Seminal Plasma Concentrations Research Articles

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.

Seminal plasma level of bisphenol A among males investigated for infertility in Benin City, Nigeria

Background: Evidence of an increasing rate of male infertility is replete in literature and has been attributed to several factors associated with environmental toxicants such as bisphenol A (BPA). It's presence in several products frequently used and the potential harmful effect on humans has raised serious concerns. Aims: The objective of this study was to determine the seminal plasma concentration of BPA among men investigated for infertility. Setting and Design: This was a cross-sectional study of men investigated for infertility in a tertiary hospital in Benin City, Nigeria. Materials and Methods: A total of 144 males (mean age: 40.55 ± 7.30) and 50 control subjects of proven fertility (mean age: 39.96 ± 5.68) were enrolled in the study. A structured questionnaire was used to obtain the sociodemographic data. Semen was collected after at least 3 days of sexual abstinence. Semen analysis was done according to the World Health Organization guidelines and seminal plasma BPA was determined using the enzyme-linked immunosorbent assay technique. Statistical Analysis Used: Chi-square, Student's t-test, and analysis of variance were used to compare measured variables. Pearson's correlation coefficient was used to determine the relationship between seminal plasma BPA and sperm indices. Results: Seminal plasma BPA was significantly higher (P &lt; 0.001) among infertile men than control subjects. The mean seminal plasma BPA concentrations increased with declining sperm count (P &lt; 0.001). Seminal plasma BPA levels strongly correlated inversely with sperm motility (r = −0.857, P &lt; 0.001) and sperm count (r = −0.821, P &lt; 0.001) and positively with abnormal morphology (r = 0.815, P &lt; 0.001). The likelihood ratios of elevated BPA concentration to cause harm to sperm count (14.562), motility (10.425), and morphology (27.838) were several folds higher among subjects being investigated for infertility. Conclusions: Seminal plasma BPA levels were significantly higher among infertile men than controls. The detection of high levels of BPA in the seminal plasma is an indication of human exposure to high concentrations of BPA, this information may be used to formulate environmental policy that may minimize human exposure and improve male reproduction.

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.

Fosfomycin Pharmacokinetic Profile in Plasma and Urine and Quantitative Estimation in Prostate and Seminal Vesicles after One and Two Consecutive Doses of Oral Fosfomycin Trometamol in Healthy Male Volunteers.

The present Phase I study investigated, for the first time, fosfomycin pharmacokinetics in humans after two 3 g doses of fosfomycin trometamol administered 27 h apart, according to the dose regimen recommended for the prophylactic indication for transrectal prostate biopsy in adult men. Plasma, urine and seminal plasma concentrations were measured after one and two consecutive doses in 24 healthy men, representative of the target population of the prophylactic indication. Prostate and seminal vesicle concentrations were estimated based on seminal plasma concentrations using a one-step regression method. The exposure to fosfomycin was very similar in rate (Cmax, tmax) after one and two doses. The AUC showed a minimal increment. On average, the apparent volume of distribution was high (>100 L), and the mean clearance had an intermediate value. The total amount and dose fraction of fosfomycin excreted in urine showed a small increment after two doses. The renal clearance was about 5 L/h. The fosfomycin concentration in the prostate and seminal vesicles showed that the antibiotic increased on average after two consecutive doses. This result confirmed the ability of fosfomycin to distribute into the prostate and into seminal vesicles after one single dose and that a two consecutive dose regimen increases the antibiotic availability inside these peripheral tissues.

Oxidative stress and iron metabolism in human sperm

Infertility is a global problem in couples. The decrease in sperm quality and function might be related to intrinsic and extrinsic factors, which ultimately converge on inducing oxidative stress (OS) in male gametes. Seminal plasma (SP) contains a large amount of antioxidants which protect sperm against OS. The iron in semen has a great influence on male fertility. Its transport, celluar uptake and storage depend on Transferrin (Tf), the cell membrane receptor for Transferrin (RTf) and Ferritin (FN), respectively. An isoform found in SP, testicular transferrin (TfT), is essential for spermatogenesis. The objective was to evaluate the antioxidant capacity of metalloproteins in SP involved in iron metabolism. A total of 328 semen samples from patients from the Urology Service of Eva Perón School Hospital (Granadero Baigorria) and Centenario Hospital (Rosario) (n=166) and healthy volunteers (n=162) were studied. Of these participants, 204 met the inclusion criteria. Semen samples were analyzed according to WHO criteria (2010-2021). Measurement of Fe and total proteins was carried out by colorimetric method, FN was measured by immunoturbidimetric method. ELISA technique was used for measuring the Tf concentration in SP. To measure OS we used 3 techniques: MOST test, (TBARs) technique and determination of total nitrite production by colorimetric method. Concentration of nitrites vs TfT showed an inverse relationship (R2=0,921; M= -0,7057; p<0,05). For abnormal MOST values (less than or equal to 0.39), the seminal Tf values were mostly below the mean value found in our population (5 mg/dL). An inverse relationship was seen between lipid peroxidation and TfT. (R2=0,4869; M=-0,0028). Our data suggest that TfT may fulfill an antioxidant role in the testis. It is necessary to study a greater number of patients and discriminate them by pathology in order to conclude which OS measurement technique is the most appropriate to assess male reproductive function.

Relation of seminal plasma trace mineral in the Arabian stallion’s semen with the semen characteristics and subsequent fertility

BackgroundSeminal plasma contains several microelements like Zn, Fe, Se, and Cu that affect sperm motility and male fertility. Biochemical evaluation of seminal plasma trace elements is important for assessing fertility and diagnosing male infertility. AimsThe present study was designed to evaluate the effect of seminal fluid trace elements on sperm parameters and fertility in Arabian horses. MethodsNinety-four ejaculates from 25 Arabian stallions (4–27 years old) were used to investigate the effect of seminal fluid trace elements on semen parameters and fertility. Data divided according to season, stallion age, and fertility of stallions. The concentrations of Zn, Fe, Se, Cu, Cr and Mo were determined using an atomic absorption spectrophotometer. Percentage stallion fertility estimated by mares that conceived on their first cycle. Data were analyzed by ANOVA using SPSS statistical software program (2013), version 22.0. ResultsThere was a significant effect of season on semen volume, pH, Fe, Se, Cu, Cr, and Mo. Stallion age had a significant effect on pH, sperm motility, concentration, total motile sperm count, sperm abnormalities, Zn, and Fe. Sperm motility was higher (P < 0.05) and sperm abnormalities were lower (P < 0.05) in group IV (>70% fertility) than in group I (infertile) and group II (<50% fertility). Sperm abnormalities were low in group IV and high in groups I and II. Seminal plasma Zn and Cu levels were higher (P < 0.05) in groups III (50_70% fertility) and IV than in group I. Fe levels were lower (P < 0.05) in group IV than in groups I, II, and III. Seminal plasma Mo concentrations were higher (P < 0.05) in group III than in group I. ConclusionsHigh seminal plasma concentrations of Zn, Se, Cu, and Mo and low Fe concentrations are associated with improved stallions' semen parameters and fertility.

Linseed oil in boar's diet improved in vivo fertility and antioxidant status.

The reactive oxygen species (ROS) which are produced during storage of boar semen are causing oxidative stress and leads to poor fertility. Also, tropical and sub-tropical weather condition adversely impacts the physicomorphological quality and fertility of boar sperm. The aim of this study was to examine the effects of feeding linseed oil to boar on its seminal attributes, sperm kinetics, biomarkers of antioxidant, fatty acid profile of seminal plasma (SP) and sperm and in vivo fertility. Six Hampshire crossbreed boars were fed with 90 ml linseed oil (LIN) whereas six Hampshire crossbreed boars were fed 90 ml canola oil (CON) for 16 weeks. Sperm quality was evaluated (60 ejaculates for each group; a total of 120 ejaculates) for motility, livability, abnormal morphology, acrosomal membrane integrity, hypo-osmotic swelling test (HOST) and sperm kinetic parameters by computer assisted semen analysis (CASA) at 0h and at 72 h of storage at 17°C. Biomarkers of antioxidant (glutathione peroxidase; GPx, catalase; CAT, total antioxidant capacity; TAC) and malondialdehyde (MDA) were measured in SP and serum. Gas chromatography-mass spectrometry (GC-MS) was used for the estimation of fatty acid composition of SP and sperm. Boars fed with linseed oil had higher semen volume (p < .01) and more total sperm numbers (p < .01). Feeding linseed oil to boar enhanced seminal attributes (p < .05) at 0h as well as at 72 h of storage. Linseed oil feeding (p < .01) improved biomarkers of antioxidants and significantly (p < .01) lowered the lipid peroxidation in serum and SP. Linseed oil feeding (p < .05) increased the proportion of alpha linolenic (ALA), arachidonic and docosahexaenoic (DHA) fatty acids in SP. The ratio of n-6 to n-3 fatty acids in sperm increased significantly (p < .01) in treatment group. Farrowing rate was significantly (p < .05) higher in treatment group. In conclusion, feeding linseed oil to boar improved the in vivo fertility, enhanced antioxidant capacity and increased the DHA content of SP and sperm.

Cryoprotection of Humanin-like Peptides in Seminal Plasma for Ejaculated Spermatozoa of Crossbred Bulls

BACKGROUND: Cryopreservation process negatively affects spermatozoa functions. Humanin, a small polypeptide encoded in the mitochondrial genome, is well known for its role in cell survival. OBJECTIVE: To quantify the endogenous levels of humanin in seminal plasma of crossbred Frieswal bulls and to study its role in cryoprotection. The presence of humanin in bull spermatozoa was also investigated. MATERIALS AND METHODS: A total of 40 semen samples were separated into two groups based on the initial progressive motility (IPM): Good (IPM ≥70%) and Poor ( IPM ≤50%) groups; and/or based on the post-thaw motility (PTM): Freezable (PTM≥50%) and Non-freezable (PTM &lt; 50%) groups. Humanin concentration in seminal plasma (SP-HN) was quantified using ELISA. RESULTS: SP-HN concentration ranged from undetectable to 67.6 pg/mL with a median level of 35.2 pg/mL. SP-HN level was significantly higher in the good quality semen group than in the poor quality semen group (p&lt;0.001), and also significantly higher in the freezable group than in the non-freezable group (p&lt;0.001). SP-HN level was positively correlated with initial progressive motility, post-thaw semen motility, viability, acrosome intactness and plasma membrane integrity, but negatively correlated the level of reactive oxygen species and malondialdehyde content. Immunochemical localization showed the presence of humanin in the proximal region of the middle piece of spermatozoa. CONCLUSION: Endogenous humanin level had significant correlation with semen quality and might protect sperm cells against freeze-induced oxidative stress.

Utilization of Aminoguanidine Prevents Cytotoxic Effects of Semen.

Studies of human semen in cell or tissue culture are hampered by the high cytotoxic activity of this body fluid. The components responsible for the cell damaging activity of semen are amine oxidases, which convert abundant polyamines, such as spermine or spermidine in seminal plasma into toxic intermediates. Amine oxidases are naturally present at low concentrations in seminal plasma and at high concentrations in fetal calf serum, a commonly used cell culture supplement. Here, we show that, in the presence of fetal calf serum, seminal plasma, as well as the polyamines spermine and spermidine, are highly cytotoxic to immortalized cells, primary blood mononuclear cells, and vaginal tissue. Thus, experiments investigating the effect of polyamines and seminal plasma on cellular functions should be performed with great caution, considering the confounding cytotoxic effects. The addition of the amine oxidase inhibitor aminoguanidine to fetal calf serum and/or the utilization of serum-free medium greatly reduced this serum-induced cytotoxicity of polyamines and seminal plasma in cell lines, primary cells, and tissues and, thus, should be implemented in all future studies analyzing the role of polyamines and semen on cellular functions.

Association of Zinc Concentration in Seminal Plasma and Blood with Several Parameters of Semen in Infertile Males

Objective: To evaluate the concentration of zinc inthe blood and seminal plasma with semen parameters between fertile and infertile males Methodology: This prospective study was carried out in the Department of Biochemistry, Hayatabad medical complex, Peshawar. Fifty (50)infertile males, not receiving any treatment and had regularintercourse for 1 yearaged 25-45 years were selected from HMC. They were divided into 35oligospermia and 15 azoospermic. 35 fertile males were nominatedamongthe overallresidents and were taken as the control group after their semen analysis was taken as normozoospermic. According to WHO criteria semen is analyzed. With the help of 5br Serum and seminal plasma zinc were estimated. Result: There was a significant difference found in serum and seminal levels of zinc in normozoospermic, oligospermia (p less than 0.05), and azoospermic (p-value less than 0.05) individuals. The correlation of seminal plasma zinc with sperm counts showed a positive association with the count of sperm and a negative association with motility of sperm in normozoospermic/oligospermia and a negative association of concentration with pH, and volume, WBC in all groups. No association was found with the morphology of sperm. Conclusion: It has been concluded that zinc interfered with fertility by having a considerable impact on many semen parameters. It suggests that measuring seminal plasma zinc could be useful for diagnosing and treating male infertility.

Common carp (Cyprinus carpio) sperm reduction during short-term in vitro storage at 4 °C

This study was designed to optimize a short-term storage protocol for common carp sperm at 4 °C under aerobic condition. Sperm from individual males were collected directly with or without extenders. The results demonstrated that in general, it was similar effect to collect sperm directly in extenders and keeping sperm for 0.5 h after collection without extenders. Sperm was diluted with eight selected extenders (sperm: extender = 2:1, 1:1 and 1:9) and undiluted sperm was used as a control. Sperm and seminal plasma parameters (sperm motility, velocity, membrane integrity, sperm concentration, osmolality and pH in seminal plasma) were evaluated in sperm stored on ice under aerobic conditions at 0, 2, 4, 6 and 8 days post stripping (DPS) using the computer- assisted sperm analysis system. Results showed that 1:1 and 2:1 dilution maintained higher sperm function and more sperm for a longer period. After 8 DPS, the best sperm quality and quantity was recorded in the common carp seminal plasma supplemented with 50 mM NaCl, Cejko extender (2 mM CaCl2, 1 mM MgSO4, 20 mM Tris, 110 mM NaCl, 40 mM KCl, pH 7.5 and 310 mOsm/kg) supplemented with/without 25 mM KCl/NaCl. The reduction of spermatozoa number with time during short-term storage but varied according to different dilution ratios and extenders. At 8 DPS, sperm count has dropped by 22.9 % in a dilution of 1:1 compared to 50.3 % in sperm without dilution. Extenders with diluted 1:1, especially Cejko solution, largely postponed sperm reduction, 21.3 % compared to 55.5 % for sperm stored without extenders.

The Bisphenols Found in the Ejaculate of Men Does Not Pass through the Testes.

Exposure to bisphenols is related to negative effects on male reproduction. The bisphenols exposure is associated with several modes of action including negative impact on the blood–testis barrier (BTB) in testes or direct effect on spermatozoa. Bisphenols have been detected in human seminal plasma, but the possible mechanism of seminal transfer of bisphenols is not clear. Some authors consider the transfer through the blood–testis barrier to be crucial. Therefore, in this work, we compared normozoospermic men and men after vasectomy who have interrupted vas deferens and their ejaculate does not contain testicular products. We measured the concentration of bisphenol A (BPA), bisphenol S (BPS) and bisphenol F (BPF) in the urine and seminal plasma of these men using liquid chromatography tandem mass spectrometry (LC/MSMS). We found that the ratio of urinary and seminal plasma content of bisphenols did not differ in normozoospermic men or men after vasectomy. From the obtained data, it can be concluded that the pathways of transport of bisphenols into seminal plasma are not primarily through the testicular tissue, but this pathway is applied similarly to other routes of transmission by a corresponding ejaculate volume ratio. To a much greater extent than through testicular tissue, bisphenols enter the seminal plasma mainly as part of the secretions of the accessory glands.

Local and Systemic Oxidative Stress Biomarkers for Male Infertility: The ORION Study.

Infertility problems occur in around 10% of all couples worldwide, with male-factor infertility as the sole contributor in 20–30% of these cases. Oxidative stress (OS) is suggested to be associated with the pathophysiology of male infertility. In spermatozoa, OS can lead to damage to the cell membrane, resulting in disruption of DNA integrity and a decrease in motility. Established biomarkers for OS include free thiols and malondialdehyde (MDA), both representing different components of the reactive species interactome (RSI). This exploratory study aimed to investigate seminal plasma-free thiol and MDA levels in relation to semen parameters as defined by the World Health Organization (WHO) to determine if these markers are adequate to define local OS status. Furthermore, this study investigated if there is a relation between systemic and local OS status by comparing seminal concentrations of free thiol (R-SH, sulfhydryl groups, representing the extracellular redox status) and MDA (lipid peroxidation product) levels to those measured in serum. Free thiol and MDA measurements in both serum and semen plasma were performed in 50 males (18–55 y) of couples seeking fertility treatment. A significant positive correlation was found between seminal plasma-free thiol levels and sperm concentration and progressive motility (r = 0.383, p = 0.008 and r = 0.333, p = 0.022, respectively). In addition, a significant positive correlation was found between MDA levels in seminal plasma and sperm concentration (r = 0.314, p = 0.031). This study supports that seminal plasma-free thiols may be promising as local OS biomarkers. No associations were observed between local and systemic OS biomarker concentrations.

Research Note: Dietary supplementation with pyrroloquinoline quinone disodium (PQQ.Na2) improves oxidative status and semen quality in aging layer breeder roosters

As the antioxidant capacity of sperm declines with age in roosters, the objective of the present study was to determine the effects of different levels of pyrroloquinoline quinone disodium (PQQ.Na2) on antioxidative and sperm quality parameters of aging layer breeder roosters. A total of ninety-six 63-wk-old Jinghong No. 1 layer breeder roosters were randomly assigned to 4 treatments (0, 0.5, 1, 2 mg/kg PQQ.Na2) for 6 wk. Antioxidant activity and semen parameters were assessed biweekly. The dietary administration of PQQ.Na2 significantly increased semen quality (semen volume, sperm motility, straightness, progressive motility, curvilinear velocity, straight-line velocity, and amplitude of lateral head displacement) and antioxidant capacity (T-SOD, GSH-Px, hydroxyl radical scavenging ability, and/or superoxide scavenging capacity) in seminal plasma in aging layer breeder roosters. Whereas, PQQ.Na2 supplementations significantly decreased malondialdehyde (MDA) concentration in seminal plasma in aging layer breeder roosters. Supplementation with 1 mg/kg dietary PQQ.Na2 as an antioxidant supplement could increase sperm quality and antioxidant activity of aging layer breeder roosters, while a higher dose (2 mg/kg) did not result in further increment.

The Association of Certain Seminal Phthalate Metabolites on Spermatozoa Apoptosis: An Exploratory Mediation Analysis via Sperm Protamine

Earlier studies have suggested that exposure to phthalates (PAEs) may induce spermatozoa apoptosis. Sperm protamine as a molecular biomarker during spermatozoa apoptotic processes may mediate the association between PAE exposure and spermatozoa apoptosis. This study aimed to explore whether sperm protamine mediates the association of PAE exposure with spermatozoa apoptosis. We determined sperm protamine levels, 8 PAE metabolite concentrations in seminal plasma, and 3 spermatozoa apoptosis parameters among 111 men from an infertility clinic. The associations of PAEs as individual chemicals and mixtures with sperm protamine were determined. The mediating roles of protamine in the associations between PAEs and spermatozoa apoptosis parameters were examined by mediation analysis. After adjusting for confounders, we observed positive correlations between seminal plasma concentrations of mono(2-ethylhexyl) phthalate (MEHP) and sperm protamine-1 and protamine ratio. Estimates comparing highest vs. lowest quartiles of MEHP concentration were 4.65% (95% CI: 1.47%, 7.82%) for protamine-1 and 25.86% (95% CI: 3.05%, 53.73%) for protamine ratio. The quantile g-computation models showed that the adjusted protamine-1 per quartile increase in PAE mixture was 9.42% (95% CI: 1.00, 20.92) with MEHP being the major contributor. Although the joint association between PAE mixture and protamine ratio was negligible, MEHP was still identified as the main contributor. Furthermore, we found that protamine-2 and protamine ratio levels in the highest quartiles exhibited a decrease of 43.45% (95% CI: 60.54%, −19.75%) and an increase of 122.55% (95% CI: 60.00%, 209.57%) in Annexin V+/PI− spermatozoa relative to the lowest quartiles, respectively. Mediation analysis revealed that protamine ratio significantly mediated 55.6% of the association between MEHP and Annexin V+/PI− spermatozoa elevation (5.13%; 95% CI: 0.04%, 10.52%). Our findings provided evidence that human exposure to PAEs was associated with increased protamine levels which may mediate the process of spermatozoa apoptosis.

Parenteral supplementation of Cu, Zn and Mn enhances boars’ sperm quality

IntroductionThe trace elements concentrations of blood might play a key role in the trace element concentration of seminal plasma, as well as in the improvement of semen volume and sperm morphology in boars. ObjectiveTo assess the effect of supplementation of Cu, Zn and Mn on their concentrations in blood serum and seminal plasma and sperm quality in boars. MethodsEighteen boars of the Duroc Jersey breed were selected and their blood and semen (54 ejaculates before supplementation) were extracted. Coinciding with the third sampling and after of blood taken, the boars were supplemented subcutaneously with 37.5 mg of Cu, 75 mg of Zn and 37.5 mg of Mn; 40 days after the supplementation the fourth sampling was taken. Cu and Zn concentrations in blood serum and seminal plasma were determined by atomic absorption spectrophotometry, while the sperm pathologies were determined by microscopy and the concentrations of Cu and Zn in blood serum and seminal plasma, and semen pathologies were compared using the t-Student test for paired samples. A simple linear correlation was made between the minerals concentration in seminal plasma with the percentage of spermatozoa with abnormal forms. ResultsAlthough the concentrations of Zn in blood serum did not show differences between sampling periods (P < 0.05), they were significantly higher (P < 0.01) in the seminal plasma after supplementation as compared to its levels before supplementation. In addition, the concentrations of Cu and Zn in seminal plasma were increased (P < 0.01), and the total spermatic pathologies were reduced; especially those of the head, neck and intermediate part of the tail (P < 0.001). Besides, macrocephaly, double head and broken acrosome were the most common pathologies (P < 0.05). Moreover, parenteral supplementation of Cu, Zn and Mn was a protective factor to the presentation of ejaculates with abnormal sperm percentages higher than 10% (χ2 = 6.1544; P = 0.0131). The prevalence of abnormal shapes of boars’ sperm before supplementation was 0.40; after supplementation the answer was 0.05 and the prevalence ratio was 0.13 with a confidence interval of 95% from 0.01 to 0.94. Moreover, Zn concentrations in blood serum were not correlated with those of the seminal plasma (P > 0.05, r = − 0.0353); however, the concentrations of Cu in both fluids were correlated (P < 0.05, r = 0.2254). In addition, the Zn values in the seminal plasma and the percentage of abnormal spermatozoa had a negative and highly significant correlation (P < 0.0001, r = − 0.5628). However, the Cu concentrations in the semen were not significantly correlated with the abnormal sperm forms (P > 0.05, r = 0.0200). ConclusionFrom the present study it can be concluded that in boars fed with diets that meeting their requirements in trace minerals according to NRC (2012) [1], parenteral supplementation of 37.5 mg of Cu, 75 mg of Zn and 37.5 mg of Mn increased the Zn concentrations in the seminal plasma and reduced the sperm pathologies, which resulted in an increase of the boars’ sperm quality.