Enzymes In Seminal Plasma Research Articles

Association of the polymorphism of the Pon-1 locus with the concentration of Paraoxonase-1 and post-thaw sperm viability in pig

The objective of the present work was to correlate the expression of the PON-1 locus polymorphism, the concentration of the enzyme paraoxonase 1 in seminal plasma and the effect on post-thawing sperm viability in the pig. Nine ejaculates from 3 boars were used, to obtain seminal plasma; part of the ejaculates was centrifuged at 800 g/10 min; the material obtained was stored at -20°C until use. To freeze the semen, the samples were diluted 1:1 in commercial diluent (Magapor®) at 37°C and the temperature was gradually lowered to 16°C. They were transported to the laboratory protected from light, and kept at the same temperature in the laboratory for 24 h. After this, they were frozen using the glycerolation technique and a portion was cryopreserved by adding 20% of seminal plasma to the conservation medium and another portion was made without seminal plasma. The concentration of PON-1 was determined by an enzyme-linked immunosorbent assay (ELISA). The functional status of the plasma membrane was evaluated by chlortetracycline staining. The amplification of a fragment of the PON-1 gene was carried out by PCR and the identification of allelic variability by the analysis of Restriction Fragment Length Polymorphism (RFLP), with the enzymes HinfI and FokI, from a DNA sample of each individual. The results were analyzed by analysis of variance (ANOVA) and Spearman correlation, with the use of statistical software (Statistica V10 for Windows). In electrophoresis of enzymatic digestion with the enzyme HinfI All the expected fragments were obtained, with the FokI enzyme a mutation was observed in one individual. No correlation was observed (R=-0.1, P>0.05) between the concentration of the paraoxonase-1 enzyme in seminal plasma and the observed mutations. As well as the concentration of the enzyme paraoxonase-1 in the ejaculates was 1.8 to 2.7 ng/mL and this had no statistically significant difference in the seminal plasma of the ejaculates analyzed (P>0.05). As well as with sperm viability in thawed semen (P>0.05).

Exploring enzymatic antioxidant activities in Asian Elephant (Elephas maximus) sperm and seminal plasma

This study explored the relationships between sperm quality and the activities of antioxidant enzymes (superoxide dismutase, SOD, and glutathione peroxidase, GPx) in sperm (S) and seminal plasma (SP) of Asian elephants, as well as the potential protective role of glutamine in semen extenders during chilled storage. In experiment 1, twenty ejaculates from 11 bulls were analyzed for percentages (%) of sperm motility (%MOT), viability (%VIA), and activities of SOD and GPx. A significant negative correlation was found between %VIA and SP-SOD (r = − 0.71), and positive correlations between %MOT and both S-GPx (r = 0.49) and SP-GPx (r = 0.57). Following these findings, glutamine, a precursor of glutathione and semen antioxidant was introduced in Experiment 2. Each ejaculate (in a total of 8 ejaculates obtained from four bulls) was diluted in Tris-fructose- (EYT) and Tris-glucose-egg yolk extender (TCG), respectively, supplemented with 0 (control), 20 and 40 mM glutamine and stored at 4 °C. Sperm parameters including %MOT, %VIA, functional membrane integrity (%sHOST), and DNA integrity (%DNA) were assessed after 24 and 48 h of storage. Results demonstrated that glutamine did not significantly affect %VIA or %DNA. A significant reduction in %MOT was observed at 24 h, but 20 mM glutamine added in extenders reversed this decline at 48 h. A 40 mM addition of glutamine in TCG markedly reduced %sHOST at 24 h. In conclusion, SOD and GPx activities in the sperm and seminal plasma are indicative of sperm quality. During chilling, using a higher level of glutamine may damage the spermatozoa of Asian elephants.

Evaluation of Marjoram Leaves (Oregano vulgar) as Feed Supplement on Quality of Semen in Awasian Pollination Rams.

This study aimed to evaluate the effect of Marjoram leaves (Oregano vulgar) as a feed supplement on sexual efficiency and semen quality in Awasian pollination rams. The study was conducted on ten rams in one of the private fields in the Babylon governorate. The rams were divided randomly into two groups. First group rams are regarded as a control group (group A) that feeds on exceptional concentric food for pollination rams. The second group consists of 5 rams (group B) provided with the same concentric food plus a supplement of 3 mg/kg of body weight of Oregano vulgar leaves (fresh) 3 times daily for 49 days before the pollination season (April and May). Semen samples were collected at the end of 49 days by artificial vagina to evaluate the volume of ejaculate, sperm number, concentration, motility, viability, and deformities, and to measure the sperm pleomorphic parameters of the head, nucleus, and acrosome. The antioxidant status of seminal plasma was evaluated by measuring superoxide dismutase (SOD), glutathione peroxidase (GPX), and total antioxidant capacity (TAC). ELISA tests were used on blood serum samples to measure the hormone levels of luteinizing hormone (LH), testosterone, and estrogen. The study demonstrated a significant increase in motility, the viability of sperm, and decreasing deformities. There was no significant variation in perimorphic parameters of the head, nucleus, and acrosome sperm after using the fresh leaves of Oregano vulgar as supplementation to Awasion sheep. However, there was an increase in the antioxidant enzymes (SOD, GPX, and TAC) in seminal plasma and an increase in LH, testosterone, and estrogen in the blood serum of experimental group B. These findings show that using fresh leaves of Oregano vulgar improved the sexual efficiency of pollinated rams and maintained the physiology and perimorphic parameters of semen and sperm.

Validation of sORP, a Measure of Oxidative Stress, in Australian Population

Background: Excessive reactive oxygen species are normally eliminated by seminal plasma free radical antioxidant scavengers and enzymes. Assessing the oxidation reduction potential of seminal fluid (mV/million sperm/mL) enables the detection of an imbalance in this biological mechanism, and identifies patients exhibiting oxidative stress (OS). A novel technology, MiOX[Formula: see text], detects static Oxidation-Reduction Potential (sORP), providing a comprehensive measure of oxidants and reductants in semen, with the advantages of rapid, simple and less sample volume compared to the conventional chemiluminescence assay. Aim: This study aims to validate the MiOX[Formula: see text] analyser for the detection of OS in seminal fluid, in the Australian population. Method: Samples were collected from consenting patients attending the Royal Women’s Hospital Andrology Unit for semen analysis (n=144); those with [Formula: see text]1 million/ml were excluded. Semen analysis was carried out as per WHO 2010 guidelines, and replicate sORP measurements taken within 60 minutes of ejaculation. Results: Replicate sORP measurements from semen samples (n=20) demonstrated no significant differences between and within operators, and across days. sORP did not vary between samples incubated at 4[Formula: see text]C, room temperature, and 37[Formula: see text]C, or samples subjected to mechanical agitation. Freeze-thaw resulted in a significant increase in sORP (p=0.031). The stability of sORP measurements was demonstrated up to 4 hours after ejaculation, however, significant increases were observed after 24 and 48 hours (p[Formula: see text]0.05). Patients grouped according to normal and abnormal total motile sperm count (TMSC) demonstrated significantly different sORP (0.75 ± 0.86, 5.02 ± 5.32, p[Formula: see text]0.001). A preliminary ROC analysis, using TMSC, revealed a cut-off value of 1.07 mV/106 sperm/mL (AUC=0.904) with 81.1% sensitivity and 81.3% specificity. Conclusion: Results from the technical validation are consistent with other studies. The clinical reference range validation requires further patient samples to better reflect the distribution of results observed in the low TMSC group.

Differences between some biochemical components in seminal plasma of first and second ejaculations in dual-purpose Simmental (Fleckvieh) bulls and their relationships with semen quality parameters.

This study aimed to evaluate differences in seminal plasma zinc (Zn), copper (Cu), iron (Fe), catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) levels in the first and second ejaculations and their relationships with semen quality parameters in Fleckvieh bulls. Repetitive ejaculates were separately collected, analyzed, and frozen from the sires. Progressive motility of frozen-thawed semen (PMFT) was considered the main factor for more data classification into three following groups: <40.00%, 40.00 - 50.00%, and >50.00%. Seminal plasma trace elements and enzymes were determined using atomic absorption spectrometry and ELISA, respectively. The results revealed significant differences between the first and second ejaculations. Semen concentration, SOD, GPx, and Fe were different in ejaculations. Although PMFT groups in different ejaculations did not show significant differences, there was significant alteration between different PMFT groups and first and second ejaculations. All frozen-thawed semen CASA parameters (except lateral head displacement) were associated with fresh motility parameters and before and after thawing sperm viability. Also, a correlation between seminal Zn concentration with fresh semen gross and progressive motility, average path velocity, and beat cross frequency, Cu with SOD and Fe and semen concentration was observed. CAT was associated with fresh and frozen-thawed sperm motility parameters except for lateral head displacement and angular displacement. Although our findings showed differences between the first and second ejaculations in some parameters, PMFT, which is the most important indicator for estimating bull fertility, was not different between them.

Effect of seasonal infertility period on boar sperm proteins and quality characteristics

Swine seasonal infertility reduces the productivity and profitability of a pig farm. The main causes of this condition are elevated environmental temperatures and long photoperiod during the summer season. The aim of this study was to investigate which sperm proteins and parameters are affected during the period of seasonal infertility. Depending on the environmental temperatures, the period from October to June was considered as cold and the period from July to September as warm season. A total of 65 ejaculates from 18 boars were collected over a year. Each semen sample was evaluated for kinetics (Computer Assisted Semen Analyzer), morphology (Sperm Blue stain), viability (Propidium Iodide - Calcein AM stain), mitochondrial membrane potential (Rhodamine 123 – Propidium Iodide stain), membrane integrity and functionality (Hypo-osmotic swelling test) and sperm DNA integrity (Acridine Orange Test). Moreover, selected proteins (HSP90, GPX5, OPN) were detected and quantified. The kinetic parameters VSL, LIN and the midpiece abnormalities were significantly higher in the warm compared to the cold season (p&lt;0.05), while a strong tendency towards higher values for HSP90 and GPX5 was observed in warm compared to cold season (p=0.07and p=0.06, respectively). In conclusion, among the boar sperm characteristics tested in our study, seasonal infertility period negatively affected VSL and LIN kinetics, while GPX5 seminal plasma enzyme and HSP90 sperm surface protein increased their sperm protective effects.

Effect of Zinc Supplementation on Urate Pathway Enzymes in Spermatozoa and Seminal Plasma of Iraqi Asthenozoospermic Patients: A Randomized Controlled Trial.

BackgroundUric acid (UA) is crucial for sperm metabolism as it protects seminal plasma against oxidative dam- age. Zinc also plays a central role in sperm metabolism. The current study was designed to investigate the role of zinc supplementation on qualitative and quantitative properties of seminal fluid, in parallel with the UA level and urate pathway enzymes in the semen of patients with asthenozoospermia.Materials and MethodsThe study was designed as a randomized controlled trial of 60 asthenozoospermic subfertile men. The current study, which was conducted during one year, involved 60 fertile and 60 asthenozoospermic subfertile men belonging to Hilla City, Iraq. Semen samples were obtained from the participants before and after treatment with zinc supplements. The levels of UA, xanthine oxidase (XO), adenosine deaminase (ADA) and 5'-nucleotidase (5'-NU) activities were determined in spermatozoa and seminal plasma of both groups.ResultsUA levels (P=0.034) and 5'-NU activity (P=0.046) were significantly lower but ADA (P=0.05) and XO (P=0.015) activities were significantly higher in infertile men than in healthy men. Treatment with zinc sulfate induced an increase in UA (P=0.001) level and 5'-NU activity (P=0.001), but a decrease in ADA (P=0.016) and XO (P=0.05) activities.ConclusionZinc supplementation restores UA levels and the activities of enzymes involved in the urate pathway (XO and ADA) in the seminal plasma and spermatozoa of patients with asthenozoospermia, to reference values. Sup- plementation of Zn compounds enhances the qualitative and quantitative properties of semen (Registration number: NCT03361618).

Activities of antioxidant seminal plasma enzymes (SOD, CAT, GPX and GSR) are higher in jackasses than in stallions and are correlated with sperm motility in jackasses

This study compared the activities of four antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPX; and glutathione reductase, GSR) in the seminal plasma of stallions and jackasses. Eighteen stallion ejaculates and 24 jack ejaculates were collected through an artificial vagina. Seminal plasma was obtained by several centrifugations at 3000×g and 4 °C for 10 min, and activities of SOD, CAT, GPX and GSR were subsequently determined. We also evaluated whether the collecting season had any influence on the activities of these four enzymes in both stallions and jackasses. Antioxidant capacity of seminal plasma was significantly higher in jackasses than in stallions (mean ± SEM, SOD: 1707.7 ± 195.9 U/mL vs. 231.9 ± 29.6 U/mL; CAT: 9094.7 ± 1292.9 U/L vs.1682.7 ± 525.9 U/L; GPX 845.4 ± 106.0 U/L vs. 469.7 ± 60.3 U/L; GSR: 50.3 ± 5.1 U/L vs. 20.7 ± 4.6 U/L). Furthermore, whereas season had no effect on the activity of these four enzymes in stallions, the activities of CAT and GPX in jack seminal plasma were significantly higher in the summer than in the other seasons. In addition, the activities of SOD and CAT were found to be significantly correlated with the percentages of progressively motile spermatozoa, and with the percentages of linearity and straightness, respectively, in jackasses. In contrast, the activities of these four enzymes were not correlated with sperm quality parameters in stallions. Finally, while SOD, CAT, and GPX activities but not those of GSR were correlated in jackasses, the activities of all four enzymes were correlated each other in stallions. We can thus conclude that the activities of SOD, CAT, GPX and GSR differ between the seminal plasma of stallions and donkeys, and vary between seasons in jackasses.

Levels of activity of superoxide dismutase in seminal plasma do not predict fertility of pig AI-semen doses

Superoxide dismutase (SOD) is a major antioxidant enzyme in boar seminal plasma (SP). This study evaluated how SP-SOD affected sperm attributes when semen of boars of various breeds, included in commercial artificial insemination (AI)-programs, was extended and liquid-stored at 17 °C for AI; as well as their in vivo fertility (farrowing rate and litter size of 10,952 AI-sows). SP-SOD-activity was assessed in 311 ejaculates (100 boars) while sperm motility (by CASA), viability and intracellular H2O2 generation in viable spermatozoa (by flow cytometry) were measured at 0 and 72 h of liquid storage. SP-SOD activity was not affected by breed but differed (P < 0.001) between boars (n = 50), ranging from 1.16 ± 0.11 to 7.02 ± 0.75 IU/mL. Semen AI-doses (n = 44) hierarchically grouped (P < 0.001) with low SP-SOD activity showed lower (P < 0.05) sperm motility and intracellular H2O2 at 72 h of liquid storage. Fertility did not differ between AI-boars (n = 39) hierarchically grouped (P < 0.001) with high or low SP-SOD activity. In conclusion, SP-SOD activity is boar dependent and positively related with sperm functionality of liquid-stored semen AI-doses. However, this positive effect is not reflected on in vivo fertility post-AI.

Evaluation of the antioxidative enzymes in the seminal plasma of infertile men: Contribution to classic semen quality analysis

ABSTRACTProtein expression/activity of antioxidative defense enzymes (AD) in seminal plasma of fertile men might be used as biomarkers of male fertility status. To test this concept, the present study examined the semen parameters of males among 14 normal idiopathic (normozoospermia) and 84 subnormal (teratozoospermia, oligoteratozoospermia, oligoasthenoteratozoospermia) infertile individuals\\. We investigated levels of protein expression/activity of Cu, Zn superoxide dismutase (CuZnSOD), manganese superoxide dismutase (MnSOD), catalase and glutathione peroxidase (GSH-Px), their association with functional sperm parameters, as well as their potential to serve as biomarkers of specific sperm pathologies. Although the activity of CuZnSOD and protein expression of catalase were significantly correlated with several sperm parameters, underlying their potential role in etiology of various sperm abnormalities, investigation of their potential usefulness as a biomarker of semen quality showed that these AD enzymes could not distinguish subtle differences between various sperm pathologies. In contrast, GSH-Px activity was decreased in all groups with sperm pathologies and was a very good indicator of aberrations in functional sperm parameters, explaining up to 94.6% of infertility cases where functional sperm parameters were affected. Therefore, assessment of GSH-Px activity showed the potential to discriminate between infertile males with normal and subnormal semen characteristics and may prove useful in the evaluation of male (in)fertility.Abbreviations: AD: antioxidative defense; Cu, Zn SOD: copper, zinc superoxide dismutase; GSH-Px: glutathione peroxidase; MnSOD: manganese superoxide dismutase; NS: normospermia; OATS: oligoasthenoteratozoospermia; OTS: oligoteratozoospermia; ROC: receiver operating characteristic; ROS: reactive oxygen species; TS: teratozoospermia; WHO: world health organization

CAT-262CT Genotype shows higher catalase activity in seminal plasma and lower risk of male infertility

Catalase (CAT), the key enzyme for high H2O2 elimination, constitutes, along with superoxide dismutase (SOD) and gluthatione peroxidase (GPX), the main enzymatic antioxidant system. Catalase enzyme plays an important role in seminal antioxidant defence and is present in seminal plasma at high concentrations. Low concentrations of catalase enzyme in seminal plasma have been related with male infertility. The objective of this study was to evaluate the correlation between the CAT C-262T polymorphism and male infertility. Semen samples from 400 infertile patients and 80 donors with proven fertility were included in the study. Standart semen parameters, DNA integrity and oxidative stress were evaluated. Genotype frequencies for the CAT C-262T polymorphism were determined by PCR-RFLP. Catalase concentration and activity in seminal plasma were evaluated by western-blot and activity gels respectively while catalase mRNA levels in mature spermatozoa were evaluated by qPCR. The CC genotype was associated with an increased risk of male infertility (OR = 1.976; CI = 1.241–3.216; P = 0.006) while the CT genotype was associated with a protective effect against infertility (OR = 0.479; CI = 0.293–0.783; P = 0.003). Functionally, the CT genotype showed higher concentrations of the enzyme in seminal plasma as well as 2.70 times higher activity levels than the CC genotype. Also, the CT genotype showed 3.5 higher levels of CAT mRNA in the mature spermatozoa than the CC genotype. This suggests that the CAT -262CT genotype correlates with a lower risk of suffering male infertility.

Sperm kinematics and subpopulational responses during the cryopreservation process in caprine ejaculates

Sperm cryopreservation in goats has been a challenge for many years due to the detrimental effects of seminal plasma enzymes produced by the bulbo-urethral glands which catalyse the hydrolysis of lecithins in egg yolk to fatty acids and lysolecithins which are deleterious to spermatozoa. This fact implies to carry out additional processing steps during sperm cryopreservation for seminal plasma removal triggering different sperm responses which may affect sperm functionality. The objective of the present study was to determine specific sperm subpopulation responses in different handling steps during the cryopreservation process by using functional sperm kinematic descriptors in caprine ejaculates. Buck ejaculates (n = 40) were analysed for sperm concentration, viability, morphology and acrosome integrity. Moreover, sperm motility was assessed using a computer-assisted sperm analysis (CASA) system after five different handling steps (fresh sperm, 1st washing, 2nd washing, cooling and frozen-thawed sperm) during a standard cryopreservation protocol for goat semen. The results were analysed using Principal Component Analysis (PCA) and multivariate clustering procedures to establish the relationship between the distribution of the subpopulations found and the functional sperm motility in each step. Except for the 1st and 4th steps, four sperm kinematic subpopulations were observed explaining more than 75% of the variance. Based on velocity and linearity parameters and the subpopulations disclosed, the kinematic response varies among processing steps modifying sperm movement trajectories in a subpopulation-specific and handling step-dependent manner (p < 0.001). The predominant motile subpopulation in freshly ejaculated buck sperm had very fast velocity characteristics and a non-linear trajectory (41.1%). Washing buck sperm twice altered the subpopulation structure as well as cooling which resulted in a dramatic reduction in sperm velocities (p < 0.01). Frozen-thawed spermatozoa showed similar characteristics to cooled sperm except there was a further increase in linearity with a large proportion of sperm attributed to new slow, linear cluster (32.5%). In conclusion, this study confirms the variability and heterogeneity of goat sperm kinematic patterns throughout the cryopreservation process and suggests that the predominant motility pattern (assayed in vitro via CASA) of high quality spermatozoa might be typified by high speed and a non-linear trajectory. The relationships among the number and distribution of sperm subpopulations and the different handling steps were particularlly relevant, specially after the cooling and the post-thawing steps, when effects derived from these critical handling steps were evident and altered drastically the sperm motion patterns.

Effects of season on boar semen parameters and antioxidant enzymes in the south subtropical region in Brazil.

Although boar semen productivity is affected by seasonality, its effects are not equal among different regions which raise concerns regarding the profitability of boar stud farms. Therefore, the goals of this study were (i) to evaluate the seasonal effect on semen production in a commercial boar stud farm located in a subtropical climate region and (ii) to verify whether the activities of superoxide dismutase and glutathione peroxidase in spermatozoa and seminal plasma were associated with seminal traits of fresh and cooled semen. Nine boars were collected twice per season, and routine seminal parameter analyses were performed together with superoxide dismutase and glutathione peroxidase activities in seminal plasma and spermatozoa. Despite a reduction in sperm concentration in spring and summer, most seminal parameters were constant year-round. Temperature-humidity index was higher in the summer compared to spring, autumn and winter (p<.05). Superoxide dismutase activity in spermatozoa was increased in summer compared to autumn and winter (p<.05). The activities of both enzymes in seminal plasma and spermatozoa glutathione peroxidase remained unaltered throughout the seasons. In conclusion, seasonality showed little influence in overall boar seminal parameters despite microclimatic differences among seasons, and spermatozoa collected during summer increased superoxide dismutase activity.

Enzymatic Activities in Fresh Seminal Plasma and Extended Refrigerated Semen in Nari Suvarna Rams

Enzyme activities in the seminal plasma are the good indicators of fertility. A study was conducted to determine the activities of certain enzymes in fresh seminal plasma and extended refrigerated semen in Nari Suvarna rams. Six rams of Nari Suvarna breed were selected and sixteen ejaculates were collected from each ram for a period of eight weeks at the rate of two collections per week by artificial vagina technique. The seminal plasma was separated by centrifugation. Enzyme activities such as alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) were estimated. There was a non-significant (P>0.05) difference between the activities of enzymes in seminal plasma except ALP activity between the rams. The semen was extended using Tris and citrate buffers and the enzymatic activities such as ALP, ALT, AST and LDH were determined after refrigeration for 24 hrs for four Nari Suvarna rams. The numerically increased activities of all the enzymes were observed after extension and refrigeration. It was concluded that the levels of ALP, ALT, AST and LDH enzymes were established in the seminal plasma of Nari Suvarna rams in the present study and the numerical increase in the activities of these enzymes after refrigeration with extension of Tris and citrate buffers for 24 hours could be due to damage to the sperm membrane and leakage of enzymes in to the seminal fluid.

Determination of enzyme activity in rabbit seminal plasma and its relationship with quality semen parameters

&lt;p&gt;The objective of this study was to determine rabbit seminal plasma enzyme activity. Furthermore, correlations between semen parameters and enzyme activity and male age were examined. The study was performed using 17 New Zealand White males from 5 to 9 mo old. Overall, 252 semen samples were collected from bucks from May to September. Semen characteristics were analysed and the seminal plasma was obtained by centrifugation. The activities of alanyl aminopeptidase (APN), aspartate transaminase (AST), alanine aminotransferase (ALT), γ-glutamyl transpeptidase (GGT), lactate dehydrogenase (LDH) and alkaline phosphatase (ALKP) in the seminal plasma fluid were measured. Significant differences between males were found in APN, GGT, LDH, ALKP and ALT activities (P&amp;lt;0.05). No significant differences between enzyme activity and male age were found. We also observed significant positive correlations between male age and sperm concentration (r=0.26), progressive motility (r=0.17) and amplitude of lateral head displacement (r=0.21), and negative ones between male age and average path velocity (r=–0.56), velocity of the sperm head along its actual curvilinear path (r=–0.61), straight line velocity (r=–0.50), linearity index (r=–0.13), and cytoplasmic droplet (r=–0.33). Furthermore, a significant negative correlation between APN activity and the status of the acrosome (r=–0.20) and significant positive correlations between APN activity and the sperm abnormalities (r=0.21), GGT activity and sperm concentration (r=0.34) and the status of the acrosome (r=0.31), and ALKP activity and sperm concentration were observed (r=0.41). In our study, APN and GGT seem to be the most predictive enzymes for rabbit semen quality.&lt;/p&gt;

Correlation between sperm parameters and protein expression of antioxidative defense enzymes in seminal plasma: a pilot study.

Background. Semen analysis is the cornerstone in the evaluation of male (in)fertility. However, there are men with normal semen tests but with impaired fertilizing ability, as well as fertile men with poor sperm characteristics. Thus, there is rising interest to find novel parameters that will help to predict and define the functional capacity of spermatozoa. Methods. We examined whether there is a correlation between semen parameters (count, progressive motility, and morphology) and protein expression/activity of antioxidative defense enzymes in seminal plasma from 10 normospermic subjects. Results. Sperm progressive motility was in positive correlation with seminal plasma protein expression of both superoxide dismutase (SOD) isoforms (MnSOD and CuZnSOD) and catalase. Also, positive correlation was observed between sperm count and MnSOD protein expression, as well as between sperm morphology and protein expression of catalase in seminal plasma. In contrast, protein expression of glutathione peroxidase was not in correlation with any sperm parameter, while its activity negatively correlated with sperm morphology and motility. Conclusions. These data suggest that evaluation of protein expression of antioxidative defense enzymes in seminal plasma might be of importance in the evaluation of male fertility status and that could be used as an additional biomarker along with classic semen analysis in assessment of semen quality.

Melatonin in sperm biology: breaking paradigms.

Melatonin is a ubiquitous molecule, present in a wide range of organisms, and involved in multiple functions. Melatonin relays the information about the photoperiod to the tissues that express melatonin-binding sites in both central and peripheral nervous systems. This hormone has a complex mechanism of action. It can cross the cell plasma membrane and exert its actions in all cells of the body. Certain melatonin actions are mediated by receptors that belong to the superfamily of G-protein-coupled receptors (GPCRs), the MT1 and MT2 membrane. Melatonin can also bind to calmodulin as well as to nuclear receptors of the retinoic acid receptor family, RORα1, RORα2 and RZRβ. The purpose of this review is to report on recent developments in the physiological role of melatonin and its receptors. Specific issues concerning the biological function of melatonin in mammalian seasonal reproduction and spermatozoa are considered. The significance of the continuous presence of melatonin in seminal plasma with a fairly constant concentration is also discussed.

Spermatozoa and seminal plasma enzyme activity in Persian sturgeon,Acipenser persicus, in relation to short-term storage

An important component of semen examination is the comparison between the characteristics of fresh and stored semen (Lahnsteiner, Berger, Weismann & Patzner 1996). The relationships among the parameters of fresh and cryopreserved semen may be determined to identify markers for prediction of the effectiveness of the cryopreservation procedure for fish semen (Lahnsteiner et al. 1996). As the quality of cryopreserved semen is significantly lower than that of fresh semen, such markers are important for successful cryobanking of fish semen (Piros, Glogowski, Kolman, Rzemieniecki, Domagala, Horvath, Urbanyi & Ciereszko 2002). The handling and storage of gametes is central to many assisted reproductive procedures and may be broadly considered as either short-term storage, at temperatures above 0°C, or long-term storage at temperatures below 0°C. Short-term storage of fish gametes may be necessary when the availability of male and female gametes for in vitro fertilization is asynchronous (Lahnsteiner, Weismann & Patzner 1997) or if the sources of gametes are spatially separated. Preservation may also be necessary during transport to a facility with the capacity to cryopreserve gametes for long-term storage, in certain instances when spermatozoa and oocytes have been removed from moribund or recentlydead animals in the field. Several factors can, however, affect the quality and viability of the stored sperm. In this regard, the assessment of the biochemical characteristics of sperm, particularly in terms of enzyme characteristics, can provide new insights into spermatozoa motility and fertilization ability, thus creating opportunities for improving artificial reproduction and germplasm resource conservation procedures (Li, Hulak & Linhart 2008). The enzymology of fish gametes is not well known, when compared with that associated with mammalian semen. Detailed results of research on the enzymes of fish spermatozoa are available with respect to aspartate aminotransferase (AST), lactate dehydrogenase (LDH), acid phosphatase (AcP), b-N-acetylglucosaminidase (b-N AGase) and arylsulfatase (AS) (Schmehl, Graham & Erdhal 1987; McNiven, Gallant & Richardson 1992; Ciereszko & Dabrowski 1994; Piros et al. 2002;). AS and b-N AGase are among the key enzymes, localized within the mammalian acrosome, that play a pivotal role during penetration of the oocyte (Nikolajczyk & O’Rand 1992; Brandon, Srivastava, Heusner & Fayer-Hosken 1997). The activity of AS and b-N AGase during fertilization in sturgeon can provide useful information regarding speciesspecific fertilization and prevention of polyspermy (when a single egg is fertilized by more than one sperm) as postulated by Cherr and Clark (1985). AcP, one of the enzymes found in sperm, has been described in mammalian spermatozoa and seminal plasma (Yousef, Diamandis, Jung & Diamandis 2001) and has also been observed in spermatozoa

Blood and seminal plasma enzyme values of Persian sturgeon, Acipenser persicus (Chordata: Osteichthyes)

An important aspect in the management and protection of endangered species from extinction is the determination of normal physiological values, including the blood and seminal plasma properties. This study was undertaken to establish a reference range for six important blood and seminal plasma enzymes. Samples of 20 Acipenser persicus Borodin, 1897 (10 female and 10 male) were analyzed. The reference ranges for the total samples and also for each sex were determined. The activities of measured enzymes in blood and semen were: alkaline phosphatase (ALP, 363.2 ± 25.24 and 12.23 ± 3.66 IU/L), acid phosphatase (ACP, 4.98 ± 0.39 and 1.86 ± 0.71 IU/L), lactate dehydrogenase (LDH, 1978.8 ± 176.5 and 472 ± 138.7 IU/L), creatine kinase (CK, 1149.6 ± 110 and 553.8 ± 119.9 IU/L), aspartate aminotransferase (AST, 51.12 ± 3.5 and 2.29 ± 1.34 IU/L) and alanine aminotransferase (ALT, 14.7 ± 0.89 and 6.9 ± 2.5 IU/L), respectively. There were significant differences between male and female fish in terms of LDH, CK and ALT activity (P < 0.05). To detect relationships among investigated parameters we used the Pearson correlation coefficient; no significant correlations were observed between these parameters in either blood or seminal plasma.

EFFECT OF ANTHELMINTICS DRUGS ON BIOCHEMICAL CHARACTERISTICS OF BULL FRIESIAN FROZEN SEMEN

Three different antihelmintic drugs were used in the present study (ivermectin, levamisole andalbendazole) on Friesian bulls .The animals were divided into four groups (control untreatedgroup, ivermectin, levamisole, and albendazole treated groups, respectively). The drugs used attwo doses with 8 weeks intervals between first and second one at 200μg/kg, 7.5mg/kg and10mg/kg, respectively. Semen samples were collected for 8th weeks after each dose.Biochemical semen characteristics were estimated. The bulls treated with ivermectin showedsignificant increase in semen- ejaculate volume, sperm- cells concentration (x109 / ml), totalsperm output ejaculate (x109 / ml), alkaline phosphatase (ALP) and prostatic acid phosphatase(ACP) activities in seminal plasma, seminal plasma total proteins, globulin, urea and zincconcentration; while significantly decreased seminal plasma alanine-aminotransferase (ALT),aspartate-aminotranferase (AST), and sodium were reported. The bulls treated with levamisoleshowed significant increase in semen-ejaculate volume, total sperm output/ejaculate, ALP andprostatic ACP activities in seminal plasma, seminal plasma total proteins, globulin, urea andzinc concentrations; while significantly decreased seminal plasma ALT, AST, and sodium wererecorded. In contrary, the bulls treated with albendazole were significantly decreased semenejaculatevolume, sperm motility, seminal plasma total proteins, globulin, creatinine andcalcium. Also, it showed significant increase in seminal plasma enzymes activity (ALT, ASTand ALP) and zinc concentrations. It is interest to note that different anthelmintics treatmentused in this study didn’t affect significantly in seminal plasma albumin and total ACP activity.In conclusion: Ivermectin treatment improved semen qualities by increasing seminal volume andconcentration. Anthelmintics treatment with levamisole should be used carefully under controlduring breeding season. Albendazole treatment should be avoided because its harmful effect insemen by decreasing semen-ejaculate volume, sperm motility, seminal plasma total proteins,globulin and increasing seminal plasma aminotransferase enzymes activity.