased after stimulations and irradiation, the thickness of ovarian epithelium and muscles was reduced in the particular parts, differently in lambing ewes and in yearling ewes.(ABSTRACT TRUNCATED AT 400 WORDS)

[Changes in the hypothalamus and adjacent ependyma after hormonal stimulation of the ovaries in ewes]. / Zmeny v hypotalame a prilahlom ependýme bahníc po hormonálnej stimulácii ovárií.

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.

[Changes in antiproteolytic activity in blood and ovarian follicular fluid in 3 breeds of sheep after stimulation of superovulation]. / Zmeny antiproteolytických aktivít krvnej plazmy a folikulárnej tekutiny ovárií oviec troch plemien po superovulacných stimuláciách.

Serine proteases help to regulate the ovarian cycle at different levels and they are subjected to the control of gonadotropic hormones and protease inhibitors. Superovulation stimulations influence the activities of trypsin inhibitors (model serine protease) in blood plasma (BP) and in follicular fluid (FF), and also in dependence on the breed. Trypsin inhibiting activities were determined from the reduced rate of trypsin hydrolysis of a chromogenic substrate (TAPA) and they were determined in percent. A change in absorbancy at 405 nm = 1.0 after 10-minute incubation at 25 degrees C and pH = 8.05 was taken as 100%. The incubation mixture as a sample contained 100 microliters blood plasma or 10 microliters follicular fluid, diluted with gammaglobulin at 1:10. The differences in the trypsin inhibiting activities (TIA) of BP in ewes of the Merino, Tsigai and Wallachian breeds were insignificant, but Agelin synchronization (20 mg chlorsuperlutin per vaginal swab) induced statistically significant differences. The lowest TIA BP was recorded in the Tsigai breed (T), P less than 0.001 in comparison with the Wallachian (W) and Merino (M) breeds. Following the administration of 1,500 IU PMSG, the TIA BP within 120 hours decreased in W (P less than 0.001), it increased in T (P less than 0.1) and in M the changes in the TIA BP were insignificant. The average numbers of ovulations increased from 2.25 +/- 2.5 to 3.0 +/- 1.2 in W; from 0.25 +/- 0.43 to 2.5 +/- 1.6 in T and from 0.00 +/- 0.0 to 2.5 +/- 2.3 in M. Following the single administration of 2,000 IU PMSG after Agelin synchronization, the changes in M ewes were also insignificant, and there were no different responses in pregnant (1st to 2nd month) and nonpregnant ewes. In pregnant T ewes the TIA BP increased after Agelin synchronization and stimulation (P less than 0.01), in nonpregnant ewes these changes were not significant. In W lambing ewes the TIA BP increased (P less than 0.001), the effects of Agelin were greater than those of PMSG. The TIA of follicular fluid (FF) of antral follicles were on average tenfold if compared with BP. After hormonal treatment of ovaries, the TIA FF mostly increased at different levels of statistical significance. The TIA FF of follicles less than 10 mm were lower than in follicles greater than 10 mm (P less than 0.001 for M and W, P less than 0.1 for T).(ABSTRACT TRUNCATED AT 400 WORDS)

[The anti-proteolytic activity of blood and ovarian follicular fluid in sheep after stimulation with serum gonadotropin (PMSG) and administration of Antisergon]. / Antiproteolytické aktivity krvnej plazmy a folikulárnej tekutiny ovárií oviec po stimulácii sérovým gonadotropínom (PMSG) a podaní Antisergónu.

The synthesis and secretion of trypsin (trypsin model serine protease) inhibitors are regulated in ovarian follicles by gonadotropins. The superovulation stimulations with 400 IU FSH, 1000 IU PMSG, 1000 IU HCG, 750 IIU PMSG + 750 IU HCG influence in a different way the trypsin inhibiting activities (TIA) of blood plasma (BP) (Figs 1 and 2) and follicular fluid (fig. 3); this points to a possibility of local effects. An increase in the average values of TIA in BP was statistically significant during the whole experiment: P less than 0.05 to P less than 0.001 (following the administration of PMSG+HCG, or PMSG, and HCG); Antisergon administered in 68 hours after PMSG reduced this increase. The changes in the fraction of low-molecular TIA in BP (after BP treatment with perchloric acid) were of converse nature; a decrease in the average values ranged from P less than 0.02 to P less than 0.001 (following PMSG or other stimulations). Antisergon did not influence this decrease. The changes observed on particular days of the trial (Figs. 1 and 2) also indicate different effects of the preparations, mainly of the component LH, which resulted in the occurrence of large nonovulating follicles (greater than 10 mm--"cystic" ones). No such follicles were observed in nonstimulated ewes and after FSH stimulation. The administration of antisergon (goat's antiserum against PMSG) 68 hours after PMSG administration did not prevent their creation. The TIA of follicular fluid (FF) of antral follicles was on average tenfold in comparison with that of blood plasma; and the TIA FF of follicles greater than 10 mm was higher (up to P less than 0.001) than the TIA FF of follicles less than 10 mm. The administration of Antisergon in shorter intervals following PMSG administration (12, 24, 48 and 58 hours) influenced the average values of TIA BP in 120 hours (since PMSG administration) in dependence on time (Tab. I). The effects of Antisergon administered in 12 and 24 hours after PMSG administration on the TIA BP were insignificant if it was administered in 48 and 58 hours the TIA BP increased (P less than 0.02; P less than 0.001) in comparison with the interval of 12 hours. The TIA FF of follicles less than 5 mm, 5-10 mm and greater than 10 mm varied in dependence on the time intervals of Antisergon administration (Fig. 4). The statistical significance of these changes in shown in Tab. II.(ABSTRACT TRUNCATED AT 400 WORDS)

[Quantitative micromorphological study of the ovaries in sheep after estrus synchronization and superovulation in the autumn mating season]. / Kvantitatívne mikromorfologické stúdium vajecníkov u oviec po synchronizácii ruje a superovulácii v jesennom pripústacom období.

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 biometry of the sex organs of sheep after estrus synchronization and superovulation in the breeding period]. / Zmeny biometrie pohlavných orgánov u oviec po synchronizácii ruje a superovulácii v pripústacom období.

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.