The aim of this study was a cytogenetic analysis of stallions semen to find sex chromosome aberrations and to determine if there was an association between stallion's age and aberration frequency for the sex chromosomes. Sperm samples were collected from 22 stallions of various age from 3 to 23 years. Multicolour FISH was performed on each sample, using probes for the sex chromosomes and EGFR gene, localized on 4p12 in domestic horse. A total of 26199 sperm cells were analysed (from 1 070 to 1 532 per animal). Among the analysed cells, there were 50.318% with X chromosome, 48.543% with Y chromosome and 1.139% with aberrant chromosomes. The frequency of aberrations was: sex chromosomes nullisomy (0.466%), XY aneuploidy (0.454%), XX disomy (0.146%), YY disomy (0.041%), diploidy (0.024%) and trisomy XXY (0.008%). Additionally there was a correlation between the age of an animal and the frequency of sex chromosome aberration and a significant positive correlation between age and disomy of XY, XX, YY, trisomy of XXY, autosomal disomy was seen. A Correlation between the age of a stallion and the level of nullisomy was negative. The present study demonstrated that FISH technique is a powerful method to identify sex chromosome aberrations in equine spermatozoa and might be very helpful for a breeder during a selection for the best stallion.

P>Reasons for performing study:Specific patterns of cytoskeletal filaments reflect a functional state of the cell. In testicular cells intermediate filaments (IFs) are of the vimentin type. Since it is known that Sertoli cells regulate the spermatogenic function in the male gonad, it became important to propose a system that could quantify the state of seminiferous tubular quality. To date, a Johnsen score system has never been used to equine testes.Objectives:To demonstrate the expression pattern of vimentin in testes of mature Arabian stallions and correlate its distribution with grade of seminiferous tubule impairment as indicated by a Johnsen score.Methods:For histological examination by the Johnsen method, routine haematoxylin-eosin staining was used. Vimentin expression and its presence in testicular sections and testicular homogenates were detected by immunohistochemistry and western blot, respectively. Both analyses were performed qualitatively and quantitatively and further validated by ANOVA tests.Results:Distinct morphology of seminiferous tubules was found in testes harvested from 3 stallions. Vimentin in IFs was immunolocalised to the cytoplasm of Sertoli, Leydig and peritubular-myoid cells. The intensity and pattern of the IFs staining was different in individual seminiferous tubules suggesting a correlation between vimentin expression and the severity of tubule degeneration. Qualitative results by immunohistochemistry and western blot were confirmed by further quantitative analyses.Conclusions:In equine testes, differential expression of vimentin was found to be correlated with the impairment of seminiferous tubules indicated by a decrease in Johnsen score.Potential relevance:The Johnsen score system may be a useful method to facilitate the identification of tubular alterations in the stallion testes. Combined histological and immunohistochemical approach may provide a detailed phenotypic classification of stallions with decreased fertility.

The objective of this study was to evaluate parthenogenetic activation of domestic cat oocytes after being exposed to either ethanol, magnetic field, calcium ionophore A23187, or cycloheximide and a combination of these agents. We also wished to evaluate the usefulness of the magnetic field for oocyte activation. In vitro matured oocytes subjected to artificial activation were randomly assigned into eight groups according to activating agents: (1) 10% ethanol; (2) the magnetic field (slow-changing, homogenous magnetic field with low values of induction); (3) 10% ethanol plus magnetic field; (4) 10 microM calcium ionophore A23187; (5) 10 microM calcium ionophore A23187 plus magnetic field; (6) 10% ethanol and 10 microg/mL of cycloheximide; (7) 10% ethanol and 10 microg/mL of cycloheximide plus magnetic field; (8) oocytes were not exposed to any of the activating agents. After activation oocytes were stained with Hoechst 33258 and parthenogenetic activation was defined as oocytes containing pronuclei and second polar bodies or two to four or six nuclei (embryonic cleavage). The total activation rate by using different activation treatments was 40%. The addition of the magnetic field to ethanol or calcium ionophore treatments resulted in increased parthenogenetic activation rates from 47% to 75%, and from 19% to 48%, respectively (P<0.001). Instead, when the magnetic field was added to ethanol and cycloheximide treatment, activation rate decreased from 48% to 30%. Oocytes activated with magnetic field only gave the lowest activation rate (12%). We concluded that a magnetic field can be used as an activating agent, and the combination of ethanol and magnetic field is an effective method for domestic cat oocyte activation.

The aim of the study was to determine the intensity of reaction of the sympatho-adrenal and hypothalamo-pituitary-adrenal systems in mares and foals during short term transportation on day 9 following parturition. Five pairs of Polish pony mares in foal heat and their foals were used in the experiment. All the animals were transported in a trailer for 20 minutes. Blood samples were collected from the jugular vein as follows: 30 minutes before transportation, in the trailer, immediately after the 20-minute-journey and 30 minutes after its conclusion. Transportation on day 9 following parturition seemed to be a stronger stress factor for the mare than for her foal (p <= 0.01) and elicited an evident and long-lasting increase of catecholamine and cortisol concentrations in mare blood. There were no significant changes in adrenaline concentration in the foals either during transportation or afterwards and only the level of noradrenalin and cortisol increased after transportation concluded. Concentrations of adrenaline, noradrenalin and cortisol were three times higher in mares than in their foals (p <= 0.01).

LIF is twice transiently expressed in the mouse uterus, first at the time of ovulation and again just prior to implantation, and studies have demonstrated a beneficial influence of this cytokine on embryo development in several species. We have investigated the effect of LIF on gametes in vitro, on the hypothesis that the ovulatory peak of LIF can exert an influence on gametes present within the oviduct. We also investigated the effect of LIF on in vitro fertilization and embryo development, in oocytes from adult sheep and from prepubertal lambs that lack the preovulatory hormone surge and that are unable to sustain early embryonic development. A higher rate of pronuclear-stage embryos derived from both, adult and prepubertal female, was obtained when in vitro fertilization was performed in the presence of LIF, and there was an improved cleavage of parthenogenetic embryos when incubated with LIF immediately following activation. In contrast, LIF was found to have no influence on the viability of ram semen. In vitro fertilized two-cell stage embryos from adult sheep and prepubertal lambs, cultured in defined medium enriched with LIF, both reached the blastocyst stage at similar rates to control embryos. However, LIF exerted a positive influence on the quality of the blastocysts as revealed by significantly higher number of ICM cells and total number of cells. Together, these data demonstrate that LIF exerts a beneficial effect on sheep oocytes and embryos in vitro, but only at stages concomitant with steroid hormones surges.

Equine Veterinary JournalVolume 36, Issue 6 p. 539-543 Immunoexpression of androgen receptors in testes of immature and mature stallions B. BILIŃSKA, Corresponding Author B. BILIŃSKA Laboratory of Endocrinology and Tissue Culture, Institute of Zoology Jagiellonian University, Ingardena 6, 30-060 Kraków, PolandLaboratory of Endocrinology and Tissue Culture, Institute of Zoology Jagiellonian University, Ingardena 6, 30-060 Kraków, PolandSearch for more papers by this authorA. HEJMEJ, A. HEJMEJ Laboratory of Endocrinology and Tissue Culture, Institute of Zoology Jagiellonian University, Ingardena 6, 30-060 Kraków, PolandSearch for more papers by this authorM. PAWLAK, M. PAWLAK Department of Animal Reproduction, University of Agriculture, Kraków, PolandSearch for more papers by this authorJ. SADOWSKA, J. SADOWSKA Laboratory of Endocrinology and Tissue Culture, Institute of Zoology Jagiellonian University, Ingardena 6, 30-060 Kraków, PolandSearch for more papers by this authorM. TISCHNER, M. TISCHNER Department of Animal Reproduction, University of Agriculture, Kraków, PolandSearch for more papers by this author B. BILIŃSKA, Corresponding Author B. BILIŃSKA Laboratory of Endocrinology and Tissue Culture, Institute of Zoology Jagiellonian University, Ingardena 6, 30-060 Kraków, PolandLaboratory of Endocrinology and Tissue Culture, Institute of Zoology Jagiellonian University, Ingardena 6, 30-060 Kraków, PolandSearch for more papers by this authorA. HEJMEJ, A. HEJMEJ Laboratory of Endocrinology and Tissue Culture, Institute of Zoology Jagiellonian University, Ingardena 6, 30-060 Kraków, PolandSearch for more papers by this authorM. PAWLAK, M. PAWLAK Department of Animal Reproduction, University of Agriculture, Kraków, PolandSearch for more papers by this authorJ. SADOWSKA, J. SADOWSKA Laboratory of Endocrinology and Tissue Culture, Institute of Zoology Jagiellonian University, Ingardena 6, 30-060 Kraków, PolandSearch for more papers by this authorM. TISCHNER, M. TISCHNER Department of Animal Reproduction, University of Agriculture, Kraków, PolandSearch for more papers by this author First published: 05 January 2010 https://doi.org/10.2746/0425164044877305Citations: 13AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Citing Literature Volume36, Issue6September 2004Pages 539-543 RelatedInformation

The purpose of the study was to estimate alterations in the haematological profile of Polish Pony foals and mares from the first hours of life up to 6 months and to compare them with the same parameters in other horse breeds. A total of 11 Polish Pony foals and mares were examined. Blood was collected at hour 1, 6, 12, 24 and on day 3, 5, 7, 14, 21, 28, 60, 90 120, 150 and 180 post partum. Red cell, haemoglobin and haematosis figures attained maximum values within the first hours of life and these values remained stable for 3 to 6 day old foals. The red cell volume was highest during the first hours after birth and stabilised onto a level of approximately 42 fl up to the age of 7 days. On the other hand, the mean values of haemoglobin volume and corpuscular haemoglobin concentration remained stable throughout the investigation. The WBC counts for foals ranged from 5.162 to 7.328 x 10(9)/1 up to day 3 of life whereas after this time the number of leucocytes ranged from 10.790 to 13.614 x 10(9)/1. The highest indices of neutrophiles were observed in the first 7 days of life while the number of lymphocytes increased after birth from 30% to 50%. The haematological profiles in suckling mares were almost stable and approximately the same as those of mature horses of other breeds.

An unpredictability of ovarian response still remains the major problem concerning ovine reproductive programs. The influence of several environmental, genetic, and ovarian cycle effects on oocyte/embryo yield from donor females has been previously reported. The present research has been designed to exclude aforementioned causes of variability, thus to verify embryogenic competence in homogenous groups of animals. for this purpose we used prepubertal ewes kept under identical conditions. Initially, we stimulated three groups of prepubertal ewes at various ages and used a number of gonadotropin treatments to assess differences in oocyte competence between individuals. The results revealed the repeatability of response within individual donor lambs throughout the study. Moreover, once the variability in both oocyte and embryo yield between homogenous groups of donors was revealed alongside the influence of age and type of gonadotropin treatment (P < 0.001), we investigated whether the individual donor effect persisted among genetically similar animals. Therefore, we compared oocyte and subsequent embryo output of sibling lambs derived from the most efficient donor. Here the genetic homogeneity of sisters kept under identical conditions substantially improved the uniformity of either follicular response or embryo production, suggesting that the genotype plays a primary role in establishing follicular recruitment and developmental capability of oocytes. This observation consents to predict the ovarian performance from a single ewe already in early prepuberty (i.e., to qualify the female to breeding programs).

Although the potential use of reproductive biotechnologies for safeguarding endangered wildlife species is undoubted, practical efforts have met with limited success to date. In those instances in which modern technologies have been adapted to rescuing rare or endangered species, procedures have been applied piecemeal, and no consistent breeding program based on reproductive biotechnologies has been undertaken. Here we describe for the first time the rescue of an endangered species, the European mouflon (Ovis orientalis musimon), by the application of an integrated package of reproductive biotechnologies. This genetic management extended from the initial collection of gametes, through the in vitro production of embryos and interspecific transfer, to the birth of healthy mouflon offspring. In addition, a genetic resource bank for the European mouflon was established, with cryopreserved sperm, embryos, and somatic cells.

Recently developed, assisted reproductive technologies (e.g., in vitro embryo production and nuclear transfer) have encountered perinatal morbidity/mortality of the offspring produced, which are likely to hinder the application of these techniques. Consequently we have sought to develop a system of hormonal stimulation that will ensure the delivery of offspring more prepared for extrauterine life. Here we examine deliveries outcome in sheep carrying in vitro-produced and nuclear transfer (NT) embryos in comparison to artificially inseminated and naturally mated control ewes. All groups (excluding NT, which received one treatment) were subjected to one of two hormonal treatments for induction of delivery, whereas the third part of each group was left without any treatment. The first (commonly used for naturally mated ewes) dexamethasone treatment did not solve a majority of parturition disturbances, and actually the number of deliveries necessitating assistance was reduced (P < 0.05) by this treatment in the control group. On the other hand, combined estradiol plus betamethasone stimulation (E + B) solved a majority of complications regarding delivery performance such as lack of the preparation of the mammary gland, low myometrial contractility, insufficient cervical ripening, and impaired maternal behavior. Moreover, substantial reduction of neonatal mortality was observed following the combined treatment. In conclusion, the E + B induction of delivery overcame the majority of physiological and behavioral intrapartum failures of sheep foster mothers and increased the survival of offspring, and thus can be recommended as a safe method for inducing delivery in foster mothers carrying in vitro-generated embryos.