Effects of supplementation of cryopreservation media with cysteine on the post-thaw quality and fertility of brown-marbled grouper (Epinephelus fuscoguttatus) spermatozoa.

The cryopreservation process is associated with the generation of excessive reactive oxygen species, which causes a series of cellular damage to spermatozoa. The objective of the current study was to investigate the effect of different concentrations of cysteine on post-thaw sperm quality of brown-marbled grouper sperm. Semen samples were frozen with cysteine supplemented at 0.5, 1, 2, 5, 10 mM and the control group (no additive). After thawing, sperm quality parameters were analyzed. In comparison to the control, cysteine treatment groups yielded relatively higher sperm total motility, progressive motility, and curvilinear velocity. Different concentrations of cysteine had no effect on average path velocity, straight linear velocity and viability (P > 0.05), while an increase in the concentration of cysteine resulted in a significant improvement in the mitochondrial membrane potential, SOD activity, and ATP content (P < 0.05). As for lipid peroxidation, the extent of which in cysteine treated spermatozoa was less than the control, although the differences were not statistically significant (P > 0.05). In terms of fertilizing capacity, a greater hatching rate (91.7 ± 1.2%) was obtained in thawed sperm treated with 2 mM cysteine, compared to the control (84.3 ± 4.2%; P < 0.05). Overall, it is concluded that the addition of cysteine is helpful in maintaining the function of frozen-thawed brown-marbled grouper sperm, which can be recommended as an effective antioxidant to improve the semen cryopreservation efficiency.

Tinospora cordifolia modulates the seminal parameters, leakage of intracellular enzymes and seminal antioxidants in equilibrated and cryopreserved semen of Sahiwal bulls.

The present study was undertaken to assess the effects of stem extract of Tinospora cordifolia (Giloy or Guduchi) in the semen extender on seminal parameters, leakage of intracellular enzymes and antioxidants in semen of Sahiwal bull. A total of 48 ejaculates from four bulls were selected for the study. Spermatozoa of 25 × 106 were incubated in 100, 300 and 500 µg of stem extract of Guduchi as Gr II, III and IV, respectively, and pre-freeze and post-thaw semen samples were analysed for seminal parameters [motility, viability, total sperm abnormality (TSA), plasma membrane integrity (PMI) and acrosomal integrity (AcI)], intracellular enzymes [aspartate aminotransferase (AST) and lactate dehydrogenase (LDH)] and seminal antioxidants [superoxide dismutase (SOD) and catalase] in comparison with an untreated control group (Gr I). The results revealed that stem extract-treated semen had significantly (p < .05) higher motility, viability, PMI, AcI, SOD and catalase and had significantly (p < .05) lower TSA, AST and LDH compared to those in untreated control group at pre-freeze and post-thaw stages. Semen treated with 100 µg stem extract/25 × 106 spermatozoa had significantly (p < .05) higher motility, viability, PMI, AcI, SOD and catalase and had significantly (p < .05) lower TSA, AST and LDH compared to those in control, 300- and 500-µg-treated groups at pre-freeze and post-thaw stages. Further, these seminal parameters and antioxidants were showing decreasing trend and TSA and leakage of intracellular enzymes were showing increasing trend from Gr II to Gr IV at pre-freeze and post-thaw stages. Thus, 100 µg/25 × 106 spermatozoa were optimum or suitable dose for cryopreservation of Sahiwal bull semen. The study concluded that T. cordifolia stem extract 100 µg/25 × 106 spermatozoa in the semen extender can be effectively utilized to reduce the oxidative stress and improve the pre-freeze and post-thaw seminal parameters in Sahiwal bull. However, further studies on effects of different concentrations of stem extract on in vitro or in vivo fertility trials are to be conducted to assess the impact of the stem extract supplementation in the semen extender on field pregnancy outcomes in bovine species.

The effect of Caulerpa sertularioides extract on bull sperm freezablity and subsequent embryo development.

Artificial insemination is a valuable and essential tool in genetic improvement programs, and its success requires proper semen collection, freezing, and thawing procedures. Nowadays, despite applying of advanced protocols for semen cryopreservation, post-thawing sperm quantitative and qualitative parameters are not satisfactorily comparable to fresh sperm. The present study was designed to evaluate the effects of the supplementation of an alcoholic extract of Caulerpa sertolarioides alga into the tris-egg yolk-based Simmental bull sperm freezing media. The pooled semen samples were divided into five groups, of which four were supplemented with 500, 1000, 1500, and 2000 ppm alga extract and one allocated as a control. Total motility, progressive motility, plasma membrane integrity, DNA integrity, apoptosis, superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities, and total antioxidant capacity (TAC) of sperm were measured. The frozen sperm from each group were used for IVF on the slaughterhouse-derived oocytes. Fertilization, cleavage, and blastocyst rates were assessed for all groups. Total motility, progressive motility, and velocity curvilinear (VCL) parameters were higher (p ≤ 0.05) in group 1000 ppm than the control group. Velocity in a straight path (VSL) was higher (p ≤ 0.05) in all treatment groups except in 500 ppm compared to the control group. Average path velocity (VAP) was higher (p ≤ 0.05) in 1000 and 1500 ppm groups than in the control group. Straightness (STR) showed a higher value (p ≤ 0.05) in 1000 and 2000 ppm than the control group. Groups 500 and 1000 ppm showed more viable sperm than the control group (p ≤ 0.05). DNA damage was lower (p ≤ 0.05) in group 1000 ppm than in the control group. HOST was higher (p ≤ 0.05) in all groups than in the control group. SOD, GPx, and TAC were higher (p ≤ 0.05) in 1000 ppm than the control and all other groups. Apoptosis was not significantly different among the treatment and control groups. In conclusion, supplementation of alcoholic extract of Caulerpa sertularioides into the Simmental bull freezing extender ameliorated the sperm parameters after the freeze-thawing process. Moreover, the results of this study indicated that the best dose to achieve the antioxidant properties of the alga extract in Simmental bull sperm freezing media was 1000 ppm. It was also evident that 1000 ppm alga extract supplementation into the bull sperm improved fertilization, cleavage, and blastocyst rates.

Protective Effects of Fisetin in the Mice Induced by Long-Term Scrotal Hyperthermia.

Exposure to heat in the male reproductive system can lead to transient periods of partial or complete infertility. The current study aimed to examine the beneficial effects of  Fisetin against spermatogenic disorders in mice affected by long-term scrotal hyperthermia. For this purpose, hyperthermia was induced daily by exposure to the temperature of 43 °C for 20 min for 5 weeks. Except for the Healthy group, six other groups were exposed to heat stress: two treated groups including Preventive and Curative which received oral administration of fisetin (10 mg/kg/day) starting immediately before heat exposure and 15 consecutive days after the end of the heat exposure, respectively. And for each treated group, two groups including Positive Control (Pre/Cur+PC group) and vehicle (Pre/Cur+DMSO group) were considered. Our results showed that the testicular volume; the density of spermatogonia, primary spermatocyte, round spermatid, and Sertoli and Leydig cells; and sperm parameters, as well biochemical properties of the testis tissue, were remarkably higher in both Preventive and Curative groups compared to the other hyperthermia-induced groups and were highest in Preventive ones. Unlike the c-kit gene transcript which was significantly increased in the  Fisetin treatment groups (specially the Preventive group), the expression of HSP72 and NF-kß genes, Caspase3 protein, and DFI in sperm cells were significantly more decreased in Preventive and Curative groups compared to other hyperthermia-induced groups and were lowest in Preventive ones. Overall,  Fisetin exerts preventive and curative effects against spermatogenic disorders induced by long-term scrotal hyperthermia.

Using Herbal dyes as an alternative staining method for sperm evaluation.

Staining, as a valuable method for sperm morphological assessment, has been used to determine sperm abnormalities, fertilization capability and sperm suitability during freezing-thawing process. Synthetic dyes have been used for sperm viability and morphological evaluation. However, most of them have been made from chemical substances and have a perilous effect on the environment. In the current study, we evaluated three different natural dyes as the natural sources of dye for sperm staining. Bull frozen semen was used and prepared on slides for staining. Aqueous extract dye of black mulberry (BM), henna (HA), safflower (SA) and eosin-nigrosine (control group) were used for sperm staining. Additionally, the effect of staining dyes on viability and some morphological parameters (head area: HR, head abnormality: HB and tail abnormality: TA) were evaluated. Although none of the natural dyes could detect viability of the sperm cells, safflower stain (HR: 26.55 µm, HB: 0% TA: 28%) and black mulberry stain (HR: 25.07 µm, HB: 2% TA: 3%) compared to control group (HR: 34.29 µm, HB: 4%, TA: 4%) provoked a strong reaction in the sperm cells, so that the sperms were observed yellow and red respectively. The reaction of sperm cells to the henna dye was very poor and it did not stain the sperm cells. Thus, the present study demonstrated that SA and BM dyes are able to stain the spermatozoa and with further modification could be used as alternative dyes for sperm staining in the study of sperm morphology, but not viability. Staining with these dyes can be an alternative to current costly chemical staining methods.

Diluente Tris suplementado com óleo de Mauritia flexuoxa sobre a qualidade do sêmen caprino após a criopreservação / Tris diluent supplemented with Mauritia flexuoxa oil on the quality of goat semen after cryopreservation

Objetivou-se verificar os efeitos, nos parâmetros espermáticos, na integridade mitocondrial, acrossomal e de membrana em células espermáticas, desencadeados pelo uso do Tris (Tris hidroximetil aminometano) suplementado com óleo de Mauritia flexuoxa como diluente para criopreservação de sêmen caprino. Quatro caprinos clinicamente saudáveis foram utilizados. Os animais eram alimentados diariamente com volumoso (Pennisetum purpureum, Schum.), concentrado (ração peletizada com teor de 20% proteína, 300 g/animal/dia) e sal mineral específico para Caprinos (Caprinofós®), à vontade. Dois ensaios foram realizados: I – Teste de toxicidade; II – Criopreservação do sêmen com concentrações ideais. No teste de toxicidade as concentrações avaliadas foram: 5%, 10%, 15% e 20% de diluente a base de óleo de Mauritia flexuoxa. Após o teste de toxicidade, foi escolhido a concentração que apresentou o melhor resultado (5%). Logo após, foram realizadas mais 32 coletas, que foram diluídas em Tris-gema-glicerol (grupo controle) ou diluente contendo óleo vegetal (Mauritia flexuoxa). As amostras foram criopreservadas com auxílio do aparelho Tk3000®. Após o período mínimo de uma semana as palhetas foram descongeladas em banho-maria a 37 °C por 30 segundos, acondicionadas em microtubos de centrifugação e homogeneizadas para a análise imediata de motilidade, vigor espermático e morfologia. Em seguida, por meio de sondas fluorescentes foram avaliadas a integridade de acrossomo, membrana plasmática (Diacetato de Carboxifluresceína e Iodeto de Propídeo) e função mitocondrial sob microscopia de epifluorescência. Quanto a motilidade e vigor, integridade mitocondrial e acrossomal, o grupo buriti foi inferior ao grupo controle. O Tris suplementado com óleo de Mauritia flexuoxa na concentração de 5% não influenciou significativamente a qualidade espermática, porém, observouse morfologia e integridade de membrana favoráveis. Dessa forma, sendo uma alternativa para substituição de diluentes a base de produtos de origem animal.

The objective was to verify the effects, sperm parameters, mitochondrial, acrosomal and membrane integrity in sperm cells, triggered by the use of Tris (Tris hydroxymethyl aminomethane) supplemented with Mauritia flexuoxa oil as a diluent for cryopreservation of goat semen. Four goats clinically healthy were used. The animals were fed daily with bulky (Pennisetum purpureum, Schum.), concentrate (pelleted feed with 20% protein content, 300 g / animal / day) and mineral salt Specific for Goats (Caprinofós®), ad libitum. Two tests were carried out: I - Toxicity test; II - Semen cryopreservation with ideal concentrations. In the toxicity test as selected were: 5%, 10%, 15% and 20% of Mauritia flexuoxa oil-based diluent. After the toxicity test, the concentration that showed the best result (5%) was chosen. Soon after, a further 32 samples were obtained, which were diluted in Tris-glycerol (control group) or diluent containing vegetable oil (Mauritia flexuoxa). The samples were cryopreserved using the Tk3000® machine. After a minimum of one week, the samples were thawed in a 37 ° C water bath for 30 seconds, packed in centrifugation microtubes and homogenized for immediate analysis of motility, sperm vigor and morphology. Then, by means of fluorescent probes, the integrity of the acrosome, plasma membrane (Carboxyflurescein diacetate and Propidium Iodide) and mitochondrial function under epifluorescence microscopy were evaluated. As for motility and vigor, mitochondrial and acrosomal integrity, the buriti group was inferior to the control group. Tris supplemented with Mauritia flexuoxa oil at a concentration of 5% did not significantly influence sperm quality, however, favorable motility, morphology and membrane integrity were observed. Thus, being an alternative to replace diluents based on products of animal origin.

A randomized placebo-controlled trial to investigate the effect of lactolycopene on semen quality in healthy males.

PURPOSE: Poor sperm quality is a major contributor to infertility in heterosexual couples, but at present there are few empirical therapies. Several studies have examined the role of dietary factors and data from randomized controlled trials suggest that oral antioxidant therapy can improve some sperm parameters. Health benefits of lycopene supplementation have been proposed for a variety of health conditions and here we examine whether it can help improve sperm quality. This study aimed to investigate the effect of 14 mg daily lactolycopene for 12 weeks on semen quality in healthy men. METHODS: Sixty healthy male participants were recruited and randomized to this double-blind, placebo-controlled parallel study and received either 14 mg/d lactolycopene or a placebo for 12 weeks. The primary endpoint was a change in motile sperm concentration. Secondary endpoints were all other aspects of sperm quality, including the level of sperm DNA damage. RESULTS: Fifty-six men completed the intervention and the level of plasma lycopene was significantly increased in the men randomized to receive lycopene supplementation. There was no significant change in the primary endpoint (motile sperm concentration) post-intervention (p = 0.058). However, the proportion of fast progressive sperm (p = 0.006) and sperm with normal morphology (p < 0.001) did improve significantly in response to lactolycopene intervention. CONCLUSIONS: Supplementation with 14 mg/d lactolycopene improves sperm motility and morphology in young healthy men. CLINICAL TRIAL REGISTRY NUMBER AND WEBSITE: ISRCTN33248724 http://www.isrctn.com/ISRCTN33248724.

Individual male dependent improvement in post-thaw dromedary camel sperm quality after addition of catalase.

Cryopreservation is stressful to sperm cells inducing an increase in the production of reactive oxygen species and subsequently reducing post-thaw sperm quality. With the present study, there was evaluation of the protective effects of two antioxidants, epigallocatechin (1 mM) and catalase (500 IU/ml), added at thawing, as well as inter-individual variation on quality of cryopreserved dromedary camel spermatozoa. Semen was collected from six males and sperm, selected using single layer centrifugation, were cryopreserved. Post-thaw sperm quality was evaluated by assessing motility variables, viability and acrosome integrity then sperm were co-incubated with or without antioxidant (control) and further assessed at 1.5 and 3 h of the incubation period. Oxidative damage was measured colorimetrically for malondialdehyde production at 3 h of the incubation period. With the use of epigallocatechin there were not promising results, however, with use of catalase there were greater total and progressive motility, and values for some kinematic variables (P<0.05) at both incubation time points, although there were some differences among males. There was no overall effect of antioxidant based on production of malonaldehyde. The capacity of thawed sperm to fertilize, with and without addition of catalase at thawing, was studied using artificial insemination (n = 10 per treatment) with no differences between treatments (10% for both). It is concluded that catalase supplementations to semen extender prolong sperm survival, however, there is no improvement of in vivo fertilization as a result of this supplementation. There was an obvious male effect, necessitating further studies to understand the mechanisms of action of catalase.

Sodium alginate potentiates antioxidants, cryoprotection and antibacterial activities of egg yolk extender during semen cryopreservation in buffalo.

The study was conducted to determine effects of sodium alginate on sperm during cryopreservation. Each ejaculate (n = 20) of five buffalo bulls (3-5 years) were divided into six equal fractions and diluted using egg yolk based extender supplemented with different concentrations of sodium alginate and cryopreserved. Frozen-thawed semen samples were evaluated using the CASA, hypo-osmotic swelling test, cervical mucus penetration capacity test, and chlortetracycline fluorescence assay (CTC). Phosphorylation of tyrosine containing proteins and malondialdehyde concentration of sperm membrane were evaluated using immunoblotting and thiobarbituric acid reactive substance assay respectively. The semen extender's anioxidative capacities were estimated by conducting 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays, metal chelating capacity by assessing ferrozine and antibacterial capacity using agar plate methods. Supplementation of sodium alginate in extender improved sperm longevity, plasma membrane integrity as well as capacity to transit through the cervical mucus. Supplementation of extender with sodium alginate minimises the phase transition of sperm membranes and phosphorylation of tyrosine containing proteins during cryopreservation. Malondialdehyde concentration of sperm was less in sodium alginate-treated sperm as compared with control samples. The results indicated that sodium alginate increased antioxidant capacity of semen extender. Supplementation with sodium alginate also improved the metal chelating capacity and antibacterial properties of the extender. In conclusion, supplementation of extender with sodium alginate enhances free radical scavenging, metal reduction and chelating capacities to protect sperm during cryopreservation.

Relación entre los antioxidantes y la calidad seminal / Relationship between antioxidants and semen quality

Los suplementos dietarios tales como vitaminas, minerales y antioxidantes mejoran la ingesta de nutrientes. Recientemente se ha descrito que, especialmente aquellos que contienen altas propiedades antioxidantes también mejoran la capacidad fértil. Se presenta el caso de un voluntario de 37 años con posible infertilidad masculina y se desea determinar el efecto del consumo de antioxidantes sobre la calidad seminal. Se realizó evaluación de los parámetros seminales convencionales y funcionales antes y después del uso del suplemento dietario Male Fertility. Se observó que el uso del suplemento dietario incrementó la concentración espermática, el potencial de membrana mitocondrial alto y la capacidad antioxidante del semen; además disminuyó la producción de 1as reactivas de oxígeno, la lipoperoxidación y la fragmentación de la cromática espermática. El suplemento dietario Male Fertility contiene altas concentraciones de vitamina A, C, E, B2, B3, B12, folato, zinc, selenio, acetil L-carnitina, coenzima Q10, L-metionina y licopeno. Se ha descrito que la ingesta de cada uno de estos compuestos tiene efectos positivos sobre la calidad seminal. El reporte de este caso permitió observar que el uso de suplementos dietarios ricos en vitaminas y antioxidantes puede mejorar la calidad seminal a través de la disminución del efecto adverso de las especies reactivas del oxígeno y por el incremento de las moléculas antioxidantes en el plasma seminal(AU)

Dietary supplements such as vitamins, minerals and antioxidants improve nutrient intake. Recently it has been described that, especially those containing high antioxidant properties also improve fertility. We report here the case of a 37-year-old volunteer with possible male infertility and we want to determine the effect of antioxidant consumption on semen quality. Evaluation of the conventional and functional seminal parameters was performed before and after the use of the Male Fertility dietary supplement. The use of this supplement was observed to increased the sperm concentration, the mitochondrial membrane potential and the antioxidant capacity of the semen. In addition, the production of oxygen reactants, lipoperoxidation and fragmentation of the spermatic chromatin decreased. The dietary supplement Male Fertility contains high concentrations of vitamin A, C, E, B2, B3, B12, folate, zinc, selenium, acetyl L-carnitine, coenzyme Q10, L-methionine and lycopene. The ingestion of each of these compounds has been described to have positive effects on seminal quality. The report of this case allowed to observe that the use of dietary supplements rich in vitamins and antioxidants can improve the seminal quality through the decrease of the adverse effect of the reactive oxygen species and by the increase of the antioxidant molecules in the seminal plasma(AU)

Correlation between sperm motility and sperm chromatin/DNA damage before and after cryopreservation and the effect of folic acid and nicotinic acid on post-thaw sperm quality in normozoospermic men.

Cryopreservation exposes sperm to physical and chemical stresses causing cell damages and impairs sperm functions. The aim of this study was to evaluate the association between motility and sperm chromatin/DNA damage before and after cryopreservation and investigate the effects of folic acid and nicotinic acid on post-thaw sperm quality. Thirty semen samples were obtained from 30 normozoospermic men, aged between 25 and 45 years old. Each sample were divided into five aliquots to form the following groups: fresh, cryopreserved with sperm-freeze only (control), with nicotinic acid (10 mM), with folic acid (50 nM), and with a combination of folic acid (50 nM) + nicotinic acid (10 mM). Sperm viability and motility in each group were assessed by eosin-nigrosine staining and computer-aided sperm analysis respectively. Sperm chromatin quality was studied by aniline blue, toluidine blue, acridine orange staining methods and sperm chromatin dispersion test. Cryopreservation led to a significant reduction in sperm quality in comparison to fresh sample groups (p < 0.05). Sperm chromatin damage was negatively correlated with the percentage of progressively motile cells. Supplementation of the cryopreservation medium with folic acid or nicotinic acid induced a significant improvement in sperm parameters and chromatin quality, compared to control groups (p < 0.05). Meanwhile, the combination of folic acid + nicotinic acid showed a significant protective effect in post thaw sperm. In conclusion, cryopreservation generated oxidative stress, inducingsperm cryodamage, reducing progressive motility and sperm quality, as an indicator of significant chromatin/DNA damage. Folic acid and nicotinic acid exhibited a potential cryoprotective effect by enhancing sperm quality.

The incorporation of cystine by the soluble carrier family 7 member 11 (SLC7A11) is a component of the redox regulatory mechanism in stallion spermatozoa†.

Oxidative stress is considered a major mechanism causing sperm damage during cryopreservation and storage, and underlies male factor infertility. Currently, oxidative stress is no longer believed to be caused only by the overproduction of reactive oxygen species, but rather by the deregulation of redox signaling and control mechanisms. With this concept in mind, here, we describe for the first time the presence of the soluble carrier family 7 member 11 (SLC7A11) antiporter, which exchanges extracellular cystine (Cyss) for intracellular glutamate, in stallion spermatozoa, as well as its impact on sperm function using the specific inhibitor sulfasalazine. Spermatozoa incubated with Cyss exhibited an increased intracellular GSH content compared with controls (P < 0.01): 50% in fresh extended stallion spermatozoa and 30% in frozen-thawed spermatozoa. This effect was prevented by the addition of sulfasalazine to the media. Cystine supplementation also reduced the oxidation-reduction potential of spermatozoa, with sulfasalazine only preventing this effect on fresh spermatozoa that were incubated for 3 h at 37°C, but not in frozen-thawed spermatozoa. While sulfasalazine reduced the motility of frozen-thawed spermatozoa, it increased motility in fresh samples. The present findings provide new and relevant data on the mechanism regulating the redox status of spermatozoa and suggest that a different redox regulatory mechanism exists in cryopreserved spermatozoa, thus providing new clues to improve current cryopreservation technologies and treat male factor infertility.

Effects of egg yolk and soybean lecithin on sperm quality determined by computer-assisted sperm analysis and confocal laser scanning microscope in chilled canine sperm.

The reduction of spermatozoa survival time is a major problem of canine chilled sperm for artificial insemination. The aim of the study was to improve the quality of canine chilled sperm during storage time. We therefore, evaluated the effects of eight treatments with different levels of soybean lecithin concentration (1, 3 and 5%) and egg yolk (20%) in Tris-citric-fructose or Tris-citric-fructose-mineral salts extender on chilled canine sperm quality during 10 days of storage. The sperm motility was analysed by computer-assisted sperm analysis (CASA), whereas plasma membrane integrity, acrosome membrane integrity and mitochondrial membrane potential parameters were determined using a fluorescent staining combination of propidium iodide (PI), Hoechst 33342 (H342), fluorescein isothiocyanate-conjugated Pisum sativum agglutinin (FITC-PSA) and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) by confocal laser scanning microscope. The results showed that egg yolk was found to be better than soybean lecithin in Tris-citric-fructose or Tris-citric-fructose-mineral salts extender for maintaining the quality of chilled canine sperm within 10 days of storage (P < 0.05). Although egg yolk in Tris-citric-fructose extender could maintain the motility better than other extenders, egg yolk in Tris-citric-fructose-mineral salts extender was the highest in intact plasma membrane, intact acrosome membrane and high mitochondrial membrane potential (P < 0.05). In contrast, the sperm quality of soybean lecithin in Tris-citric-fructose-mineral salts extender was lower than that of soybean lecithin in Tris-citric-fructose extender, and soybean lecithin 1% was greater than soybean lecithin 3% and 5% in plasma membrane integrity, acrosome membrane integrity and mitochondrial membrane potential (P < 0.05). In conclusion, soybean lecithin cannot replace egg yolk in Tris-citric-fructose or Tris-citric-fructose-mineral salts extenders, and egg yolk in Tris-citric-fructose-mineral salts extender is superior to other extenders in chilling canine sperm.

Adverse Effects of Common Sports and Recreational Activities on Male Reproduction.

CONTEXT: Male factor infertility plays a significant role in infertility. Many factors have been associated with male infertility; however, the link between many sports and recreational factors and male reproduction remains poorly characterized. OBJECTIVE: To evaluate the current literature regarding the impact of many common sports and recreational factors on male reproduction. EVIDENCE ACQUISITION: A comprehensive PubMed and Embase search for relevant articles published between 1970 and 2017 was performed by combining the following search terms: male, sports (including individual sports), traumatic brain injury, sauna, hot tub, fertility, erectile dysfunction, varicocele, environment, cell phone, and laptop computer. EVIDENCE SYNTHESIS: Hypogonadism and erectile dysfunction can be associated with sports with high rates of head injuries, such as American football. Although early reports linked other sports, such as bicycling, to erectile dysfunction, subsequent studies isolated these associations to sports cycling rather than recreational cycling. Certain sports (football, basketball, handball, and volleyball) were linked to increasing prevalence and severity of varicocele, offering a potential link to male infertility. In addition, recreational activities such as sauna, hot tubs, Jacuzzis, heated car seats, and laptop use were associated with high testicular temperature, which can impair spermatogenesis. Radio frequency electromagnetic waves from cell phones and laptops have also been shown to have deleterious effects on sperm viability and motility. CONCLUSIONS: Many common sports and daily activities represent potential sources of male infertility. Clinicians should be aware of these associations in explaining idiopathic infertility in males. PATIENT SUMMARY: Male infertility is an often overlooked component of a couple's inability to conceive. We outline many common and often overlooked sports and recreational exposures that have been associated with male infertility.

Semen Uranium Concentrations in Depleted Uranium Exposed Gulf War Veterans: Correlations with Other Body Fluid Matrices.

Environmental metal exposure, as well as dietary metals, may adversely affect semen quality even as others play an essential role in normal spermatogenesis and fertility. Measures of seminal fluid metals have therefore been of high interest in the last several decades but have shown inconsistent results in correlations with some semen quality parameters. As well, environmental metal measures across various body fluid matrices have not been consistently correlated contrary to what one might hypothesize based on a systemic body burden of metal. This may be due to the body fluid matrices assessed and to other differences in laboratory methods and sample preparation. Measures of uranium, a potentially toxic metal in humans, have not previously been reported in the semen of environmentally metal-exposed populations. We report here uranium seminal fluid results and the high correlation of uranium concentrations across several body fluid matrices in a cohort of military veterans exposed to depleted uranium in combat events during the Iraqi Gulf War. These results inform the risk communication conversation for exposed populations and broaden the public health assessments from various exposure scenarios.

Hypospermia Improvement in Dogs Fed on a Nutraceutical Diet.

Male dog infertility may represent a serious concern in the canine breeding market. The aim of this clinical evaluation was to test the efficacy of a commercially available nutraceutical diet, enriched with Lepidium meyenii, Tribulus terrestris, L-carnitine, zinc, omega-3 (N-3) fatty acids, beta-carotene, vitamin E, and folic acid, in 28 male dogs suffering from infertility associated with hypospermia. All dogs received the diet over a period of 100 days. At the end of the evaluation period, no adverse effects, including head and tail anomalies percentage onset, were reported. Interestingly, motility percentage, semen volume and concentration, and total number of sperms per ejaculation significantly increased. Further investigations on a wider cohort of dogs might be useful to better correlate the presence of oxytetracycline in pet's diet and the onset of infertility and clearly assess the action mechanism of an oxytetracycline-free nutraceutical diet.

Supplementation of cryopreservation medium with TAT-Peroxiredoxin 2 fusion protein improves human sperm quality and function.

OBJECTIVE: To investigate the potential effects of TAT-PRDX2 protein supplementation to the cryopreservation medium on post-thaw sperm quality and function. DESIGN: In vitro prospective study. SETTING: Medical university hospital. PATIENT(S): Fifty normozoospermic, 50 asthenozoospermic, and 50 oligoasthenozoospermic men undergoing semen analysis for couple infertility. INTERVENTION(S): Each semen sample was divided into three aliquots: fresh, cryopreserved control (without additive), and cryopreserved with TAT-PRDX2 protein. MAIN OUTCOME MEASURE(S): Sperm motility, viability, mitochondrial potential, and DNA damage as well as reactive oxygen species (ROS) levels and lipid peroxidation were analyzed. Acrosome reaction and zona-free hamster oocyte penetration tests were performed to assess the fertilization ability of cryopreserved spermatozoa. RESULT(S): In normozoospermic and asthenozoospermic groups, the addition of 150 µg/mL TAT-PRDX2 significantly reduced intracellular ROS and malondialdehyde levels and enhanced post-thaw sperm motility and viability when compared with the cryopreserved control of the respective groups but did not produce any significant protective effect in the oligoasthenozoospermic group. Mitochondrial potential was significantly increased, whereas DNA fragmentation was significantly decreased, after TAT-PRDX2 supplementation only in the asthenozoospermic group when compared with the cryopreserved control. Although the penetration rate and the penetration index were not markedly improved, TAT-PRDX2 supplementation obviously reduced spontaneous acrosome reaction and increased calcium ionophore-induced acrosome reaction in the normozoospermic and asthenozoospermic groups. CONCLUSION(S): TAT-PRDX2 protein effectively exerted cryoprotective effects on spermatozoa by reducing intracellular ROS level and thereby improved post-thaw sperm quality and function, especially for asthenozoospermic samples. TAT-PRDX2 protein is a promising additive for developing a new and highly efficient semen cryoprotectant.

Diet and men's fertility: does diet affect sperm quality?

Male contribution to a couple's fecundity is important, and identifying the dietary factors that can influence male fertility potential is of high importance. Despite this importance, there are currently no clear clinical guidelines for male patients seeking fertility treatment. In this review, we present the most up-to-date evidence about diet and male fertility in humans. We focus on the dietary factors necessary for production of healthy functioning sperm with high fertility potential. Based on this review, men may be encouraged to use antioxidant supplements and to follow dietary patterns favoring the consumption of seafood, poultry, nuts, whole grains, fruits, and vegetables. Evidence is strongest for recommending the use of antioxidant supplements to men in couples undergoing infertility treatment-although the specific antioxidants and doses remain unclear-and increasing consumption of omega-3 fatty acids from fish and nuts.

Comparison of soy lecithin, coconut water, and coconut milk as substitutes for egg-yolk in semen cryodiluent for black rhinoceros (Diceros bicornis) and Indian rhinoceros (Rhinoceros unicornis).

Semen cryopreservation for the black rhinoceros (Diceros bicornis) and Indian rhinoceros (Rhinoceros unicornis) relies on extenders containing egg-yolk (EY). Use of such media is not ideal as inter-batch composition varies and there is risk of pathogenic contamination. The goal of this study was to test animal protein-free extenders. Semen collected via electroejaculation from 10 rhinoceros (6 black, 4 Indian) was diluted with extender containing EY, 1% or 2% soy lecithin (1%SL; 2%SL), coconut water (CW), or coconut milk (CM), cryopreserved and evaluated for sperm motility, viability, morphology, progression, and acrosomal integrity at 0, 1, 3, 6 and 24 h post-thaw. Mean ±â€¯SD fresh ejaculate motility was 84.5 ±â€¯7.6%, progression: 3.6 ±â€¯0.6 (scale 0-5), viability: 83.4 ±â€¯7.1%, intact acrosomes: 71.3 ±â€¯6.9%, and morphologically normal: 78.8 ±â€¯13.6%. Motility and progression decreased in all groups post-thaw, were greatest in EY, and decreased over time (P ≤ 0.05). Motility and progression did not differ (P > 0.05) between 1%SL and 2%SL, but were lower (P ≤ 0.05) in CM and CW, and acrosomal integrity was higher (P ≤ 0.05) in EY, 1%SL and 2%SL than in CM and CW. Post-thaw viability was greatest in EY and 2%SL followed by 1%SL, then CM and CW (P ≤ 0.05). Morphology did not differ among treatments (P > 0.05). Morphology, acrosomal integrity, and viability were maintained over time (P > 0.05). Although some rhinoceros sperm survived cryopreservation in SL treatments, reduced post-thaw motility rendered all treatments inadequate substitutes for EY-based extenders.

Selenium and vitamin E supplementation enhances the antioxidant status of spermatozoa and improves semen quality in male dogs with lowered fertility.

Studies showed a beneficial effect of supplementation with selenium (Se) and vitamin E on semen quality. The aim of the study was to investigate the effect of Se and vitamin E supplementation on the antioxidant status of spermatozoa and semen quality in dogs with lowered fertility. Ten dogs were supplemented daily with Se (6 µg/kg organic Se yeast) and vitamin E (5 mg/kg) per os for 60 days. Control group consisted of 10 males without the supplementation. Semen was collected on day 0, 30, 60 and 90. Sperm quality parameters were evaluated using CASA and a microscope. Concentrations of Se and vitamin E in blood as well as glutathione peroxidase (GSH-Px) activity and total antioxidant capacity (TAC) in the spermatozoa were determined. After 60 days of supplementation the concentration of spermatozoa, the majority of motility indicators and the percentage of normal morphology and live spermatozoa increased significantly (p < .05). An increase (p < .05) in concentration of Se and vitamin E in blood and GSH-Px-activity and TAC in the spermatozoa was detected. The study results indicate that Se and vitamin E supplementation for 60 days enhances the antioxidant status of spermatozoa and improves the quality of the semen in dogs with lowered fertility.