Seminal Plasma Antioxidant Research Articles

Effect of Dietary Copper and Zinc Supplementation on Semen Quality of Murrah Bulls

The effect of dietary supplementation of Cu and Zn on semen quality parameters and certain bio-chemicals parameters were evaluated in Murrah bulls. Twelve mature Murrah bulls (4-6 years of age) were divided into three groups (n=4) T1, T2, and T3 based on semen volume and concentration and were fed as per ICAR standard (2013). However, the animals were supplemented with 0%, 25%, and 50% Cu and Zn above the basal diet in T1, T2 and T3 groups, respectively for 180 days. Quantitative and qualitative characteristics of semen, blood and seminal plasma antioxidant status, blood and semen minerals (Cu, Zn, Ca and Mn) were determined in experimental Murrah bulls. Semen ejaculate volume (mL) increased in T2 and T3 while sperm concentration (million/mL), intact acrosome (%), HOST reacted spermatozoa (%) increased in T3 group compared to T1 and T2 groups. No difference was observed in mass motility, pH, live spermatozoa (%) in semen sexual behaviour except dismounting time which was decreased in T3 than T1 and T2 (plessthan0.05). SOD activity in blood plasma and LPO activity decreased in seminal plasma in both T2 and T3 groups than T1, whereas catalase activity did not show any significant difference. Cu and Zn supplementation in T2 group improved plessthan0.05) Zn level in blood and seminal plasma and Cu level in blood plasma only, but not in seminal plasma. Therefore, it can be concluded that supplementation of Zn and Cu at 50% above the recommended levels of ICAR (2013) improved the qualitative and quantitative attributes of semen in Murrah bulls.

Specific Activity of Superoxide Dismutase in Stallion Seminal Plasma Is Related to Sperm Cryotolerance.

While the removal of seminal plasma is a routine practice prior to equine sperm cryopreservation, this fluid contains the main source of antioxidant enzymes able to scavenge these reactive oxygen species. Therefore, stallion seminal plasma components may have an impact on ejaculate freezability. Against this background, this study was designed to investigate whether the activities of the main stallion seminal plasma antioxidant enzymes are related to sperm cryotolerance. With this purpose, 16 ejaculates were collected from 14 healthy stallions, and each ejaculate was split into two aliquots. The first one was used to evaluate the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), and glutathione reductase (GSR) in seminal plasma. The second aliquot was extended and then processed for cryopreservation. Sperm motility and viability were evaluated before and after cryopreservation, and ejaculates were classified as of good (GFE) or poor freezability (PFE) based on total motile and viable spermatozoa at post-thaw. We observed that, while the specific activities of CAT, GPX, and GSR were similar between GFE and PFE, that of SOD was significantly (p < 0.05) higher in GFE than in PFE. We can thus conclude that, in stallions, the specific activity of SOD in the seminal plasma of a given ejaculate might be related to its freezability.

Effect of Zinc Supplementation on Semen Quality, Sperm Antioxidant Ability, and Seminal and Blood Plasma Mineral Profiles in Cashmere Goats.

This study evaluated the effects of the different dietary zinc (Zn) levels on semen quality, on spermatozoa and seminal plasma antioxidant status, and on the seminal and blood plasma mineral status in mature male Cashmere goats during the breeding season. Twenty-eight mature male Liaoning Cashmere goats were divided into four groups based on body weight (56.2 ± 2.45 kg) and semen characteristics; these goats were fed with basal diet supplemented with 0, 20, 40, or 80 mg Zn/kg DM (zinc sulfate) for 3 months. Results showed that the Zn-supplemented diets linearly increased the semen volume (0.98, 1.04, 1.27, and 1.17 ml for the 0, 20, 40, and 80 mg Zn/kg DM supplementation, respectively) (P < 0.05) and the total sperm output (3.87, 4.52, 5.73, and 5.33 × 109/ml for the 0, 20, 40, and 80 mg Zn/kg DM supplementation, respectively) (P < 0.05); by contrast, Zn supplementation exerted no effect on sperm concentration, motility, and abnormal sperms rate. The activities of copper zinc superoxide dismutase (CuZn-SOD) (linear P < 0.05) and glutathione peroxidase (GSH-Px) (linear P < 0.05; quadratic P < 0.01) were highest in the intermediate supplementation (40 mg Zn/kg DM). Moreover, the malondialdehyde (MDA) content of spermatozoa decreased linearly (P < 0.01) with the increase in Zn supplementation. In seminal plasma, the highest GSH-Px activity was observed in 20 mg Zn/kg DM supplementation (P < 0.05). Catalase (CAT) activities both in the spermatozoa and seminal plasma showed no difference in all treatments. Seminal plasma Zn level was highest in 40 mg Zn/kg DM (linear P = 0.068), and K increased linearly (P = 0.001) with increasing Zn level. Furthermore, blood plasma Zn (linear P < 0.01; quadratic P < 0.05), Fe (linear P < 0.05; quadratic P < 0.05), and Mg (linear P < 0.05) increased with increasing Zn supplementation. These results indicated that dietary Zn supplementation in Cashmere goats during the breeding season improved the semen quality and quantity, elevated the antioxidative indices and Zn concentration, and decreased the MDA content both in spermatozoa and seminal plasma.

Combined addition of superoxide dismutase, catalase and glutathione peroxidase improves quality of cooled stored stallion semen

During cold storage stallion spermatozoa experience undergo oxidative stress, which can impair sperm function and fertilizing capacity. Superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) are the main endogenous enzymatic antioxidants in stallion seminal plasma, and counteract reactive oxygen species. Semen dilution reduces the endogenous antioxidant concentrations. The aim of this study was to investigate whether addition of 15 IU/mL each of SOD, CAT, and GPX to diluted stallion semen would ameliorate a reactive oxygen-mediated decrease in semen quality during 72 h of storage at 5 °C. Ejaculates (n = 7) were divided in two aliquots and diluted in INRA 96 without (control) or with addition of antioxidants. Semen analysis was performed at the time of dilution and every 24 h during chilled storage. Antioxidant supplementation completely inhibited the storage-dependent increase in activated caspase 3 (P < 0.05). Concomitantly, the antioxidant-supplemented samples had a greater percentage of viable, motile and rapidly moving sperm than control samples after 72 h storage (P < 0.05). The DNA damage, as evaluated by TUNEL assay and SCSA, increased with storage time (P < 0.05). Antioxidant supplementation did not prevent, but did significantly reduce the increase in DNA strand breakage. The results indicate part of the intrinsic apoptotic pathway leading to effector caspase activation was inhibited, although an activation of molecules with endonuclease activity still occurred. In conclusion, adding equal concentrations of SOD, CAT and GPX to a semen extender suppressed caspase-3 activation and improved preservation of stallion sperm motility and viability during 72 h of storage at 5 °C.

Antioxidant defence system in protein fractions of common carp (Cyprinus carpio) seminal plasma

There are growing evidences that the damage which is caused to the spermatozoa by the reactive oxygen species (ROS) plays a key role in male infertility. The seminal plasma is endowed with many enzymatic and non-enzymatic antioxidants which protect the spermatozoa against oxidative stress. The present study was undertaken to compare the level of superoxide dismutase (SOD), glutathione reductase (GR) and glutathione peroxidase (GPx) in separated protein fractions of whole common carp Cyprinus carpio seminal plasma. Seminal plasma from six male common carp was separated into four fractions. The fractions differed from each other in their total seminal plasma protein content and antioxidant status. The highest value of SOD (2.81 ± 0.11 mU/mg protein) was found in fraction 4, and the lowest (1.83 ± 0.18 mU/mg protein) in fraction 1. There were significant variations (ANOVA; P = 0.05) in GR activity, mainly between fractions 1, 2, and 4. The lowest value of GPx activity (14.9 ± 0.4 mU/mg protein) was found in fraction 2; however, the GPx activity showed low variability among fractions. We conclude that distinct seminal plasma fractions differ in antioxidant capacity. The inclusion of fractions with high levels of SOD, GR, and GPx in sperm storage medium may enhance protection against oxidative stress and promote sperm survival during the in vitro storage.

Molecular and subcellular cryoinjury of fish spermatozoa and approaches to improve cryopreservation

AbstractThe quality of frozen/thawed fish sperm is generally lower than that of fresh sperm. Extremely low temperatures are associated with damage to fish spermatozoon subcellular compartments and associated molecules such as DNA and proteins. Cryodamage can negatively affect DNA integrity, spermatozoon metabolism and motility, consequently impairing fertilization and embryo development. To preserve sperm efficiently, the addition of proteins including seminal plasma proteins, antioxidants, antifreeze proteins and antifreeze glycoproteins, and bovine serum albumin before and/or after freezing is suggested to minimize cryoinjury. An appropriate quantity of seminal plasma components in cryopreservation medium can prevent spermatozoon membrane damage by maintaining antioxidant enzymes and their proper distribution on the spermatozoon surface. Under storage conditions, the semen antioxidant system is ineffective in protecting spermatozoa from reactive oxygen species. The addition of antioxidants can provide protection for spermatozoa and reduce cryoinjuries during storage, but the effect of a given antioxidant is species‐specific. Antifreeze proteins and antifreeze glycoproteins can stabilize the cell membrane via interaction with phospholipid components and inhibit ice‐crystal growth to reduce damage related to osmotic stress and ice crystallization. However, they may exhibit a cytotoxic effect. This review summarizes the sources and characteristics of subcellular spermatozoon cryoinjury and discusses approaches to improve outcome of fish sperm cryopreservation.

Effects of Low Seminal Plasma Antioxidant Potential on Semen Quality and Male Fertility

It is very important to identify the factors which affect normal sperm functions. New Bio-chemical parameters in seminal plasma were analyzed to determine the biochemical factors that affect normal sperm function. The research groups consists of infertile groups Asthenozoospermia (n=31) and Normozoospermia (n=27) with a healthy men as a control (n=24). The patients have been selected and examined according to the World Health Organization (2010). In addition the seminal fluid analysis the biochemical parameters were analyzed in the seminal plasma of each sample including; Total antioxidant capacity, Catalase, Glutathione Reductase , glutathione and Malondialdehyde.Results showed a decrease in seminal plasma total antioxidant capacity of Astheno-zoospermia and Normozoospermia , and a significance decrease in catalase level for both infertile groups, Malondialdehyde level significantly increased in both infertile groups and Glutathione level significantly decreased in Asthenozoospermia group as compared to control group. Antioxidants have an important role in sperm protection via disturbing the balance in ROS production which destroys sperm plasma membrane causing loss sperm activity and it's ability to fertilize the egg.

The effect of dietary doum supplementation on- productive and reproductive performance of male rabbits in Upper Egypt

The objective of this work was conducted to study the effect of Doum (Hyphaenethebaica) supplementation on productive and reproductive performance of rabbit bucks in Upper Egypt during spring season. Thirty- two males16 California (CAL) and 16 Newzland (NEZ) rabbits 8 months old with average initial body weight 3.409±0.05 kg were randomly divided into four groups of 4 bucks each/breed (CAL and NEZ). Group 1 served as control fed a basal diet. Groups 2, 3 and 4 fed basal diets supplemented with 0.3, 0.6 and 0.9 g doum/kg diets, respectively for 12 weeks. The results revealed that doum supplementation significantly (p≤0.01) increased bucks live body weight compared with control group. Ejaculate volume, mass motility, sperm concentration, total sperm output, total motile sperms, live normal sperm, total functional sperm fraction and initial fructose were significantly increased, while reaction time significantly decreased in rabbit bucks received doum compared to control group. Administration of doum increased (P < 0.05) seminal plasma total proteins, globulins, alkaline phosphatase, acid phosphatase and lactate dehydrogenase. Conversely, seminal plasma aspartate aminotransferase and alanine aminotransferase were significantly decreased compared to the control group. Seminal plasma lipid peroxidation as indicated by thiobarbituric acid-reactive substances was significantly decreased while, seminal plasma antioxidant enzymes were significantly increased due to doum supplementation Also, doum supplementation significant increased serum testosterone level .In conclusion, doum supplementation improved semen quality, and seminal plasma antioxidant status of rabbit bucks. Thus, the doum could help to improve productive and reproductive efficiency of rabbit bucks under Upper Egypt condition.

Impact of Smoking and Alcohol Consumption on Oxidative Status in Male Infertility and Sperm Quality

The most common unhealthy lifestyle factors like cigarette smoking and drinking alcohol have controversial reports on how these impact male fertility. The present study assessed the potential effects of these habits on sperm quality, male reproductive hormones, and blood plasma oxidative profile in 133 infertile men with 124 fertile, healthy controls. Among the participants, 28 (10.8 %), 29 (11.2 %), 33 (12.8 %), 34 (13.2 %) fertile men were non-alcoholics and non-smokers (total abstainers), smokers, alcohol drinkers, and smokers plus alcohol drinkers, respectively. On the other hand, 32 (12.4 %), 30 (11.6 %), 34 (13.2 %), 37 (14.3 %) infertile men were total abstainers, smokers, alcohol drinkers, and smokers plus alcohol drinkers, respectively. The conventional sperm parameters and reproductive hormone levels were found to be significantly different in the subsets of infertile men than those of fertile men (p<0.001). In these subsets of infertile men, blood plasma malondialdehyde, nitric oxide concentrations were significantly higher and superoxide dismutase, reduced glutathione activities were significantly lower when compared to fertile men and infertile men total abstainers group (p<0.001). In infertile subjects, particularly smoking-alcohol drinking group was more affected than only smoking or alcohol drinking groups. Furthermore, these oxidative markers were correlated with sperm concentration, motility, and morphology, except nitric oxide concentration, which was only associated with sperm motility. These findings highlighted that blood plasma antioxidants are as efficient as seminal plasma antioxidants in male fertility evaluation.

Oxidative stress-induced alterations in seminal plasma antioxidants: Is there any association with keap1 gene methylation in human spermatozoa?

Kelch-like ECH-associated protein 1 (keap1)-nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway is one of the master regulators of cellular defence against oxidative stress. Epigenetic alterations like hypermethylation of keap1 gene impair keap1-Nrf2 system in several oxidative stress-associated diseases. The objective of this study was to evaluate the epigenetic status of keap1 in sperm DNA of normozoospermic subjects, having different levels of reactive oxygen species (ROS) in seminal plasma. Semen samples were obtained from 151 apparently healthy male partners of couples who attended the Avicenna infertility clinic. Samples were categorised into four groups according to their ROS levels: group A (n=39, ROS<20RLU/s per 106 spermatozoa), group B (n=38, 20≤ROS<40RLU/s per 106 spermatozoa), group C (n=31, 40≤ROS<60RLU/s per 106 spermatozoa) and group D; (n=43, ROS≥60RLU/s per 106 spermatozoa). Keap1 methylation status was assessed using methylation-specific PCR along with seminal total antioxidant capacity. The results showed no significant alterations in keap1 methylation in any groups, whereas the total antioxidant capacity enhanced with increasing levels of ROS exposure. These results indicate that keap1 was not methylated during ROS elevation and oxidative stress, suggesting that the cells have adopted other mechanisms to elevate antioxidant level.

Oxidative stress induced alterations in seminal plasma antioxidants, is there any association with Keap1 gene methylation in human spermatozoa

Oxidative stress induced alterations in seminal plasma antioxidants, is there any association with Keap1 gene methylation in human spermatozoa

Cadmium effects on sperm morphology and semenogelin with relates to increased ROS in infertile smokers: An in vitro and in silico approach

Smoking releases cadmium (Cd), the metal toxicant which causes an imbalance in reactive oxygen species level in seminal plasma. This imbalance is envisaged to impair the sperm DNA morphology and thereby result in male infertility. In order to correlate this association, we performed in vitro and in silico studies and evaluated the influence of reactive oxygen species imbalance on sperm morphology impairments due to smoking. The study included 76 infertile smokers, 72 infertile non-smokers, 68 fertile smokers and 74 fertile non-smokers (control). Semen samples were collected at regular intervals from all the subjects. Semen parameters were examined by computer assisted semen analysis, quantification of metal toxicant by atomic absorption spectrophotometer, assessment of antioxidants through enzymatic and non-enzymatic methods, diagnosis of reactive oxygen species by nitro blue tetrazolium method and Cd influence on sperm protein by in vitro and in silico methods. Our analysis revealed that the levels of cigarette toxicants in semen were high, accompanied by low levels of antioxidants in seminal plasma of infertile smoker subjects. In addition the investigation of Cd treated sperm cells through scanning electronic microscope showed the mid piece damage of spermatozoa. The dispersive X-ray analysis to identify the elemental composition further confirmed the presence of Cd. Finally, the in-silico analysis on semenogelin sequences revealed the D-H-D motif which represents a favourable binding site for Cd coordination. Our findings clearly indicated the influence of Cd on reactive oxygen species leading to impaired sperm morphology leading to male infertility.

Evaluation of Seminal Plasma Antioxidants and Serum Male Hormones Status in Infertile Patients with Unbalanced Chromosomal Abnormalities

Background: Male infertility appears to be a major clinical problem among men of reproductive age in all societies. Idiopathic male infertility is considered to be a multifactorial disorder affected by genetic, environmental, and hormonal factors. Oxidative stress seems to stand out as one of the underlying mechanisms. In this context, we aimed to evaluate seminal plasma antioxidants SOD (superoxide dismutase), GPx (glutathione peroxidase), CAT (catalase) and zinc levels, hormone levels, and semen parameters in fertile donors and patients with unbalanced chromosomal abnormalities.Methods: Semen samples from 119 patients with unbalanced chromosomal abnormalities and 30 fertile men were analyzed according to World Health Organization guidelines (2010). All patients underwent a measurement of testosterone (T), follicle-stimulating hormone (FSH) and luteinizing hormone (LH). Seminal plasma antioxidants activities of SOD, GPx, CAT and zinc concentration were measured using colorimetric methods.Results and conclusion: Hormonal analysis showed statistically significant increase of FSH and LH levels in all patients (p<0.001). However, a significant decrease of serum testosterone level was observed in patients compared to fertile group (p<0.05). Seminal antioxidant system evaluation revealed a significant decrease of CAT, GPx, SOD and zinc activities in infertile group compared to fertile group (p<0.05). Our study showed that patients with unbalanced chromosomal abnormalities had significant alterations of antioxidant system and a significant perturbation of hormonal levels which interfere with fertility. These alterations may be due to a loss or derangement of chromosomal segments containing genes involved in the regulation of these systems.

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.

Seminal plasma antioxidants are directly involved in boar sperm cryotolerance

Boar ejaculates are ejected in fractions with a specific composition in terms of sperm numbers and seminal plasma (SP), which is reflected in the varying sperm cryotolerance observed among different fractions. As boar sperm are particularly sensitive to oxidative stress, this study evaluated the role of SP antioxidants in the observed differences in sperm cryotolerance among ejaculate fractions. Ten ejaculates from five boars were manually collected in fractions: the first 10 mL of the sperm-rich fraction (SRF), the rest of the SRF and the post-SRF. Semen samples comprising the entire ejaculate (EE) were created by proportionally mixing the three fractions described above. Each of the 40 resulting semen samples was split into two aliquots: one was used for sperm cryopreservation following a standard protocol utilizing 0.5-mL straws, and the other was used to collect SP for antioxidant assessment. Frozen-thawed (FT) sperm from the SRF (the first 10 mL of the SRF and the rest of the SRF) and those from post-SRF were of the highest and worst quality, respectively, which was measured in terms of total and objective progressive motility and viability (P < 0.01). Viable FT sperm from the post-SRF generated more reactive oxygen species and experienced more lipid peroxidation than those from the SRF (both the first 10 mL and the rest of the SRF) (P < 0.01). The percentage of FT sperm exhibiting fragmented nuclear DNA did not differ among ejaculate fractions and the EE. Catalase, glutathione peroxidase and glutathione peroxidase 5 (GPx-5) were lowest in SP from the first 10 mL of the SRF (P < 0.001), whereas superoxide dismutase (SOD) and paraoxonase 1 (PON-1) were highest in SP of the SRF (both the first 10 mL and the rest of the SRF) (P < 0.01). Trolox-equivalent antioxidant capacity (TEAC) and the ferric-reducing ability of plasma (FRAP) were highest in SP from the first 10 mL of the SRF and lowest in the post-SRF (P < 0.001), whereas cupric-reducing antioxidant capacity was lowest (P < 0.05) in SP from the first 10 mL of the SRF. Regression analyses indicated that certain SP antioxidants had good predictive value for post-thaw recovery rates of total motility (R2 = 54.8%, P < 0.001; including SOD, TEAC and FRAP) and viability (R2 = 56.1%, P < 0.001; including SOD, PON-1, GPx-5 and TEAC). These results demonstrated that certain SP antioxidants are positively involved in boar sperm cryotolerance, minimizing the oxidative stress imposed by cryogenic handling.

Revisiting The Relationship between The Ejaculatory Abstinence Period and Semen Characteristics.

Variation in the ejaculatory abstinence period suggested by different guidance bodies have resulted in a growing concern among researchers and clinicians over what the precise period of ejaculatory abstinence ought to be for an optimal semen sample. Several studies have thus been undertaken to examine the association between the length of sexual abstinence and semen characteristics. Not all studies, however, have arrived at the same conclusions. This study aims to review all existing literature published during the past few decades pertaining to the influence of ejaculatory abstinence on semen quality. For the purpose of this systematic review, all data related to sexual abstinence duration and seminal parameters were re-analysed to homogenize the current data. Thorough PubMed, MEDLINE and Google Scholar, a literature search was conducted using the keywords “sexual abstinence”, “ejaculatory abstinence”, “semen”, “spermatozoa”, “semen analysis”, “sperm parameters”, “motility”, “reactive oxygen species (ROS)” and “DNA fragmentation”. After carefully reviewing all the literature, 30 relevant papers, both written in English and published between January 1979 and December 2016, were included in this review. The weight of the evidence suggests that the decline in semen volume and sperm concentration with shorter abstinence periods is accompanied by a substantial improvement in sperm motility characteristics, especially progressive motility and velocity. Nevertheless, available data are insufficient to support definitive conclusions regarding the influence of the ejaculatory abstinence period on advanced semen parameters (ROS, DNA fragmentation and seminal plasma antioxidant capacity) and pregnancy rates. In conclusion, taking all data into account, shortening of the abstinence period may be beneficial to sperm quality. Furthermore, we recommend that the current guidelines regarding the prescribed abstinence period should be revisited.

Effects of parsley supplementation on the seminal quality, blood lipid profile and oxidant status of young and old male rabbits

&lt;p&gt;The high unsaturation levels of spermatozoal membrane make it very susceptible to oxidative damage and this problem increases with advancing age. In this study, the aim is to investigate whether parsley seed (PS) has a protective effect on semen quality, serum lipid profile and antioxidative status of old and young bucks. Male rabbits (n= 36) (18 young 9-12 mo old and 18 old 36-42 mo old) were each assigned to 3 dietary treatments (a control and 2 levels of PS: 0.3 and 0.6 kg/100 kg diet) to evaluate the ability of parsley to enhance bucks’ reproductive status. Most of the studied traits were adversely affected by age of rabbit bucks. On the other hand, the inclusion of PS significantly boosted ejaculate volume and improved mass motility concentration and total sperm output. Seminal plasma and blood serum total antioxidant capacity increased, while serum lipid peroxidase decreased with parsley treatments. In conclusion, dietary supplementation of parsley seed alleviates most semen quality parameters and counteracts oxidative stress, especially with the advance of age (seminal plasma and blood serum of total antioxidant capacity and malondialdehyde.&lt;/p&gt;

Comparison of Oxidative Stress Intensity in the Seminal Plasma and Serum of Infertile Patients with Different Varicocele Grades and Subjects with No Varicocele: A Cross-Sectional Study

Background: Varicocele is the leading cause of male infertility throughout the world. Objectives: This study aimed to identify the effects of varicocele on oxidative stress, total antioxidant capacity (TAC), and semen parameters. Methods: In this cross-sectional study, serum and semen samples were collected from subjects with no varicocele (n, 35) and patients with varicocele (n, 86), who were referred to infertility and in vitro fertilization (IVF) centers of Babol Clinic Hospital, Babol, Iran during 2015 - 2016. The semen samples were analyzed according to the world health organization (WHO) guidelines. After the ultrasound and physical examinations, varicoceles were classified into three grades: G1, G2, and G3. TAC and malondialdehyde (MDA) levels were measured in the serum and seminal plasma of subjects, using the ferric reducing ability of plasma (FRAP) and thiobarbituric acid reaction (TBAR) methods, respectively. Nitric oxide (NO) was also measured using Nitric Oxide assay kit. Results: The mean total sperm count in the G1, G2, and G3 groups were significantly lower than the control group (P = 0.037; P = 0.003, and P = 0.044, respectively). A trend of lower semen volume was observed in the G3 group, compared with the control group (P = 0.06). A significant positive correlation was observed between elevated serum MDA level and varicocele degree (P G2 > G1 > controls. There was also a significant negative correlation between the serum and seminal plasma TAC levels (P < 0.05). Varicocele patients with G3 degree had a significantly lower mean TAC level in the serum and seminal plasma, compared with the control group (P = 0.001 and P = 0.008, respectively). No significant difference was found in the mean Nitric Oxide level between the groups. However, an increasing trend for the mean Nitric Oxide level was observed in the serum and seminal plasma of varicocele patients. Conclusions: The reduced level of seminal plasma antioxidants, which is associated with increased lipid peroxidation, is one of the main reasons for low sperm quality in patients with varicocele. Antioxidant therapy may be useful in decreasing oxidative stress intensity and improving the condition of varicocele patients.

55 SEMINAL PLASMA COMPONENTS AND THEIR RELATIONSHIP WITH STALLION SEMEN FREEZABILITY

Oxidative stress has been identified as a major cause of low seminal fertility. Among the components of stallion seminal plasma, some enzymatic and non-enzymatic antioxidants have been identified, which protect sperm from injurious effects of reactive oxygen species (ROS); however, the characterisation of these components is still in preliminary stages, as well as their relationship with freezability. The aim of this study was to evaluate the effect of some components of seminal plasma (SP) on stallion semen freezability. Semen of 30 Colombian Creole horses, and a total of 60 ejaculates, were collected. Semen was centrifuged to recover the SP. It was lyophilized and some components were assayed: total protein concentration (TP) by Bradford assay, CRISP3 protein concentration by ELISA, vitamin C (CVIT), vitamin E (EVIT) and vitamin A (AVIT), by HPLC; content of iron (Fe), copper (Cu), magnesium (Mg) and Zinc (Zn) by atomic absorption spectroscopy flame. Semen was supplemented with 10% stallion lyophilized SP and cryopreservation was performed. Post-thaw, total motility (TM), progressive motility (PM), straight line velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF), were assessed using a computer-assisted sperm analysis (SCA®, Microptic SL, Barcelona, Spain). Sperm viability (SV) was determined by the Live/Dead kit (Molecular Probes Inc., Eugene, OR, USA). Normal sperm morphology (NM) was performed by the supravital technique and plasmatic membrane integrity (MI) was evaluated by the hypo-osmotic test. For statistical analysis, completely randomised mixed models were fitted. Levels according to the concentration of components of SP (high, medium, and low) were established. Comparisons of the means between levels were done with Tukey’s test. The significance level used for all assessments was P &lt; 0.05. Means for TP of 0.35 mg BSA/g, CRISP3 of 55.22 ng/mg, CVIT of 2.66 mg/g, EVIT of 72.36 µg/g, AVIT of 37.37 µg/g, Fe of 17.37 mg/kg, Cu of 33.64 mg/kg, Mg of 109.08 mg/kg, and Zn of 0.49 g/100 g of SP were found. We found that a high level of CRISP3, AVIT, Cu, and Fe had higher results for post-thaw TM, PM, NM; medium levels of TP and Mg showed higher post-thaw TM, PM, NM, and MI; and lower levels of Zn had better results for post-thaw TM, PM, VCL, and VAP. In contrast, high and medium levels of CVIT had a deleterious effect on post-thaw TM, PM, SV, NM, and MI. We concluded that there is a relationship between concentrations of seminal plasma components and stallion semen freezability.

Evaluation of enzymatic and non-enzymatic antioxidants in seminal plasma of men with genitourinary infections, varicocele and idiopathic infertility.

This study was aimed to assess the antioxidant enzymatic and non-enzymatic compounds in semen of infertile men. Seventy-four infertile patients were grouped according to their clinical diagnosis: genitourinary infection, varicocele, idiopathic infertility. Semen samples of fertile men represent the control. Semen characteristics were evaluated by light and transmission electron microscopy (TEM). TEM data was quantified with a mathematical formula, which provides numerical scores. Spectrophotometric and HPLC methods were used to measure the amount of reduced (GSH), oxidised glutathione (GSSG), ascorbic acid (AA) and malondialdehyde (MDA, marker of lipid peroxidation) and the activity of glutathione reductase, catalase (CAT), glutathione peroxidase. Infertile groups showed significantly decreased values of sperm parameters vs. In infection and varicocele groups, the seminal MDA levels were significantly increased when compared to controls (p<0.001), indicating an alteration of oxidative status and a peroxidative damage. In infection and varicocele groups, AA levels were reduced (p<0.05) vs. control; in the varicocele group, the GSH levels were also decreased (p<0.05). Significantly higher CAT activity was observed in infection and varicocele groups vs. fertile men (p<0.001 and p<0.05 respectively). The GSH/GSSG ratio was significantly decreased in varicocele and idiopathic infertility groups vs. control (p<0.01). The study of the alteration of a single parameter of oxidative stress or of the antioxidant system may not have a relevant clinical value to estimate male fertilising potential and the background of infertility causes, since complex and multifactorial mechanisms are involved in different pathologies. In our study, each pathology is characterised by a definite pattern of markers such as MDA and enzymatic and non-enzymatic antioxidant compounds. In the different pathologies related to infertility, the identification of the complex of involved parameters could be useful in the diagnosis, prognosis and in the choice of a possible treatment such as specific antioxidant supplements.