Effect of donkey seminal plasma on sperm movement and sperm-polymorphonuclear neutrophils attachment in vitro.

To evaluate the effect of seminal plasma in endometrial inflammation in donkeys, samples from fresh pure, fresh diluted and frozen-thawed semen of three different jackasses were co-incubated in water bath at 37°C with uterine Jennie's secretions collected 6h after artificial insemination with frozen-thawed donkey semen. Individual sperm movement parameters using the computerised sperm analysis system (CASA) and sperm-polymorphonuclear neutrophils (sperm-PMN) attachment observed in Diff-Quick stained smears were evaluated at 0, 1, 2, 3 and 4h of co-incubation. Controls consisted of incubating diluted or frozen-thawed sperm in the absence of uterine secretions.

Comparative effects of adding β-mercaptoethanol or L-ascorbic acid to culture or vitrification-warming media on IVF porcine embryos.

The aims of the present study were to; (1) determine the effects of supplementation with two antioxidants during in vitro culture (IVC) on embryo development and quality; and (2) test the effects of adding the antioxidants to vitrification-warming media on the cryotolerance of in vitro-produced (IVP) porcine blastocysts. In Experiment 1, presumptive zygotes were cultured without antioxidants, with 50 µM β-mercaptoethanol (β-ME) or with 100 µM L-ascorbic acid (AC). After culture, blastocyst yield, quality and cryotolerance were evaluated in each treatment group.

Adenosine monophosphate-activated kinase, AMPK, is involved in the maintenance of the quality of extended boar semen during long-term storage.

Boar semen preservation for later use in artificial insemination is performed by diluting semen in an appropriate medium and then lowering the temperature to decrease spermatozoa metabolism. The adenosine monophosphate-activated kinase, AMPK, is a key cell energy sensor that controls cell metabolism and recently has been identified in boar spermatozoa. Our aim was to investigate the role of AMPK in spermatozoa functional parameters including motility, mitochondrial membrane potential, plasma membrane integrity, acrosome integrity, and cell viability during long-term boar semen storage at 17 °C in Beltsville thawing solution.

Acrosin-binding protein (ACRBP) and triosephosphate isomerase (TPI) are good markers to predict boar sperm freezing capacity.

Sperm cryopreservation is the most efficient method for storing boar sperm samples for a long time. However, one of the inconveniences of this method is the large variation between and within boars in the cryopreservation success of their sperm. The aim of the present work was thus to find reliable and useful predictive biomarkers of the good and poor capacity to withstand the freeze-thawing process in boar ejaculates.

How do different concentrations of Clostridium perfringens affect the quality of extended boar spermatozoa?

Bacteriospermia in boar fresh and extended semen is a frequent finding that produces alterations on sperm quality and, consequently, causes economic losses in artificial insemination (AI) centres. The present study sought to evaluate the effect of different infective concentrations of Clostridium perfringens on boar sperm quality, assessed as sperm motility (CASA), morphology and viability, through 11 days of storage at 15°C (experiment 1), and after 96h of incubation at 37°C (experiment 2). With this purpose, different seminal doses were artificially inoculated with different infective concentrations of C.

Acrosin activity is a suitable indicator of boar semen preservation at 17 °C when increasing environmental temperature and radiation.

The effect of increasing environmental temperature and radiation on the sperm quality and the field fertility of refrigerated seminal doses from AI boars (N = 30) was analyzed throughout four experimental months (from March through June). In each experimental month, analyses of sperm quality were performed at days 0, 1, 3, 5, 7, and 9 of refrigeration of seminal doses; pregnancy rate and litter size were evaluated using double monospermic inseminations of multiparous female animals using seminal doses at Days 1 to 2 and Days 3 to 4 of refrigeration. Sperm quality was assessed from the evaluation of conventional parameters of sperm concentration, sperm motility, sperm morphology, and sperm viability, and capacitation parameters of membrane lipid disorder, intracellular calcium content, and acrosin activity.

Embryo development and sex ratio of in vitro-produced porcine embryos are affected by the energy substrate and hyaluronic acid added to the culture medium.

In the present study, the effects of replacing glucose with pyruvate-lactate and supplementing these in vitro culture (IVC) media with hyaluronic acid (HA) on porcine embryo development and sex ratio were examined. The in vitro-produced (IVP) porcine embryos were cultured in NCSU-23 medium with 0.0, 0.

Good and bad freezability boar ejaculates differ in the integrity of nucleoprotein structure after freeze-thawing but not in ROS levels.

The main aim of the present study was to determine whether differences in the amounts of free cysteine residues in sperm nucleoproteins, which are a direct marker of the integrity of the disulfide bonds between nucleoproteins, existed between good (GFE) and poor boar freezability ejaculates (PFE) during the different steps of the freeze-thawing process. The analyzed steps were: (1) immediately before starting cryopreservation (17 °C), (2) at the end of the cooling step (5 °C), and (3) 30, and (4) 240 minutes after thawing. In addition, the present study also sought to determine whether GFE and PFE differed in the amounts of peroxides and superoxides generated during freeze-thawing as an overall measure of the boar sperm reactive oxygen species (ROS) accumulation rate.

Reduced glutathione and procaine hydrochloride protect the nucleoprotein structure of boar spermatozoa during freeze-thawing by stabilising disulfide bonds.

One important change the head of boar spermatozoa during freeze-thawing is the destabilisation of its nucleoprotein structure due to a disruption of disulfide bonds. With the aim of better understanding these changes in frozen-thawed spermatozoa, two agents, namely reduced glutathione (GSH) and procaine hydrochloride (ProHCl), were added at different concentrations to the freezing media at different concentrations and combinations over the range 1-2mM. Then, 30 and 240 min after thawing, cysteine-free residue levels of boar sperm nucleoproteins, DNA fragmentation and other sperm functional parameters were evaluated.

Predictors of outcome in stable outpatients with peripheral artery disease.

Patients with peripheral artery disease (PAD) are at increased risk for subsequent ischemic events. We used data from the FRENA Registry to find predictors of subsequent myocardial infarction (MI), ischemic stroke, and limb amputation in stable outpatients with PAD. As of January 2012, 1,270 patients with PAD were recruited, of whom 1,042 (82 %) had Fontaine stage II; 113 (8.

Cryotolerance of in vitro-produced porcine blastocysts is improved when using glucose instead of pyruvate and lactate during the first 2 days of embryo culture.

The objective of the present study was to determine the effects of replacing glucose with pyruvate and lactate during the first 48 h of in vitro culture (IVC) in NCSU-23 medium on embryo development, embryo quality and survival of porcine blastocysts after vitrification. To this end, in vitro-produced (IVP) porcine oocytes were cultured with either glucose for 6 days (IVC-Glu) or pyruvate-lactate from Day 0 to Day 2 and then with glucose until Day 6 (IVC-PyrLac). Blastocysts were vitrified on Day 6 using the Cryotop device and, after warming, survival rate and the apoptosis index were evaluated after 24 h incubation in NCSU-23 medium.

A proper assessment of boar sperm function may not only require conventional analyses but also others focused on molecular markers of epididymal maturation.

In swine, predicting the fertilizing ability of boar ejaculates before using seminal doses for artificial insemination purposes is very important for pork breeders. Routinely, semen quality is evaluated by means of sperm concentration, viability, motility and morphology. However, in some cases, these spermiogram parameters may not be precise enough to detect altered/non-functional spermatozoa within boar ejaculates that may yield lower reproductive performance.

Direct binding of boar ejaculate and epididymal spermatozoa to porcine epididymal epithelial cells is also needed to maintain sperm survival in in vitro co-culture.

The aim of the present study was to compare the influence of cultured epididymal epithelial cells (EEC) from corpus, caput or cauda, oviductal epithelial cells (OEC) and non-reproductive epithelial cells (LLC-PK1) on function and survival of epididymal and ejaculated spermatozoa, in the latter case to determine whether such influence differed between morphologically normal and abnormal spermatozoa. For this purpose, either spermatozoa were directly co-cultured with EEC from caput, corpus, or cauda, OEC and LLC-PK1 cells (experiment 1) or a membrane-diffusible insert was included in these co-cultures (experiment 2). EEC cultured from the three epididymal regions did not differently affect the sperm parameters.

Boar sperm thawing practices: the number of straws does matter.

The number of straws thawed has been largely neglected in reports of boar sperm cryopreservation. Whereas previous studies confirm the effect of sperm concentration on function and survival of thawed boar spermatozoa, it is still unknown whether, for a same concentration, total number of sperm in the thawing solution affects its mechanics. The present trial sought to define good boar sperm thawing practices by checking if a minimal number of straws as well as the percentage of air volume in the thawing tube should be stated or not to decrease variability from one trial to another.

Effects of different concentrations of enterotoxigenic and verotoxigenic E. coli on boar sperm quality.

The presence of bacteria in boar semen causes economic losses in artificial insemination (AI) centers, as a consequence of alterations on boar sperm quality. For this reason, the effects of different concentrations of enterotoxigenic Escherichia coli (ETEC) and verotoxigenic E. coli (VTEC) on boar sperm quality were determined in this study, by conducting two experiments.

Study of the proacrosin-acrosin system in epididymal, ejaculated and in vitro capacitated boar spermatozoa.

The present study aimed to develop a set of sensitive assays to evaluate the presence of different isoforms, the activity degree, and the immunolocalisation of proacrosin-acrosin in sexually mature boars. The goal was to determine the proacrosin-acrosin status of boar spermatozoa throughout epididymal maturation, during ejaculation and after in vitro capacitation. In epididymal samples, proacrosin expression was high in all regions studied.

A PCR technique to detect enterotoxigenic and verotoxigenic Escherichia coli in boar semen samples.

In semen, bacteria's isolation from a pure culture is complex, laborious and easily alterable by the presence of antibiotics and inhibitors. We developed a PCR technique to detect the presence of the enterotoxigenic (ETEC) and verotoxigenic Escherichia coli (VTEC) (strains with high prevalence in the swine industry) in semen by adapting the protocols developed by Zhang et al. (2007) and Yilmaz et al.

Impact of epididymal maturation, ejaculation and in vitro capacitation on tyrosine phosphorylation patterns exhibited of boar (Sus domesticus) spermatozoa.

Mammalian spermatozoa acquire functionality during epididymal maturation and ability to penetrate and fertilize the oocyte during capacitation. The aim of this study was to investigate the impact of epididymal maturation, ejaculation and capacitation on phosphotyrosine content of sperm proteins. Western blot, immunocytochemical and flow cytometry analyses demonstrated that epididymal maturation in vivo is associated with a progressive loss of phosphotyrosine residues of the sperm head followed by a subtle increase after in vitro capacitation.

Alcohol consumption and outcome in stable outpatients with peripheral artery disease.

Background: The influence of alcohol consumption on outcome in patients with peripheral artery disease (PAD) has not been thoroughly studied.

Methods: Factores de Riesgo y ENfermedad Arterial (FRENA) is an ongoing, multicenter, observational registry of consecutive stable outpatients with arterial disease. We compared the mortality rate and the incidence of subsequent ischemic events in patients with PAD, according to their alcohol habits.

The effects on boar sperm quality of dietary supplementation with omega-3 polyunsaturated fatty acids differ among porcine breeds.

The present study was undertaken to shed light on the relationship between boar sperm quality and dietary supplementation with omega-3 polyunsaturated fatty acids, which has been reported inconsistently in the literature. With this aim, such effects were evaluated and compared among three different porcine breeds: Duroc, Large-White, and Pietrain. Animals were randomly separated into two groups and fed either with a control diet or with a diet supplemented with omega-3.

The HSP90AA1 sperm content and the prediction of the boar ejaculate freezability.

In a previous study we reported that the immunolabelling of GLUT3, HSP90AA1, and Cu/ZnSOD proteins on boar sperm did not show differences between good and poor freezability ejaculates, in terms of a qualitative analysis based on location and reactivity of these proteins at 17 degrees C and at 240 min post-thaw. Since predicting the ejaculate freezability is considerably important in sperm cryopreservation procedures, the objective of the present study was to quantify the expression of these three proteins in good and poor freezability ejaculates. For this purpose, 10 ejaculates from 9 Piétrain boars were cryopreserved and their sperm quality assessed in the three main steps of the freezing process (17 degrees C, 5 degrees C, and 240 min post-thaw).

The osmotic tolerance of boar spermatozoa and its usefulness as sperm quality parameter.

Predicting the fertility outcome of ejaculates is very important in the field of porcine reproduction. The aims of this study were to determine the effects of different osmotic treatments on boar spermatozoa and to correlate them with fertility and prolificacy, assessed as non-return rates within 60 days (NRR(60d)) of the first inseminations, and litter size (LS), respectively. Sperm samples (n=100) from one hundred healthy Piétrain boars were used to assess 48 treatments combining different osmolalities (ranged between 100 and 4000 mOsm kg(-1)), different compounds used to prepare anisotonic solutions, and two different modalities: return and non-return to isotonic conditions.

A diet supplemented with L-carnitine improves the sperm quality of Piétrain but not of Duroc and Large White boars when photoperiod and temperature increase.

It has been reported that a diet supplemented with L-carnitine can improve sperm quality in some mammalian species. Against this background, the current study seeks to determine the effects of feeding L-carnitine (625 mg day(-1)) on boar semen characteristics (ejaculate volume, sperm concentration, sperm viability, acrosome and mitochondrial sheath integrity, sperm motility, sperm morphology, and osmotic resistance of spermatozoa) in three different porcine breeds (Sus domesticus) (Piétrain, Duroc, and Large White) exposed to natural environmental changes in temperature and photoperiod over a 20-wk period (February to July 2007). One hundred twenty boars (40 per breed) were randomly separated into two groups (60 boars each): the first (20 boars per breed) was fed a control diet and the second (also 20 males per breed) the same diet supplemented with L-carnitine (625 mg day(-1)).

Freezability prediction of boar ejaculates assessed by functional sperm parameters and sperm proteins.

The objective of this work was to look for useful predictive indicators of the potentially "good" or "poor" ability of a boar ejaculate to sustain cryopreservation by assessing both the conventional sperm quality parameters (Study 1) and the immunolabeling of three proteins involved in the physiology of the sperm cell: GLUT3, HSP90AA1 and Cu/ZnSOD (Study 2). Study 1 was carried out in three different steps during the cryopreservation process of the sperm-rich fraction of 29 Piétrain boar ejaculates (17 degrees C, 5 degrees C, and 240min postthaw). These ejaculates were clustered based on sperm quality parameters analyzed at 240min postthaw, obtaining 16 good freezability ejaculates (GFEs) and 13 poor freezability ejaculates (PFEs).

Fertility after post-cervical artificial insemination with cryopreserved sperm from boar ejaculates of good and poor freezability.

This study compared the field fertility outcomes in frozen-thawed (FT) sperm from boar ejaculates with different freezability (good, GFE/poor, PFE) while testing the reliability of the post-cervical artificial insemination (post-CAI) in FT sperm. The assay was conducted over eight months with 86 weaned sows being inseminated by post-CAI. Every ejaculate in a total of 26 from 15 Piétrain boars was divided into a refrigerated semen portion (FS; control treatment) and a cryopreserved portion (FT sperm), and the ejaculates were in turn classified as GFE or PFE in function of the sperm progressive motility and viability at 240min post-thaw.