Collected Semen Samples Research Articles

Association of Functional Genetic Variations in Uric Acid Transporters with the Risk of Idiopathic Male Infertility: A Genetic Association Study and Bioinformatic Analysis.

Uric acid plays an important role in sustaining and improving sperm morphology, viability, and motility. It is known that SLC2A9 and ABCG2 protein are the main urate transporter and genetic variations in these genes could be associated with the levels of serum uric acid. This study aimed to investigate the association between single-nucleotide polymorphisms (SNPs) SLC2A9-rs16890979, SLC2A9-rs3733591, ABCG2-rs2231142, and ABCG2-rs2231137 with male infertility. Additionally, the correlation of these SNPs with the uric acid level in seminal plasma of infertile men was examined. Subsequently, an in silico analysis was performed. In a case-control study, 193 infertile and 154 healthy controls were recruited. After semen sample collection, the uric acid level of seminal plasma was measured by a commercial kit. After genomic DNA extraction from sperm samples, SNPs genotyping was performed by PCR-RFLP method. Lastly, the effects of SNPs on the SLC2A9 and ABCG2 gene function were evaluated by bioinformatics tools. The genetic association study revealed that there are significant associations between rs16890979, rs3733591, rs2231142, and rs2231137 genetic variations and increased risk of male infertility. Also, these variations were associated with oligozoospermia and teratozoospermia, and sometimes with asthenozoospermia. Also, we found that four studied SNPs could be associated with a decreased level of uric acid of seminal plasma in teratozoospermia and asthenozoospermia. Bioinformatic analysis revealed that the mentioned polymorphisms could affect molecular aspects of SLC2A9 and ABCG2 genes. In this preliminary study, the rs16890979, rs3733591, rs2231142, and rs2231137 genetic variations could be considered as genetic risk factors for male infertility by interfering with the uric acid level of seminal plasma.

O-015 Enabling laboratory-quality male fertility analysis with 72-hour post ejaculation sample viability

Abstract Study question Can SPX72-treated semen samples sustain diagnostic viability and integrity for up to 72 hours, enabling comparable analysis of mail-in samples to fresh samples? Summary answer SPX72 preserves sperm viability, motility, and integrity up to 72 hours post-collection, equalling the diagnostic precision of fresh samples and enhancing at-home testing capabilities. What is known already Challenges with laboratory-based fertility assessments stem from logistical burdens and the necessity for timely sample delivery, often deterring male patient participation. Semen samples typically need to be assessed within 60 minutes of collection to ensure accuracy. Pre-existing solutions have failed to sufficiently prolong sample viability while matching the diagnostic accuracy of immediate laboratory analyses, an issue Sapyen's SPX72 medium addresses effectively. Study design, size, duration An exhaustive controlled study with over 100 semen samples compared SPX72-treated samples at 72 hours against fresh controls. The study incorporated actual postal logistics to emulate mail-in scenarios. Participants/materials, setting, methods A predetermined quantity of SPX72 per millilitre was added to the collected semen samples, with key indicators, including progressive and total motility, morphology, and DNA fragmentation, measured at intervals of 0, 24, 48, and 72 hours, contrasted with untreated, fresh samples. Main results and the role of chance SPX72 application preserved notable semen motility, evidenced by a mean progressive motility of 37.12% after 72 hours, a slight decrease from the initial 41.14%. This minor 4.02% reduction underscores SPX72's proficiency in maintaining diagnostic-grade motility over an extended duration. In terms of DNA integrity, SPX72 maintained DNA fragmentation at 14.78%, juxtaposed with 12.44% in control samples, a statistically insignificant variance (p-value of 1.15037E-08). This consistency in DNA fragmentation levels is pivotal for the accuracy of fertility diagnostic metrics. Additionally, SPX72 demonstrated its efficacy in stabilising the pH level of semen samples throughout the 72-hour period, ensuring an optimal environment for sperm preservation. This maintenance of pH is crucial as deviations can significantly affect sperm functionality and overall sample integrity. These results substantiate SPX72's capacity to preserve diagnostic sperm quality for up to three days, potentially redefining the logistics of male fertility testing by enabling accurate, laboratory-level assessment through a mail-in format. Limitations, reasons for caution Despite the controlled study and real-world postal validation, ongoing surveillance of sample stability in diverse global postal environments will be crucial to ensure consistent results across all geographies. Wider implications of the findings The data indicate that SPX72 sets a new standard for at-home fertility testing, matching clinical quality without the immediacy constraint. For the first time, home-based sperm diagnostics do not compromise clinical quality, revolutionising how male fertility is approached and managed. Trial registration number not applicable

P-040 The characteristic of human seminal plasma extracellular vesicle (SPEVs) profiles with different sperm features in a large cohort of 309 Taiwanese men

Abstract Study question To investigate human seminal plasma extracellular vesicle profiles in different sperm characteristics and their potential clinical application. Summary answer Normozoospermic semen has significantly higher SPEV concentration than asthenozoospermic or oligozoospermic samples. Moreover, SPEV concentration was highly correlated with total motile sperm count. What is known already Seminal plasma extracellular vesicles (SPEVs) are cell-derived membrane structures that carry proteins, nucleic acid, and lipids, which can modify the constitution and biological function of sperm. However, the characteristics of SPEVs in different sperm features remained unclear. Study design, size, duration From January 2021 to September 2023, 309 men who visited the NCKUH andrology clinic agreed to collect semen samples for this study, which the Institutional Review Board has approved. Participants/materials, setting, methods Semen parameters were measured by the X1 CASA system (Bonraybio, Taiwan). SPEVs were isolated by size exclusion chromatography column and ExoQuick™. Next, the SPEVs’ concentration and size distribution were performed with nanoparticle tracking analysis. The Kruskal-Wallis test calculated comparison in multiple groups, and Dunn’s multiple comparison tests accessed group-group comparison. The Pearson correlation coefficient r of exosome concentration with individual sperm parameters was calculated. A p-value less than 0.05 was set as statistical significance. Main results and the role of chance Among them, 59 semen samples were presented with normozoospermia, 36 were oligozoospermia, 73 were asthenozoospermia, and 141 samples had no sperm in semen. Among these 141 azoospermic men, 89 men are characterized by non-obstructive azoospermia (NOA), while the other 52 men had a vasectomy. Our data showed that normozoospermic semen has significantly higher SPEV concentration than asthenozoospermic or oligozoospermic semen samples (26 x109 [IQR 17.25 x109-56.25 x109] versus 12 x109 [IQR 5.6 x109-19.5 x109] versus 15.5 x109 [IQR 8.575 x109-26.75 x109], p = 0.0001). Nevertheless, azoospermic semen has the lowest SPEV concentrations. The concentration of SPEVs from infertile men diagnosed with NOA is slightly lower than vasectomized men (3.9 x109 [IQR 1.875 x109-7.675 x109] versus 5.4 x109 [IQR 2.9 x109-8 x109], p = 0.0001). The Pearson r coefficient of SPEV concentration with sperm concentration, progressive motility, and total motile sperm count was 0.238, 0.303, and 0.451, respectively. (P < 0.0001) Limitations, reasons for caution Because our data were collected from Taiwanese men, it is not yet known whether SPEV profiles differ among races. Wider implications of the findings Human SPEV concentration and its bio-content may be a potential non-invasive biomarker to predict the male reproductive outcome. Trial registration number not applicable

Recent decline in sperm motility among donor candidates at a sperm bank in Denmark.

Has there been variation in semen quality among men applying to be sperm donors (i.e. donor candidates) in Denmark in recent years (2017-2022)? The motile sperm concentration and total motile sperm count (TMSC) in ejaculates-both measures of sperm quality-declined by as much as 22% from 2019 to 2022. Questions remain about whether human semen quality has declined in recent years. Whilst some studies provide evidence for a decline in human semen quality, these findings have been disputed owing to potential biases in the populations studied or in the methods used to measure semen quality. Resolution of this issue has important implications for human fertility, as well as for those involved in the recruitment of sperm donors for use in medically assisted reproduction. We obtained data on the semen quality of ejaculates previously collected from 2017 to 2022 at sperm bank locations in four cities in Denmark: Aarhus, Aalborg, Copenhagen, and Odense. Our study focuses on the single semen samples provided by 6758 donor candidates aged between 18 and 45 years old to determine whether their sperm quality met a minimum criterion for them to be accepted as sperm donors. All ejaculates were analyzed within 1 hour of production. Semen volume (ml) was estimated by weight and both the concentration (106/ml) of sperm as well as the concentration of motile sperm (World Health Organization grades a and b) were measured using the same protocols and computer-assisted semen analysis system across all years at each site. Statistical analyses of the semen variables were controlled for age and donation site, as well as the average monthly high temperature when the ejaculate was produced. From 2017 to 2019, semen volume, sperm concentration, and total sperm count in the ejaculates of donor candidates increased by 2-12%. Then, from 2019 to 2022, sperm concentration and total sperm count changed by 0.1-5% from year to year, but none of those changes were statistically significant. In contrast, both motile sperm concentration and TMSC declined significantly, by 16% and 22%, respectively, between 2019 and 2022. Thus, the concentration of motile sperm in donor candidates declined from 18.4 [95% CL: 17.0, 20.0] million/ml in 2019 to 15.5 [14.4, 16.7] million/ml in 2022, and TMSC declined from 61.4 [55.8, 67.5] million per ejaculate in 2019 to 48.1 [44.1, 52.4] million in 2022. We cannot determine from the available data the causes of the decline in semen quality of donor candidates from 2019 to 2022. However, as this period coincides with lockdowns and changes in work patterns during the coronavirus disease 2019 pandemic, it is possible that changes in motile sperm concentration and TMSC were the result of changes in the lifestyles of the men whose semen was analyzed. Men providing initial semen samples at sperm banks, when applying to be sperm donors, are a useful population in which to monitor changes in human semen quality over time. Our results have implications for human fertility and the recruitment of sperm donors for medically assisted reproduction, where motile sperm concentration is an essential selection criterion because it influences fertility. We suggest that gathering health and lifestyle data on donor candidates at sperm banks might help to identify causal factors for the decline of sperm quality that could be addressed and intervention, if desired, could be personalized for each accepted donor. No external funding was obtained for this study. E.L. and A.-B.S. are employees of Cryos International. AP reports paid consultancy for Cryos International, Cytoswim Ltd, Exceed Health, and Merck Serono in the last 2 years of this study, but all monies were paid to the University of Sheffield (former employer). AP is also an unpaid trustee of the Progress Educational Trust (Charity Number 1139856). RM declares support from Cryos International to present results of this research at ESHRE 2023. None of the authors were directly involved in the collection or physical analysis of semen samples. N/A.

Variations in seminal microbiota and their functional implications in chickens adapted to high-altitude environments

Seminal fluid, once believed to be sterile, is now recognized as constituting a complex and dynamic environment inhabited by a diverse community of micro-organisms. However, research on the seminal microbiota in chickens is limited, and microbiota variations among different chicken breeds remain largely unexplored. In this study, we collected semen samples from Beijing You Chicken (BYC) and Tibetan Chicken (TC) and explored the characteristics of the microbiota using 16S rRNA gene sequencing. Additionally, we collected cloacal samples from the TC to control for environmental contamination. The results revealed that the microbial communities in the semen were significantly different from those in the cloaca. Firmicutes and Actinobacteriota were the predominant phyla in BYC and TC semen, respectively, with Lactobacillus and Phyllobacterium being the dominant genera in each group. Additionally, the seminal microbiota of BYC exhibited greater richness and evenness than that of TC. Principal coordinate analysis (PCoA) indicated significant intergroup differences between the seminal microbiotas of BYC and TC. Subsequently, by combining linear discriminant analysis effect size and random forest analyses, we identified Lactobacillus as the predominant microorganism in BYC semen, whereas Phyllobacterium dominated in TC semen. Furthermore, co-occurrence network analysis revealed a more intricate network in the BYC group than in the TC group. Additionally, unique microbial functional characteristics were observed in each breed, with TC exhibiting metabolic features potentially associated with their ability to adapt to high-altitude environments. The results of this study emphasized the unique microbiota present in chicken semen, which may be influenced by genetics and evolutionary history. Significant variations were observed between low-altitude and high-altitude breeds, highlighting the breed-specific implications of the seminal microbiota for reproduction and high-altitude adaptation.

Prevalence and implications of microplastic contaminants in general human seminal fluid: A Raman spectroscopic study

Microplastics are ubiquitous environmental contaminants that have been detected in human semen from polluted areas, yet their prevalence and effects in the general population remain largely unexplored. To examine microplastic presence, abundance, polymer types, and associations with semen quality parameters in individuals without occupational exposures, this study was conducted by collecting semen samples from 40 participants undergoing premarital health assessments in Jinan, China. Raman microspectroscopy was employed to identify, quantify, and categorize microplastic polymers, sperm motility was assessed via computer-assisted analysis, and morphology was evaluated through Diff-Quik staining. Correlations between demographics, semen parameters, and microplastic content were examined by statistical analysis. We found that microplastics were detected in all semen samples, with 2 particles per sample (ranging from 0.72 to 7.02 μm). Eight distinct polymers were identified, with polystyrene (31 %) being most prevalent. Semen exposed to polystyrene demonstrated higher sperm progressive motility as compared to polyvinyl chloride exposure group (43.52 ± 14.21 % vs 19.04 ± 13.46 %). Sperm morphological abnormalities were observed but not significantly associated with specific plastic types. In conclusion, this study reveals microplastic contamination in semen from individuals without occupational exposure, with PS, PE, and PVC being the most prevalent and exhibiting differential correlations with sperm progressive motility, and highlight the need for further research into the potential reproductive impacts of microplastic exposure.

Effects of shilajit addition to honey bee diet on semen freezing

This study aimed to investigate the effects of adding shilajit to the diet of honey bees on semen freezing. A total of 5 groups were formed in the research, one of which was a control group (SH-0) and the other four were an experimental group (SH-1, SH-2, SH-3, SH-4). A total of 25 study colonies were used, 5 in each group. While the SH-0 group was formed without using any additives, the experimental groups were fed with the addition of shilajit (1/1 sugar/water) in different doses (5, 10, 15, 20 mg/L) to the honey bee diet. The collected semen samples were frozen in liquid nitrogen vapor and then stored in liquid nitrogen at -196°C. Then, semen samples were thawed at 37°C and evaluated to determine spermatological parameters (motility, acrosome integrity, plasma membrane integrity, spermatozoa concentration). It was also examined in terms of total oxidant and total antioxidant. Compared to the control group, it was determined that all shilajit doses significantly increased spermatological parameters such as motility, hypoosmotic swelling test (HOST), acrosome integrity (p<0.001), but did not significantly affect the spermatozoa concentration value (p >0.05). Although there is no statistical difference between the groups in terms of semen TAS (Total Antioxidant Status) and TOS (Total Oxidant Status) values, which are oxidative stress parameters, the numerical increase in TAS values in SH-2 and SH-3 groups; striking. As a result, it was determined that adding shijajit to the honey bee diet positively affected the post-thawing spermatological parameters of frozen bee semen.

Epigallocatechin-3-gallate affects the quality of fresh and frozen-thawed semen of Simmental bull by two different cryopreservation methods.

During the freezing process of semen, due to the generating of significant amounts of free radicals, the quality of sperm changes. Epigallocatechin-3-gallate (EGCG) is a green tea catechin, which in this study was applied to investigate its effect on the quality of bulls' sperm. We collected semen samples with an artificial vagina from 12 Simmental bulls to evaluate the effect of EGCG (10.00 and 20.00 µmol) in two cryopreserving methods on the quality parameters of semen. We designed six groups including two control groups (method one and two) and four treatments (EGCG 10.00 µmol + method one; EGCG 20.00 µmol + method one; EGCG 10.00 µmol + method two; EGCG 20.00 µmol + method two). The 20.00 µmol EGCG and a method two significantly affected the amending oxidative conditions as well as an increase in total antioxidant capacity and a decrease in malondialdehyde. The effect of EGCG in both concentrations was more on method two. The desired impact on sperm motility, viability, inhibition of lipid peroxidation and sperm DNA damage was observed in EGCG groups compared to control groups. Among the two methods, the method two had fewer adverse effects on the plasma membrane, motility parameters, viability and DNA of sperm. The EGCG in the semen extender yielded a favorable impact on thawed sperm. This effect was prompted in combination with the method two.

A Possible Role for Nerve Growth Factor and Its Receptors in Human Sperm Pathology.

Nerve growth factor (NGF) signalling affects spermatogenesis and mature sperm traits. In this paper, we aimed to evaluate the distribution and the role of NGF and its receptors (p75NTR and TrKA) on the reproductive apparatus (testis and epididymis) and sperm of fertile men (F) and men with different pathologies, namely varicocele (V) and urogenital infections (UGIs). We collected semen samples from 21 individuals (31-40 years old) subdivided as follows: V (n = 7), UGIs (n = 7), and F (n = 7). We submitted the semen samples to bacteriological analysis, leucocyte identification, and analysis of sperm parameters (concentration, motility, morphology, and viability). We determined the seminal plasma levels of NGF, interleukin 1β (IL-1β), and F2-isoprostanes (F2-IsoPs), and the gene and protein expression of NGF receptors on sperm. We also used immunofluorescence to examine NGF receptors on ejaculated sperm, testis, and epididymis. As expected, fertile men showed better sperm parameters as well as lower levels of NGF, F2-IsoPs, and IL-1β compared with men with infertility. Notably, in normal sperm, p75NTR and TrKA were localised throughout the entire tail. TrKA was also found in the post-acrosomal sheath. This localisation appeared different in patients with infertility: in particular, there was a strong p75NTR signal in the midpiece and the cytoplasmic residue or coiled tails of altered ejaculated sperm. In line with these findings, NGF receptors were intensely expressed in the epididymis and interstitial tissue of the testis. These data suggest the distinctive involvement of NGF and its receptors in the physiology of sperm from fertile men and men with infertility, indicating a possible role for new targeted treatment strategies.

Alterations in sperm DNA methylation may as a mediator of paternal air pollution exposure and offspring birth outcomes: Insight from a birth cohort study

Paternal exposure to environmental risk factors influences the offspring health. This study aimed to evaluate the association between paternal air pollution exposure mediated by sperm DNA methylation and adverse birth outcomes in offspring. We recruited 1607 fertile men and their partners from 2014 to 2016 and collected semen samples to detect sperm DNA methylation. Multivariate linear regression and weighted quantile sum regression models were used to assess the associations between paternal air pollution exposure and offspring birth outcomes. A critical exposure window was identified. Reduced representation bisulfite sequencing was used to detect sperm DNA methylation. The results demonstrated that high paternal exposure to PM2.5 (β = −211.31, 95% CI: (−386.37, −36.24)), PM10 (β = −178.20, 95% CI: (−277.13, −79.27)), and NO2 (β = −84.22, 95% CI: (−165.86, −2.57)) was negatively associated with offspring's birthweight, especially in boys. Additionally, an early exposure window of 15–69 days before fertilization was recognized to be the key exposure window, which increased the risk of low birth weight and small for gestational age. Furthermore, paternal co-exposure to six air pollutants contributed to lower birthweight (β = −51.91, 95% CI: (−92.72, −11.10)) and shorter gestational age (β = −1.72, 95% CI: (−3.26, −0.17)) and PM2.5 was the most weighted pollutant. Paternal air pollution exposure resulted in 10,328 differentially methylated regions and the IGF2R gene was the key gene involved in the epigenetic process. These differentially methylated genes were predominantly associated with protein binding, transcriptional regulation, and DNA templating. These findings indicate that spermatogenesis is a susceptible window during which paternal exposure to air pollution affects sperm DNA methylation and the birth outcomes of offspring.

Experiences and Outcomes of Fertility Testing in Male Adolescents and Young Adults with Sickle Cell Disease

Introduction: Most adolescents and young adults (AYAs) with sickle cell disease (SCD) desire future biological parenthood. Recent research shows abnormal semen parameters in men with SCD, yet few studies have been conducted to elucidate fertility related impacts of SCD and its treatments among male AYAs with SCD. AYAs face many barriers to receiving care that could also impact their ability to complete fertility testing (i.e., via semen analysis). Thus, the aims of this study were to: 1) assess the feasibility of obtaining semen samples at a reproductive diagnostic center from male AYAs with SCD, 2) describe AYAs' and caregivers' experiences with testing, and 3) examine semen parameters among AYAs who completed testing. Methods Male AYAs with SCD (14-22 years, any genotype) and their caregivers were recruited to a single site study from November 2022-April 2023; the protocol involved completion of a novel web-based reproductive health program (i.e., FUTURES) describing potential fertility-related aspects of SCD and its treatments. AYA participants ≥18 years of age and AYA-caregiver dyads (if the AYA was <18 years old) were subsequently informed of the option for the AYA to obtain a semen analysis at an off-site reproductive diagnostic center (with costs of testing and transportation covered). Up to three attempts were made to contact AYA participants about their interest in testing. If interested, an order was sent to the testing center who then coordinated testing with the AYA. Research staff made up to three attempts to follow up with AYAs who reported interest but did complete testing. AYAs (and caregivers, if <18) who completed testing were notified of their results by a hematologist and invited to complete a facilitators interview. AYAs (and caregivers, if <18) who did not complete testing were invited to complete a barriers interview. Research staff conducted, transcribed, and analyzed interview transcripts for themes that demonstrated strong interrater reliability (κ=.94). Testing was considered feasible if ≥25% of AYAs completed testing. Demographic and treatment characteristics were abstracted from the medical record. Results Thirty-three AYAs completed FUTURES; five (15%) completed testing (Table 1) and a facilitators interview. Ten AYAs completed a barriers interview. Of the 18 enrolled caregivers, one completed a facilitators interview and seven completed a barriers interview. Facilitators included: 1) interest in learning about reproductive health [“ I told him ... sometimes people don't know if it is too late {to have a biological child} and if we checked we could have done something about it.” - Caregiver], 2) assistance coordinating testing appointments [“ Helping to coordinate {made testing easier} because I kept on missing the appointments.” -AYA] 3) alleviating costs [“ It {the funding} helped a lot because they {research team} had to pay and we didn't have to worry about it.” - Caregiver]. Barriers included: 1) discomfort [“ I don't think anything could have helped with the decision... he was just a little uncomfortable.” -Caregiver], 2) time [“ It is something I'm interested in, but not at the moment. I'm catching up on a lot of things.” -AYA], 3) transportation [e.g., “ I don't have a car. I didn't have a way to get there.” -AYA], and 4) forgetting the appointment [“ We just forgot about the appointment.” -Caregiver]. Conclusions Despite providing reproductive health education, compensation, and access to a local testing center, obtaining semen samples at an offsite center from male AYAs with SCD was not feasible. Abnormal semen parameters were noted across genotypes. While azoospermia and cryptozoospermia were noted in those on prolonged hydroxyurea treatment, oligospermia was also found in an individual with less severe disease and no hydroxyurea exposure. These findings underscore the importance of larger, longitudinal studies to examine semen parameters in male AYAs with SCD, ideally starting in mid-puberty. Further, qualitative findings highlight the need for: 1) developmentally and culturally appropriate fertility education, 2) innovative methods to collect semen samples (e.g., mail-in testing), and 3) advocacy to increase access to affordable reproductive healthcare. This is crucial to elucidate the fertility impacts of SCD and hydroxyurea and because the curative options that can also affect fertility are expanding for this underserved minority population.

Phosphatidylserine Levels in Seminal Plasma of Iraqi Infertile Men Using Direct and Indirect Swim Up Sperm Preparation Techniques

Phosphatidylserine (PS), exposed on viable sperm, is required for fertilization as it is recognized by specific receptors located on the microvilli of the oocyte to promote sperm: egg fusion. The heads of viable and motile sperm exhibit PS. With (PS) exposure progressively increasing during sperm transit through the epididymis. PS asymmetry on the leaflets of plasma membrane is arranged in a complex way, via multiple proteins functioning in opposite directions; further, PS is also essential for many intracellular trafficking events. The aim of this study was to evaluate the phosphatidylserine PS level in seminal plasma of infertile men before and after activation using Direct Swim Up (DSU) and Indirect Swim Up (IDSU) activation method and make a comparison between them. Forty infertile men were included in this study during their attendance to the infertility clinic at High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University. Normozoospermic or mild asthenozoospermic semen samples were used to choose the cases, according to WHO manual standards (1999 -2010). The collected semen samples obtained were divided into three parts: The first part prepared was as an in vitro sperm characterization before activation while the second and third parts were after activation using direct and indirect swim up techniques respectively. The level of seminal plasma phosphatidylserine was increased after the activation techniques from the baseline level before activation. Comparison of Seminal plasma phosphatidylserine levels between direct and indirect sperm activation technique were significantly higher in direct swim up activation technique than indirect one. Phosphatidylserine (PS) in semen plasma of infertile men was significantly increased after activation using Direct Swim Up DSU preparation method.

SUMO1 and Defective Spermatozoa Correlate with Endogenous Hydrogen Peroxide and Live Birth Outcome in Intrauterine Insemination Cycles for Unexplained Infertility.

This study aimed to investigate the correlation between hydrogen peroxide (H2O2), small ubiquitin-like modifier molecules (SUMO), and pregnancy outcomes in couples with unexplained infertility (UI) undergoing intrauterine insemination (IUI) treatment. We prospectively collected semen samples from 56 couples with UI and divided the spermatozoa into motile and immotile fractions by density gradient centrifugation (DSC). Immunofluorescence staining was used to examine the immunostaining and localization of nuclear pore complex (NPC), SUMO1, and SUMO2/3 in spermatozoa. We detected H2O2 levels by chemiluminescence methods. We found that H2O2 levels correlated with NPC (neck) (r = 0.400) and NPC (tail) (r = 0.473) in motile sperm fractions. In immotile fractions, H2O2 positively correlated with NPC (tail) (r = 0.431) and SUMO1 (neck) (r = 0.282). Furthermore, the positive NPC (tail) group had a significantly lower live birth rate than the negative NPC group (17.9% = 5/28 vs. 42.9% = 12/28). In conclusion, H2O2 positively correlated with SUMO1 (neck) and NPC (tail) in human spermatozoa. The DSC may partially eliminate defective spermatozoa (positive NPC staining); however, if defective spermatozoa remain in the motile fraction, this scenario is associated with a low live birth rate following IUI treatment.

Relevance of PUFA-derived metabolites in seminal plasma to male infertility.

This study aims to investigate the biological effects of polyunsaturated fatty acid (PUFA)-derived metabolites in seminal plasma on male fertility and to evaluate the potential of PUFA as a biomarker for normozoospermic male infertility. From September 2011 to April 2012, We collected semen samples from 564 men aged 18 to 50 years old (mean=32.28 years old)ch., residing in the Sandu County, Guizhou Province, China. The donors included 376 men with normozoospermia (fertile: n=267; infertile: n=109) and 188 men with oligoasthenozoospermia (fertile: n=121; infertile: n=67). The samples thus obtained were then analyzed by liquid chromatography-mass spectrometry (LC-MS) to detect the levels of PUFA-derived metabolites in April 2013. Data were analyzed from December 1, 2020, to May 15, 2022. Our analysis of propensity score-matched cohorts revealed that the concentrations of 9/26 and 7/26 metabolites differed significantly between fertile and infertile men with normozoospermia and oligoasthenozoospermia, respectively (FDR < 0.05). In men with normozoospermia, higher levels of 7(R)-MaR1 (HR: 0.4 (95% CI [0.24, 0.64]) and 11,12-DHET (0.36 (95% CI [0.21, 0.58]) were significantly associated with a decreased risk of infertility, while higher levels of 17(S)-HDHA (HR: 2.32 (95% CI [1.44, 3.79]), LXA5 (HR: 8.38 (95% CI [4.81, 15.24]), 15d-PGJ2 (HR: 1.71 (95% CI [1.06, 2.76]), and PGJ2 (HR: 2.28 (95% CI [1.42, 3.7]) correlated with an increased risk of infertility. Our ROC model using the differentially expressed metabolites showed the value of the area under the curve to be 0.744. The PUFA-derived metabolites 7(R)-MaR1, 11,12-DHET, 17(S)-HDHA, LXA5, and PGJ2 might be considered as potential diagnostic biomarkers of infertility in normozoospermic men.

Collecting semen samples at home for fertility assessment: time for a new standard?

In fertility clinics the standard approach to semen collection involves a private room close to the laboratory to avoid fluctuations in temperature and to control the time between collection and processing. There are still no firm conclusions whether collecting semen at home has any influence on sperm quality and reproductive competence. The purpose of this study was to assess whether the site of semen collection affects semen parameters. This retrospective cohort study performed at a tertiary level public fertility center included 8634 semen samples from 5880 men undergoing fertility assessment from 2015 to 2021. The impact of sample collection site was evaluated using a generalized linear mixed model. A subgroup analysis comparing clinic to home collection within the same patient was performed on 1260 samples from 428 men by paired t-test or Wilcoxon Signed Rank Test. Samples collected at home (N.=3240) had significantly higher semen volume, sperm concentration and total sperm count respect to samples collected at clinic (N.=5530) (median (range): 2.9 (0.0-13.9) mL versus 2.9 (0.0-11.5) mL, P=0.016; 24.0 (0.0-252.0) million/mL versus 18.0 (0.0-390.0), P<0.0001; 64.6 (0.0-946.0) million versus 49.3 (0.0-1045.0), P<0.0001, respectively). There was no difference in abstinence period and sperm motility. Paired comparisons of semen characteristics in 428 patients with home-collected (N.=583) and clinic-collected (N.=677) samples confirmed a no negative effect on volume and total sperm count. Our data provide evidence for a not disadvantage with collection at home.

Comparative Studies on the Effect of Tris-Citric AcidFructose Egg Yolk Glycerol and Tris-Egg Yolk-Glucose based Semen Extender on various Semen Parameters for Effective Long Term Cryopreservation

Aim: To evaluate the suitable Semen extender for a long duration of cryopreservation among Tris citric acid fructose egg yolk glycerol extender and Tris-egg yolk-glucose extender,. Materials and Methods: The sample size has been calculated by the clinCalc with an alpha error-threshold at 0.05, 95 % confidence interval (CI), G power at 80%, and enrolment ratio 1. Our study has been repeated 14 times which is (N=14) and it was measured using statistical Independent Samples Test. The study involved 28 sample sizes with 2 different groups with each group containing 14 samples. The collected semen samples are diluted with the Tris-citric acid-fructose egg yolk glycerol extender and Tris-egg yolk-glucose extender, based novel semen extenders. A small quantity of the diluted semen sample was taken to a counting device known as makler counting chamber used for analyzing the sperm count and motility of the semen samples. Results: The Tris-citric acid-fructose egg yolk glycerol extenders were better than Tris-egg yolk-glucose extender. So the estimated sperm count and sperm motility were found to be P &lt;0.05, revealing the statistical insignificance. The characteristics after cryopreservation were decreased than the characteristics before cryopreservation. Conclusion: The sperm parameters were preserved well in Tris-citric acid-fructose egg yolk glycerol semen extender when compared with Tris-egg yolk-glucose based semen extender with respect to sperm count and sperm motility.

Comparative Studies on the Effect of Tris -Egg Yolk and Soya Milk Based Semen Extender on various Semen Parameters for Effective Long Term Cryopreservation

Aim: To analyze the most effective Novel semen extender for long time cryopreservation among Tris egg yolk extender and Soya milk extender. Materials and Methods: The sample size was calculated using clinCalc.com with an alpha error-threshold at 0.05, 95 % confidence interval, G power at 80 %, and enrolment ratio 1. The study was repeated 14 times (N=14) and was measured using statistical Independent Samples Test. The study contained a total sample size of 28 with 2 different groups. Tris egg yolk and soya milk based novel semen extenders were added to the collected semen samples and few drops of the sample was placed on a makler counting chamber which was used for analyzing the concentration, sperm motility and sperm count of the semen samples. Results: The Tris egg yolk based semen extenders were better than the soya milk based semen extenders. Estimated sperm motility and sperm count were found to be P&gt;0.05, revealing statistical insignificance. Conclusion: The sperm parameters were preserved well in Tris egg yolk based semen extender when compared with soya milk based semen extender with respect to sperm count and sperm motility.

Effects of photobiomodulation therapy on human sperm function

IntroductionSperm motility is a crucial factor in male infertility and it depends on mitochondrial tail movements. Photobiomodulation light therapy allows the cells to produce their energy through activation of the mitochondria. The aim of the present study was to examine the impact of photobiomodulation on sperm motility in astenozoospermic individuals. Materials and methodsFollowing semen analyses of 20 astenozoospermic individuals, collected semen samples were centrifuged. Pellet was obtained and homogenized through mixing with culture media in 1:1 ratio. Each semen samples were divided into 3 groups. In the first group, control samples were not exposed to laser irradiation. The Group 2 and Group 3 were exposed to 650nm wavelength of photobiomodulation from 10cm distance in dark environment via a 36cm2 aperture sizer with 200mW output power for 30 and 60min duration, respectively. Sperm motilities were evaluated and chromatin condensation of sperms was determined. ResultsSperm motilities were significantly increased in photobiomodulation groups compared with the controls. Sperm motilities tended to be different between the 30 and 60min red light exposure groups; however, it was not statistically significant. When the motility grades were compared, no significant difference was observed in non-progressive motility sperms. While immotile sperms decreased significantly in the photobiomodulation groups compared to the control group, progressive sperms increased. ConclusionsThe results of the present study demonstrated that the photobiomodulation is an efficient method to increase the sperm motility of astenozoospermic individuals independent of the duration of exposure.

Pilot Study: Next-generation Sequencing of the Semen Microbiome in Vasectomized Versus Nonvasectomized Men

BackgroundApproximately half a million vasectomies are performed every year in the USA. There is a paucity of literature on the impact of male sterilization on the semen microbiome and whether it prompts microbiota dysbiosis. ObjectiveTo investigate if vasectomy induces changes in the seminal microbiome via comparison of semen samples from men before and after vasectomy, and if the seminal microbiome profiles for vasectomized men follow a particular pattern with respect to diversity and abundance. Design, setting, and participantsFrom July 2021 to February 2022, we prospectively collected and analyzed semen samples from 58 men at one outpatient clinic. Eighteen men provided a semen sample before and 3 mo after vasectomy. We also collected semen samples from 22 fertile nonvasectomized men and from a further 18 vasectomized men at 3 mo after vasectomy. Outcome measurements and statistical analysisSemen microbiome α-diversity, beta-diversity, and relative abundance were compared initially between paired and then between unpaired vasectomized and nonvasectomized samples. Analysis of variance (ANOVA), permutational multivariate ANOVA, and analysis of the composition of microbiomes with bias correction were used to assess differences. Results and limitationsIn both paired and unpaired sets of samples, a decreasing trend for α-diversity in semen after vasectomy was observed. Shannon diversity, the relative abundance of species with an abundance >2%, and composition were not significantly changed. Sphingomonas, Brevundimonas, and Paracoccus abundance decreased after vasectomy, while Corynebacterium abundance increased. The results may be limited by the sample size and lack of demographic heterogeneity. ConclusionsVasectomy is followed by a decrease in α-diversity and changes in the relative abundance of bacterial species in the semen microbiome. Further investigation is necessary to understand the clinical significance of these changes after vasectomy. Patient summaryWe evaluated changes in the bacteria species in semen after vasectomy. We found that vasectomy decreased the richness and evenness of bacteria species in semen, but the overall bacterial community remained similar. Further studies are needed to assess the implications of changes in semen bacteria after vasectomy.

Masturbation in a Free-ranging Male Long-tailed Macaque Macaca fascicularis (Raffles, 1821) on Mindanao Island, Philippines

Long-tailed macaques in the Philippines are understudied in terms of ecology and behavior, which includes their sexual behaviors, in their natural habitats. Masturbation is a natural, auto-erotic behavior rarely observed in the wild in most non-human primates. In this paper, we report an act of masturbation by a free-ranging male long-tailed macaque in a beach forest at an eco-tourism site in Tagum City, Mindanao Island, Philippines. The masturbation was performed by a sub-adult male resting on the handrail of a wooden staircase. A few seconds after masturbation, ejaculation was achieved. Although more detailed observations are needed, it is likely that limited access to female mates contributed to the male’s self-stimulation. This further calls for more scientific explorations on reproductive and non-reproductive sexual behaviors of long-tailed macaques in the wild. Moreover, this can be crucial for conservation purposes such as its application to the non-invasive collection of semen samples in wild populations.