Computer-aided Semen Analysis Research Articles

Alpha-lipoic acid improves cryopreservation of rooster semen by reducing oxidative stress

Inhibiting oxidative stress is key for ensuring sperm motility during semen cryopreservation. The aim of this study was to investigate the effect of adding alpha-lipoic acid (ALA) as an extender in rooster semen cryopreservation. Different concentrations of ALA were added to the frozen diluent of rooster semen; subsequently, computer-aided semen analysis was used to determine membrane functional integrity, acrosome integrity, antioxidant capacity (based on T-AOC, GSH-Px, SOD, CAT, and MDA contents), and mitochondrial integrity. The frozen sperm ultrastructure was observed using transmission electron microscopy. The results showed that the addition of different concentrations of ALA partially to greatly improved the quality of frozen sperm; in particular, 8 μg/mL ALA significantly improved multiple parameters of sperm quality, including sperm motility and antioxidant enzyme activity, after freeze-thaw. The results of this study provide empirical and theoretical support for effective rooster semen cryopreservation and can inform the development of new protective agents in the field of livestock reproduction.

CASA Based Assessment of Kangayam bull semen processed with different extenders and Conception rate following AI

Semen extenders are the essential components of cryopreservation of semen. Identification of suitable and ready to use semen extender is the need of hour for the rapid propagation of Kangayam cows through artificial insemination. Kangayam bull, aged 5 years was utilized for the study. The semen was collected twice in a week and each time two ejaculates were collected. A total of 24 ejaculates were utilized for this study. After assessing the general macroscopical examination the semen was subjected to computer aided semen analysis (CASA) for assessing the kinematic parameters in fresh diluted semen, pre freeze and post thawed such as sperm motility, progressive individual motility, VAP, VSL, ALH and LIN were analyzed using CASA. Among all the three extenders, the sperm kinematic parameters during fresh diluted, pre freeze and post thawed semen stages were higher in group III (liposome based extender) followed by group I (TEYG) and in group II (soya bean lecithin based extender). Among the three extenders used, the components of liposome based extender was better and it was able to maintain the structural integrity, plasma membrane integrity and motility in the sperms of the frozen thawed semen than the other two extenders (TEYG and soya bean lecithin based extenders). Further the fertility rate was evaluated by the in vivo insemination of 60 Kangayam cows after 12 hours of onset of estrum. The fertility rate observed was 45.00, 40.00 and 65.00 per cent in groups I, II and III, respectively. The overall conception rate recorded in Kangayam cow was 50.00 per cent. Based on the findings, it was concluded that liposome based extender may be used for cryopreservation of Kangayam bull semen in order to maximize the fertility rate.

Analysis of 2 men with t(8;22)(q13;q13) and t(8;14)(q13;q22) chromosomal translocation karyotypes.

Male infertility is a multifactorial condition that is closely associated with chromosomal abnormalities. Reciprocal chromosomal translocation (RCT) is a significant structural genetic abnormality. The specific mechanisms of forms of RCT affecting male infertility include the product of chromosomally unbalanced gametes, thereby disrupting the structure and function of important genes responsible for spermatogenesis. RCT breakpoints have been found to disrupt gene structure and function in many medical fields However, the relationship between RCT breakpoints and male infertility remains to be determined. The purpose of this study is to describe 2 male carriers of RCTs 46,XY,t(8;22)(q13;q13) and 46,XY,t(8;14)(q13;q22). Both patients were collected from the second hospital of Jilin University. Semen parameters were detected using the computer-aided semen analysis system. Cytogenetic analysis was performed using standard operating procedure. Related genes on chromosomal breakpoints were searched using Online Mendelian Inheritance in Man. One man had semen parameters within the normal range, but the couple was infertile after 5 years of marriage. The other man showed normal semen parameters, and his wife had experienced 2 spontaneous miscarriages. Using a literature search, the association between chromosome 22q13 breakpoint and fertility were investigated. The results suggest that physicians should focus on the clinical phenotype of the patients and the breakpoints of RCT in genetic counseling. An important gene related to human male infertility is clearly located in chromosome region 22q13, and its function is worthy of further study.

Association between chromosome 22q11.2 translocation and male oligozoospermia.

Chromosomal aberrations in peripheral blood are a major cause of reproductive disorders for the infertile couples. Reciprocal translocation is closely related to male infertility. The breakpoint of translocation may disrupt or dysregulate important genes related to spermatogenesis. The relationship between some breakpoints of chromosome and male infertility has been paid attention. Chromosome 22q11.2 translocation has not been reported with male infertility. The purpose of this study is to evaluate the relationship between chromosome 22q11.2 translocation and male infertility. All patients were collected from the second hospital of Jilin University. Semen parameters were detected using the computer-aided semen analysis system. Cytogenetic analysis was performed using standard operating procedure. Related genes on chromosomal breakpoints were searched using online mendelian inheritance in man (OMIM). The association between this breakpoint and spermatogenesis is also discussed. We report 6 cases of translocation in chromosome 22. Of 7 breakpoints involved in these translocations, the common feature is that they all included chromosome 22q11.2 translocation and presented with oligozoospermia. The analysis of breakpoint related genes showed testis-specific serine/threonine kinase 2 (TSSK2) gene is associated with human spermatogenesis impairment. Overall, these results suggest that the breakpoint involved in translocation deserves attention from physicians in genetic counseling. The breakpoint rearrangement has the possibility of disrupting spermatogenesis. The relationship between 22q11.2 breakpoint and male infertility deserves further study.

Computer aided semen analysis (CASA) to determine the quality and fertility of frozen thawed sumba ongole sperm supplemented with amino acids

<p><strong>Objective</strong>: The objective of this research was to investigate the quality and reproductive potential of frozen-thawed Sumba Ongole (SO) bull spermatozoa supplemented with specified amino acids using computer-assisted semen analysis (CASA).</p><p><strong>Methods</strong>: A total of 18 ejaculates were collected once a week from three Sumba Ongole bulls and diluted in tris-citric-fructose-egg yolk (TCFY) extender and various concentrations of selected amino acids. The post-thaw spermatozoa were examined for motility, head behavior, and swimming pattern using CASA technique. Data were analysis using SPSS 24 ebvaluated by ANOVA.</p><p><strong>Results</strong>: Total motility (72.56%±3.20) and progressive motility (71.11%±3.31) the addition of Glutamine 5% significantly different (p<0.05) than other treatments. The addition of 7mM cysteine resulted in significantly (p<0.05) in velocity average path (VAP), velocity curve line (VCL), velocity straight line(VSL) parameters also have higher in distance average path (DAP), distance curve line (DCL), dan distance straight line (DSL) parameters significantly (p<0,05). </p><p><strong>Conclusion</strong>: In conclusion, the introduction of CASA systems enabled quantification of individual spermatozoa motility patterns and CASA colud be used to estimate the fertilizing capacity of sperm based on the characteristics of sperm velocity.</p>

P-008 Microgravity exposure significantly decreases sperm motility and vitality. Can we consider human reproduction outside the Earth?

Abstract Study question Are fresh human sperm samples affected by different gravitational conditions than on Earth? Summary answer Motility and vitality of fresh human sperm samples are significantly decreased under microgravity conditions obtained by parabolic flight. What is known already Microgravity effects on the male reproductive system have mainly been studied in the animal model with diverse results and discouraging extrapolation in humans. While an increased motility was reported in bulls, mice models showed a decrease. Although preliminary data from the Micro-11 experiment presented by NASA reported human sperm alterations after microgravity exposure, our first study failed to show any significant effect of microgravity on “frozen” samples, suggesting that human sperm could be safely shipped outside the earth if important aspects related with cryopreservation were solved. Study design, size, duration Prospective study carried out in collaboration between the ART centre, a Technical University, and an Aviation Club specializing in parabolic flights.Two parabolic flights were conducted between 2020-2021, each consisting of 20 parabolic maneuvers, which means 160 seconds of microgravity exposure per sample. Fifteen sperm samples obtained from healthy men were included in the study in order to analyse the effects of microgravity and compare the results with those obtained in Earth gravity. Participants/materials, setting, methods Fresh sperm samples were checked pre-flight to evaluate vitality, concentration, motility and morphology. Samples were split into two to compare the effects of different gravity exposure: microgravity (flight) and Earth gravity (ground). After the flight, the same analysis were repeated, plus kinematics, DNA fragmentation by sperm chromatin dispersion, apoptosis by magnetic activated cell sorting, and oxidative stress by colorimetric test (Halosperm-Halotech). Computer Aided Semen Analysis (SCA-Scope) was used for cell counting. Main results and the role of chance On comparison of the mean values between fresh samples exposed to microgravity and those maintained on Earth gravity, statistical significant differences (p < 0,05) were found in the following parameters: vitality (69,7 ± 9,9 vs 72,4 ± 9,7 %), motile sperm concentration (23,7 ± 15,3 M/ml vs 31,5 ± 25,1 M/ml), grade “a” sperm concentration (8,7 ± 6,5 vs 11,7 ± 9,9 M/ml), percentage of spermatozoa with progressive motility (30 ± 12,9 vs 36 ± 14,3 %), curvilinear motility-VCL (45,7 ± 12,8 vs 47,7 ± 13,3 μm/s). Under the study conditions, non-statistically significant differences were observed in the other kinematic parameters: Lineal Velocity (VSL), Average Path Velocity (VAP), Straightness (STR), Amplitude of Lateral Head displacement (ALH), Linearity (LIN), Wobble (WOB), Beat-Cross Frequency (BCF), total sperm concentration (81,7 ± 112,1 vs 79,7 ± 89,8 M/ml), morphology (11,3 ± 6,3 vs 10,6 ± 5,3%), DNA fragmentation (14,6 ± 9,6 vs 15,7 ± 9,4), apoptosis (2,8 ± 2,8 vs 3,8 ± 4,4) and oxidative stress, since all samples maintained the same stress level in both splits. Limitations, reasons for caution Parabolic flight is an accepted ground-based method for obtaining microgravity conditions, but provides a short period of elapsed exposure to microgravity. Therefore, the results obtained need to be confirmed by using other platforms that provide a much longer time of exposure. More cases must be analysed to confirm the results. Wider implications of the findings Short exposure to microgravity significantly decreases sperm motility and vitality. Such an effect is likely to be stronger with longer exposure. These findings should be taken into account since this may eventually affect sperm fertilizing capacity and therefore natural conception or ART with fresh/frozen sperm, outside of the Earth. Trial registration number NCT03760783

P-047 The effects of dietary branched-chain amino acids on mice semen parameters and plasma amino acids profile

Abstract Study question Are there any differences between the effect of each member of the branched-chain amino acids (BCAAs) on sperm parameters and plasma amino acids profile? Summary answer Valine supplementation had destructive effects on sperm parameters and viability. However, blood amino acids profile were similar among treatments. What is known already BCAAs, a group of three essential amino acids, have a crucial role on protein synthesis, besides being strongly associated with impaired glucose and fat metabolism. Although BCAAs supplements are one of the most demanding products for the fitness industry to increase the lean body mass and boost exercise performance, little information exists on the advantages and disadvantages of dietary BCAAs on male fertility. Study design, size, duration Fifty NMRI mature male mice were divided into five groups and were fed diets as follow: Control (CTR): 2.4% Beta-alanine, Leucine (Leu): 1.18% Leu + 1.2% Beta-alanine, Isoleucine (Ile): 0.58% Ile + 1.8% Beta-alanine, Valine (Val): 0.65% + 1.7% Beta-alanine, and BCAAs (Leu:Ile:Val ratio of 2:1:1): 2.4% BCAA: 1.18% Leu + 0.58% Ile + 0.65% Val. After 5 weeks, testes and epididymides were dissected, and sperm parameters and plasma amino acids profile were assessed. Participants/materials, setting, methods Semen parameters were evaluated by a Computer-Aided Semen Analysis (CASA) system. Plasma amino acids concentrations were determined by HPLC. Data were analyzed using SPSS and statistical differences among various groups were determined by ANOVA and Tukey’s post-hoc test. Main results and the role of chance Higher BCAAs intake, particularly Val, increased the destructive mechanisms on sperm viability. The results showed no significant difference on body weight after 40 days. Sperm concentration (8.4 vs. 4.5 ×106), progressive motility (51.5 vs. 27.2 %), and viability (67 vs. 56 % for CTR and Val group, respectively) decreased dominantly in the Val group compared to CTR (p≤0.05). The immotile parameter was significantly higher in the Val group compared to the CTR (p≤0.05). In addition, there were no significant differences on plasma amino acids profile. Limitations, reasons for caution The current study's limitation is that only one level of BCAAs was used. Given the potential role of Val on sperm cell death, further data on different levels of all BCAAs, especially Val administration and evaluation of the related mechanisms are needed. Wider implications of the findings The findings of our study show, for the first time, that individual levels of Ile and Leu have different effects on sperm parameters compared with all BCAAs or Val. More attention should be given to BCAAs supplementation and semen quality that was previously ignored. Trial registration number not applicable

Influence of Fruit and Vegetable Consumption on Antioxidant Status and Semen Quality: A Cross-Sectional Study in Adult Men.

The influence of fruit and vegetable consumption on semen quality by reducing oxidative stress is inconsistent. Thus, the association between the consumption of these products, antioxidant status, and semen quality was investigated in 90 men aged 18–40. The consumption of fruit and vegetables was collected using the 3-day food record method. Antioxidant status: total antioxidant capacity in semen (TAC-s) and blood (TAC-b), blood superoxide dismutase (SOD-b), glutathione reductase (GR-b), glutathione peroxidase (GPx-b), catalase (CAT-b) activity, and malondialdehyde concentration in blood (MDA-b) were measured. Sperm concentration, leukocytes in the ejaculate, vitality, motility, and sperm morphology were examined using computer-aided semen analysis (CASA). The consumption of fruit and vegetables was positively correlated with sperm concentration, vitality, motility, TAC-s, TAC-b, and SOD-b activity. The TAC-s and TAC-b were positively related to motility, TAC-s was inversely correlated with sperm tail defects. The SOD-b activity was positively correlated with vitality, motility, sperm morphology, and inversely with sperm tail defects and leukocytes in the ejaculate. Compared to the men in the first quartile of fruit and vegetable consumption (<318 g/day), those in the highest quartile (>734 g/day) had the highest sperm concentration, vitality, motility, TAC-s, TAC-b, GPx-b activity, and the lowest MDA-b concentration (based on multivariate regression models). A high consumption of fruit and vegetables may positively influence selected sperm quality parameters by improving the antioxidant status of semen and blood.

P–048 Effects of bisphenol S and bisphenol F on human spermatozoa: an in vitro study

Abstract Study question Are plasticizers bisphenol S (BPS) and bisphenol F (BPF) safer alternatives to bisphenol A (BPA) for human sperm function? Summary answer Unlike BPA, the analogues BPS and BPF do not significantly affect human sperm mitochondrial functions, motility and viability What is known already The widespread distribution of BPA, along with its reputation to be an endocrine disruptor has generated concerns about possible adverse effects for human health, thus prompting the European Food Safety Authority and the Food and Drug Administration to ban the use of this chemical in many plastic products. Following such restrictions, several substitutes have been developed, with BPS and BPF representing the main replacements to BPA. While it has been demonstrated that BPA promotes oxidative damages in spermatozoa from different species, including human, the possible effects exerted by BPS and BPF on human sperm, have not yet been investigated. Study design, size, duration We explored the effect of 4 h in vitro exposure to scalar concentrations of BPS and BPF (from 10 to 400 μM), and to 400 μM BPA on sperm motility, viability, mitochondrial membrane potential (Δm) and mitochondrial generation of reactive oxygen species (ROS). In a set of experiments, the effect of a combination of both BPF (400 μM) and BPS (400 μM) on ΔΨm and mitochondrial ROS generation was also assessed. Participants/materials, setting, methods Sperm Δm was analyzed by flow cytometry with the fluorescent lipophilic cationic dye JC–1. Flow cytometric assessment of mitochondrial generation of ROS was carried out using the lipid soluble cation MitoSOX red (MSR). Sperm motility and viability were evaluated by Computer-Aided Semen Analysis (CASA) and eosin assay, respectively. Main results and the role of chance The exposure to scalar concentration of BPS did not significantly affect sperm motility and viability with respect to untreated controls. A lower, albeit not significant, sperm motility was registered in samples exposed to the highest concentrations of BPF (300 μM and 400 μM). As expected, 400 μM BPA produced a complete sperm immobilization along with a dramatically loss in sperm viability. No significant differences were observed in sperm Δm and ROS generation after exposure to scalar concentration of BPS compared to untreated controls and the trend towards lower Δm and higher mitochondrial ROS generation at the highest concentrations of BPF did not reach statistical significance. On the contrary, after 4 h exposure to 400 μM BPA a significant lower Δm and higher mitochondrial ROS generation were observed. Finally, the exposure to a combination of BPF and BPS at high concentrations (400 μM) did not significantly affect sperm Δm, or mitochondrial ROS generation, when compared to 400 μM BPA, used as positive control. Limitations, reasons for caution: The present study only evaluated BPS and BPF effects, but in daily-life people are exposed to several plasticizers containing different bisphenols at different concentrations. Therefore, adverse effects of synergic exposure to BPA analogues other than BPS and BPF, alone or in combination with BPA, cannot be ruled out. Wider implications of the findings: The analogues BPS and BPF, alone or in combination, appeared to be safer alternatives to BPA on sperm biology as they exert a neutral effect on sperm motility, viability, and mitochondrial functions even at high concentrations. These results could be useful to identify more secure plasticizer components. Trial registration number Not applicable

Effects of bisphenol S and bisphenol F on human spermatozoa: An in vitro study

Bisphenol A (BPA), the main chemical monomer of epoxy resins and polycarbonate plastics, has generated concerns about its endocrine disruptor properties, along with the reported possible links with several human health disorders. Accordingly, some restrictions on its use have been recommended. Bisphenol S (BPS) and bisphenol F (BPF) are the main replacements to BPA, with which they share homologies in chemical structure. However, to date, little is known about their possible adverse effects for human reproduction. As the in vitro exposure of human spermatozoa to BPA induces oxidative/pro-apoptotic effects, the aim of the present study was to verify whether BPS and BPF could represent safer compounds for human sperm functions.The exposure of motile sperm suspensions to scalar concentrations of BPS or BPF for 4 h did not significantly reduce sperm motility (as assessed by computer-aided semen analysis) and viability. At flow cytometry, no changes in mitochondrial membrane potential, or mitochondrial generation of reactive oxygen species were detected by using the JC-1 and MitoSOX red probes, respectively. Interestingly, it nor even the combination of both BPS and BPF at the highest concentrations impaired sperm mitochondrial functions.In conclusion, BPS and BPF seem to be safer alternatives to BPA for sperm biology, as they do not affect mitochondrial functions, sperm motility and viability. These findings could help regulatory agencies to identify more secure chemicals to replace BPA in industrial production of plastics.

Melatonin ameliorates cisplatin-induced asthenozoospermia via reducing oxidative stress

Background and objective: Cisplatin is an alkylating agent that has become a first-line therapy for some tumors. However, overproduction of reactive oxygen species (ROS) by cisplatin can cause male infertility, which can affect patients' quality of life. Melatonin, which has the ability to provide resistance against oxidation, is a potential therapy for male infertility caused by cisplatin.Material and methods: Normal human spermatozoa samples were divided into (i) control group incubated with physiological saline solution; (ii) cisplatin group incubated with cisplatin; (iii) melatonin group incubated with melatonin; and (iv) melatonin + cisplatin group incubated with melatonin and cisplatin. Spermatozoa motility was measured using a computer-aided semen analysis system. Additionally, we determined spermatozoa apoptosis through terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling. The mitochondrial membrane potential (∆Ψm) and levels of ATP, ROS, and malondialdehyde, as a metabolite of oxidative stress, were measured to study the mechanism by which melatonin can protect human spermatozoa.Results: Melatonin improved sperm motility, and decreased apoptosis and increased the ∆Ψm value and level of ATP in sperm. More importantly, the overproduction of ROS induced by cisplatin was reduced by melatonin. Furthermore, western blotting revealed that melatonin increased the expression of antioxidant pathway components nuclear factor erythroid 2-related factor (Nrf2) and heme oxygenase-1 (HO-1) that was reduced by cisplatin.Conclusion: Our studies suggested that melatonin might protect human spermatozoa against the effects of cisplatin through Nrf2/HO-1.

Comparison of commercially available chamber slides for computer-aided analysis of human sperm

ABSTRACT Despite the increasing use of computer-aided sperm analysis (CASA) in clinical practice, there is still no golden standard for the type of slide to be used with these systems. Counting chamber depth and loading method, can profoundly influence motility and concentration estimates, thereby undermining the validity and accuracy of CASA. To contribute toward standardized sperm analysis, this study compared different commercially available capillary-filled slides including 10 and 20 µm deep Leja slides (Leja10 and Leja20); 10, 16 and 20 µm deep CellVision slides (CV10, CV16 and CV20); and drop-loaded slides including slide and coverslip (SCS) with a depth of 20.1 µm and the Makler chamber with a depth of 10 µm for sperm analysis when using CASA. The Sperm Class Analyzer (SCA) CASA system was used to assess concentration, motility, and detailed kinematic parameters of 20 normozoospermic human samples using the different chamber slides. Results were evaluated by the repeated measures ANOVA and Intraclass correlation coefficients. The Makler chamber showed significantly (P < 0.05) higher concentrations than other slides. However, there was no significant difference in the percentage of sperm in different motility groups among the slides. CV10, Leja10 and Makler showed significantly higher curvilinear-, average path- and straight-line velocity (VCL, VAP, VSL) values than other slides. In conclusion, despite the objectiveness of the assessments by CASA systems, there are still some discrepancies in the results of sperm concentration, motility and other kinematic parameters when using different commercially available slides. The possible negative influence of the sperm quality misdiagnosis on the selection of treatment strategy in a clinical setting, emphasizes the need for further standardization and quality control of the commercially available chamber slides for use with CASA. Furthermore, this study found more consistent results for capillary-filled chambers compared to drop-loaded slides, suggesting a superior method when using CASA. Abbreviations: DNA: Deoxyribonucleic acid; CASA: Computer-aided semen analysis; SCA: Sperm Class Analyzer; WHO: World Health Organization; CV: CellVision; SCS: Slide and Coverslip; SD: Standard deviation; ANOVA: Analysis of Variance; PR: Progressive; NP: Non-progressive; IMT: Immotile; VCL: Curvilinear velocity; VAP: Average path velocity; VSL: Straight-line velocity; Lin: Linearity; STR: Straightness; WOB: Wobble; ALH: Amplitude of lateral head displacement; BCF: Beat cross frequency

Exposures to air pollution and endocrine disruptors and measures of semen quality in a population based cohort of US men

Background: Multiple studies have identified adverse associations between exposures to air pollution and endocrine disrupting chemicals on semen quality. However, few studies have assessed these associations among men from the general population, as opposed to clinical settings. Our objective was to assess these associations in a subset of participants in the US nationwide Growing Up Today Study (GUTS).Methods: We aim to enroll 200 men from GUTS living in the conterminous US, without a vasectomy, are who have not undergone chemotherapy, and are not taking anabolic steroids. For 90 days, participants use a smartphone application to track GPS location, run an air pollution sampler to collect real-time indoor measurements of NO2, and wear 3 separate passive wristband samplers each for 30 days. After 90 days of monitoring, participants produce a semen sample, which is analyzed for concentration and morphology at the Massachusetts General Hospital Andrology Laboratory via computer-aided semen analysis. For the first 45 participants, we assessed the interclass correlation (ICC) of exposures and assessed associations between exposures and outcomes using Spearman correlations and linear regressions.Results: The average NO2 concentration was 18.11 (SD=10.2) ppb and 21 chemicals were detected in at least 20% of wristbands measured. The ICC for daily NO2 was 0.71 (95%CI: 0.61, 0.79), the ICCs for the 21 chemicals varied widely, ranging from 0.04 (0.00, 0.97) for dicyclohexyl phthalate to 0.88 (0.78, 0.94) for galaxolide. Ninety-day average exposures to NO2 were not associated percent normal morphology, but were associated with decreased sperm concentration (-12.09 M/ml per IQR (14.49 ppb) NO2). There were no clear patterns observed between the EDC exposures and semen parameters.Conclusions: In an initial subsample of general population men, exposures to NO2, but not endocrine disrupting chemicals, were associated with lower sperm concentrations.

Supplementing post-wash asthenozoospermic human spermatozoa with coenzyme Q10 for 1hr in vitro improves sperm motility, but not oxidative stress.

We investigated the effect of supplementing post-wash asthenozoospermic spermatozoa with coenzyme Q10 (CoQ10) in vitro, which may reduce oxidative stress and improve sperm motility. Semen samples were collected from 39 men with asthenozoospermia, and their spermatozoa were isolated by two-layer Percoll density-gradient centrifugation. Kinetic parameters of the isolated spermatozoa (baseline before intervention) were determined immediately by computer-aided semen analysis. Total anti-oxidant capacity and protein carbonyl levels, as markers of oxidative stress, were also measured in the baseline spermatozoa. The baseline spermatozoa suspension was divided equally into two portions, one for CoQ10 supplementation (50µg/ml for 1hr) and the other as an un-supplemented vehicle control. The total motility of the CoQ10-supplemented spermatozoa was significantly higher than in the control (p=.009) and progressive motility tended to be higher (p=.053). Immotile sperm concentration in the CoQ10-supplemented spermatozoa was significantly lower than in both the baseline (p=.026) and control (p=.009). Total anti-oxidant capacity and protein carbonyl levels between the baseline, CoQ10-supplemented and control spermatozoa were not significantly different. Our data suggest that CoQ10 treatment reactivated sperm motility. We propose short-term supplementation of post-wash asthenozoospermic spermatozoa with CoQ10 before intrauterine insemination.

Processed meat intake in relation to semen quality indicators among men attending a fertility clinic

AbstractIt is estimated that in Central and Eastern Europe 8–12% of men are infertile, and in the case of 20% of infertile couples in the region, indicates a 56% part of the malefactor in the occurrence of infertility. Numerous studies have shown that a healthy diet with a high intake of fish and seafood, poultry, vegetables, fruits, legumes, through the high content of antioxidants are associated with better parameters of semen quality and reduced risk of asthenozoospermia. Men's diets may affect spermatogenesis as reflected in semen quality indicators, but the literature on the relation between meat intake and semen quality is limited. Our objective was to prospectively examine the relation between meats including red, poultry, proceesed and fish and indicators of semen quality. Men in subfertile couples presenting for evaluation at the Olsztyn Hospital Fertility Center were invited to participate in an ongoing study of environmental factors and fertility. This cross-sectional study was carried out in 208 men aged 18–55 years from Northern-Eastern Poland. Semen samples were collected at the clinic and analyses were performed by an experienced technician. Semen microscopic measurements were determined with the use of computer-aided semen analysis (CASA) and evaluated according to the World Health Organization (WHO) guidelines. A total of men completed a validated food-frequency questionnaire and subsequently provided semen samples. We used linear mixed regression models to examine the relation between meat intake and semen quality indicators (total sperm count, sperm concentration, progressive motility, morphology, and semen volume) while adjusting for potential confounders. After adjustment for age, BMI, physical activity, residence place and education, one serving increase in intake of processed meat was associated with a 56% higher risk of abnormal progressive sperm motility (OR = 1.56 95%CI:1.09–2.21). Consuming processed meat may have a negative impact on sperm counts and morphology. Potentially, these products may be a significant source of saturated and trans fatty acids, xenobiotics, mainly xenoestrogen, and steroids. Is need to clarify the potential effect of meat products, especially processed meat, on male reproductive potential in RCTs or longitudinal studies. Future findings may be useful to establish a more precise recommendation connected with red and processed meat intake in the context of reproductive health.

Association of the Dietary Approaches to Stop Hypertension, Physical Activity, and Their Combination with Semen Quality: A Cross-Sectional Study.

The influence of individual lifestyle factors is widely described in studies on semen quality. However, their synergistic effect is often neglected. The aim of the study was to examine the association between semen quality and dietary approaches to stop hypertension (DASH) diet, physical activity (PA), and the two separately and in combination. A cross-sectional study was carried out among 207 men aged 20–55. Dietary data were collected by a food frequency questionnaire (FFQ) and analysed according to the DASH scoring index. Physical activity was evaluated by the International Physical Activity Questionnaire. Semen parameters were assessed via the computer-aided semen analysis (CASA). Adherence to the DASH diet was associated with higher sperm count (ΔT3-T1 = 82.1 mln/ej) and concentration (ΔT3-T1 = 24.6 mln/mL). Higher PA was related to higher sperm count (ΔT3-T1 = 69.4 mln/ej), total (ΔT3-T1 = 11.9%), and progressive motility (ΔT3-T1 = 8.5%) and morphology (ΔT3-T1 = 2.8%) in the crude model and remained significant after adjustment. The combination of the DASH diet and PA, was significantly positively associated with sperm count (ΔT3-T1 = 98.1 mln/ej), sperm concentration (ΔT3-T1 = 17.5 mln/mL), total (ΔT3-T1 = 11.8%), and progressive motility (ΔT3-T1 = 10.0%) and morphology (ΔT3-T1 = 3.3%) in both models. Adherence to the DASH diet was related to higher sperm count and concentration, whereas after its combination with physical activity it was also positively correlated with sperm motility and morphology.

140 The relationship between morphometric characteristics and semen parameters in Bapedi rams

Bapedi is a fat-tailed, hardy, and disease-tolerant breed of sheep that is indigenous to South Africa. The Bapedi sheep breed is facing genetic degradation due to nonselective cross-breeding and irregular mating; therefore, breeding strategies for urgent conservation of Bapedi sheep are crucial. The aim of the present study was to investigate the relationship between body measurements and semen traits of Bapedi rams conserved in situ (conserved in their natural habitat) and ex situ invivo (live animals conserved outside their natural habitat). Semen was collected twice per week from 33 rams aged 2-4 years (four ejaculates per ram) using an electroejaculator. Body measurements (rump height, body length, scrotum circumference (SC), scrotal width, and scrotum length (cm)), body condition score (BCS; scale of 1-5), bodyweight (BW; kg), rectal temperature (°C), semen volume (mL), spermatozoa concentration (billionsmL−1), and motility parameters were measured using a computer-aided semen analysis system. Data were analysed using analysis of variance and the general linear models of SAS (SAS Institute Inc.). The Pearson correlation coefficient in SAS was used to determine the relationship among variables. There was uniformity in all body measurements of Bapedi sheep regardless of the conservation method (P&amp;gt;0.05). There were no significant differences in body temperature during semen collection, SC, semen volume, semen pH, semen concentration, spermatozoa total motility, and kinematics in Bapedi rams between methods of conservation (P&amp;gt;0.05). Pearson correlation revealed moderate significant relationships between semen volume and BW, BCS, and SC (r=0.315, 0.638, and 0.381, respectively). Rump length had a moderate correlation (0.566) with normal spermatozoa (P&amp;lt;0.05). It was concluded that Bapedi sheep are still a uniform breed regardless of their decreasing numbers. The BW, BCS, and SC must be considered in the selection criteria for improving the reproductive performance of Bapedi breeding rams because they were positively correlated with semen volume. It is recommended that more studies be done to correlate body measurements, testicular morphometry, and semen parameters in this breed.

Effect of ureaplasma urealyticum infection on sperm quality and sperm cytokine concentration

Objective To investigate the effects of ureaplasma urealyticum(Uu) infection on male sperm quality, nitric oxide(NO) concentration and cytokine interleukin (IL)-17, IL-18 and tumor necrosis factor alpha(TNF-α) concentrations, the mechanism of its action in male infertility was analyzed at the molecular level. Methods From September 2015 to December 2017, 53 male infertility patients who admitted in the outpatient clinic of the Second People's Hospital of Lianyungang were selected.According to whether the patients were infected Uu, they were divided into two groups: male infertility without Uu infection group (B group) and male infertility with Uu infection group (C group, Uu≥107CCU/L). The content of NO in seminal plasma was determined by nitrate reductase method.The contents of cytokines IL-17, IL-18 and TNF-α were detected by flow cytometry microsphere capture chip technique (CBA), and the spermate-related parameters were analyzed in conjunction with the computer-aided semen analysis system(CASA). Results The concentration of NO in the male infertility group with Uu infection group[(145.98±32.28)μmol/L] was significantly higher than that in the male infertility without Uu infection group [(60.93±16.62)μmol/L] and the control group [(35.92±1.73)μmol/L](F=200.987, P<0.01). The semen pH(6.09±0.14), sperm concentration [(26.64±8.34)×106/mL], and forward movement rate(a+ b) [(21.46±6.88)%], sperm survival rate [(28.92±9.17)%] and sperm morphology normal rate [(10.12±4.97)%] in the male infertility group with Uu infection were significantly lower than those in the male infertility without Uu infection group and the control group(all P<0.01). And the concentration of NO in the semen of males waspositively correlated with the concentrations of IL-17, IL-18 and TNF-α(R2=0.916 8, 0.863 0, 0.878 9, all P<0.01). The concentrations of IL-17 and IL-18 were positively correlated(R2=0.934 4, P<0.01). Conclusion Uu infection in infertility patients can cause high NO concentration.The levels of cytokines IL-17, IL-18 and TNF-α are positively correlated with the concentration of NO.The imbalance of various cytokines may be one of the causes of male infertility. Key words: Male infertility; Infection; Ureaplasma urealyticum; Interleukin-17; Interleukin-18; TNF-α; Cytokine; Nitric oxide

Human Semen Quality, Sperm DNA Damage, and the Level of Urinary Concentrations of 1N and TCPY, the Biomarkers of Nonpersistent Insecticides

The aim of the study was to evaluate the association between environmental exposure to nonpersistent insecticides and semen quality (concentration, motility, morphology, computer-aided semen analysis [CASA] parameters, and sperm DNA damage). Urine samples (n = 315) collected from men who attended the infertility clinic with normal semen concentration of 15 to 300 mln/ml and age under 45 years were analyzed for two metabolites (1-naphthol [1N] and 3,5,6-trichloro-2-pyridinol [TCPY]) of nonpersistent insecticides. Participants provided semen, blood, and saliva samples; additionally, men filled a detailed questionnaire. The results identified that urinary TCPY concentration was significantly associated with a decrease in motility; also there was a positive association between TCPY and DNA fragmentation index (DFI). 1N concentration was negatively associated with a percentage of sperm with normal morphology and positively with one of the CASA parameters (curvilinear velocity [VCL]). The results suggest that environmental exposure to nonpersistent insecticides may have an impact on semen quality parameters and sperm DNA damage.

THE INTEREST OF HYPEROSMOLAR EXTENDERS IN ROOSTER SPERM CRYOPRESERVATION

The aim of the present study was to investigate thepotential benefit of hyperosmolar extender during the freezing-thawing process in rooster sperm. The goal was to minimize the amount of intracellular water and reduce the impact of intracellular icecrystal during the freezing process. A total of five 45-week old Hubbard commercial broilers were subjected to bi-weekly semen collections. Collected sperm was pooled and divided in three aliquots. The control aliquot was diluted with Tris-extender without further supplementation at 300 mOsm (Control). The two (2) other aliquots were diluted with Trisextender at 300 mOsm containing vitamin E (Vit E) or with Tris-extender without Vitamin E but at osmolarity of 450 mOsm (Hyper). After incubation at 22 °C for 15 min, all aliquots were cooled and then frozen in liquid nitrogen. A Computer Aided Semen Analysis (CASA) was used to investigate the impact on different motility parameters. After thawing, Hyperosmolar (Hyper) and vitamin E extenders showed the highest values in terms of sperm motility preservation. Hyperosmolar extender (Hyper) showed particularly the highest values in terms of VSL velocities and progressive motile spermatozoa, known as indicators of sperm quality. In conclusion, the present results revealed that a significant impact was observed when using extenders at osmolarity of 450 mOsmin rooster sperm, in the same manner and even better then when using vitamin E. The positive impact is probably related to the reduction of intracellular ice formation.