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Цель исследований состояла в определении влияния фолликулярной жидкости и альбумина сыворотки скота на расход кислорода и подвижность живчиков. Семя быка подвергали капацитации in vitro, применяя жидкости, содержащие разные концентрации альбумина либо фолликулярной жидкости скота. Во время инкубации определяли расход кислорода, подвижмость живчиков и активность глютаматаспортаттрансаминазы. Фолликулярная жидкость в значительной степени усиливала расход кислорода живчиками после ее прибавления к суспензиям живчиков. Иной эффект можно было заметить в пробах, содержащих 10 мг/мл альбумина. Фолликулярная жидкость вызывала внезапный значительный рост активности, AspAT, освобождаемой в плазму семени. В пробах семени с альбумином отметили тенденцию к уменьшенному освобождению AspAT в инкубаты. Как альбумин, так и фолликулярная жидкость, стимулировали в течение 4 часов инкубации, а даже дольше подвижность живчиков.
Twenty five Holstein-Friesian heifers, clinically normal and with regular oestrous cycles, were used for induction of superovulation (PMSG-PGF2alphalpha-Neutralpha-PMSG). Animals were divided into 5 groups receiving: I - detomidine (40 µg/kg b.w.), II - doxazosin (0.2 mg/kg b.w.), III - yohimbine HCL 1% (1 ml/50 kg b.w.), IV - carazolol (0.01 mg/ kg b.w., i.v.), and V – physiological saline (1 ml/50 kg b.w.). The heifers with PGF2alphalpha-induced cycles were treated with the substances 88 hrs after being given a single i.m. injection of 2500 IU PMSG. All animals were examined by ultrasonography, and by the number and size of ovarian follicles > 3 mm in diameter. The follicles were divided into 3 groups according to the diameter. Blood plasma was stored at -20°C until LH, P4, E2 and PGFM analyses. In the control (V) group, two waves of follicle growth were observed. Yohimbine produced a significant blockage of ovulation. The mean number of corpora lutea in the group III was significantly lower than that in the control group (p< 0.02). No significant differences in the number of corpora lutea were observed between the groups I, II and III. The increase in E2 concentrations could be the response to the PMSG treatment with two waves of growth of large follicles before and after ovulation. Pulsatile LH release was altered by yohimbinum injection, however, the greater amplitude of pulses immediately following yohimbinum administration are suggestive of a positive influence of the alpha-2 adrenergic receptors antagonist. Yohimbinum administration did not affect plasma concentration of examined hormones. There was a difference between the plasma levels of LH after the doxazosin injection. Single injection of the stimulators and blockers of adrenergic receptors did not affect superovulatory response in terms of the numbers of CL, unruptured follicles and embryos recovered. The affectivity of artificial insemination was not significantly different between the control group and the detomidinum groups, while in the yohimbinum group it was significantly lower.
The aim of this study was to compare the efficiency of in vitro maturation in relationship to pathomorphology of uterus and oocytes quality. Morphological evaluation of the uterus helped to classify the queens into one of three groups: those with normal uteri, those with transformed uteri and pregnant queens. Cumulus-oocyte complexes were separated at recovery into three types according to pigmetation, uniformity and smoothness of ooplasm, compactness and number of layers of cumulus cells, as well as integrity of zona pellucida. Oocytes were maturated for 36 hours at 38°C in M199 medium containing cysteine and 17b oestradiol. After IVM, the meiotic status of oocytes was determined and the percentage of matured oocytes was calculated in relationship to COCs quality and uterus condition. In the group with normal uterus, oocytes of class A maturated (63.93%) better than oocytes class B and C, although the differences in maturation of oocytes class A and B were not statistically significant. The worst results of IVM were for oocytes class C. In the group of transformed uterus, 55.17% of oocytes class A maturated, while in only 7.84% of oocytes class C metaphase II had been observed. Significant differences were established in availability to IVM between oocytes class A and C as well as class B and C. In the group with pregnant uteri, the highest percentage of matured oocytes was in class A (46.27%), though there are no statistically significant differences in IVM between oocytes class A and B (28.8%), and the oocytes class C maturated in only 1.82%. Only oocytes of class A and B should be qualified for IVM procedure. Pregnant queens as well as queens with transformed uteri can also serve as oocyte donors for in vitro maturation.
The aim of this study was to determine the influence of alpha- and ß-stimulators (alpha-stimulator: detomidinum HCl) as well as blockers (alpha1-blocker: doxazosin, alpha2- blocker: yohimbinum HCL, ß-blocker: carazolol) on bovine granulosa cells culture from preovulatory follicles. The cell culture was passed in TCM 199 medium with 10% FCS and antibiotics. Tested substances were added to the culture medium in different concentrations. The experiment began when at least 80% of the wells were covered (in four well culture dish of NUNCK-DK). The culture medium was collected every 24 h for hormone analysis. Hormone levels of T, E2, and P4 were determined. The culture was used up to 120 hours. Our results showed a decrease in P-4 secretion after detomidinum addition for all tested concentrations. A slight testosterone level increase was seen in the first 24 hours and then its concentration remained at a constant low level. A slight increase in 17-ß estradiol secretion was also observed. After yohimbinum addition, a statistically significant decrease of progesterone was observed for all concentrations tested. No significant changes were observed at other hormones levels when compared with the control. Doxazosin, when added into the culture medium, did not cause any statistically significant changes in hormone secretions. The addition of carazolol caused a significant decrease in progesterone secretion after culturing for 48 hours. Changes observed in other hormones levels did not differ statistically from the control. These results seem to support the hypothesis that drugs stimulating and blocking adrenergic receptors may play some role in ovarian steroidogenesis in cows.
The experiments have been carried out with a modified Tyrode’s liquid (TALP), PBS, TALP and PBS with the addition of 1 uM of epinephrin and 10 uM of hypotaurine (TALP+, PBS+) and Ringer’s solution. There were assessed a progressive movement, morphological changes of the acrosoma and the ability of spermatozoons to penetrate the eggs of the hamster. The results showed that the solutions of TALP+ and PBS+ maintained a progressive motion of spermatozoons and increased the percentage of morpho­logical changes in the acrosoma. Following incubation in the TALP+ there was found the highest percentage of penetrated oocytes. The findings point to epinephrin and hypotaurine' as the drugs maintaining the mobility of bull spermatozoons. No correlation was found between the per­centage of changes in the acrosoma and the percentage of penetrated eggs. Such correlation was observed in case of a high percentage of spermatozoons with a progressive movement.
The purpose of the study was to compare flow cytometric and haematologic variables in dogs with spontaneous endometritis pyometra complex (EPC) treated with aglepristone to healthy controls. Peripheral blood mononuclear cells: CD3, CD4, CD8, and B lymphocytes (CD21) were analysed by flow cytometry and white blood cell count. Significant differences were observed (P≤0.01) between control (C) and study (S) group in the total number of leukocytes, monocytes and granulocyte populations (without lymphocytes) on beginning, after 7 d, and return to reference value after 14 d of the treatment. The percentage of the T-cell (CD3+) at the beginning was 47.22 ±9.64% of total lymphocytes, in contrast to B lymphocytes (CD21+) that represented the smallest percentage of 14.24 ±7.74% (P≤0.01). The percentage of the lymphocytes CD4+ was 27.42 ±5.53% and CD8+ was 25.18 ±4.36%. The percentage of CD3+ lymphocytes was increasing throughout the experiment in group C and gradually decreased in group S from 14th to 28th d of dioestrus. No differences in the number of CD3+, CD4+, CD8+, and CD21+ lymphocytes between group C and S on the 14th and 28th d of dioestrus (P≥0.05) were observed. The number of CD8+ cells in group S decreased gradually from day 14 to 28 but no statistical differences were noted. Treatment of pyometra with aglepristone decreased the number of leukocytes, monocytes, and granulocytes to referential value but statistically significant influence on the level of subpopulations of T and B lymphocytes was not observed. The results enabled to estimate for the first time the number of lymphocyte subpopulations in dioestrus in both healthy bitches as well as in those suffering from pyometra.
This study was aimed at investigating the use of intra-testicular calcium chloride (CaCl₂) and 4-vinylcyclohexene 1,2-monoepoxide (VCM) injections as a side effect-free alternative method for the control of reproduction in guinea pigs. Fifty male guinea pigs were randomly assigned to five groups. In all groups, the chemical agents were injected into both testes in 1% lidocaine hydrochloride. While Groups I, II and III were administered with a single dose (0.25 mL) of sterile physiological saline, 15 mg/100 g CaCl₂, and 240 mg/kg VCM, respectively, Group IV and V received a daily dose of 15 mg/100 g CaCl₂, and 240 mg/kg VCM for 3 days, respectively. On day 90 post-administration, all animals were weighed and later decapitated under ether anaesthesia. Blood and tissue (testis, liver, hypophysis and adrenal gland) samples were taken. Sperm samples from the cauda epididymis were examined for spermatological parameters. Blood was used for hormone analyses and tissue samples were examined histopathologically (haematoxylin-eosin) and immunohistochemically (Tunel staining). The epididymal sperm count decreased in all treatment groups. Excluding 2 animals, Group V displayed azoospermia. When compared to the control group, Group V displayed the highest prolactin and lowest testosterone levels, and Group III showed the highest testosterone level. Histopathological examination revealed no intoxication finding. Chemical castration with VCM may be a good alternative to surgical castration as it enables mass sterilization without postoperative risks in guinea pig.
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