Seminal Plasma Concentrations Research Articles

Concentration of fatty acid binding protein as a new indicator of ejaculate fertility

Objective. To determine the marker function of fatty acid binding protein (FABP) concentration in seminal plasma (SP) in order to assess ejaculate fertility used in in vitro fertilization (IVF) treatment of infertility in couples. Materials and methods. The study involved semen samples of 96 men of reproductive age: the study group (n=63) – patients with a decrease in concentration and/or total content of spermatozoa in ejaculate, the comparison group (n=33) – men with normal concentration and number of spermatozoa in ejaculate. The content of FABP in SP was determined by enzyme-linked immunosorbent assay using the test system “FABP – ELISA – BEST” (A-9102, Vector-Best, Russia). In order to determine the informative value of using the concentration of FABP in SP as a criterion of ejaculate fertility of men of the study group, the predictive value of positive and negative IVF results was determined by calculating the diagnostic sensitivity, specificity and efficiency. Results. The content of FABP in SP accounted for 1.29 ± 0.24 ng/mL, the median and interquartile range comprised 1.23 [1.13–1.35] ng/mL, ranging from 1.08 to 2.79 ng/mL. The study revealed statistically significant intergroup differences (Mann-Whitney test U=79.00; p=0.000016), a weak correlation between the level of FABP and the concentration of spermatozoa (R=0.578008) and their number in ejaculate (R=0.583599). The diagnostic sensitivity of the test for FABP in SP of the men of the study group accounted for 81.82%, specificity – 78.95%, efficiency – 80.95%. Conclusion. Seminal plasma FABP can act as a marker of spermatogenesis disorders. The study of this protein in ejaculate provides the accuracy of predicting the outcome of in vitro fertilization.

Glyphosate presence in human sperm: First report and positive correlation with oxidative stress in an infertile French population

Environmental exposure to endocrine disruptors, such as pesticides, could contribute to a decline of human fertility. Glyphosate (GLY) is the main component of Glyphosate Based Herbicides (GBHs), which are the most commonly herbicides used in the world. Various animal model studies demonstrated its reprotoxicity. In Europe, GLY authorization in agriculture has been extended until 2034. Meanwhile the toxicity of GLY in humans is still in debate. The aims of our study were firstly to analyse the concentration of GLY and its main metabolite, amino-methyl-phosphonic acid (AMPA) by LC/MS-MS in the seminal and blood plasma in an infertile French men population (n=128). We secondly determined Total Antioxidant Status (TAS) and Total Oxidant Status (TOS) using commercial colorimetric kits and some oxidative stress biomarkers including malondialdehyde (MDA) and 8-hydroxy-2′-deoxyguanosine (8-OHdG) by ELISA assays. We next analysed potential correlations between GLY and oxidative stress biomarkers concentration and sperm parameters (sperm concentration, progressive speed, anormal forms). Here, we detected for the first time GLY in the human seminal plasma in significant proportions and we showed that its concentration was four times higher than those observed in blood plasma. At the opposite, AMPA was undetectable. We also observed a strong positive correlation between plasma blood GLY concentrations and plasma seminal GLY and 8-OHdG concentrations, the latter reflecting DNA impact. In addition, TOS, Oxidative Stress Index (OSI) (TOS/TAS), MDA blood and seminal plasma concentrations were significantly higher in men with glyphosate in blood and seminal plasma, respectively. Taken together, our results suggest a negative impact of GLY on the human reproductive health and possibly on his progeny. A precaution principle should be applied at the time of the actual discussion of GLY and GBHs formulants uses in Europe by the authorities.

Possible Impact of Human β-defensin 1 on sperm motility in infertile men with abnormal sperm parameters

Human β-defensins and interleukins may be auxiliary in sperm maturation. This cross-sectional study aimed to evaluate the expression of Human β-defensins 1 and 2, interleukins (ILs)− 10 and -18 genes in sperm, as well as seminal plasma levels of these two cytokines in subfertile men with different types of sperm abnormalities compared to those with normozoospermic men. Participants were separated into two experimental groups: the control group (n = 25) and the group with sperm abnormalities (SA) (n = 45). SA participants were further subdivided into the following groups with n = 15 individuals each: Teratozoospermia (T), Asthenoteratozoospermia (AT), and Oligoasthenoteratozoospermia (OAT) groups. The quantitative real-time polymerase chain reaction was used to quantify the mRNA levels of hBDs 1 and 2, IL-10, and IL-18 in sperm. The seminal plasma concentrations of IL-10 and IL-18 were measured by using the enzyme-linked immunosorbent assay technique. The mRNA expression of hBD-1 and IL-10 showed a significant decrease in the OAT compared to the controls (P < 0.0001 and P = 0.02, respectively). The lowest seminal plasma concentration of IL-10 belonged to the OAT (P = 0.04). ROC curve analysis showed a sensitivity, specificity, and cutoff value of 82.35%, 86.67%, and 0.63 for hBD-1 levels, respectively. A positive and significant correlation was found between hBD-1 expression and sperm motility and IL-10 expression rate and normal sperm morphology.Therefore, hBD-1 could be considered as the alternative biomaterial to pre-treatments of infertile men with abnormal sperm parameters, specifically OAT men, which led to improving the assisted reproduction success rate.

EFFECT OF SPERMS FREEZABILITY ON SOME SEMEN ATTRIBUTES AND AMINO ACID CONCENTRATIONS IN SEMINAL PLASMA OF HOLSTEIN BULLS

This study investigated the effect of different sperm freezability on some semen characteristics and amino acid concentrations in the seminal plasma of Holstein bulls cryopreserved for two years. The semen straws (0.25 ml) that cryopreserved for two years were divided into three equal groups (7 bulls per group) according to the sperm’s freezability: medium (73.02%; F1), good (83.76%; F2), and high (91.20%; F3). The F1 bulls were superior (P≤ 0.05) in sperm’s cell individual motility (SCIM) for fresh semen as compared with F2 and F3 bulls. The F2 and F3 groups were superior (P≤ 0.01) to the F1 group in live sperm percentage at cryopreservation. The sperm’s DNA damage percentage was lesser (P≤ 0.05) in the F1 and F3 groups as compared with the F2 group. The differences among groups in normal sperm percentage, sperm’s plasma membrane and acrosome integrity percentage, malondialdehyde, and total antioxidant concentrations lacked significance. The essential and non-essential amino acid concentrations in seminal plasma did not differ among groups. In conclusion, sperm freezability influenced some semen attributes but did not affect amino acid concentrations in the seminal plasma of Holstein bulls.

FRESH AND CRYOPRESERVED SEMEN CHARACTERISTICS OF BUFFALO BULLS AS INFLUENCED BY MELATONIN IMPLANTATION

This study explored the effect of melatonin implantation on the fresh and cryopreserved semen quality of Iraqi buffalo bulls (Bubalus bubalis). Nine adult bulls were used and divided equally into three groups. The first group was left without treatment and regarded as control (T1), whereas the second (T2) and third (T3) groups were subcutaneously implanted with 72 and 90 mg of melatonin respectively on the left ear base and repeated one month post the first implantation. Semen was collected for 12 weeks and evaluated weekly for fresh, cooling, and post-cryopreserved (PC) protocols. The T3 group exhibited a higher (P≤0.05) sperm cell individual motility percentage, whereas, the T1 and T2 groups recorded better (P≤0.05) sperm acrosome integrity percentage for fresh semen. A lesser (P≤0.05) total sperm abnormality (TSA) percentage in the T2 and T3 groups and a greater (P≤0.05) sperm acrosome integrity (SAI) percentage in the T3 group were noticed during the cooling preservation. Moreover, the T3 group exhibited a lesser (P≤0.05) TSA percentage whereas, the T2 and T3 groups revealed a leaser (P≤0.05) SAI percentage during the PC period. Concomitantly, lower (P≤0.05) PC malondialdehyde concentration in seminal plasma was observed in the T2 and T3 groups than control group. In conclusion, the melatonin implantation (72 and 90 mg) ameliorated some fresh and PC semen attributes of the Iraqi buffalo bulls.

Effects of the Neuropeptides Pituitary Adenylate Cyclase Activating Polypeptide and Vasoactive Intestinal Peptide in Male Fertility.

Background and Objectives: The neuroendocrine system plays a crucial role in regulating various bodily functions, including reproduction, with evidence suggesting its significant involvement in male fertility and sperm development. Vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase activating polypeptide (PACAP) are expressed in both male and female reproductive tissues, influencing penile erection and regulating steroidogenesis in males. Therefore, our study aimed to compare the protein levels of VIP and PACAP in seminal plasma between healthy controls and sub-fertile patients. Additionally, we sought to correlate the levels of these biomarkers with clinical, functional, and laboratory findings in the participants. Materials and Methods: The study included a total of 163 male participants for analysis. The participants were further stratified into subgroups of fertile and sub-fertile men of four subgroups according to the 2021 WHO guidelines. Seminal plasma concentrations of the neuropeptides VIP and PACAP were measured using human enzyme-linked immunosorbent assay technique. Results: The findings showed statistically significant differences in total sperm count, sperm concentration, total motility, and vitality (p < 0.001) between the fertile group and the sub-fertile group. Specifically, significant differences found between healthy males and oligoasthenospermic patients (p = 0.002), and between asthenospermic and oligoasthenospermic patients (p = 0.039). An ROC analysis showed associated sensitivity and specificity values of 62.2% and 55.6%, respectively, to PACAP seminal levels differentiated between sub-fertile patients from fertile males (p = 0.028). No significant difference in seminal levels of VIP was found between the sub-fertile and fertile groups. Conclusions: Previous research leads to the point of PACAP active involvement in spermatogenesis. In accordance to our study, in human semen samples, we have seen a significance change in PACAP levels amongst patients with low sperm count or with both low sperm count and low motility, hinting at its contribution and acting as a possible factor in this complex process. Thus, alterations in the levels or actions of these neuropeptides have been associated with certain reproductive disorders in males.

Oxidative stress on alpaca epididimal sperm frozen with different concentrations of seminal plasma

Objective. To assess the oxidative stress on alpaca epididimal sperm frozen with different concentration of seminal plasma. Materials and methods. 29 post mortem alpaca epididymis were used. Epididymal sperm were obteined thought cuts and suspended in a diluent based on skim milk, egg yolk and fructose and dimethylacetamine. Samples were separated into four aliquots which were supplied with seminal plasma (v/v) in concentrations of 0, 10, 25 and 50% respectively; then, sperm motility was assessed before freezing; then, motility was assessed before freezing with an automatic freezer machine. After thawing, mitochondrial membrane activity, sperm viability, lipid peroxidation and sperm apoptosis were assessed using flow cytometry with images. Results. Sperm exposed to 50% seminal plasma showed the lowest post-thaw motility (3.5±3.7%) and the highest percentages of apoptotic cells (71.43±20.6%). No significant differences were found in the rest of parameters. Conclusions. This study constitutes non-conclusive evidence that presence of seminal plasma does not affect viability, mitochondrial activity, or lipid peroxidation of alpaca epididymal sperm during cryopreservation processes; however, when it is absent or added up to 25% in the cryopreservation process, it may be possible to obtain better results in terms of sperm motility and sperm apoptosis.

Bisphenol A Negatively Impacts Human Sperm MicroRNA and Protein Profiles

AbstractBisphenol A (BPA), a widely prevalent endocrine disruptor, has detrimental effects on human health and male reproduction. Elevated BPA levels have been linked to reduced sperm quality and production, while paternal exposure may result in poor reproductive outcomes and offspring health. For many years, the effects of BPA exposure in human reproduction were mainly attributed to the deregulation of the hypothalamic–pituitary–gonadal (HPG) axis. In the past decades, other mechanisms of action have been proposed, and studies highlighting epigenetic alterations linked to BPA in sperm emerged. Nevertheless, the impact of BPA on human sperm miRNA and protein profiles and its implications for fertilization and post-fertilization events remain unexplored. This study aimed to investigate the association between BPA concentration in seminal plasma and seminal quality and to examine the alterations in human sperm miRNA and protein expression among men with varying BPA levels in seminal plasma. BPA levels were measured in seminal plasma from 102 Portuguese men, and it was detected in 88% of the samples. No correlation was observed between BPA levels and age or seminal parameters. The small RNA content of 15 normozoospermic samples was evaluated using small RNA sequencing, revealing that 15 miRNAs exhibited a correlation with BPA levels. Gene ontology analysis of their target genes indicated their involvement in embryonic development and response to stress. The proteome of 20 normozoospermic human sperm samples, categorized into four groups based on BPA levels, was accessed by liquid chromatography–mass spectrometry. Data are available via ProteomeXchange with the identifier PXD047280. Sixty-two differentially expressed proteins were identified across groups. The biological processes most affected in samples with higher BPA levels were “protein sumoylation” and “cytoplasmic translation”. The altered miRNAs and proteins identified in spermatozoa from men environmentally exposed to BPA represent potential biological markers of exposure to this endocrine disruptor and may help elucidate cases of idiopathic male infertility, fertilization failure, and abnormal embryo development.

Association of urinary and seminal plasma vanadium concentrations with semen quality: A repeated-measures study of 1135 healthy men

Although animal studies have shown the reproductive toxicity of vanadium, less is known about its effects on semen quality in humans. Among 1135 healthy men who were screened as potential semen donors, we investigated the relationships of semen quality with urinary and seminal plasma vanadium levels via inductively coupled plasma-mass spectrometry (ICP-MS). Spearman rank correlation tests and linear regression models were used to assess the correlations between average urinary and within-individual pooled seminal plasma vanadium concentrations (n = 1135). We utilized linear mixed-effects models to evaluate the associations of urinary and seminal plasma vanadium levels (n = 1135) with repeated sperm quality parameters (n = 5576). Seminal plasma vanadium concentrations were not significantly correlated with urinary vanadium concentrations (r = 0.03). After adjusting for possible confounders, we observed inverse relationships of within-individual pooled seminal plasma vanadium levels with total count, semen volume, and sperm concentration (all P values for trend < 0.05). Specifically, subjects in the highest (vs. lowest) tertile of seminal plasma vanadium concentrations had − 11.3% (−16.4%, −5.9%), − 11.1% (−19.1%, −2.4%), and − 20.9% (−29.0%, −11.8%) lower sperm volume, concentration, and total count, respectively; moreover, urinary vanadium levels appeared to be negatively associated with sperm motility. These relationships showed monotonically decreasing dose-response patterns in the restricted cubic spline analyses. Our results demonstrated a poor correlation between urinary and seminal plasma levels of vanadium, and elevated vanadium concentrations in urine and seminal plasma may be adversely related to male semen quality.

Using Bayesian and weighted regression to evaluate the association of idiopathic oligoastenoteratozoospermia with seminal plasma metal mixtures

Idiopathic oligoastenoteratozoospermia (iOAT) affects 30% of infertile men of reproductive age. However, the associations between Cr, Fe, Cu, Se or Co levels and iOAT risk have not been determined. This research aimed to assess the associations between Cr, Fe, Cu, Se and Co levels as well as their mixtures in seminal plasma and the risk of iOAT and severe iOAT. Therefore, a case‒control study including 823 participants (416 iOAT patients and 407 controls) recruited from October 2021 to August 2022 at the reproductive medicine center of the First Affiliated Hospital of Anhui Medical University was conducted in Anhui, China. The concentrations of Cr, Fe, Cu, Se and Co in seminal plasma were detected via inductively coupled plasma‒mass spectrometry. Binary logistic regression models were used to assess the associations between the levels of Cr, Fe, Cu, Se and Co and the risk of iOAT and severe iOAT; additionally, Bayesian kernel machine regression (BKMR) and weighted quantile sum (WQS) regressions were performed to evaluate the joint effect of seminal plasma levels of Cr, Fe, Cu, Se and Co on the risk of iOAT and explore which elements contributed most to the relationship. We found significant associations between the concentrations of Fe, Cu and Se in seminal plasma and iOAT risk after adjusting for covariates (Fe, lowest tertile vs. second tertile: aOR = 1.86, 95% CI = 1.31, 2.64; Cu, lowest tertile vs. second tertile: aOR = 1.95, 95% CI = 1.37, 2.76; Se, lowest tertile vs. second tertile: aOR = 1.65, 95% CI = 1.17, 2.35). A lower Se concentration in seminal plasma (lowest tertile vs. second tertile: aOR = 1.84, 95% CI = 1.10, 3.10) was positively associated with the risk of severe iOAT. Additionally, we also observed an association between the concentration of Cr in seminal plasma and the risk of iOAT before adjusting for covariates (Cr, third tertile vs. lowest tertile: OR=1.44, 95% CI: 1.03, 2.02). According to the BKMR analyses, the risk of iOAT increased when the overall concentrations were less than the 25th percentile. The results from the WQS regression indicated that a negative WQS index was significantly associated with the iOAT risk, while a positive WQS index was not. Se and Fe had significant weights in the negative direction. In conclusion, lower Cu, Fe and Se levels in seminal plasma were positively associated with iOAT risk, while higher Cr levels in seminal plasma were positively associated with iOAT risk according to the single element model, and lower levels of Se were related to a greater risk of severe iOAT; when comprehensively considering all the results from BKMR and WQS regression, Fe, Se and Cr levels contributed most to this relationship.

Seminal Plasma Bisphenol A Concentrations Among Men with Secondary and Primary Infertility

Background: The declining trend of male infertility have been attributed to some factors including environmental, lifestyle behaviors, and dietary habits. Bisphenol A (BPA) which is preponderance in the environment and in various products frequently used has been implicated in male infertility. It is not known whether the seminal fluid BPA concentrations differ among men with primary infertility and secondary infertility. This study was aimed at determining the seminal plasma BPA concentrations among men with secondary infertility and primary infertility and to associated their levels with duration of infertility. Materials and Methods: This was a cross‑sectional study of 145 men clinically diagnosed with primary infertility (n=80) and secondary infertility (n=65), and men with proven fertility (n=60) as controls. Semen analysis was done manually according World Health Organization guidelines and seminal plasma BPA was assayed using an Elisa kit. Chi square and Student’s t-test were used to analyze continuous data and discrete variables respectively. Pearson’s correlation coefficient was used to determine the association between seminal plasma BPA and duration of infertility, and a p&lt;0.05 was considered statistically significant. Results: The seminal plasma BPA was significantly higher (p&lt;0.001) among subjects with secondary infertility than those with primary infertility. Sperm count, total motility, progressive motility and normal morphology were lower among secondary infertility than primary infertility, but the mean differences were not significant (p&gt;0.05) except viability which was significantly higher among primary infertility than secondary infertility (p&lt;0.05). The multiple regression model indicates that, the ages of men (OR=3.26 95% CI 2.23-7.26) was independently associated with BPA concentrations among secondary infertility. The finding of higher BPA concentrations in seminal plasma of men with secondary infertility than primary infertility may be associated with age and not duration of marriage.

Can evaluation of seminal plasma and serum anti-Müllerian hormone be useful in the diagnosis of male infertility? (Literature review)

Background. Anti-Müllerian hormone (AMH) is secreted in men by Sertoli cells and plays an important role in the development of the male reproductive organs. High expression of AMH during embryonic development contributes to the regression of the Müllerian (paramesonephric) ducts; in the absence of AMH, the Müllerian ducts develop into the female internal genital organs. In adulthood, the clinical value of measuring AMH in serum and seminal plasma remains controversial. It is assumed that AMH, as a marker of the functional state of Sertoli cells, can be useful in the diagnosis of male infertility.Aim. To determine the clinical value of measuring serum and seminal plasma AMH levels in the diagnosis of male infertility by examining the current evidence base in the world literature.Materials and methods. A search was made for publications devoted to the study of the role of AMH in the diagnosis of male infertility in the international database PubMed, in the scientific electronic library eLIBRARY.RU and in the Google Scholar search engine. There were no restrictions regarding the status and period of publication.Results and conclusion. The results of the analysis indicate the absence of a significant relationship between the parameters of the ejaculate and the serum concentration of AMH, which does not allow using this indicator to assess male fertility. Serum AMH is of no value in the differential diagnosis of obstructive and non-obstructive azoospermia, the prediction of assisted reproductive technologies outcomes, and the prediction of testicular biopsy outcomes in the general non-obstructive azoospermia patient population. It is likely that serum AMH may be useful in predicting testicular biopsy results in patients with Klinefelter’s syndrome and idiopathic non-obstructive azoospermia.The clinical application of measuring the level of AMH in seminal plasma is limited by the high variability of this indicator, probably due to the dependence on the volume of the ejaculate and exposure to semen proteolytic enzymes. This indicator may be useful for the differential diagnosis of obstructive and non-obstructive azoospermia. In obstructive azoospermia, AMH is not detected in seminal plasma, but undetectable AMH concentrations can also be observed in patients with non-obstructive azoospermia, which reduces the differential value of this indicator. A promising direction is the use of AMH concentration in seminal plasma to predict the recovery of sperm motility after cryopreservation.

Inverse relationship between transfer efficiencies into reproductive system and exposure concentration for organic pollutants: Implications for hazard assessment

Organic pollutants impact fertility and reproductive outcomes. Assessing their risk required understanding the great variation of their parental transfer efficiency. To determine whether the proportion of organic pollutants transferred to the reproductive system varies with the level of parental contamination, we analyzed large-scale datasets on organic environmental contaminants found in parental blood, cord blood, and semen. Our results demonstrated that although chemical concentrations in seminal plasma and cord blood significantly increased with those in parental blood, the proportion of parental contaminants transferred to the reproductive system decreased as the chemical concentration in parental blood increased. Using a hierarchical model, a negative association between transfer efficiencies and parental contamination levels was revealed. It is estimated that 41.78% and 46.23% of the variance in transfer efficiencies can be explained by paternal and maternal blood concentrations, respectively. In comparison, chemical class accounted for 25.22% and 18.95% of the variance, while the remaining inconsistencies between studies contributed only 3.38% and 2.56% of the variance for paternal and maternal transfers, respectively. We observed no significant interaction between parental blood concentration and chemical properties. Thus, our findings implied that the transfer efficiency of organic pollutants is not just an intrinsic chemical property and the inverse association has significant implications for health risk assessments and benchmark dose estimations related to environmental pollutants.

Effect of Electroejaculation Protocols on Semen Quality and Concentrations of Testosterone, Cortisol, Malondialdehyde, and Creatine Kinase in Captive Bengal Tigers

The Bengal tiger (Panthera tigris tigris) is critically endangered, so assisted reproductive technologies, including artificial insemination, are important conservation tools. For wild and domestic felids, electroejaculation (EE) is the most common semen collection method, with protocols optimized to obtain sufficient amounts of viable sperm for artificial insemination. However, less attention has been paid to ensuring animal wellbeing during the process. This study examined the effects of three EE protocols (Low, 2–5 volts; Medium, 3–6 volts; High, 4–7 volts) on semen quality, testicular size, serum testosterone, creatine kinase (CK), and malondialdehyde (MDA) concentrations, and serum cortisol as a proxy for stress. Blood samples were collected before, during, and after each EE series. Seminal plasma pH, and sperm motility, viability, and morphology were evaluated after each procedure. Seminal plasma and sperm pellet MDA concentrations were also determined. Primary sperm abnormalities and seminal plasma MDA were higher in the Low compared to Medium and High voltage groups (p &lt; 0.05). Serum CK in the High voltage group increased during the EE series (p &lt; 0.05), suggesting the potential for muscle damage. However, no significant changes were observed for serum cortisol, testosterone, or MDA concentrations. Results suggest the Medium voltage protocol produced good quality samples at lower voltages than the High protocol with no negative effect on muscle function, which might be better for animal welfare.

Effect of post-thaw addition of seminal plasma to cryopreserved epididymal equine spermatozoa

Seminal plasma (SP) has been reported to increase the immediate motility of frozen-thawed epididymal sperm and appears to be important for plasma membrane remodeling, capacitation, acrosomal reaction and fertilization. The objective of this study was to evaluate the effect of incubating equine epididymal frozen-thawed sperm with different concentrations of seminal plasma. Sperm samples were obtained from testes after castration of 40 young Silla Argentino stallions (24 to 36 months of age). Testicles were cooled (5°C) and after 24 h, the cauda epididymis were dissected and flushed (retrograde) with Kenney extender. Epididymal sperm were cryopreserved in EDTA-glucose with 5% DMF, 11% lactose, 20% egg-yolk and Equex, using a rapid temperature-descent curve. The samples were thawed at 37°C for 1 min and then incubated 10 minutes with different concentrations of SP (10%, 50%, 100%) vs. a control (0% SP). To create a pool of SP, Silla Argentino stallions of proven fertility were used. Seminal plasma was separated and stored at -18 °C until use. Semen parameters evaluated were sperm motility (Computer Assisted Semen Analysis; ISAS v1, Proiser), morphology (Diff Quick stain), viability and membrane function (combined hypoosmotic test and Coomassie blue stain). Data were analyzed using Kruskal Wallis test with P<0.05considered significant. Post-thaw incubation with increasing concentrations of seminal plasma significantly decreased sperm total and progressive motility (0%: 33.06 ± 2.02; 26.71 ± 1.90; 10%: 24.06 ± 1.74; 18.85 ± 1.59; 50%: 19.44 ± 1.71; 14.92 ± 1.53; 100%: 14.97 ± 1.57; 11.66 ± 1.48 respectively. Values are mean +/- SEM). However, the values of Beat-Cross Frequency, Linearity and Straightness were significantly higher when the spermatozoa were incubated with SP (BCF: 0%: 10.24 ± 1.47a; 10%: 10.67 ± 3.13a,b; 50%: 10.63 ± 2.31b; 100%: 11.45 ± 2.73b; STR: 0%: 0.59 ± 0.07a; 10%: 0.62 ± 0.09a,b; 50%: 0.64 ± 0.09b; 100%: 0.64 ± 0.12b; LIN: 0%: 0.23 ± 0.05a; 10%: 0.25 ± 0.06b; 50%: 0.26 ± 0.07b; 100%: 0.26 ± 0.07b). There was no effect on membrane function, acrosome status and sperm morphology, regardless of the concentration of SP used (P>0.05). The results indicate that there is no evidence of a beneficial effect of post-thaw supplementation with SP at the concentrations used in this study. It has to be considered that the main role of SP is not associated with sperm physiology but as a modulator in the female reproductive tract. Hence we are continuing in vivo studies to assess whether SP addition can improve pregnancy rates after artificial insemination with frozen-thawed epididymal sperm.

A different approach to the quantification of human seminal plasma metabolites using high-resolution NMR spectroscopy

In this study, a reliable method was established for the absolute quantification of metabolite concentrations in human seminal plasma using ERETIC2, a quantification tool developed by Bruker based on the PULCON principle. The performance of the ERETIC2 was examined using an AVANCE III HD NMR spectrometer (600 MHz) equipped with a triple inverse 1.7 mm TXI probe in terms of some experimental parameters that may affect the accuracy and precision of the quantitative results. Then, the accuracy, precision, and repeatibility of ERETIC2 were determined using L-asparagine solutions at different concentrations. And it was evaluated by comparing it with the classical internal standard (IS) quantification method. The relative standard deviation (RSD) values for ERETIC2 were calculated in the range of 0.55–1.90% and the minimum recovery value was 99.9%, while the RSD values for the IS method were calculated in the range of 0.88–5.83% and recovery value was minimum 91.0%. Besides, the RSD values of the inter-day precisions for the ERETIC2 and IS methods were obtained to be in the range of 1.25 − 3.03% and 0.97 − 3.46%, respectively. Finally, the concentration values of seminal plasma metabolites were determined using different pulse programs with both methods for samples obtained from normozoospermic control and azoospermic patient groups. The results proved that this quantification method developed using NMR spectroscopy is easy to use in complex sample systems such as biological fluids and is a good alternative to the classical internal standard method in terms of accuracy and sensitivity. In addition, the improvement of the spectral resolution and sensitivity with the microcoil probe technology and the possibility of analyzing with minimum sample quantities has contributed positively to the results of this method.

Effect of Adding Different Levels from Melatonin in Some Characteristics Semen of Broiler Breeder Male Ross308

The aim of the present study was to evaluate the effect of adding different levels of melatonin in some of semen Characteristics and antioxidant status in male broiler breeder Ross308, from16/11/2017 to 30/1/2018.Atotal of 25 bird of Ross 308 broiler breeder males ,30 weeks' old were used in this study. The birds were randomly distributed into five group with five replicates each. Each treatment group constituted of 5 birds (1bird each replicate). the broiler breeder males administrated orally with capsules contain melatonin. Treatment groups were as following: T1: Birds fed the basal diet without any addition (control),T2: Birds fed diet supplemented with 15mg/kg of diet,T3: Birds fed diet supplemented with 30 mg/kg of diet,T4: Birds fed diet supplemented with 40 mg/kg of diet and T5: Birds fed diet supplemented with 60 mg/kg of diet. The result of the study revealed a significant decrease in the ejaculated volume, sperm's mass and individual motility and sperm concentration, With a significant increase in dead and abnormal sperm ratio. However, administration with melatonin at (60) mg/kg diet resulted in significant increase (p&lt;0.05) in glucose and total protein concentration of seminal plasma. Also adding melatonin with (15) mg/kg diet had significant improved in the antioxidant status. &#x0D; &#x0D;

Metabolomic and Transcriptomic Changes Underlying the Effects of L-Citrulline Supplementation on Ram Semen Quality.

This study examined the effects of L-Cit supplementation on ram semen quality through metabolomics and transcriptomics. A total of 16 rams were randomly categorized into two groups. The control group was fed a basic diet, whereas the experimental group received feed supplemented with 12 g/d of L-Cit. Semen and blood were collected from the rams on days 0 and 72 to measure sugar, pyruvate, amino acid, and nontargeted metabolite contents. Additionally, hypothalamic and testicular tissues were collected for a transcriptomic analysis. We found 27 differential metabolites between the control and experimental groups, of which 21 were downregulated (p &lt; 0.05) and 6 were upregulated (p &lt; 0.05). Compared with the control group, xylose and pyruvate contents in seminal plasma increased by 43.86% and 162.71%, respectively (p &lt; 0.01). Additionally, the levels of 11 amino acids showed a significant increase in seminal plasma (p &lt; 0.01). Furthermore, 961 and 715 differentially expressed genes were detected in the hypothalamic and testicular tissues, respectively. The pathways of significant enrichment in the hypothalamus and testes were protein digestion, absorption, glycolysis/gluconeogenesis, and amino as well as nucleotide sugar metabolisms. In the present study, L-Cit improved protein synthesis and blood metabolism, consequently increasing the contents of most amino acids in ram seminal plasma. Specifically, the hypothalamus controlled the expression of glycolysis/gluconeogenesis-related genes in the testes through its metabolites released into the serum, thereby providing energy for sperm production, which led to a decrease in the sugar content of seminal plasma.

The association between reactive oxygen species and α-tocopherol treatment in a sample of infertile Iraq male

Male infertility estimated about 20-70% of couples or >30 million men worldwide. The goal of this study was to elucidate the interrelations between oxidative stress (O.S) in seminal fluid and antioxidant inducing disruption of spermatogenesis. Twenty four infertile patients aged (23-41) years were involved in this study. Which were received α- tocopherol (vitamin E). This group (vit E group) was sub classified according to the WHO criteria (2010) into three different subgroups: oligoasthenoteratozoospermia (OAT) Asthenoteratozoo- spermia (A.T), and Asthenozoospermia (Astheno.). The control group consist of samples obtained from healthy donors of proven fertility (n=23). The main parameters were determined in seminal fluid of infertile patients and healthy men are the sperm and seminal plasma Malondialdehyde (MDA) concentration, and vitamin E concentration in seminal plasma. The results showed that, there was a high significant decrease (p≤0.001) in sperm parameters (concentration, progressive motility and normal morphology), and vitamin E concentration in infertile group compared with that fertile group.After analyzing the statistically difference in the different measured parameters between the vit E group after treatment, in comparison with that of before treatment; the following was observed A high significant increase (p≤0.001) were found in progressive motility, sperm [MDA] (MDAp), vitamin E concentration, and significant increase (P ≤0.05) in the sperm concentration. From all of these observations, it can be concluded that O.S contributes most likely to sperm damage and may be responsible for poor seminal fluid characteristic of the infertile patients involved in the present study. Antioxidants α-tocopherol (vitamin E) plays an important role in the improvement of seminal fluid parameters.

Evaluation of metabolomic profiling and oxidative stress markers of bull seminal plasma and their relationship with sperm motility before and after thawing

Mechanisms underlying differentiation of good or poor freezability ejaculates in bull are still poorly understood. This study aims at investigating oxidative stress parameters and metabolomic profiling of seminal plasma (SP) in relation to bull semen motility before and after cryopreservation. Semen samples were collected by artificial vagina from 20 bulls aged from 10 to 12 mo. An aliquot of semen was centrifuged to obtain SP (stored at -80°C), another was diluted 1:1 with a pre-warmed extender and cryopreserved. Motility assessment (CASA) was performed on fresh and thawed semen (0 and 3 h after thawing). Semen samples were divided into good freezers (GF) and bad freezers (BF) groups according to thawed semen motility, with cutoff values as follows: TM ≥ 60% and PM ≥ 30%. SP was used for oxidative stress markers analysis by spectroscopy techniques and metabolomic profile by nuclear magnetic resonance (NMR). The relationship between NMR spectra and semen quality was evaluated by the PCA/PLS-DA analysis. General linear models and mixed linear models were used to evaluate the relationship between seminal plasma oxidative stress parameters and semen quality. A clear separation between GF and BF ejaculates was observed on the first two principal components of the variance (PC1 vs PC2) of NMR spectra of SP. The increase of advanced oxidative protein products (AOPP) and thiols concentrations on SP were significantly related with higher straightness (STR), beat cross frequency (BCF) and linearity (LIN) and the decrease of amplitude of lateral head displacement (ALH) of fresh semen. Thiols and AOPP concentrations on SP were positively related to semen freezability. In addition, higher AOPP concentration was significantly related to higher TM, PM and LIN of thawed spermatozoa, whereas higher levels of thiols in SP concentration were positively related to the increment of sperm PM after thawing. These findings suggested that SP composition can modulate bull semen freezabilty. Further analyses could elucidate which metabolites and oxidative stress parameters could be used as biomarkers of good semen quality after cryopreservation.