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Sperm Viability (sperm + viability)
Selected AbstractsRelationship between Sperm Response to Glycosaminoglycans in vitro and Non-return Rates of Swedish Dairy AI BullsREPRODUCTION IN DOMESTIC ANIMALS, Issue 5 2000A Januskauskas Contents In this study, the relations between fertility (56-day non-return rates, 56-day NRR) after artificial insemination (AI) and bull sperm characteristics post-thaw, after swim-up and after co-incubation with heparin (Hep) and hyaluronan (HA), respectively, were determined, attempting to determine if such a procedure could be of value to evaluate the potential fertilizing ability of frozen-thawed AI bull spermatozoa. Spermatozoa from 20 semen batches derived from 20 Swedish Red and White AI bulls ranging widely in their field fertility after AI (55,79% 56-day NRRs) were evaluated with regards to post-thaw motility, membrane integrity, and migration through a simple swim-up procedure. Sperm viability and capacitation status were evaluated by two different vital staining procedures and chlortetracycline hydrochloride staining. Sperm motility and membrane integrity post-thaw (e.g. indicators of sperm viability) were significantly correlated (r = 0.53, p < 0.05 and r = 0.59, p < 0.01, respectively) with fertility. Heparin (5 µg/ml) significantly (p lt; 0.001) increased the frequencies of capacitation and acrosome-reaction (AR) among swim-up separated spermatozoa, whereas HA at a concentration of 50 ng/ml did not have any significant capacitating effect. The incidences of capacitated or AR-spermatozoa following Hep-treatment were not correlated with fertility. On the other hand, the percentage of viable spermatozoa was significantly (p < 0.001) lower in Hep-treated samples than in control and HA-treated samples and was significantly (r = 0.49, p < 0.05) correlated with fertility after AI (56-day NRR). The results indicate that the percentage of viable spermatozoa after swim-up separation and heparin-exposure from a selected population of AI bulls were significantly and positively related to the AI fertility of the donors and thus could be used as a parameter to determine the fertilizing ability of frozen,thawed AI bull spermatozoa. [source] Liposome-mediated uptake of exogenous DNA by equine spermatozoa and applications in sperm-mediated gene transferEQUINE VETERINARY JOURNAL, Issue 1 2008B. A. BALL Summary Reasons for performing study: Sperm-mediated gene transfer has been reported as a method for production of transgenic animals in a variety of species, and this technique represents a possible method for production of transgenic equids. Objectives: To evaluate the uptake of exogenous DNA (enhanced green fluorescent protein; pEGFP) by equine spermatozoa and to assess the ability of transfected spermatozoa to introduce this transgene into early equine embryos. Methods: To evaluate incorporation of pEGFP into equine spermatozoa, washed spermatozoa were incubated with 32P-pEGFP, with or without lipofection. Spermatozoa were also transfected with fluorescently-labelled DNA (Alexa647 -pEGFP) and changes in sperm viability and DNA uptake were assessed. Mares were inseminated with pEGFP-transfected spermatozoa and embryos recovered. Expression of pEGFP was assessed by epifluorescence microscopy of embryos, and the presence of pEGFP DNA and mRNA was assessed by PCR and RT-PCR, respectively. Results: Liposome-mediated transfection increased the incorporation of 32P-pEGFP into spermatozoa compared to controls. Flow cytometric evaluation of spermatozoa after transfection with Alexa647 -pEGFP revealed a linear increase in the proportion of live, Alexa647+ spermatozoa with increasing DNA concentrations. After insemination with transfected spermatozoa, 8 embryos were recovered. There was no evidence of EGFP expression in the recovered embryos; however, PCR analysis revealed evidence of the pEGFP transgene in 2 of 5 embryos analysed. Conclusions: The incorporation of exogenous DNA by equine spermatozoa was enhanced by liposome-mediated transfection and this did not adversely affect sperm viability, acrosomal integrity or fertility. Although the EGFP transgene was detected in a proportion of Day 7,10 embryos, there was no evidence of expression of EGFP in these embryos. Potential relevance: Sperm-mediated gene transfer offers a potential technique for the generation of transgenic equids. [source] A histochemical study of the reproductive structures in the flatworm Dugesia leporii (Platyhelminthes, Tricladida)INVERTEBRATE BIOLOGY, Issue 2 2006Gavina Corso Abstract. The functional morphology and the topographic distribution of tissues in the reproductive system of specimens of Dugesia leporii, an endemic Sardinian free-living planarian, are investigated. Data are provided on the nature of epithelial and glandular secretions, spermatophores, and cocoons by histochemistry, light microscopy, and scanning electron microscopy. All secreting epithelial cells produce strongly acidic sulfated glycoproteins. Glandular cells secrete strongly acidic sulfated glycoproteins or keratohyalin-like material in the penis bulb, and prekeratin-like material in atrial glands. Secretions of the bursa copulatrix may be involved in the activation of sperm while material produced by the bursa canal and oviducts probably serves to propel spermatophores or sperm and eggs. Mucous secretion of the seminal vesicle may serve to dilute and activate sperm before copulation. The viscous secrete of the ejaculatory duct and vasa deferentia may play a protective role to maintain sperm viability. Materials produced by the penis papilla and atrium probably lubricate the epithelial surface. The bilayered wall of spermatophore made of keratohyalin-like material and strongly acidic sulfated glycoproteins is produced by two gland types of the penis bulb. The bilayered shell of cocoon made of prekeratin-like and keratohyalin-like materials is secreted by both atrial glands and vitelline cells. The cocoon stalk is made of keratohyalin-like material produced by cement glands. Shell glands, producing GAG, are not involved in cocoon formation, but they may be implicated in the dilution and activation of seminal material to favor sperm movement toward the oviducts. [source] Ferulic acid: pharmaceutical functions, preparation and applications in foodsJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 11 2004Shiyi Ou Abstract Ferulic acid (4-hydroxy-3-methoxycinnamic acid), an effective component of Chinese medicine herbs such as Angelica sinensis, Cimicifuga heracleifolia and Lignsticum chuangxiong, is a ubiquitous phenolic acid in the plant kingdom. It is mainly conjugated with mono- and oligosaccharides, polyamines, lipids and polysaccharides and seldom occurs in a free state in plants. Ferulic acid is a phenolic acid of low toxicity; it can be absorbed and easily metabolized in the human body. Ferulic acid has been reported to have many physiological functions, including antioxidant, antimicrobial, anti-inflammatory, anti-thrombosis, and anti-cancer activities. It also protects against coronary disease, lowers cholesterol and increases sperm viability. Because of these properties and its low toxicity, ferulic acid is now widely used in the food and cosmetic industries. It is used as the raw material for the production of vanillin and preservatives, as a cross-linking agent for the preparation of food gels and edible films, and as an ingredient in sports foods and skin protection agents. Ferulic acid can be prepared by chemical synthesis and through biological transformation. As polysaccharide ferulate is a natural and abundant source of ferulic acid, preparation of ferulic acid from plant cell wall materials will be a prospective pathway. Copyright © 2004 Society of Chemical Industry [source] Effect of exogenous DNA on bovine sperm functionality using the sperm mediated gene transfer (SMGT) techniqueMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 8 2010Sebastian Canovas Sperm mediated gene transfer (SMGT) could provide the opportunity to carry out transgenesis on a mass scale using spermatozoa as vectors for exogenous DNA. However, the efficiency of sperm-mediated DNA transfer is still questionable, and the mode of transmission to the egg has not yet been well understood. Our aim was to investigate the capacity of bovine spermatozoa to carry exogenous DNA and its relationship to sperm functionality. We studied these parameters using flow cytometry to measure viability (necrosis and apoptosis) and capacitation status, computer-assisted semen analysis (CASA) to measure motility parameters and in vitro fertilization (IVF) to assess fertilizing capacity. Furthermore, we studied the effect of capacitation status on interaction with exogenous DNA, and the role of heparin supplementation in this process. Bull spermatozoa showed a high capacity to bind DNA quickly and reached a maximum after 30,min, with approximately half of the DNA-bound spermatozoa being viable. Incubation with exogenous DNA induced a decrease in sperm viability and motility and increased the proportion of apoptotic cells, but did not affect the cleavage rate in IVF assay. Heparin increased high-lipid disorder and the number of sperm with DNA bound (viable and dead). In conclusion, this study shows that live spermatozoa can bind exogenous DNA with a slight negative effect in some parameters of sperm function that in our opinion, would not drastically compromise fertility. Mol. Reprod. Dev. 77: 687,698, 2010. © 2010 Wiley-Liss, Inc. [source] Effects of 2-hydroxypropyl-,-cyclodextrin and cholesterol on porcine sperm viability and capacitation status following cold shock or incubationMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 5 2006Hannah L. Galantino-Homer Abstract Porcine sperm are extremely sensitive to the damaging effects of cold shock. It has been shown that cholesterol-binding molecules, such as 2-hydroxypropyl-,-cyclodextrin (HBCD), improve post-cooling porcine sperm viability when added to an egg yolk-based extender, but also enhance sperm capacitation in other species. The objective of this study was to determine the effects of HBCD and cholesterol 3-sulfate (ChS) on porcine sperm viability and capacitation following cold shock or incubation under conditions that support capacitation using a defined medium. We report here that porcine sperm incubated in medium containing both HBCD and ChS have significantly improved viability following cold shock (10 min at 10°C) when compared to sperm incubated without HBCD or ChS, or with either component alone. Treatment with HBCD plus ChS also completely inhibited the increase in protein tyrosine phosphorylation induced by the cold shock treatment or by incubation for 3 hr under conditions that support capacitation. Two assays of sperm capacitation, the rate of calcium ionophore-induced acrosome reactions and chlortetracycline (CTC) staining, were not significantly altered by HBCD and ChS following cold shock. However, 3-hr incubation with HBCD plus ChS or with 1 mM ChS alone decreased the percentage of sperm undergoing the induced acrosome reaction without significantly affecting viability when compared to the control. These results indicate that the manipulation of sperm plasma membrane cholesterol content affects porcine sperm viability and capacitation status and could therefore be useful to protect sperm from cold shock during cryopreservation by improving viability without promoting premature capacitation. Mol. Reprod. Dev. © 2006 Wiley-Liss, Inc. [source] Qualitative aspects of sperm stock in males and females from Eupelmus orientalis and Dinarmus basalis (Hymenoptera: Chalcidoidea) as revealed by dual fluorescencePHYSIOLOGICAL ENTOMOLOGY, Issue 2 2002David Damiens Abstract The quality of a sperm population can be characterized physiologically and its fecundity predicted by its viable : non-viable sperm ratio. To improve the knowledge of reproductive strategies in two ectoparasitoid hymenopteran species, Eupelmus orientalis Crawford (Hymenoptera: Eupelmidae) and Dinarmus basalis Rondani (Hymenoptera: Pteromalidae), the assessment of sperm viability using the dual fluorescence staining procedure SYBR-14 : propidium iodide was developed. The aim of the study was to provide a comparative test in vitro applicable to both sexes to study the evolution of sperm quality at various stages of the reproductive processes. The reliability of propidium iodide to detect non-viable sperm (stained in red) was confirmed in both species on the basis of two stress tests (ethanol and Triton X-100) but our study also revealed that propidium iodide concentrations must be adequately adjusted for each single species. This experiment also demonstrated the physiological heterogeneity of sperm populations in E. orientalis and D. basalis males and females. In both species, 40% of the sperm in the seminal vesicles was found to be non-viable. By contrast with E. orientalis, the populations of non-viable sperm estimated from the seminal vesicles of D. basalis were found to be strongly different from those observed in the spermatheca. From the present results, the population of viable sperm detected in the spermatheca of females from both species proved a reliable predictor of fertilization achieved in ovipositing females. [source] The seminal fluid proteome of the honeybee Apis melliferaPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 8 2009Boris Baer Dr. Abstract Ejaculates contain sperm but also seminal fluid, which is increasingly recognized to be of central importance for reproductive success. However, a detailed biochemical composition and physiological understanding of seminal fluid is still elusive. We have used MS to identify the 57 most abundant proteins within the ejaculated seminal fluid of the honeybee Apis mellifera. Their amino acid sequences revealed the presence of diverse functional categories of enzymes, regulators and structural proteins. A number have known or predicted roles in maintaining sperm viability, protecting sperm from microbial infections or interacting with the physiology of the female. A range of putative glycoproteins or glycosylation enzymes were detected among the 57, subsequent fluorescent staining of glycolysation revealed several prominant glycoproteins in seminal fluid, while no glycoproteins were detected in sperm samples. Many of the abundant proteins that accumulate in the seminal fluid did not contain predictable tags for secretion for the cell. Comparison of the honeybee seminal fluid proteins with Drosophila seminal fluid proteins (including secreted accessory gland proteins known as ACPs), and with the human seminal fluid proteome revealed the bee protein set contains a range of newly identified seminal fluid proteins and we noted more similarity of the bee protein set with the current human seminal fluid protein set than with the known Drosophila seminal fluid proteins. The honeybee seminal fluid proteome thus represents an important addition to available data for comparative studies of seminal fluid proteomes in insects. [source] Effects of Matrix Filtration of Low-Quality Boar Semen Doses on Sperm QualityREPRODUCTION IN DOMESTIC ANIMALS, Issue 3 2009E Bussalleu Contents The aim of this work was to develop a method to enhance the sperm parameters of ejaculates with low sperm quality from Piétrain boars. Seminal doses were filtered through columns of DEAE Sephadex (length 2.5 ± 0.5 cm), CM Sephadex (length 5 ± 0.5 cm), glass wool (length 2 ± 0.5 cm) or glass bead (length 10 ± 0.5 cm), with an exit flow rate of 1 ml/40 s in all cases. For each male, 10 ml of the sperm cell-rich fraction diluted at 1 : 6 were filtered. Sperm quality was assessed before and after filtration. Sperm morphology, sperm motility and sperm concentration were determined using the computer program sca® 2002 Production, and sperm viability was evaluated by fluorescence multistaining. Osmotic resistance test and hyperosmotic resistance test were used to determine the osmotic resistance of spermatozoa, whereas l -lactate production estimated the metabolic activity. Results showed a decrease of sperm concentration and osmotic resistance of spermatozoa after filtration in the four matrixes. However, an increase in the frequency of viable spermatozoa with intact acrosome after filtration in glass bead columns and an increase of morphologically normal spermatozoa after filtration in Sephadex CM-50, glass wool and glass bead columns were observed. Despite the decrease in the frequency of progressive motile spermatozoa, l -lactate production and mitochondrial sheath integrity maintained constant after filtration. Our findings indicate that column filtration is an effective method to enhance the sperm quality by selecting viable and morphologically normal spermatozoa without altering DNA, plasma membrane, mitochondrial sheath integrity or inducing premature acrosome reaction. [source] Post-thaw Survival and Longevity of Bull Spermatozoa Frozen with an Egg Yolk-based or Two Egg Yolk-free Extenders after an Equilibration Period of 18 hREPRODUCTION IN DOMESTIC ANIMALS, Issue 3 2007R Muiño Contents The aim of the present study was to determine the suitability of using two egg yolk-free commercial extenders, Andromed® and Biociphos Plus®, as compared with the Tris-egg yolk based diluent Biladyl®, for the cryopreservation of bull spermatozoa when the freezing protocol involved holding the extended semen at 4°C for 18 h before the freezing. Six ejaculates from each of 10 Holstein bulls were collected by using artificial vagina. The ejaculates were evaluated for volume, sperm concentration and motility, divided in to three equal volumes, and diluted, respectively, with the three extenders as specified above. Extended semen was equilibrated for 18 h at 4°C and frozen in 0.25-ml straws. After thawing, 100- ,l aliquots of semen were labelled with SYBR-14, PI and PE-PNA (Phycoerythrin-conjugated Peanut agglutinin) and analysed by flow cytometry at 0, 3, 6 and 9 h after incubation at 37°C. A General Lineal Model procedure for repeated measures was used to determine the effects of extender, bull, replicate and the interaction between them, on sperm viability and acrosomal integrity. Semen samples frozen with Biladyl® showed higher (p < 0.001) sperm survival after 0 h (47.9%) and 9 h (30.3%) of incubation than those frozen with Andromed® (38.5% and 17.3%, after 0 and 9 h respectively) or Biociphos Plus® (34.9% and 21.6%, after 0 and 9 h respectively). The bull and replicate had significant effects (p < 0.001) on both sperm viability and acrosomal integrity, but the interactions between bull and extender and between replicate and extender were not significant. It was concluded that, when holding the semen overnight before freezing, the use of Biladyl® results in higher sperm survival and longevity than the use of Andromed® or Biociphos Plus®. [source] Multivariate Cluster Analysis Regression Procedures as Tools to Identify Motile Sperm Subpopulations in Rabbit Semen and to Predict Semen Fertility and Litter SizeREPRODUCTION IN DOMESTIC ANIMALS, Issue 3 2007A Quintero-Moreno Contents Computerized motility analysis (CASA) shows that four separate subpopulations of spermatozoa with different motility characteristics co-exist in rabbit ejaculates. There were significant (p < 0.01) differences in the distribution of these subpopulations among separate genetic lines, total sperm abnormalities and the percentage of altered acrosomes. Furthermore, logistic and linear multivariate regressions among several parameters of rabbit semen quality analysis were tested for use as predictive tools for the fertilizing ability of a specific artificial insemination semen sample. Logistic regression analysis rendered two mathematical, significant (p < 0.01) models: one between sperm viability and conception rate and the other between total sperm abnormalities and conception rate. Multiple linear regression analyses also yielded some significant relationships between both fertility (p < 0.001) and litter size (p < 0.05), with respect to some semen characteristics. Our results support the hypothesis that the predictive in vivo fertility use of the standard rabbit semen quality analysis coupled with a CASA determination could be reasonably achieved by applying linear and logistic regression analyses among several parameters of rabbit semen quality analysis. [source] Relationship between Sperm Response to Glycosaminoglycans in vitro and Non-return Rates of Swedish Dairy AI BullsREPRODUCTION IN DOMESTIC ANIMALS, Issue 5 2000A Januskauskas Contents In this study, the relations between fertility (56-day non-return rates, 56-day NRR) after artificial insemination (AI) and bull sperm characteristics post-thaw, after swim-up and after co-incubation with heparin (Hep) and hyaluronan (HA), respectively, were determined, attempting to determine if such a procedure could be of value to evaluate the potential fertilizing ability of frozen-thawed AI bull spermatozoa. Spermatozoa from 20 semen batches derived from 20 Swedish Red and White AI bulls ranging widely in their field fertility after AI (55,79% 56-day NRRs) were evaluated with regards to post-thaw motility, membrane integrity, and migration through a simple swim-up procedure. Sperm viability and capacitation status were evaluated by two different vital staining procedures and chlortetracycline hydrochloride staining. Sperm motility and membrane integrity post-thaw (e.g. indicators of sperm viability) were significantly correlated (r = 0.53, p < 0.05 and r = 0.59, p < 0.01, respectively) with fertility. Heparin (5 µg/ml) significantly (p lt; 0.001) increased the frequencies of capacitation and acrosome-reaction (AR) among swim-up separated spermatozoa, whereas HA at a concentration of 50 ng/ml did not have any significant capacitating effect. The incidences of capacitated or AR-spermatozoa following Hep-treatment were not correlated with fertility. On the other hand, the percentage of viable spermatozoa was significantly (p < 0.001) lower in Hep-treated samples than in control and HA-treated samples and was significantly (r = 0.49, p < 0.05) correlated with fertility after AI (56-day NRR). The results indicate that the percentage of viable spermatozoa after swim-up separation and heparin-exposure from a selected population of AI bulls were significantly and positively related to the AI fertility of the donors and thus could be used as a parameter to determine the fertilizing ability of frozen,thawed AI bull spermatozoa. [source] Metabolic evaluation of cooled equine spermatozoaANDROLOGIA, Issue 2 2010A. B. Vasconcelos Summary Microscopy has been used in the routine evaluation of sperm metabolism. Nevertheless, it has limited capacity to preview male fertility. As calorimetry may be used to evaluate directly the metabolic activity of a biological system, the aim of this study was to use microcalorimetry as an additive method for sperm metabolism evaluation of cooled equine semen. Two ejaculates of four stallions were collected and motility, viability (eosin 3%) and membrane functional integrity (hyposmotic swelling test) of spermatozoa were evaluated. Sperm samples were processed following different protocols and the metabolism of these samples was accessed by calorimetry. Centrifugation is part of some of these processing protocols and although this procedure has been deleterious for sperm viability and plasma membrane integrity, no decrease in sperm motility was observed. Microcalorimetry was capable of detecting the positive effect of re-suspending the sperm pellet with Kenney extender. Thus, the use of microcalorimetry offered additional information for equine sperm metabolism evaluation and was efficient in detecting important information from sperm cell metabolism. [source] Effect of norethisterone and its A-ring reduced metabolites on the acrosome reaction in porcine spermatozoaANDROLOGIA, Issue 5 2002G. Martinez Summary. The synthetic progestin, norethisterone (NET), has been reported as a contragestational postcoital agent in humans, rodents and rabbits. The effect and molecular mechanisms of NET and its A-ring reduced metabolites, 5,-NET and 3,5,-NET, on the acrosome reaction (AR) are unknown. The aim of this study was to assess the effect of these compounds on an in vitro progesterone-induced AR in porcine spermatozoa. The spermatozoa were obtained from semen ejaculated by proven fertile adult pigs. Seminal plasma removed and incubated under capacitating conditions was performed in TALP-Hepes medium for 4 h. Progesterone (P4) and three different progestins: norethisterone (NET), 5,-norethisterone (5,-NET) and 3,5,-NET were then added at equimolar doses, and the spermatozoa were incubated for 15 min. Double-staining with PSA-FITC and Hoechst-33258 assessed the AR and sperm viability. Both P4 and NET induced the AR, while 5,-NET not only did not induce this process, but was able to block the effect of P4 on the spermatozoa. 3,5,-NET was not able to inhibit P4 action. These results suggest that NET and its A-ring reduced metabolites act in different ways on the progesterone-induced AR in porcine spermatozoa. [source] Impairment of human sperm motility and viability by quercetin is independent of lipid peroxidationANDROLOGIA, Issue 5 2001K. L. Khanduja Summary., Human sperm motility, viability and lipid peroxidation were assessed in Ringer-Tyrode supplemented with different concentrations of quercetin. An irreversible and dose-dependent fall in sperm motility was observed with quercetin (5,200 µm). However, sperm viability was decreased only at higher concentrations (50,100 µm) of quercetin. This inhibition in sperm motility and viability has been linked with decreased Ca2+ -ATPase activity, as there was no effect of quercetin on sperm lipid peroxidation. [source] Semen cryopreservation in the Salmonidae and in the Northern pikeAQUACULTURE RESEARCH, Issue 3 2000F. Lahnsteiner The present paper summarizes the data on a semen cryopreservation method for the Salmonidae (Oncorhynchus mykiss, Salmo trutta f. lacustris, Salvelinus fontinalis, Salvelinus alpinus, Salmo trutta f. fario, Hucho hucho, Coregonus lavaretus, Thymallus thymallus) and for the Northern pike (Esox lucius) published during recent years. It describes (1) methods used for the determination of sperm viability; (2) the protective efficiency of substances specifically for protection of internal and external parts of cells and the process of extender development; (3) the freezing, thawing and fertilization conditions; and (4) the tolerable deviations from the freezing protocol for more easy application. Finally, biomarkers are reported that predict the suitability of semen for cryopreservation and the quality of frozen,thawed semen. [source] | ||||||||||||||||