Seminal Plasma Catalase Research Articles

Coenzyme Q10 improves the quality and invitro fertility of post-thawed buffalo (Bubalus bubalis) semen via its antioxidative effect.

This study aimed to determine the effects of Coenzyme Q10 (CoQ10) in the freezing medium on functional and oxidative stress parameters and invitro fertilization (IVF) rate of buffalo sperm. Collected samples were relocated to the laboratory for initial evaluation, gentle dilution in extenders, cooling (4°C, 2 h), equilibration (4°C, 4 h), packaging (straws, 0.5 mL), programmable freezing, and thawing (37°C, 30 s). Statistical analysis depicted that adding CoQ10 (100 μM) in a freezing medium caused a significant augmentation in total motility (%), average path, and straight-line velocities (μm/sec) of buffalo sperm than control. Adding CoQ10 (100 μM) improved sperm progressive motility, rapid velocity, and functional parameters (%) compared to the control and 10 μM of CoQ10. Moreover, CoQ10 in a freezing medium caused a significant augmentation in seminal plasma catalase (U/mL) and glutathione reductase (GSH; nmol/109 ) at 100 μM than control and other treatments. CoQ10 inclusion (100 μM) ameliorates seminal plasma superoxide dismutase (U/mL), glutathione-S-transferase (GST; nmol/mL/min) fructose (μg/mL), and ATP (nmol/million) than control. Furthermore, CoQ10 at 100 μM improved seminal plasma glutathione peroxidase (μM) levels than control, 10 μM, and 20 μM. Lastly, hydrogen peroxide (H2 O2; nM) production was significantly lower at 100 μM than at control and 10 μM. CoQ10 (100 μM) caused a significant augmentation in the un-capacitated pattern followed by a reduction in the capacitated pattern, and apoptosis-like changes (%) than control, and other treatments, whereas viability was increased than control and other treatments. CoQ10 (100 μM) significantly improved the IVF rate in comparison with control, CoQ10 at 10 μM, and 20 μM groups. In conclusion, the addition of CoQ10 (100 μM) in the freezing medium can improve the quality and invitro fertility of post-thawed buffalo semen via its antioxidative effect. Further studies are needed to evaluate the effect of CoQ10 on the invivo fertility of buffalo bull semen.

P-566 Is the seminal oxidative stress the mirror of psychological stress perceived by infertile men?

Abstract Study question To assess the association between anxiety and depression scores and the levels of oxidative stress in seminal plasma of Tunisian infertile men. Summary answer Depression in hypofertile men is associated to higher levels of catalase in seminal plasma and thus to oxidative stress. What is known already Within the last decades, the knowledge concerning the link between psychological and oxidative stress in infertile men has been surging. In a limited number of studies aiming to elucidate the psychological aspect of male infertility, data related to increased incidence of depression and anxiety have been reported. It has also been reported that anxiety and depression may trigger the production of reactive free oxygen radicals leading to disruption of the balance between free radicals and antioxidants semen properties. Study design, size, duration This was a cross-sectional study performed in the Laboratory of Cytogenetics and Reproductive Biology of Fattouma Bourguiba University Teaching Hospital (Monastir Tunisia). A total of 282 patients were assessed for levels of anxiety and depression and evaluated for semen parameters. Among these, 105 patients were assessed for levels of superoxide dismutase (SOD) and 51 patients were evaluated for the levels of catalase in seminal plasma from September 2022 to January 2023. Participants/materials, setting, methods Were included patients addressed for semen quality assessment. All patients with a previously diagnosed psychological disorder, those who presented severe depression and/or anxiety symptoms or had a stressful life event were not included. Informed consent was obtained from all the participants. The subjects were evaluated for anxiety and depression symptoms using the valid Arab version of the HAD (Hospital Anxiety and Depression) scale. Semen analysis and results interpretation were performed according to 2021 WHO guidelines. Main results and the role of chance The mean HAD-D (depression) and HAD-A (anxiety) scores were of 6.56 ± 3.07 (IIQ [4-8]) and 7.94 ± 3.73 (IIQ [5-10]) respectively. The mean levels of SOD and catalase were of 42.32 ± 24.08 and 19599.82 ± 12745.36 respectively. The results showed that patients exhibiting elevated HAD-D scores have higher levels of catalase in seminal plasma compared to those with normally ranging HAD-D scores (29856.07±15904.51 VS 17968.14±11564.79 respectively; p= 0.02). However, SOD levels were similar between the two groups and no correlation was found between seminal oxidative stress as assessed by catalase and SOD levels and both HAD-D and HAD-A scores. Limitations, reasons for caution The main limitation reason could be related to the limited number of patients evaluated for levels of catalase in seminal plasma (N = 51). Furthermore, assessing hormone’s levels (LH, FSH and testosterone) would be of great interest in elucidating the implicated pathways leading to oxidative stress and impaired semen quality. Wider implications of the findings Our results shed the light on the elevated levels of the catalase in patients with high depression scores. The absence of correlation between antioxidant enzymes and psychological well-being of infertile patients is a reassuring finding. However, awareness and recognition of depressive symptoms in infertile men is crucial when managing infertility. Trial registration number not applicable

Enhancing semen quality, antioxidant status and sex hormones of V-line rabbit bucks fed on supplemented diets with dried moringa leaves

This study was conducted to evaluate the impact of dried moringa leaves (DML) on semen quality, immunoglobulin measurements, blood metabolic analysis, antioxidant status and sex hormones of V-line rabbit bucks. Twenty-four mature rabbit bucks of 7-8 months-old of proven fertility with an average weight (2.8 ± 0.4 kg) were classified into four treatments (basal diet with 0, 750, 1500 and 3000 mg DML/kg diet) and 6 bucks per treatment were used. Seminal plasma cholesterol (p = 0.013) and triglycerides (p = 0.0001) of all moringa supplemented rabbits were higher than the control. Moringa leaves enhanced (p < 0.05) serum testosterone, follicle stimulating hormone and luteinizing hormone. Total antioxidant capacity, superoxide oxidase, glutathione peroxidase and catalase in seminal plasma of moringa rabbits were higher (p < 0.05) than the control. Serum total lipid (p = 0.002), cholesterol (p = 0.008), triglycerides (p = 0.019) and blood urea (p < 0.05) were lower in DML rabbits. Moringa leaves improved total antioxidant capacity (p = 0.0001), glutathione peroxidase (p = 0.0001) and superoxide oxidase (p = 0.037) in rabbits versus control. Rabbits consuming DML had higher (p < 0.05) immunoglobulin IgG and IgM than control. Results suggest that DML improved rabbits’ antioxidant, and immunological status, semen quality, and sex hormones, but DML (750 mg/kg diet) is recommended because it was more effective than the other two high doses.

Differentially expressed microRNAs in biochemically characterized FrieswalTM crossbred bull semen

In addition to the transmission of paternal genome, spermatozoa also carry coding as well as noncoding microRNAs (miRNAs) into the female oocyte during the process of biological fertilization. Based on RNA deep sequencing, a total 28 number of differentially expressed miRNAs were cataloged in categorized FrieswalTM crossbred (Holstein Friesian X Sahiwal) bull semen on the basis of conception rate (CR) in field progeny testing program. Validation of selected miRNAs viz. bta-mir-182, bta-let-7b, bta-mir-34c and bta-mir-20a revealed that, superior bull semen having comparatively (p < .05) lower level of all the miRNAs in contrast to inferior bull semen. Additionally, it was illustrated that, bta-mir-20a and bta-mir-34c miRNAs are negatively (p < .01) correlated with seminal plasma catalase (CAT) activity and glutathione peroxidase (GPx) level. Interactome studies identified that bta-mir-140, bta-mir-342, bta-mir-1306 and bta-mir-217 can target few of the important solute carrier (SLC) proteins viz. SLC30A3, SLC39A9, SLC31A1 and SLC38A2, respectively. Interestingly, it was noticed that all the SLCs were significantly (p < .05) expressed at higher level in superior quality bull semen and they are negatively correlated (p < .01) with their corresponding miRNAs as mentioned. This study may reflect the role of miRNAs in regulating few of the candidate genes and thus may influence the bull semen quality traits.

Effect of heavy metals on oxidative markers and semen quality parameters in HF crossbred bulls

The study was conducted to assess the effect of toxic heavy metals on seminal antioxidants, trace minerals and semen quality parameters in crossbred breeding bulls. Semen samples were collected from breeding bulls (82) maintained at bull rearing unit of the institute. Toxic heavy metals (Pb and Cd), trace minerals (Zn, Cu, Co and Fe) and oxidative stress markers (SOD, catalase and MDA) were determined in semen samples of breeding bulls. Sperm motility and concentration were measured in fresh ejaculates. Significant levels of Pb (0.23±0.006 μg/ml) and Cd (0.11±0.005 μg/ml) were detected in semen samples of breeding bulls. On analysis of the data between good and poor bull categories, significantly higher concentration of Pb and Cd was present in semen samples of poor bulls as compared to good bulls. There was significantly higher malondialdehyde (MDA) and low superoxide dismutase (SOD) and catalase in seminal plasma of poor bulls compared to good bulls. A significant decrease in Zn and Cu was observed in poor bulls as compared to good bulls. Cobalt and iron values did not show any significant variation between good and poor quality bulls. On correlation analysis, lead and cadmium showed significant negative correlation with Zn, Cu, SOD, catalase, motility and sperm concentration while significant positive correlation was seen with MDA respectively. The study concludes that increased Pb and Cd in bull semen may increase the risk of seminal oxidative stress development and a subsequent reduction in male fertility.

Human Sperm Characteristics with Regard to Cobalt, Chromium, and Lead in Semen and Activity of Catalase in Seminal Plasma.

We analyzed cobalt (Co), chromium (Cr), and lead (Pb) concentrations in human semen and catalase CAT activity in seminal plasma and the effects of their relations on the sperm quality. We obtained semen samples from men (n = 168) undergoing routine infertility evaluation. Studies included two groups based on the ejaculate parameters: I (n = 39; normal ejaculate; normozoospermia); II (n = 129; pathological spermiogram). We examined relationships and differences between Co, Cr, and Pb concentrations in seminal plasma, CAT activity, and semen parameters. We did not establish differences in Co, Cr, and Pb concentrations and CAT activity from men between normozoospermic and those with pathological spermiogram. We found a significantly lower Co concentration and CAT activity in males with normal sperm motility than in asthenozoospermic males. We found significantly lower Co and a higher Pb concentration in males with normal morphology of spermatozoa than in teratozoospermic males. We found a significantly higher Pb concentration in the individuals with consumption of alcohol than in those without consumption. There were significant correlations between Co and Pb concentrations, sperm progressive motility (A + B, i.e., fast and slow progressive motility; Co-negatively; Pb-positively), and normal morphology of spermatozoa (Co-negatively; Pb-positively). We found a significant negative correlation between Cr concentration and slow progressive motility, and between CAT activity and volume of ejaculate. Co, Cr, and Pb levels and CAT activity were related to sperm characteristics and male fertility. The impact of alcohol may be manifested by a disturbance in Pb equilibrium in the body. Co and Pb influence progressive motility and normal morphology of human spermatozoa. Thus, Co and Pb levels in semen may be a useful diagnostic in male infertility. Most of the results of this study are in contrast to expectations. Namely, Pb is a toxic element and its harmful effects (poor semen quality) may be expected already at relatively low level of Pb exposure and are particularly visible with increasing of Pb. Co and Cr(III) are essential elements and harmful effects may be expected at their deficiency and/or overexposure.

Age-related differences of semen quality, seminal plasma, and spermatozoa antioxidative and oxidative stress variables in bulls during cold and warm periods of the year

The aims of this study were to determine the presence and quantities of antioxidative status and oxidative stress (OS) variables in the seminal plasma and spermatozoa of bulls of varying age during cold and warm periods of the year, and to establish the correlation of these variables with semen quality parameters. The study was conducted on two groups each comprising nine Simmental bulls: one group contained younger animals (aged 2 to 4 years) and the second older animals (aged 5 to 10 years). Semen samples were collected using an artificial vagina for biochemical analysis. Seminal plasma and spermatozoa activities of total superoxide dismutase (TSOD), manganese superoxide dismutase (MnSOD), copper–zinc superoxide dismutase (CuZnSOD), catalase (CAT), selenium-dependent glutathione peroxidase, reduced glutathione and concentrations of total protein (TP), thiobarbituric acid reactive substances (TBARS) and protein carbonyl content (PCC) were determined. Several antioxidants in seminal plasma were also determined: total glutathione peroxidase (TGSH-Px), selenium-independent glutathione peroxidase (Non-SeGSH-Px), uric acid, albumins (ALB) and alkaline phosphatase (ALP). Significantly higher spermatozoa motility was observed during the cold v. warm period, and a significantly higher volume and total number of spermatozoa per ejaculate was observed in older than in younger bulls. Significantly higher values of ALP, TP and ALB were found in seminal plasma of older bulls than in younger bulls during the warm period. The seminal plasma of younger bulls showed significantly higher activities of TSOD, MnSOD, CuZnSOD, TGSH-Px and Non-SeGSH-Px. Younger bulls had significantly higher PCC concentration and activity of CAT in seminal plasma than older bulls during the cold period. Significantly higher concentrations of PCC and TBARS, and activities of TSOD, MnSOD and CuZnSOD were established in spermatozoa of the younger than in older bulls during the warm period. It could be concluded that antioxidative and OS variables differ significantly depending on bull age and time of year. Younger bulls were more sensitive to elevated ambient temperatures during the warm period, when the higher enzymatic antioxidative protection in seminal plasma and spermatozoa were insufficient to counteract the intensive oxidative processes in spermatozoa, which eventually resulted in decreased spermatozoa motility. The estimation of antioxidative and OS variables in seminal plasma and spermatozoa may have practical value for the assessment of bull semen quality.

Association of CAT–262C/T with the concentration of catalase in seminal plasma and the risk for male infertility in Algeria

ABSTRACTCatalase (CAT) plays a central role in the protection of different cell types against the deleterious effects of hydrogen peroxide. In human, CAT is implicated in many physiological and pathological conditions including idiopathic male infertility. In this study we examined the association between CAT levels in seminal plasma with different sperm parameters and with CAT–262 C/T polymorphism and their risk for idiopathic male infertility in Algeria. Semen and blood samples were obtained from 111 infertile males and 104 fertile controls from the region of Eastern Algeria following informed consent. Standard semen parameters, DNA integrity, and CAT concentration in seminal plasma were evaluated. CAT-262C/T genotypes were screened using allele specific PCR. Seminal CAT activity was significantly different (p<0.0001) between infertile males and controls, it was also markedly decreased in oligo-astheno-teratozoospermia (p<0.0001), azoospermia (p<0.0001), and normozoospermia (p=0.045) subgroups compared to controls. Positive correlations between CAT activity and semen parameters (volume, motility, concentration, and morphology) were detected, but not with sperm DNA integrity. There was no direct association between CAT-262C/T polymorphism and general male infertility. However, the results presented in this study showed that CAT activity is remarkably associated with the CAT-262T allele (p=0.001) and the different CAT-262C/T genotypes. This study highlighted the major differences in the seminal plasma CAT content between infertile and fertile males and the differences of CAT concentration between different CAT-262C/T genotypes carriers.

45 ACROSOME REACTION AND HETEROLOGOUS ZONA BINDING ASSAY OF FROZEN STALLION SPERM AFTER HYPERACTIVATION

During cryopreservation and due to the large portion of seminal plasma removal, there is a decrease in equine sperm antioxidant protection. Lactoferrin and catalase in seminal plasma play an antioxidant role. The fertilizing ability of equine sperm has been analysed in vitro using sperm-zona binding assays with heterologous oocytes. The results have been correlated with in vivo fertility by means of acrosome reaction (AR) and the number of attached sperm to the zona pellucida (ZP). The aim of the present work was to estimate the potential fertilizing ability of stallion sperm frozen with INRA82 extender (Battelier et al. 1997) with lactoferrin and catalase, and after hyperactivation with procaine and calcium ionophore A-23187 (Ca-I) by determining the AR rate and number of attached sperm to the bovine ZP. Semen from 6 stallions was frozen with 3 extenders: (T1) control, INRA 82; (T2) T1 + 500 μg mL–1 lactoferrin; and (T3) T1 + 200 IU mL–1 catalase. After semen thawing, the sperm were selected by swim-up and distributed in 3 aliquots according to the hyperactivation treatments: (H1) control, after thawing; (H2) capacitating Whitten’s medium + 5 mM procaine chloride; and (H3) capacitating Whitten’s medium + 5 μM Ca-I. To the zona binding assays, bovine oocytes derived from abattoir ovaries were incubated at 38.5°C with 5% CO2 (1 h), and 5 oocytes were poured into each treatment droplet under mineral oil. Sperm were stained with Hoechst 33342 dye (35 μg mL–1), and after 2 h co-culture, the number of sperm attached to the ZP was determined with epi-fluorescent microscopy. The rate of sperm AR was determined after freezing-thawing (control) and hyperactivation treatments with propidium iodide and fluorescein isothiocyanate/peanut agglutinin dies with a flow cytometer. The green fluorescent (peanut agglutinin+) and not red stained (propidium iodide) sperm were considered acrosome reacted. Means of ZP attached sperm and percentage of AR sperm were analysed by ANOVA and Tukey test. A probability of P &lt; 0.05 was considered significant. The mean of ZP attached sperm (4.2 ± 3.5) and AR sperm rate (4.4 ± 3.7%) did not differ among the extenders (P &gt; 0.05). The rate of sperm AR after hyperactivation with procaine (5.2 ± 2.4%) did not differ to the Ca-I (6.1 ± 3.7%); however, they were higher than the spontaneous AR rate (1.1 ± 0.5%, P &lt; 0.05). Lower number of ZP attached sperm was observed by the Ca-I induced hyperactivation protocol (1.9 ± 2.1) compared with the procaine (5.9 ± 3.7; P &lt; 0.05), although they did not differ to the control (3.3 ± 2.7). In conclusion stallion frozen sperm were better hyperactivated with procaine than with Ca-I, and therefore, it is a more suitable sperm hyperactivation inductor to study equine IVF protocols with frozen semen. Acknowledgments are extended to CAPES, Brazil, for the financial support.

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.

Blood catalase and haematocrit values in a breeding colony of Dutch-belted rabbits

Rabbit seminal plasma catalase is much higher than in the semen of other mammals, and differences appear to be inherited. Because of the scarcity of information on rabbit blood catalase and haematocrit in Dutch-belted rabbits, an investigation of possible effects of gender, age and genetics on these variables was undertaken. There were 191 rabbits sampled at 2-3 months, 130 at 12 months and 61 at 18-24 months of age. There was no age effect on the haematocrit values and on blood catalase activity. At 12 months of age males had an average haematocrit value of 44% compared with 40% for females (P < 0.05). Corresponding average catalase values were 431 and 356 units/ml of blood (P < 0.05). Also catalase was measured in the semen and blood of 34 males, and males differed in both their blood and semen catalase activity (P < 0.05). The correlation between the two traits was r = 0.44. Heritability (h2) estimates, based on 231 rabbits were 0.40 for blood catalase activity, and 0.26 for haematocrit. The genetic correlation between the two variables was 0.83 (P < 0.05). These studies are consistent with the literature in that female rabbits have a slightly lower haematocrit value than males, and this is associated with a lower catalase activity. This appears to be the first report of a study that compares rabbit blood catalase in males and females of different ages. Preliminary evidence that differences may have a heritable basis is consistent with previous studies on rabbit semen catalase.