Seminal Plasma Concentrations Research Articles

Relative content of Niemann-Pick C2 protein (NPC2) in seminal plasma, but not that of spermadhesin AQN-1, is related to boar sperm cryotolerance

Variation between and within boar ejaculates in terms of their ability to withstand freeze-thawing is a limitation for sperm cryopreservation. Consequently, searching for freezability markers not only in sperm but also in seminal plasma (SP) is imperative. The present study aimed to evaluate the relationship between cholesterol content, relative levels of NPC2 and AQN-1 at two different holding times (0 h: HT0 and 24 h: HT24) at 17 °C, and boar sperm freezability. Forty-five ejaculates were cryopreserved and subsequently classified as of good (GFE) or poor (PFE) freezability according to their post-thaw sperm viability and total motility. Prior to cryopreservation, relative abundances of two SP proteins (NPC2 and AQN-1) and cholesterol content in sperm and SP were determined through immunoblotting and colorimetric methods, respectively. These determinations were made after ejaculation (HT0) and after 24 h of storage at 17 °C (HT24). Two bands for NPC2 protein (16 kDa and 19 kDa) were identified. Relative amounts of the 16 kDa-band were significantly (P < 0.05) higher in poor (PFE) than in good (GFE) freezability ejaculates both at HT0 and HT24, whereas those of the 19 kDa-band were significantly (P < 0.05) higher in PFE than in GFE at HT24 only. In the case of AQN-1, no significant differences between GFE and PFE were observed. In addition, no variations in the cholesterol content of sperm and SP were observed either between HT0 and HT24 or between GFE and PFE. We can conclude that the content of two NPC2 isoforms in SP, but not of that of spermadhesin AQN-1, may be involved in the sperm resilience to withstand freeze-thawing procedures and may predict ejaculate freezability. While a possible mechanism through which NPC2 during HT could affect boar sperm cryotolerance is suggested to be related to its ability to bind the plasma membrane cholesterol, further research is warranted.

Effects of TLR4 overexpression on sperm quality, seminal plasma biomarkers, sperm DNA methylation and pregnancy rate in sheep

Genetic modification provides a means to enhancing disease resistance in animals. In this study, the first generation of genetically modified (GM) sheep overexpressing TLR4 was produced by microinjection for better disease resistance. To compare semen characteristics including sperm quality, seminal plasma biochemical index, sperm DNA methylation and pregnancy rate of three-year old transgenic sheep with TLR4 overexpressed (toll like receptor 4, TLR4) and non-transgenic ram. Sixteen transgenic ram of F0 generation were produced by microinjection of the TLR4 plasmid into the pronucleus of fertilized ova. Seven transgenic sheep of F1 generation was produced by breeding F0 transgenic founders with non-transgenic sheep of the same breed. There were no significant differences between transgenic and control rams for all semen quality parameters, including semen volume, sperm concentration, sperm viability, and percentages of sperm with an intact plasma membrane, acrosomal integrity, and viable sperm with high mitochondrial membrane potential in both F0 and F1 generation. Furthermore, no significant differences were found for seminal plasma concentrations of zinc, neutral alpha-glucosidase, acid phosphatase or fructose, nor for levels of H19 and IGF2R methylation in sperm DNA. In addition, pregnancy rate was also similar between these two groups. In conclusion, there was no evidence that TLR4 overexpression altered the sperm quality, seminal plasma or sperm DNA of transgenic sheep.

Variability of Seminal Plasma Cadmium Concentrations and Its Associations with Semen Quality among Healthy Chinese Men Screened as Potential Sperm Donors

TPS 631: Metals and health 1, Johan Friso Foyer, Floor 1, August 27, 2019, 3:00 PM - 4:30 PM Background: Cadmium (Cd) is a well-documented toxic metal, which has demonstrated adverse effects on male semen quality in animal studies. However, the results from human studies are inconsistent, probably due to the high within-subject variability in measurements of Cd in biological specimens and semen quality parameters. Objectives: To characterize the variability of Cd concentrations in seminal plasma and assessed its associations with repeated measurements of semen quality among healthy men who came to the Hubei Province Human Sperm Bank for donations screening. Methods: The reproducibility of Cd concentrations in 232 seminal plasma samples collected from 93 men was assessed using the intraclass correlation coefficient (ICC). The associations of Cd in within-subject pooled seminal plasma (n=1156) with repeated measurements of semen quality parameters (n = 5619) were assessed using linear mixed models with an individual-specific random intercept. Semen analyses were conducted by qualified laboratory technologists following the guidelines of the World Health Organization, and Cd concentration was determined by inductively coupled plasma-mass spectrometry. Results: Cd was detected in > 85% of the seminal plasma specimens. Fair to good reproducibility (ICC = 0.71) was achieved for serial measurements of Cd. Compared with men in the lowest Cd quartile, men in the highest quartile of Cd had a reduced semen volume and total motility of 3.25% (95% CI: -6.29%, -0.20%; p for trend = 0.027) and -1.88% (95% CI: -3.82%, 0.10%; p for trend = 0.083), respectively. Similar results were achieved when modeling Cd concentration (log-transformed) as a continuous variable in cubic spline models. Conclusion: Cd in seminal plasma were relatively stable. Exposure to high Cd may adversely affect semen volume and motility. Future studies in a larger and general population are needed to verify our findings.

Analysis of seminal plasma biochemical parameters and sperm cryostability in different age groups of stallions

We studied sperm of stallions of different age groups of the Arab breed at the age of 4 to 21 (n=36). The concentrations of total protein (TP), albumin, globulins, glucose, urea, phospholipids, ionized calcium (Ca2+), and enzyme activity of creatine kinase (CK), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), amylase, lactate dehydrogenase (LDH) were determined in seminal plasma. After sperm freezing and thawing the progressive sperm motility and viability at +4 °C were studied. We found that progressive motility and viability of spermatozoa after freezing and thawing in stallions at the age of 6-10 years was statistically significantly higher than that of stallions at the age of 4-5 (p=0.004 and p=0.02), 11-15 (p=0.04 and p=0.04) and 16-21 (p=0.01 and p=0.01, respectively). The concentration of sperm in the ejaculates of older stallions (16-21 years old) is significantly lower than in the ejaculates of stallions aged 4-5 (p=0.04), 6-10 (p=0.003), 11-15 (p=0.04). The level of urea in seminal plasma of older stallions (16-21 years old) was statistically significantly higher than in the group of stallions aged 6-10 (p=0.01). There is a negative correlation between progressive sperm motility after cryopreservation and urea concentration in stallion seminal plasma (r=-0.48; p<0.05).

Features of seminal plasma cytokine profile and an occurrence of pregnancy when carrying out a program of assisted reproductive technologies

Aim. To asses an effectiveness of assisted reproductive technologies (ART) program given characteristics of the cytokine profile of seminal plasma (SP) entering the female reproductive tract during sexual intercourse. Outcomes and methods. 33 married couples who applied for a treatment of infertility by means of in vitro fertilization /ICSI (Intracytoplasmic Sperm Injection) were included in the prospective study. Patients were recommended to have sexual intercourse with no restrictions during treatment and to have the last sexual intercourse 3 days before an intended transvaginal puncture (ITP). Testing of cytokines (transforming growth factor (TGF)-b1, interferon (IFN)-g, interleukin (IL)-33, IL-6, IL-8, IL-23, IL-10, tumor necrosis factor (TNF)-a, IL-18, IL- 17A, IFN-a, IL-12, monocyte chemotactic protein-1) levels in samples of partners’ SP obtained on the day having ITP were carried out using multiplex analysis with LegendPlex kits (BioLegend, USA). Results. When comparing a cytokine profile of SP in couples who did not become pregnant (n=25) and couples who become pregnant (n=8) increased IL-18 and reduced IL-10 levels (p=0,017 and p=0, 01 respectively) were revealed in the group which got pregnant. To assess a clinical relevance of cytokine content in SP ROC (Receiver Operating Characteristics) curve was used. It was established that determining of IL-18 concentration in SP has the greatest diagnostic significance (the area under a curve was 0.792±0.107, test sensitivity - 62.5%, test specificity - 95.24% at threshold concentration&gt;210.43 pg/ml). Incidence of pregnancy at IL-18 concentration above threshold levels was 83,3% while at lower concentrations of IL-18 pregnancy occurred only in 13.0% of women. Conclusions. Elevated IL-18 levels and decreased IL-10 levels in SP of female patients’ partners who don’t have restriction of sexual life when treating infertility with ART are favorable factors for a pregnancy to occur.

Белок, связывающий жирные кислоты, в сыворотке крови и сперме

a composition of seminal plasma reflects a state of reproductive organs that are involved in sperm production. to study a concentration of fatty acid-binding protein (FABP) in blood serum and sperm samples with normal and altered characteristics. a total of 82 men with mean age of 33.1+/-4.7 years old who were examined to clarify the cause of infertility were included in the study. The main group (n=21) consisted of men with oligozoospermia, while control group (n=33) consisted of healthy men with normal sperm analysis. In addition, patients with high viscosity of sperm were included in the comparison group (n=28). There were no changes in complete blood count and biochemical blood panel in all participants. The content of FABP in blood serum and seminal plasma was determined. the average content of FABP in seminal plasma was 1.347+/-0.26 ng/ml and exceeded the serum concentration of this compound (p<0.000001), which was 0.305+/-0.193 ng/ml. The concentration in seminal plasma did not depend on its serum concentration (R=0,068194), as well as the sperm volume and viscosity, but it was correlated with sperm concentration (R=0,66387). The concentration of FABP in seminal plasma was highest in the control group (1.47+/-0.33 ng / ml) and it was higher compared to the main group (1.22+/-0.09 ng/ml; p=0.000057) and the comparison group (1.29+/-0.19 ng/ml; p=0.010822). There was no difference between groups in serum concentration of FABP (p=0.9814). the obtained data suggest that a reduced level of FABP in seminal plasma can be considered as unfavorable criterion indicating a reduced fertility.

Effect of heavy metals arsenic, cadmium, and lead on the semen variables of dromedary camels (Camelus dromedarius)

In this study, there was investigation of the effect of heavy metals on the fertility of dromedary camels. Fourteen camels at the Camel Research Center, King Faisal University, and 41 infertile dromedaries admitted to the Veterinary Teaching Hospital were used for semen evaluation during the breeding season. Seminal plasma and blood serum were collected from all males until analysis. Concentrations of three heavy metals [arsenic (As), cadmium (Cd), and lead (Pb)] were determined in the seminal plasma and serum using an atomic absorption spectrophotometer. The results indicate there are differences (P < 0.05 – P < 0.01) in pH, sperm motility, sperm concentration, and sperm abnormalities between the fertile and infertile male camels. In seminal plasma, there were marked differences (P < 0.01– P < 0.0001) between the control and infertile male camels in As, Cd, and Pb concentrations. In serum, there were differences (P < 0.01 – P < 0.001) between the fertile and infertile camels in serum As, Cd, and Pb concentrations. There was a positive correlation (P < 0.05; r = 0.77 and r = 0.94, respectively) between serum and seminal plasma concentrations of both As and Cd in the infertile dromedaries. In the control group, there was a positive correlation (P < 0.05; r = 0.70) between seminal plasma concentrations of Cd and percent sperm abnormalities. In conclusion, relatively greater seminal plasma and serum concentrations of As, Cd, and Pb are associated with lesser values for semen quality variables and infertility in dromedary camels.

Short Communication: Effect of Seminal Plasma on Functions of Monocytes and Granulocytes.

Most HIV-1 transmissions occur at mucosae and involve exposure to semen. Semen contains immunomodulatory factors, which inhibit anti-HIV-1 natural killer cell and T cell responses. We demonstrate high concentrations (1:2 dilution) of seminal plasma (SP) inhibit monocyte phagocytosis and anti-HIV-1 Fc-dependent functions of both neutrophils and monocytes. In addition, slightly lower SP concentrations (1:2-1:10 dilutions) inhibit granulocyte phagocytosis and oxidative burst of both monocytes and granulocytes. These observations may have implications for HIV-1 infectivity after mucosal exposure.

Population Modeling Highlights Drug Disposition Differences Between Tenofovir Alafenamide and Tenofovir Disoproxil Fumarate in the Blood and Semen.

Understanding antiretroviral disposition in the male genital tract, a distinct viral compartment, can provide insight for the eradication of HIV. Population pharmacokinetic modeling was conducted to investigate the disposition of tenofovir disoproxil fumarate (TDF), tenofovir alafenamide (TAF), and emtricitabine and their metabolites in blood and semen. Blood plasma and seminal plasma (SP) concentrations of tenofovir and emtricitabine were measured, as were tenofovir-diphosphate and emtricitabine-triphosphate concentrations in peripheral blood mononuclear cells (PBMCs) and seminal mononuclear cells. Sequential compartmental modeling described drug disposition in blood and semen. Our modeling suggests slower elimination of apparent tenofovir-diphosphate PBMC and faster elimination of tenofovir SP after administration of TAF compared with TDF, likely reflecting flip-flop kinetics. Additionally, TAF metabolism to tenofovir appeared slower in semen compared with blood; however, SP elimination of TAF-derived tenofovir appeared faster than its blood plasma elimination. These findings provide valuable insight for further mechanistic study of cellular entry and drug metabolism in the male genital tract.

A novel functional role of nickel in sperm motility and eukaryotic cell growth

BackgroundMetal ions are essential for numerous life processes. This study aims to investigate the relationship between seminal quality and ion levels in seminal plasma. Basic proceduresA total of 205 semen samples were collected and seminal plasma ion levels were examined with inductively-coupled plasma-mass spectrometry. The nickel function was demonstrated by in vitro assay and cell growth. Main findingsThe low sperm motility group showed distinctively reduced nickel concentration in seminal plasma compared with the normal sperm motility group. However, arsenic, sulfur, selenium, magnesium and zinc were negatively associated with sperm quality. No significant relationship between other examined cations and semen quality was observed. In vitro assay suggested low concentration of nickel significantly increased sperm total motility and progressive motility. Cell growth assay further confirmed nickel promoted eukaryotic yeast cell growth. Nickel level in seminal plasma may play important functions to determine sperm quality. Principal conclusionsOur study reveals a strong correlation between S, Mg, Se, Zn, As, Ni and seminal quality as well as discovers a novel functional role of nickel in sperm motility and eukaryotic cell growth. These findings may provide a potential avenue for assessment of sperm quality and treatment of reproduction disorders.

Assessment of influence of “BESTFertil” supplement on semen analysis parameters, oxidative stress markers and sperm DNA fragmentation index in infertile males

Background. Idiopathic male infertility remains an important problem in andrology and reproduction, despite steady advances in diagnostics and treatment of conditions associated with impaired fertility in men. Only available option of conservative treatment for idiopathic male infertility is an empiric antioxidant therapy. Materials and methods. Fifty five men from infertile couples without any identifiable cause of male infertility who received “BESTFertil” supplement were enrolled in this study. Semen analysis parameters (sperm density, percentage of sperm with progressive motility (categories A and B), sperm morphology, leukocyte concentration), MAR-test results, reactive oxygen species concentration as measured by chemiluminescence assay and sperm DNA fragmentation index evaluated using TUNEL method were registered and compared with baseline values after 90 days of treatment. Patient compliance was assessed by a number of capsules taken throughout the study period. Results. There was a significant difference in percentage of sperm with normal morphological features (median 2 % vs 4 %; p &lt;0.001) and in percentage of A + B motile sperm (medium 31.2 % vs 41.15 %; p &lt;0.001). Also, a statistically significant decrease in sperm DNA fragmentation index (median 17.8 % vs 15 %; p &lt;0.001) and reactive oxygen species concentration in seminal plasma (median 0.7 mV/s vs 0.5 mV/s; p = 0.002). Forty-three (78.2 %) patients were fully compliant, 11 (20 %) patients had compliance over 90 %, and 1 patient was 89 % compliant. No treatment-related adverse events were observed. Conclusion. “BESTFertil”supplement may be used in empiric therapy of idiopathic male infertility and has a good safety profile. Separate intake of morning and evening supplement doses increases efficacy of multicomponent antioxidant therapy by preventing possible interaction between biochemically incompatible components. Treatment with “BESTFertil”supplement leads to a decrease in reactive oxygen species seminal concentration and sperm DNA fragmentation index.

The association between iron, calcium, and oxidative stress in seminal plasma and sperm quality.

The present study aimed to determine the level of iron and calcium in the seminal plasma of men with different fertility potentials and to examine its relationship with oxidative stress. Seventy-nine sub-fertile patients with asthenoteratozoospermia (AT), n 27; teratoleucozoospermia (TL), n 20; teratozoospermia (Terato), n 32; and 29 healthy donors were included. The ability of spermatozoa to produce reactive oxygen species (ROS) was evaluated by using nitroblue tetrazolium (NBT) staining. The lipid peroxidation end product, malondialdehyde (MDA), and the trace element levels (iron and calcium) were measured spectrophotometrically. Iron and calcium concentrations in seminal plasma of the patient groups were significantly more elevated than the normal group. Nevertheless, both calcium and iron showed strong negative correlations with the total sperm motility and normal sperm morphology, but only iron was positively and significantly associated with multiple anomalies index and seminal leucocyte concentration. On the other hand, the rates of MDA and ROS production in semen were significantly higher in the three abnormal groups than in controls. These two oxidative stress biomarkers were significantly associated with the percentage of atypical forms in semen. However, only semen ROS level was significantly associated with the decreased sperm motility and the sperm leucocytes concentration. Meanwhile, there are positive correlations between seminal iron and calcium content and the studied oxidative stress biomarkers. Oxidative stress and trace element excess are implicated in low sperm quality. Iron and calcium might be the mediators of the effects of oxidative damage and induces lipid peroxidation.

Effect of Cryopreservation on Lipid Peroxidation, Antioxidant Potential, Chromatin Integrity, and Mitochondrial Activity of Indian Red Jungle Fowl (Gallus gallus murghi) Semen.

Aim: The study elucidates the impact of cryopreservation on lipid peroxidation (LPO), antioxidant potential, DNA integrity, and mitochondrial activity of Indian red jungle fowl sperm. Materials and Methods: Semen from eight mature cocks was pooled and diluted at 37°C using a specific medium for red fowl species. The diluted semen samples were immediately refrigerated at 4°C for 2 hours (0.275°C min-1). Glycerol was then added to a final concentration of 20%, and the samples were equilibrated for 10 minutes at 5°C before loading into 0.25 mL French straws. These straws were then frozen by placing them in liquid nitrogen vapor for 10 minutes and plunged into liquid nitrogen. Motility, viability, DNA integrity, antioxidant activity, and LPO were assessed before dilution (fresh semen), after equilibration (processed semen), and post-thawing (frozen semen). Results: Sperm motility, viability, DNA integrity, and mitochondrial activity decreased (p < 0.05) in processed and frozen semen compared with fresh semen. Nevertheless, the concentration of malondialdehyde (MDA) in sperm and seminal plasma was greater (p < 0.05) in frozen-thawed and processed semen compared with fresh semen. Multivariate regression analysis showed a negative impact of MDA concentration in sperm (R2 = 0.90, Wilk's λ = 0.003, p < 0.001) and seminal plasma (R2 = 0.84, Wilk's λ = 0.02, p < 0.001) on motility, viability, DNA integrity, and mitochondrial activity. Nonetheless, total antioxidant capacity (TAC) had a positive impact on sperm variables (R2 = 0.82, Wilk's λ = 0.096, p < 0.001). Conclusions: The decrease in motility, viability, DNA integrity, and mitochondrial activity of Indian red jungle fowl sperm was associated with an increase in LPO during cryopreservation. Furthermore, TAC was reduced during the freeze-thaw process, which was insufficient in protecting the sperm against high reactive oxygen species levels.

In vitro aging of boar spermatozoa: role of sperm proximity and seminal plasma.

Knowledge on the effect of seminal plasma on sperm function in extended semen during invitro storage is lacking. The aim was to examine the interactive role of sperm concentration and seminal plasma concentration on boar sperm function during invitro aging. Experiment 1: Twenty-one boar ejaculates were aliquoted with Beltsville Thawing Solution into five semen doses containing between 32.5 and 8.5×106 sperm/mL. Experiment 2: Semen samples (n=8) containing 18×106 or 10×106 sperm/mL with their natural amount of seminal plasma and 10×106 sperm/mL substituted with autologous seminal plasma to the same concentration as in doses with 18×106 sperm/mL were prepared. Experiment 3: Four variants of semen doses containing 18×106 or 10×106 sperm/mL with either 10% or 0.5% (v/v) seminal plasma were used. Lipid peroxidation was assessed using Bodipy 581/591 in samples (n=8) with two different sperm concentrations. Semen was examined during 144-h storage at 17°C by computer-assisted semen analysis and flow cytometry. Experiment 1: 3D kinematic patterns revealed a concentration- and time-dependent loss of sperm kinematics in samples with <23×106 sperm/mL (p<0.05). Percent viable spermatozoa with high mitochondria membrane potential were lower (p<0.05) in samples with <15×106 sperm/mL. Experiment 2: Seminal plasma supplementation in samples with 10×106 sperm/mL did not restore the loss of sperm kinematics (p>0.05). Experiment 3: At 144h, motility was lowest in samples containing 10×106 sperm/mL and 10% (v/v) seminal plasma (p<0.05). Sperm lipid peroxidation did not differ between samples with different sperm concentration. Long-term exposure to seminal plasma has a negative impact on invitro-aged boar spermatozoa. Reduced sperm-to-sperm proximity but not the reduction of seminal plasma limits sperm function in long-term stored boar semen.

Melatonin-improved buffalo semen quality during nonbreeding season under tropical condition

The role of melatonin in protecting spermatozoa from different kinds of injury has been widely reported. The present study aimed to test whether treatment of buffalo bulls with melatonin could ameliorate sperm function during nonbreeding season under tropical condition. Ten Murrah buffalo bulls were randomly allocated into control and treated groups of equal numbers of bulls to study the effect of melatonin on semen characteristics, seminal plasma constituents, blood plasma hormonal levels, and antioxidant enzyme activities during nonbreeding season. Treated bulls were implanted with melatonin (18 mg/50 kg of body weight) for a period of 2 mo. During this period, semen was collected twice a week, and blood samples were collected weekly to determine plasma concentration of melatonin and LH and activities of antioxidant enzymes. During nonbreeding season, melatonin implantation improved semen characteristics by increasing (P < 0.05) percentages of sperm with forward motility, viability, total motile sperm, and rapid motility, average path, curvilinear, and straight-line velocity and amplitude of lateral head displacement and decreasing (P < 0.05) percentages of abnormal sperm and linearity index as compared to the control group. Furthermore, melatonin implantation increased (P < 0.05) seminal plasma concentrations of total protein, albumin, and cholesterol and decreased (P < 0.05) seminal plasma aspartate aminotransferase activity. In addition, melatonin-implanted bulls exhibited an increase (P < 0.05) in red blood cells superoxide dismutase activity compared to untreated bulls. In conclusion, melatonin implantation successfully improved semen quality of buffalo bulls during nonbreeding season under tropical condition.

Kisspeptin Protein in Seminal Plasma Is Positively Associated with Semen Quality: Results from the MARHCS Study in Chongqing, China.

Objectives To study the associations between kisspeptin levels in seminal plasma and blood plasma and semen quality. Materials and Methods We conducted a male reproductive health survey in June 2014. A total of 666 volunteers were recruited from colleges in Chongqing, China. All volunteers completed a questionnaire including information on domestic characteristics and some potential confounders. We tested the kisspeptin levels in both blood and seminal plasma. Total seminal kisspeptin was calculated as the concentration of kisspeptin in seminal plasma multiplied by semen volume. Semen samples were tested according to the 2010 World Health Organization's (WHO) guidelines. Spearman correlation and multivariate linear regression were used to explore the association between kisspeptin concentrations in seminal plasma and blood plasma and semen quality. Potential confounders that were adjusted for included age, abstinence time, body mass index (BMI), grade, and smoking. Results The median of kisspeptin levels in seminal plasma was 60,000 times higher than kisspeptin in blood plasma (28.0 × 106 pg/ml versus 448.9 pg/ml). Each interquartile range (IQR) of kisspeptin in seminal plasma was associated with a 4.6% (95% confidence interval [CI]: 1.6%–7.6%) increase in sperm concentration. Each IQR of total kisspeptin was associated with a 7.7% (95% CI: 4.4%–11.0%) increase in total sperm number and a 7.8% (95% CI: 4.0%–11.7%) increase in total motile sperm count. Kisspeptin levels were further classified into quartiles and Q1 was set as the reference level. Subjects in the high total kisspeptin group had 57.5% (95% CI: 33.2%–86.2%) higher total sperm number than the reference group. Conclusion The positive association between kisspeptin levels in seminal plasma and semen quality supported an important role for the KISS1/GPR54 system in male reproductive health. Kisspeptin may be a potential marker of male reproductive health and an alternative strategy for treating infertility.

Seminal plasma amino acid profile in different breeds of chicken: Role of seminal plasma on sperm cryoresistance.

Seminal plasma is a key biological fluid that modulates sperm function in the reproduction process. However, its role in sperm biotechnologies is scarce in poultry. The aims of the present study were to study the amino acids profile and total proteins of seminal plasma in 12 Spanish chicken breeds and to investigate the role of seminal plasma on cryoresistance of rooster sperm. To investigate the role of seminal plasma on cryoresistance, diluted pooled semen samples were cryopreserved in the presence and absence of seminal plasma. Glutamic acid was the most abundant free amino acid in seminal plasma, followed by alanine, serine, valine, and glycine. There was an influence of breed (P<0.05) on the percentage of viable sperm after freezing-thawing of samples with seminal plasma. Cluster analysis revealed that White Prat, Black Castellana, Blue Andaluza, Quail Castellana, and Red-Barred Vasca returned the best freezing-thawing response (good freezers). There was a positive correlation between seminal plasma concentrations of valine, isoleucine lysine, leucine and post thaw viability. The evaluation of fertilization capacity of frozen-thawed semen from the breeds White Prat (‘good freezer’) and Black-Red Andaluza (‘bad freezer’) showed that good freezer had higher fertility (20/68, 29.4%) compared to bad freezer breed (14/76, 18.4%), even if the difference was not significant (P = 0.08). The TUNEL assay revealed that freezing/thawing procedures in presence of seminal plasma provoked higher DNA fragmentation in most of the breeds, with a positive correlation between seminal alanine, valine, isoleucine, methionine, leucine, tyrosine, phenylalanine concentrations and DNA integrity. DNA fragmentation was lower in absence of seminal plasma and the breed effect on sperm viability was highly reduced. It is concluded that specific seminal plasma amino acids were associated with post-thaw percentage of viable sperm and DNA integrity. The removal of seminal plasma decreases the variability of the results and DNA fragmentation damages.

Protein and lipid species in seminal plasma of fertile Holstein-Friesian bulls

Protein and lipid molecules in seminal plasma (SP) collected from fertile bulls were investigated. Semen was collected from 10 bulls (2 ejaculates each) and examined for standard semen analysis. Raw SP was recovered by centrifugation and total protein (TP) concentration was determined using a refractometer. Raw SP was desalted using a Sephadex G-25 desalting column then both raw and desalted SP was subjected to SDS-PAGE. Neutral lipids and phospholipids of raw and desalted SP were separated by thin-layer chromatography (TLC). The results revealed that, all bulls had normal semen characteristics and TP concentration in SP ranged from 7.0 to 10.4 g/dL except bull No. 6 had a relatively low concentration of 4.9 to 6.8 g/dL. Neither proteins nor lipids species were different between raw and desalted SP. Seventeen proteins were detected ranging from 8.5 to 185.8 kDa, and those of 12, 13.5, 15, 21, 23 and 38 kDa were predominant. Notably, proteins of 10, 17.5, 19, 21, 80 and 185.8 kDa might be new candidates of SP proteins (SPPs). The detected neutral lipid spots corresponded to cholesterol, 1,2-dimyristoyl glycerol, 1,2-dioleoyloglycerol, 1,3-dimyristoyl glycerol and 1,3-dioleoyloglycerol. The detected phospholipids spots corresponded to non-migrating phospholipids, sphingomyelin (SM), phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidylethanolamine (PE), cerebroside and polyglycerol phosphatide. Cholesterol represents the major molecule of neutral lipids, whereas SM, PC, PI and PE represent the major phospholipids. Noteworthy, there were 2 species of diacylglycerol (DAG) and 3 species of PI in bovine SP. In conclusion, this study gave a general picture of SP protein and lipid species in fertile bull semen, which might serve as fundamental knowledge for either semen analysis or prediction of male fertility.

Differential extracellular, but similar intracellular, disposition of two tenofovir formulations in the male genital tract.

The male genital tract (MGT) is a viral sanctuary and likely HIV reservoir; understanding MGT pharmacokinetics (PK) of antiretrovirals (ARVs) used for curative strategies is critical to eradication and cure. Tenofovir alafenamide (TAF) is a tenofovir (TFV) formulation designed to maximize efficacy/minimize toxicity with unknown MGT PK. HIV-positive and HIV-negative men receiving TFV-based regimens provided six paired blood plasma (BP) and semen samples. Extracellular (TFV, TAF, emtricitabine [FTC]) drug concentrations in BP and seminal plasma (SP), and intracellular metabolite (IM) and endogenous nucleotide (EN) concentrations were measured in peripheral blood mononuclear cells (PBMCs) and seminal mononuclear cells (SMCs). Exposure ratios for SP:BP, SMC:PBMC and IM:EN were calculated from PK parameters generated by noncompartmental analysis. HIV viral load was measured in BP and SP. Sixteen HIV-positive (n=8, TDF/FTC; n=8, TAF/FTC) and eight HIV-negative (TDF/FTC) men provided samples. Median TFV SP:BP ratios differed between TDF and TAF (1.5 versus 7.4), due to lower TFV BP concentrations with TAF coupled with TFV SP concentrations similar to TDF. BP ratios were approximately 3. SMC concentrations of IMs and ENs were a fraction of PBMC concentrations (1-22%), though IM:EN ratios exceed a suggested protective threshold. TAF SP PK was unexpected. IM SMC concentrations were low relative to PBMC, as were EN concentrations, suggesting differences in cell phenotype and lineage in the MGT; these differences in phenotype and pharmacology may have an impact on selecting and dosing ARVs used in cure strategies.

EFFECTS OF PARSLEY SEEDS AT DIFFERENT LEVELS AS NATURAL ADDITIVES ON SEMEN QUALITY IN RABBIT MALES

This study is designed to evaluate the effect of using parsley seeds (PAS) at different levels as a natural on semen quality and reduce the effect of lipidperoxidation of bucks. Forty New Zealand rabbit bucks 8 month old were divided into four groups (each 10 rabbit bucks). The groups were divided as follows: Control diet (free from PAS), T1: Control diet + 5g/kg PAS, T2: Control diet + 10g/kg PAS; T3: Control diet + 15g/kg PAS. Semen sample were collected twice a week; from each rabbit buck by artificial vagina. The first semen collection was used to evaluate semen quality parameters. However, the second semen collection was used for analysis semen plasma. Results revealed that feeding diets containing different levels of PAS significantly (P&lt;0.05) increased semen volume, mass activity, individual motility, PSV, sperm concentration, total sperm and seminal plasma activity of ALP enzyme and significant (P&lt;0.05) decrease in dead sperm and seminal plasma concentrations of total lipids, triglycerides, cholesterol and MDA and seminal plasma activities of AST and ALT enzymes and TAC as compared with the control group. In conclusion, dietary supplementation with different levels of PAS, especially at the level of 15 g/kg (T3) resulted in significant improvement in semen quality, antioxidant status and reproductive traits of rabbit bucks.