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ObjectiveEvaluate reduction in progressively motile sperm per high power field (HPF) in midcycle cervical mucus after intercourse with Ovaprene: an investigational monthly non-hormonal vaginal contraceptive consisting of a vaginal ring and mechanical barrier, releasing spermiostatic ferrous gluconate.Study designOpen-label, multicenter study enrolling heterosexually-active women with previous permanent contraception. Participants underwent a baseline postcoital test cycle with no device to confirm the presence of sperm, followed by one diaphragm postcoital test cycle, one Ovaprene safety cycle, and two Ovaprene postcoital test cycles. In each postcoital test cycle, participants underwent a midcycle cervical mucus evaluation to confirm an Insler score ≥10 and absence of sperm, and then returned two to four hours after vaginal intercourse for repeat cervical mucus evaluation. We considered

The ex situ population of the endangered black-footed ferret (Mustela nigripes) has been experiencing declines in reproductive success over the past 30 years of human-managed care. A potential cause may be environmental-dependent inbreeding depression with diet being one of the contributing factors since ferrets are not fed their natural diet of prairie dogs. Here, we generated and analyzed semen proteome and transcriptome data from both wild and ex situ ferrets maintained on various diets. We identified 1757 proteins across all samples, with 149 proteins unique to the semen of wild ferrets and forming a ribosomal predicted protein-protein interaction cluster. Wild ferrets also differed from ex situ ferrets in their transcriptomic profile, showing enrichment in ribosomal RNA processing and potassium ion transport. Successful fertility outcomes documented for ex situ ferrets showed the strongest association with the semen transcriptome, with enrichment in genes involved in translation initiation and focal adhesion. Fertility also synergized with the effect of diet on differentially expressed transcriptomes, mainly affecting genes enriched in mitochondrial function. Our data and functional networks are important for understanding the causes and mechanisms of declining fertility in the ex situ ferret population and can be used as a resource for future conservation efforts.

CRISPR-Cas9 short guide RNA (sgRNA) library screening is a powerful approach to understand the molecular mechanisms of biological phenomena. However, its in vivo application is currently limited. Here, we developed our previously established in vitro revival screening method into an in vivo one to identify factors involved in spermatogenesis integrity by utilizing sperm capacitation as an indicator. By introducing an sgRNA library into testicular cells, we successfully pinpointed the retinal degeneration 3 (Rd3) gene as a significant factor in spermatogenesis. Single-cell RNA sequencing (scRNA-seq) analysis highlighted the high expression of Rd3 in round spermatids, and proteomics analysis indicated that Rd3 interacts with mitochondria. To search for cell-type-specific signaling pathways based on scRNA-seq and proteomics analyses, we developed a computational tool, Hub-Explorer. Through this, we discovered that Rd3 modulates oxidative stress by regulating mitochondrial distribution upon ciliogenesis induction. Collectively, our screening system provides a valuable in vivo approach to decipher molecular mechanisms in biological processes.

Aminergic signaling is known to play a critical role in regulating female reproductive processes in both mammals and insects. In Drosophila, the ortholog of noradrenaline, octopamine, is required for ovulation as well as several other female reproductive processes. Two octopamine receptors have already been shown to be expressed in the Drosophila reproductive tract and to be required for egg-laying: OAMB and Octβ2R. The Drosophila genome contains 4 additional octopamine receptors-Octα2R, Octβ1R, Octβ3R, and Oct-TyrR-but their cellular patterns of expression in the reproductive tract and potential contribution(s) to egg-laying are not known. In addition, the mechanisms by which OAMB and Octβ2R regulate reproduction are incompletely understood. Using a panel of MiMIC Gal4 lines, we show that Octα2R, Octβ1R, Octβ3R, and Oct-TyrR receptors are not detectable in either epithelium or muscle but are clearly expressed in neurons within the female fly reproductive tract. Optogenetic activation of neurons that express at least 3 types of octopamine receptors stimulates contractions in the lateral oviduct. We also find that octopamine stimulates calcium transients in the sperm storage organs and that its effects in spermathecal, secretory cells, can be blocked by knock-down of OAMB. These data extend our understanding of the pathways by which octopamine regulates egg-laying in Drosophila and raise the possibility that multiple octopamine receptor subtypes could play a role in this process.

There has recently been an explosion of studies implicating the human microbiome in playing a critical role in many disease and wellness states. The etiology of abnormal semen analysis (SA) parameters is not identified in 30% of cases; investigations involving the semen microbiome may bridge this gap. Here, we explore the relationship between the semen microbiome and alterations of sperm parameters. We recruited men presenting for fertility evaluation or vasectomy consultation with proven biological paternity. SA and next generation sequencing was performed. Differential abundance testing using Analysis of composition of Microbiota with Bias Correction (ANCOM-BC) was performed along with canonical correlational analysis for microbial community profiling. Men with abnormal (N = 27) sperm motility showed a higher abundance of Lactobacillus iners compared to those with normal (N = 46) sperm motility (mean proportion 9.4% versus 2.6%, p = 0.046). This relationship persisted on canonical correlational analysis (r = 0.392, p = 0.011). Men with abnormal sperm concentration (N = 20) showed a higher abundance of Pseudomonas stutzeri (2.1% versus 1.0%, p = 0.024) and Pseudomonas fluorescens (0.9% versus 0.7%, p = 0.010), but a lower abundance of Pseudomonas putida (0.5% versus 0.8%, p = 0.020), compared to those with normal sperm concentration (N = 53). Major limitations are related to study design (cross-sectional, observational). Our results suggest that a small group of microorganisms may play a critical role in observed perturbations of SA parameters. Some of these microbes, most notably Lactobacillus iners, have been described extensively within other, fertility-related, contexts, whereas for others, this is the first report where they have potentially been implicated. Advances in our understanding of the semen microbiome may contribute to potentially new therapeutic avenues for correcting impairments in sperm parameters and improving male fertility.

Primordial germ cells (PGCs) are the embryonic precursors of sperm and eggs. They transmit genetic and epigenetic information across generations. Given the prominent role of germline defects in diseases such as infertility, detailed understanding of human PGC (hPGC) development has important implications in reproductive medicine and studying human evolution. Yet, hPGC specification remains an elusive process. Here, we report the induction of hPGC-like cells (hPGCLCs) in a bioengineered human pluripotent stem cell (hPSC) culture that mimics peri-implantation human development. In this culture, amniotic ectoderm-like cells (AMLCs), derived from hPSCs, induce hPGCLC specification from hPSCs through paracrine signaling downstream of ISL1. Our data further show functional roles of NODAL, WNT, and BMP signaling in hPGCLC induction. hPGCLCs are successfully derived from eight non-obstructive azoospermia (NOA) participant-derived hPSC lines using this biomimetic platform, demonstrating its promise for screening applications.

Despite rapid evolution across eutherian mammals, the X-linked MIR-506 family miRNAs are located in a region flanked by two highly conserved protein-coding genes (SLITRK2 and FMR1) on the X chromosome. Intriguingly, these miRNAs are predominantly expressed in the testis, suggesting a potential role in spermatogenesis and male fertility. Here, we report that the X-linked MIR-506 family miRNAs were derived from the MER91C DNA transposons. Selective inactivation of individual miRNAs or clusters caused no discernible defects, but simultaneous ablation of five clusters containing 19 members of the MIR-506 family led to reduced male fertility in mice. Despite normal sperm counts, motility, and morphology, the KO sperm were less competitive than wild-type sperm when subjected to a polyandrous mating scheme. Transcriptomic and bioinformatic analyses revealed that these X-linked MIR-506 family miRNAs, in addition to targeting a set of conserved genes, have more targets that are critical for spermatogenesis and embryonic development during evolution. Our data suggest that the MIR-506 family miRNAs function to enhance sperm competitiveness and reproductive fitness of the male by finetuning gene expression during spermatogenesis.

Background: Glutamine is a nonessential amino acid and the most abundant amino acid found in the seminal plasma and sperm-rich fraction of boar semen. Glutamine plays an important role in enhancing glutathione (GSH) synthesis. It acts as an effective antioxidant in semen and provides intracellular defense to sperm against oxidative stress. This study aimed to improve the quality of frozen-thawed boar semen by using glutamine supplementation in a short-term semen extender during the holding time at 17 °C before cryopreservation. Results: The results indicate that the total motility, progressive motility, LIN, STR, and WOB were the highest in the 20 mM supplementation group at the 2 h timepoint after thawing. Thus, the optimal concentration for glutamine supplementation in short-term boar semen extender during the holding time at 17 °C was 20 mM. Interestingly, at all of the time points after thawing, 20 mM glutamine supplementation exhibited the highest level of sperm viability and membrane integrity when compared to the CONTROL (0 mM) and other experimental dilution groups. Moreover, the acrosome integrity, mitochondrial activity, and capacitation status (F pattern) were significantly greater in the 20 mM supplementation group than the other groups at the 2 h timepoint after thawing. Conclusion: Supplementation of glutamine at a concentration of 20 mM in a short-term semen extender (Bio Pig®) during the 17 °C holding time before cryopreservation, which had a standard freezing extender (9.0% glycerol and 1.9% Equex paste), could enhance the post-thaw sperm motility and quality parameters of cryopreservation. [ABSTRACT FROM AUTHOR]

Male infertility has become an important issue of global concern. Semen analysis is the cornerstone of male fertility assessment. External quality assessment (EQA) of sperm concentration, motility, and morphology is widely recognized in the world. However, over the past 34 years, the implementation of EQA for semen analysis has varied across different countries, and there is no global consensus. The goal of this paper is to first explore the overall development of EQA during this period. Secondly, it aims to discuss the extent of difference of participating laboratories in different countries. Finally, the paper examines the differences in EQA programs developed by various EQA providers in order to seek a global standard. In total, 29 papers met the inclusion criteria and were included in this review. There is inconsistent in the implementation of EQA across different countries, and there is no global consensus. Policies for EQA of semen analysis vary from country to country. Some countries mandate laboratory participation, while others permit voluntary involvement. Different EQA organizers choose different ways to calculate assigned value and acceptance limits. The coefficient of variation (CV) for each EQA item was large. The CVs of concentration, motility, morphology, and viability were 12.7–138.0 %, 17.0–127.0 %, 7–375 %, and 6–41.1 %, respectively. The results of the semen analysis varied considerably among the participating laboratories. The collaborative efforts of national policymakers, EQA organizers, and all participating laboratories are essential to improving the current situation. [ABSTRACT FROM AUTHOR]

Background: Due to global warming seasonal heat stress is an increasing problem in temperate zones. Heat stress not only decreases fertility in females, but can also be detrimental to male fertility. Objectives: We studied the effects of natural summer heat stress during spermatogenesis in Holstein bulls on semen quality parameters and on fertilization performance in vitro and possible intergenerational transmission of effects on the next male generation. Materials and Methods: Semen samples from young Holstein breeding bulls, referred to as F0 founders during this study, were collected during summer (F0 "summer" semen) and the following winter (F0 "winter" semen). Parameters such as ejaculate volume, sperm density, motility, thermoresistance, and in vitro blastocyst rates from these F0 semen samples were determined. In addition, after generation of offspring by artificial insemination, semen samples from F1 male offspring were collected and tested for the same quality and performance parameters to capture intergenerational effects. F1 bulls were raised together under identical conditions and semen was collected at about 1 year after birth. Results: The data showed that in vitro blastocyst rates of F0 "summer" semen samples were lower compared with "winter" semen, whereas blastocyst rates of F1 semen samples did not show significant differences. However, whereas F0 semen samples did not indicate significantly different quality parameters we found that motility of F1 semen samples showed significant differences with higher values when collected from bulls generated with F0 "winter" semen. Discussion and Conclusion: From our data, we conclude that (i) natural summer heat stress during spermatogenesis can affect in vitro fertility parameters and (ii) the observed effects on sperm motility of F1 semen samples suggest intergenerational paternal transmission. [ABSTRACT FROM AUTHOR]

Background: Carriers of reciprocal translocations often have more unbalanced spermatozoa and higher DNA fragmentation rates, elevating reproductive risk. The simple swim‐up method (SSUM) can decrease the amount of spermatozoa with abnormal chromatin structure and fragmented DNA, however, it has limited efficacy in eliminating chromosomally unbalanced sperm. Methods: The spermatozoa of eight Robertsonian translocation (Rob) carriers were split into three groups: original raw semen group (control group); SSUM and swimming trapper method group (STM) processed semen samples. After different semen preparation procedures, semen qualities, sperm chromosomal aneuploidy, and sperm fragmented DNA were evaluated. Results: Although spermatozoa with higher motility was obtained by both SSUM and STM, the population of faster forward moving sperm was greater with STM as compared to SSUM. While the rates of DNA fragmentation were statistically much lower in both groups than ejaculated semen sample, our data showed better effect on the decrease of DNA fragmentation index (DFI) after selection by STM for patients who have high DFI (>20%) in neat semen. For all patients, significant decrease in the frequency of chromosomally unbalanced spermatozoa was observed after selection using STM. Although similar trends can be seen in the SSUM group, a significant difference was identified in one patient only. Conclusions: Use of swimming trapper (STM) is superior for enriching high‐motile and genetically competent sperm in comparison with SSUM. [ABSTRACT FROM AUTHOR]

Background: Socioeconomic differences are present within the population of men who experience infertility and seek treatment. Objective: To study the association of socioeconomic status with semen parameters in a group of men using mail‐in semen analyses. Materials and methods: The records of 11,134 men that used mail‐in semen analyses from a fertility company were identified. Their demographic information, semen parameters, and ZIP codes were collected. Area deprivation index (ADI) was used as a proxy for socioeconomic status and was calculated for each individual using their ZIP codes in order to measure their level of socioeconomic deprivation. A higher ADI signifies a more deprived area. The association between ADI and the semen parameters of this group was measured using linear regression analysis adjusted for age. Results: 11,134 men were included in the study with a median age of 35 years (interquartile range (IQR): 32–40) and a median ADI of 83 (IQR: 68–97). The cohort had a median sperm concentration of 31 million/mL (IQR: 14–59), median total sperm count of 123 million (IQR: 57–224), median total motile sperm of 35 million (IQR: 9–95), median total motility of 32% (IQR: 15–52), progressive motility of 22% (IQR: 9–38), and morphology percent normal of 4% (IQR: 2–7). Higher ADI, indicating lower socioeconomic status, was negatively associated with various semen parameters, including sperm concentration, total sperm count, total motile sperm, and total and progressive motility. Discussion and conclusion: Men who live in more deprived areas are more likely to have worse semen parameters. Further research is needed to thoroughly examine the impact of socioeconomic status on male fertility. A comprehensive approach that targets upstream social, economic, and healthcare factors can possibly alleviate the negative association of socioeconomic status with fertility and semen parameters. [ABSTRACT FROM AUTHOR]

Background: Severe acute syndrome coronavirus 2 can invade a variety of tissues, including the testis. Even though this virus is scarcely found in human semen polymerase chain reaction tests, autopsy studies confirm the viral presence in all testicular cell types, including spermatozoa and spermatids. Objective: To investigate whether the severe acute syndrome coronavirus 2 is present inside the spermatozoa of negative polymerase chain reaction‐infected men up to 3 months after hospital discharge. Materials and methods: This cross‐sectional study included 13 confirmed moderate‐to‐severe COVID‐19 patients enrolled 30–90 days after the diagnosis. Semen samples were obtained and examined with real‐time polymerase chain reaction for RNA detection and by transmission electron microscopy. Results: In moderate‐to‐severe clinical scenarios, we identified the severe acute syndrome coronavirus 2 inside spermatozoa in nine of 13 patients up to 90 days after discharge from the hospital. Moreover, some DNA‐based extracellular traps were reported in all studied specimens. Discussion and conclusion: Although severe acute syndrome coronavirus 2 was not present in the infected men's semen, it was intracellularly present in the spermatozoa till 3 months after hospital discharge. The Electron microscopy (EM) findings also suggest that spermatozoa produce nuclear DNA‐based extracellular traps, probably in a cell‐free DNA‐dependent manner, similar to those previously described in the systemic inflammatory response to COVID‐19. In moderate‐to‐severe cases, the blood–testes barrier grants little defence against different pathogenic viruses, including the severe acute syndrome coronavirus 2. The virus could also use the epididymis as a post‐testicular route to bind and fuse to the mature spermatozoon and possibly accomplish the reverse transcription of the single‐stranded viral RNA into proviral DNA. These mechanisms can elicit extracellular cell‐free DNA formation. The potential implications of our findings for assisted conception must be addressed, and the evolutionary history of DNA‐based extracellular traps as preserved ammunition in animals' innate defence might improve our understanding of the severe acute syndrome coronavirus 2 pathophysiology in the testis and spermatozoa. [ABSTRACT FROM AUTHOR]

Background: Dietary factors, including high sugar intake, may have adverse effects on male reproduction. Studies of the association between sugar‐sweetened beverage (SSB) intake and semen quality have reported inconsistent results. Objective: We estimated the effects of SSB consumption on semen quality in a North American preconception cohort study. Methods: We analyzed baseline data from 690 males (n = 1,247 samples) participating in Pregnancy Study Online (PRESTO) during 2015–2022. Participants aged ≥21 years completed a baseline questionnaire on which they reported information about intake of SSBs, including sodas, energy drinks, sports drinks, and fruit juices. After enrollment, we invited U.S. participants to a semen testing substudy, in which they collected and analyzed two samples using an at‐home semen testing kit. We used linear regression models to estimate adjusted percent differences (%D) and 95% confidence intervals (CI) for associations of SSB intake with semen volume, sperm concentration, total sperm count (TSC), motility, and total motile sperm count (TMSC). We used modified Poisson regression models to estimate adjusted risk ratios (RRs) and 95% CIs for the association of SSB intake with World Health Organization semen parameter cut points. Results: Relative to non‐consumers of SSBs, those who consumed ≥7 SSBs/week had lower semen volume (%D = −6, 95% CI: −13, 0), sperm concentration (%D = −22, 95% CI: −38, 0), TSC (%D = −22, 95% CI: −38, −2), motility (%D = −4, 95% CI: −10, 2), and TMSC (%D = −25, 95% CI: −43, −2). High SSB consumers also had greater risks of low sperm concentration (≤16 million/mL; RR = 1.89, 95% CI: 1.11, 3.21), low TSC (≤39 million; RR = 1.75, 95% CI: 0.92, 3.33), low motility (≤42%; RR = 1.23, 95% CI: 0.87, 1.75) and low TMSC (≤21 million; RR = 1.95, 95% CI: 1.12, 3.38). Associations were stronger among participants with body mass index ≥ 25 kg/m2. Conclusion: Greater SSB consumption was associated with reduced semen quality in a North American preconception cohort. [ABSTRACT FROM AUTHOR]

Background: Clinical palpable varicocoeles in conjunction with isolated teratozoospermia are rarely observed. Therefore, the effects of varicocoelectomy on sperm morphology are not clear. The aim of this meta‐analysis is to compile studies that assess the effectiveness of varicocoelectomy in isolated teratozoospermia to reach a more consistent and reliable conclusion. Material‐Methods: The present meta‐analysis was registered to PROSPERO (CRD42023467933). We utilized the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta‐Analyses) guideline to report the outcomes. Articles published before 1 October 2023 were included in the study. The search terms used were teratozoospermia, isolated teratozoospermia, varicocoelectomy for isolated teratozoospermia, and semen analysis after varicocoelectomy in isolated teratozoospermia. Results: We identified 1,013 studies in full publications or abstracts using the methodology and the search terms. Five studies were included for systematic review, while four were included for meta‐analysis. The five studies (10–14) included 348 patients aged 18–44 years. The pooled analysis revealed a significant improvement in sperm morphology in isolated teratozoospermia patients undergoing varicocoelectomy (Q = 199.42, p < 0.0001; I2 = 98.49%). The pooled analysis revealed no significant improvement in sperm concentration in isolated teratozoospermia patients undergoing varicocoelectomy (Q = 5.69, p = 0.058; I2 = 64.85%). Three of the examined studies provided information regarding pregnancy rate and it was high in all studies. According to the Newcastle–Ottowa scale (NOS) assessment, the total quality score of all studies was 7. The funnel plot test demonstrated a visible asymmetry, and Begg and Mazumdar's rank correlation test confirmed the publication bias (p = 0.04). Discussion: Varicocoelectomy can be an effective and reliable treatment option in patients with isolated morphology abnormalities and clinically palpable varicocoele. Conclusion: This meta‐analysis reported that varicocoelectomy may increase pregnancy rates by improving semen parameters in infertile men with isolated teratozoospermia, although this conclusion requires further evidence. [ABSTRACT FROM AUTHOR]

The purpose of the study is to explore the impact of dietary habits, including daily intake of nutrients and dietary patterns, on the semen characteristics of South-Asian men. This cross-sectional study included 162 South-Asian men aged 21–40 years who were undergoing infertility evaluation. Based on semen analysis, men were grouped into categories of normal and abnormal semen characteristics. The participants' dietary habits were captured using '24-hour dietary recall' and 'Food Frequency Questionnaire (FFQ)'. Daily dietary intake of various nutrients and adherence to prevalent dietary patterns were compared between men with normal and abnormal semen characteristics. Six dietary patterns were identified following factor analysis of FFQ data. Men with normal semen characteristics showed significantly higher consumption of "coastal diet" rich in seafood, fish, cereals, beans, and coconut oil than those with abnormal semen characteristics. Further, the multivariable-adjusted model, adjusted for clinically relevant confounders, like age, body mass index (BMI), physical activity, smoking and sexual abstinence, also showed a lower risk of abnormal semen characteristics among men consuming "coastal diet" (odds ratio [OR] = 0.56, 95% confidence interval [CI] 0.37–0.82). In terms of daily nutrient intake, men with normal semen characteristics showed significantly lower consumption of monounsaturated fatty acids (MUFA) compared to men with abnormal semen characteristics (p = 0.008). Our findings suggest that higher consumption of "coastal pattern diet" lowers the risk of abnormal semen characteristics. Additionally, a negative association of MUFA consumption with semen quality was also identified, which needs cautious interpretation until replicated in future studies with biochemical validations. [ABSTRACT FROM AUTHOR]

Around 7% of the male population in the world are entangle with considerable situation which is known as male infertility. Photobiomodulation therapy (PBMT) is the application of low-level laser radiation, that recently used to increase or promote the various cell functions including, proliferation, differentiation, ATP production, gene expressions, regulation of reactive oxygen spices (ROS), and also boost the tissue healing and reduction of inflammation. This systematic review's main idea is a comprehensive appraisal of the literatures on subjects of PBMT consequences in four light ranges wavelength (blue, green, red, near-infrared (NIR)) on sperm cell characteristics, in vitro and in vivo. In this study, PubMed, Google Scholar, and Scopus databases were used for abstracts and full-text scientific papers published from 2003–2023 that reported the application of PBM on sperm cells. Criteria's for inclusion and exclusion to review were applied. Finally, the studies that matched with our goals were included, classified, and reported in detail. Also, searched studies were subdivided into the effects of four ranges of light irradiation, including the blue light range (400–500 nm), green light range (500–600 nm), red light range (600–780 nm), and NIR light range (780–3000 nm) of laser irradiation on human or animal sperm cells, in situations of in vitro or in vivo. Searches with our keywords results in 137 papers. After primary analysis, some articles were excluded because they were review articles or incomplete and unrelated studies. Finally, we use the 63 articles for this systematic review. Our category tables were based on the light range of irradiation, source of sperm cells (human or animal cells) and being in vitro or in vivo. Six% of publications reported the effects of blue, 10% green, 53% red and 31% NIR, light on sperm cell. In general, most of these studies showed that PBMT exerted a positive effect on the sperm cell motility. The various effects of PBMT in different wavelength ranges, as mentioned in this review, provide more insights for its potential applications in improving sperm characteristics. PBMT as a treatment method has significant effectiveness for treatment of different medical problems. Due to the lack of reporting data in this field, there is a need for future studies to assessment the biochemical and molecular effects of PBMT on sperm cells for the possible application of this treatment to the human sperm cells before the ART process. [ABSTRACT FROM AUTHOR]

In this study, high‐performance liquid chromatography‐evaporative light scattering detection (HPLC‐ELSD) and HPLC‐mass spectrometry (HPLC‐MS) were employed for the quantitative analysis of steroidal saponins of Semen Allii Fistulosi. The HPLC‐ELSD method was simple, accurate, and repeatable, and the recoveries were 97.91%∼99.77%. Similarly, the HPLC‐MS method was rapid, sensitive, accurate, and reproducible, and the recoveries ranging from 95.61% to 99.70% in the control samples. The sensitivity of the HPLC‐MS method was several times higher than that of the HPLC‐ELSD method. These results collectively demonstrated that these methods are suitable for the content determination of steroidal saponins of Semen Allii Fistulosi and the identification of its plant source. [ABSTRACT FROM AUTHOR]

This study aimed to assess the ten-year trend in semen quality among couples referred to the Infertility Center in Kerman between 2008 and 2017. The study included 2952 semen samples from men 18 to 60 years old referred to the infertility center as infertile couples living in Kerman province, Iran, whether they had normal or abnormal semen analysis. A total of 2952 sperm samples were included. Statistically significant changes were observed in semen parameters. Particularly, significant changes were observed for volume (−0.08 mL/year), sperm concentration (−2.34 (mio/mL)/year), total sperm count (−13.17 (mio/ejaculate)/year), progressive motility (−2.62%/year), non-progressive motility (−0.59%/year), immotile sperm (2.49%/year), and normal morphology (−0.134%/year). In bivariate analysis, the prevalence of oligozoospermia in this study showed a statistically significant association with age (OR = 1.019; 95% CI = 1.007−1.032; p = 0.003). Likewise, there was a statistically significant association with the year (OR = 1.087; 95% CI = 1.050−1.125; p = 0.000). Semen quality parameters showed a downtrend during the last 10 years in this study, emphasizing the importance of male reproductive health monitoring and warning public health coordinators to pay more attention to this important issue. [ABSTRACT FROM AUTHOR]

The study investigated selenium’s (Se) efficiency in preserving South African Zulu rams’ sperm quality and testicular parameters when they were exposed to heat stress. Indigenous Zulu rams (20) between 2 and 5 years old were allocated into four groups, namely the Se, testicular heat stress (THS), selenium plus testicular heat stress (SeTHS), and control. Each group comprised five rams; the groups were balanced according to the rams’ body weight and scrotal circumference. The Se and SeTHS groups received sodium selenite orally bi-weekly for 5 months. To induce heat stress, testicular heat insulation bags were wrapped around the testes of the rams receiving the THS and SeTHS treatments for 49 days. Semen was collected from the rams weekly from the third month onward; the first two months were for Se & THS acclimatization. In addition, testicular measurements were taken bi-weekly. Analysis of variance was used to analyze the sperm quality data. Duncan’s multiple range test was used to compare the groups’ data for significant differences. The results showed that the Se-supplemented rams’ scrotal circumference was smaller (p < 0.05) compared with the other groups. The Se, SeTHS, and control groups demonstrated similar total sperm motility; in contrast, the THS and SeTHS groups recorded low and high total sperm motility, respectively, compared with other treatment groups (p < 0.05). The semen from the rams that received THS without Se displayed a significantly higher number of immotile sperm cells (p < 0.05) and poor sperm quality, including total and progressive motility, and kinematic parameters when compared with other treatments, suggesting that Se protects sperm against THS. We concluded that selenium protected some sperm parameters (TSM, PSM, MV, VCL, VSL) of THS- treated rams while others did not improve (RV, NSM, C, STR). [ABSTRACT FROM AUTHOR]

Introduction: Triclosan is an antibacterial and antifungal compound that is frequently found in personal care and consumer products, and its its impact on male reproductive health is a growing concern. Despite existing experimental studies demonstrating its potential threats to male fertility, reports on its effects on human semen quality remains limited and inconsistent. Therefore, this study presents a systematic review and meta-analysis assessing the relationship between triclosan exposure and semen quality. Methods: This study was registered with PROSPERO (CRD42024524192) and adhered to PRISMA guidelines. Results: The study analyzed 562 screened studies, out of which five articles including 1,312 male subjects were finally included in the study. The eligible studies were geographically diverse, with three from China, one from Belgium, and one from Poland. More so, the eligible studies were both case-control and cross-sectional. The meta-analysis revealed that triclosan exposure significantly reduced sperm concentration (Standard Mean Difference (SMD) -0.42 [95% CI: -0.75, -0.10], P = 0.01) and sperm total motility (SMD -1.30 [95% CI: -2.26, -0.34], P = 0.008). Mechanistic insights from animal and in vitro studies showed that oxidative stress may mediate the adverse effects of triclosan on semen quality. Discussion: This meta-analysis is the first comprehensive evaluation of the impact of triclosan on human semen quality, highlighting its potential to impair male fertility through reductions in sperm concentration and motility. However, the high heterogeneity among the included studies underscores the need for further high-quality research to establish more definitive conclusions regarding the effects of triclosan exposure on human reproductive health. [ABSTRACT FROM AUTHOR]

Background: Ejaculatory abstinence (EA) duration influences semen parameters. However, the impact of penultimate ejaculatory abstinence (PEA) on conventional and functional sperm parameters remains underexplored. Method: A cross-sectional study recruited 1,503 men from a reproductive center between November 2023 and July 2024. Each participant underwent a physical examination, completed clinical questionnaires, and provided a semen sample for analysis. Generalized linear models were adjusted for potential confounders such as EA to investigate the association between PEA and various sperm parameters. Logistic regression was used to evaluate the relationship between PEA and the risk of high sperm DNA fragmentation index (DFI), oligozoospermia, asthenozoospermia, and necrozoospermia. Results: Participants were categorized into four quartiles based on PEA duration. (Q1: 1-3 days; Q2: 4-5 days; Q3: 6-9 days; Q4: > 9 days). After adjusting for potential confounders, a significant positive linear association was found between PEA and DFI, while a significant negative linear association was observed with progressive sperm motility. The longest PEA duration (Q4) correlated positively with semen concentration (P = 0.025), total sperm count (P < 0.001), and sperm vitality (P < 0.001). Compared to Q1, a PEA of > 9 days (Q4) was associated with higher risks of sperm DFI > 30% (adjusted odds ratio [OR] = 4.25; 95% confidence interval [CI]: 2.37-7.62), asthenozoospermia (adjusted OR = 1.45; 95% CI: 1.07-1.96), and necrozoospermia (adjustedOR = 1.99; 95% CI: 1.07-3.69). Moreover, the risk of sperm DFI > 15% was higher in Q2, Q3, and Q4 compared to Q1. Conclusion: Prolonged PEA adversely affects sperm DFI, progressive motility, and sperm vitality, increasing the likelihood of asthenozoospermia, necrozoospermia, and elevated DFI levels. These findings suggest that both EA and PEA should be considered in fertility assessments, with shorter PEA durations potentially yielding higher quality sperm, thereby enhancing male fertility evaluation and outcomes. [ABSTRACT FROM AUTHOR]

Background: Mycoplasma bovis is a global pathogen of cattle but was detected for the first time in New Zealand in 2017, triggering a response under their Biosecurity Act as an "unwanted organism". Following a lengthy eradication campaign, the Ministry of Primary Industries (MPI) now requires all bovine semen destined for export to New Zealand to be screened with an M. bovis-specific real-time PCR (rtPCR) compliant with amended import health standard (IHS) test requirements aimed at preventing the accidental importation of M. bovis. The standard stipulates that semen samples cannot be centrifuged prior to DNA extraction. To comply with these strict requirements, one of the listed tests was validated together with different DNA preparation steps and compared with existing in-house procedures. DNA was extracted from semen straws using the current in-house semi-automated platform procedures for processing culture, tissue and body fluid sample submissions and was compared with the stipulated test requirements. DNA from centrifuged and unspun semen samples spiked with M. bovis was also compared. Results: The rtPCR had a sensitivity and specificity of 100% (95% confidence interval = 79–100% and 74–100%, respectively) when testing DNA from other Mycoplasma species or bovine semen spiked with the latter, with a high level of repeatability for within- and between- run replicates. The consistent limit of detection was 0.001 pg/µl M. bovis DNA and between 5.3 × 102 and 7.5 × 102 CFU/ml M. bovis when artificially spiked in semen. DNA extracted using the KingFisher Flex was detected with lower Cq values than the Maxwell 16, but the comparable improvements in sensitivity were mainly associated with non-centrifuged samples (p < 0.001). None of the procedures tested impeded the detection sensitivity of M. bovis in the presence of competitor organisms Acholeplasma laidlawii, Mycoplasma bovigenitalium and Ureaplasma diversum, confirming M. bovis specificity of the polC target. Conclusions: Under the experimental conditions applied, this rtPCR test efficiently detected M. bovis in extended bovine semen straw samples from DNA extracted using both semi-automated extraction platforms, irrespective of prior centrifugation of extended semen. [ABSTRACT FROM AUTHOR]

Microfluidics provides unique opportunities for the high throughput selection of motile sperm with improved DNA integrity for assisted reproductive technologies (ARTs). Here, through a parametric study on dimensions and geometrical angles, a butterfly-shaped chip (BSC) is presented to isolate sperm with high progressive motility and intact DNA at a separation rate of 1125 sperm per minute. Using finite element simulations, the flow field and shear rates in the device were optimized to leverage the inherent motility characteristics of sperm for maximum selection throughput. The device incorporates a triple selection mechanism in series, initially activating sperm rheotaxis by rotation against the semen flow, penetrating the counter buffer flow and swimming against the direction of the buffer flow, leaving dead cells and debris behind, and subsequently leveraging boundary-following behavior to direct progressively motile sperm to swim along the walls and reach the device outlet. The device selects over 4.1 million sperm per mL within 20 minutes, with 29.2%, 68.2%, and 57.3% improvement in total motility, DNA integrity, and velocity parameter (VCL), as compared with the conventional swim-up method, respectively. Overall, the performance of the device to separate sperm with approximately 95.9% total motility, 97.8% viability, and 96.6% DNA integrity at high concentrations demonstrates its potential for enhancing the efficiency of conventional treatment methods. [ABSTRACT FROM AUTHOR]

Our research focuses on semen collection from 42 African penguin males, grouped by age, habituation levels, and reproductive season timing. We assess behavioral and physiological responses to dorso-abdominal massage, evaluating species-specific seminal traits using conventional and advanced methods. Positive behavioral responses corresponded with successful semen collection. Young and human-reared males exhibited more favorable behaviors, and samples containing spermatozoa were more likely collected during the reproductive season. Age did not influence sample collectability or spermatozoa presence, though mature males produced higher semen volumes. Young males exhibited more sperm morphological abnormalities, especially outside the breeding season. Sperm concentration and number per ejaculate showed no significant differences across age or seasonal groups. Young males had higher motile and progressive sperm percentages, while mature males had more static sperm. Additionally, percentages of live sperm and intact acrosomes were higher in mature males. Young males and samples from non-reproductive season presented more dead sperm with damaged acrosomes. Plasma membrane integrity positively correlated with age for live sperm and negatively for dying sperm. This research demonstrates the feasibility of semen collection from African penguins regardless of habituation level, fills the knowledge gap by describing sperm morphological abnormalities, and provides insights into using flow cytometry in Sphenisciformes. [ABSTRACT FROM AUTHOR]

Background: The rate of male infertility has increased in the last decades. One factor that may improve sperm quality and testosterone is fish consumption, which supplies the organism with essential nutrients. Aim: To determine evidence of a relationship between the frequency of fish consumption and sperm quality. Methods: Seventy‐four healthy men, aged 25–45 years, were involved in this observational study. Semen quality was evaluated by spermogram analyses and testosterone was measured in blood. Volunteers were classified as low, moderate, and high fish consumption groups according to the replies indicated in a questionnaire they had to fill out. Results: The moderate fish consumption group showed better semen quality with all spermogram parameters in the reference range and the low fish consumption group manifested the worst semen quality with sperm morphology below the reference range. The low fish consumption group had lower testosterone levels (3.6 ± 0.9 vs. 4.3 ± 1.5; p = 0.04), decreased nonprogressive spermatozoa (11.9 ± 6.2 vs. 15.8 ± 8.2; p = 0.04), and lower semen volume (3.1 ± 2.0 vs. 3.6 ± 1.3; p = 0.06) compared to the moderate fish consumption group. Positive associations were detected between the frequency of fish consumption, the semen volume, and nonprogressive spermatozoa, respectively. Conclusions: Fish consumption at moderate level such as once a week leads to improvement in sperm quality and testosterone levels. [ABSTRACT FROM AUTHOR]

Theobromine (TBR) is a methylxanthine known for its bronchodilatory and stimulatory effects. This research evaluated the vitality, capacitation patterns, oxidative characteristics, microbial profile and expression of capacitation-associated proteins (CatSper1/2, sodium bicarbonate cotransporter [NBC], protein kinases A [PKA] and C [PKC] and adenylate cyclase 10 [ADCY10]) in cryopreserved bovine spermatozoa (n = 30) in the absence (cryopreserved control [CtrlC]) or presence of different TBR concentrations (12.5, 25, and 50 µM) in egg yolk extender. Fresh ejaculate served as a negative control (CtrlN). Significant post-thaw maintenance of the sperm motility, membrane and DNA integrity and mitochondrial activity (p < 0.001) were recorded following the administration of 25 μM and 50 μM TBR, then compared to CtrlC. All groups supplemented with TBR exhibited a significantly lower percentage of prematurely capacitated spermatozoa (p < 0.001) than CtrlC. Significantly decreased levels of global reactive oxygen species (ROS), hydrogen peroxide and hydroxyl radicals were observed in the presence of 25 μM and 50 μM TBR (p < 0.01). Western blot analysis revealed that supplementation with 50 μM TBR significantly prevented the loss of NBC and ADCY10 (p < 0.01), while all TBR doses stabilized the levels of PKC (p < 0.05 at 50 μM TBR; p < 0.001 at 12.5 μM and 25 μM TBR). In summary, we suggest that TBR is effective in protecting the spermatozoa during the cryopreservation process through its potential to stimulate energy synthesis while preventing ROS overproduction and the loss of proteins involved in the sperm activation process. [ABSTRACT FROM AUTHOR]

Simple Summary: The standardization of the poultry semen cryopreservation technique is necessary for the conservation of genetic resources; however, during this process, a series of cellular lesions are generated as a consequence of the increase in reactive oxygen species (ROS). The objective of the present study is to determine which antioxidants are most commonly used in roosters to mitigate this effect after their addition to the cryopreservation diluent by studying different characteristics of seminal quality. A discriminant canonical analysis (DCA) was performed, and the variables hypo-osmotic swelling test (HOST), viability, and total motility were reported to be the ones that provided the most information, as they were the most discriminant variables between groups. According to the descriptive statistics, antioxidants catalase and serine were the ones that gave the best results in most of the variables studied. In conclusion, given that catalase had the most beneficial effects on the seminal quality parameters that proved to have the greatest discriminating power, this antioxidant could be the one of choice. The standardization of the semen cryopreservation technique could be an effective tool in poultry for the conservation of genetic resources. During this process, the production of reactive oxygen species increases, leading to oxidative stress that causes damage to the spermatozoa. To reduce this effect, the addition of exogenous antioxidants in the cryopreservation diluent has been reported to be effective. Multiple antioxidants such as catalase, vitamin E, cysteamine, ergothioneine, and serine have been studied in roosters. Therefore, the present investigation aims to perform a meta-analysis to determine if the use of the aforementioned antioxidants added to the cryopreservation extender produces an improvement in semen quality parameters in roosters after thawing. After collecting the data, a discriminant canonical analysis was performed to determine which of the selected semen quality traits provided the most information, with hypo-osmotic swelling test (HOST), viability, and total motility variables showing the highest discriminatory power. However, according to the descriptive statistics, catalase and serine are the antioxidants that improve a greater number of seminal quality parameters, and since catalase gives the most favorable results for most of the discriminating variables, it could be the antioxidant of choice. [ABSTRACT FROM AUTHOR]

Male infertility is one of the most common reproductive dysfunctions. Despite oligospermia being a cause of infertility, few studies have been conducted on it. This study aimed to investigate differences in semen metabolic patterns in patients with oligospermia and to identify potential biomarkers associated with oligospermia. Semen samples from oligospermia patients (20 cases) and healthy controls (20 cases) were detected by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), and 72 and 89 metabolites were identified as potential markers in positive and negative ion modes, respectively. In addition, the results identified multiple metabolic pathways in patients with oligospermia, such as glycine serine and threonine metabolism, Synthesis and degradation of ketone bodies, Valine, leucine, and isoleucine degradation. These results described unique metabolic characteristics of semen in patients with oligospermia and provided novel insights into the mechanism of the semen disorder. [ABSTRACT FROM AUTHOR]

The rising incidence rate of depression presents a substantial threat to human well‐being. Semen Trigonellae (ST), the dried mature seeds of Trigonella foenum‐graecum L., has a long‐standing traditional reputation for alleviating anxiety and hopelessness. However, the anti‐depressant mechanism of ST remains poorly understood. This study aimed to assess ST's anti‐depressant, as well as explore its potential mechanism from a gut microbial aspect. The Kunming mice were challenged by lipopolysaccharide (LPS) to induce depression‐like behavior and then orally administrated with aqueous extract of ST. The behavioral test, and hippocampal and serum biochemical indicators were detected to assess anti‐depressant effect of ST. We utilized full‐length 16S rRNA gene sequencing to investigate how ST influences gut microbiota modulation. Administration of ST mitigated LPS‐induced depression‐like behaviors. ST reversed the decrease in hippocampal 5‐hydroxytryptamine (5‐HT) and brain‐derived neurotrophic factor (BDNF) levels while reducing serum levels of tumor necrosis factor α (TNF‐α), interleukin 1β (IL‐1β), and interleukin 6 (IL‐6). Microbial analysis revealed that administration of ST markedly shifted the gut microbiota structure, dramatically and dose‐dependently increased the abundance of Ligilactobacillus murinus and Ligilactobacillus animalis. Experimentally, oral administration of live L. murinus and L. animalis to LPS‐challenged mice yielded similar effects to ST in ameliorating depression, elevating 5‐HT and BDNF, and reducing proinflammatory cytokines. These findings provide evidence that ST is a promising medical food for the management of depression, in which modulation of the gut microbiota, particularly enhancing the abundance of Ligilactobacillus plays an important role. [ABSTRACT FROM AUTHOR]

Biological fluid stains can be instrumental in solving crimes. Identification of semen can help reconstruct events in sexual assault cases and identify suspects via DNA profiling. Current methods for semen identification suffer from limitations, including destruction of the sample and potential false positives. One of the main unsolved issues is the elimination of underlying substrate interference. In this paper, chemometric approaches were developed to isolate and identify a biofluid stain on interfering substrates using Raman spectroscopy. The first approach, called Multivariate Curve Resolution with the Addition Method, combines the standard addition method with multivariate curve resolution. The second one uses a criterion based on reducing the spectrum complexity when a spectral component is removed from a Raman spectrum of a multi-component sample entirely. The results demonstrate the superiority of the first approach relative to the second for both small volume fraction of the fluid stain compared to the substrate and random noise. [ABSTRACT FROM AUTHOR]

Purpose: Chromosomal abnormalities play an important role in male infertility, which is becoming a significant issue in human fertility. Aim of this study was to evaluate the incidence of spermatic aneuploidies and diploidies in human sperm, according to semen parameters. Methods: We performed semen analysis according to the 6th edition of WHO criteria in 50 male subjects; samples were divided into normozoospermic (n = 23) or those with altered seminal parameters (n = 27). To assess chromosomal numerical alterations of sperm, fluorescence in situ hybridization (FISH) was used. Result: A significant increase in aneuploidies and diploidies was observed in samples with altered seminal parameters. Furthermore, stratifying this group, we observed a significant increase in aneuploidies and total abnormalities in oligozoospermic, asthenoteratozoospermic (AT), and oligoteratoasthenozoospermic (OAT) samples compared to normozoospermic. Conclusion: Our results showed the correlation between altered seminal parameters and numerical chromosomal abnormalities, confirming that sperm FISH analysis could be an additional clinical tool to assess reproductive potential in infertile males. Moreover, our results point to the importance of updating the normality ranges for detecting chromosomal aneuploidies using FISH. [ABSTRACT FROM AUTHOR]

The present experiment was carried out to investigate the role of Oxyrase in preserving the in vitro quality, redox status and in vivo fertility of crossbred boar spermatozoa. A total of 24 ejaculates from 6 crossbred (n = 4 from each boar) boars were collected and extended in Beltsville Thawing Solution (BTS) in 1:2 ratio and divided into three aliquots. The first aliquot served as a control (without Oxyrase). Rest of the two aliquots were supplemented with 0.125 (T1) and 0.25 IU/mL Oxyrase (T2). Semen samples were preserved at 15°C for 5 days and kinematics of spermatozoa by CASA, semen quality parameters and oxidative stress status were evaluated at 0, 72 and 120 h of storage. The findings of studies revealed that supplementation of Oxyrase at 0.25 IU/mL resulted in higher (p < 0.05) total motility, progressive motility, plasma membrane integrity, acrosome integrity and functional integrity of plasma membrane at 72 and 120 h in comparison to the control group. Mitochondrial membrane potential (MMP) was higher (p < 0.05) at 72 and 120 h, whereas higher (p < 0.05) DNA integrity was observed at 120 h in T2. The lipid peroxidation (LPO) was lower (p < 0.05) and superoxide dismutase activity (SOD) and total antioxidant capacity (TAC) were higher (p < 0.05) in the T2 group at 120 h as compared to control. In vivo fertility trials indicated a higher (p < 0.05) litter size in T2 in comparison to other groups. The study concluded that the inclusion of Oxyrase at 0.25 IU/mL in the extender protects the crossbred boar spermatozoa against oxidative damage and improves the in vivo fertility. [ABSTRACT FROM AUTHOR]

Premature acrosomal exocytosis in cryopreserved semen is one of the reasons attributed to low fertility among livestock. In the present study, we attempted to enhance the cryopreserved semen quality by selective removal of acrosome‐reacted spermatozoa using FITC‐PNA conjugated iron magnetic nanoparticles (MNPs). Further, the effect of nano purification on other sperm functional attributes was also assessed. Iron MNPs were prepared using co‐precipitation method and dextran‐coated MNPs were conjugated with FITC‐PNA (0.04 mg/mL). A preliminary experiment was conducted to standardise the dose of FITC‐PNA conjugated iron MNPs (0.02, 0.05, 0.1, 0.15, 0.2, 0.4 and 0.6 mg). Among the different doses used, 0.6 mg FITC‐PNA conjugated iron MNPs significantly (p < 0.05) removed higher acrosomal reacted spermatozoa from the semen, and therefore, this dose was used in further experiments. Cryopreserved semen from Holstein Friesian breeding bulls (n = 6) were thawed and washed using Sperm‐TALP to remove residual extender. Washed spermatozoa (2 × 106) were exposed to 0.6 mg of FITC‐PNA conjugated iron MNPs for 10 min at 37°C. The nano purified semen was assessed for various vital sperm parameters viz., viability, intracellular calcium, apoptosis, mitochondrial ROS and mitochondrial membrane potential using flow cytometry. We found that nanopurification using FITC‐PNA conjugated iron MNPs significantly (p < 0.05) improved the sperm quality. The proportion of viable non‐apoptotic spermatozoa with low intracellular calcium levels was significantly (p < 0.05) enriched in nano purified semen. Nano purification did not affect sperm mitochondrial membrane potential and ROS production. In conclusion, these preliminary findings indicate that FITC‐PNA coated iron MNPs effectively removed acrosome reacted spermatozoa and significantly improved sperm functional attributes in the purified fraction. [ABSTRACT FROM AUTHOR]

The main goals of this study were to document and compare the normal ranges of testicular haemodynamic parameters in pre‐ (aged 8–12 months; n = 4) and postpubertal (aged 24–259 months; n = 16) Miranda donkeys in the breeding season, and to correlate animal biometric data and testicular Doppler indices with basic semen quality parameters of sexually mature jacks. Colour and pulsed‐Doppler ultrasonography were employed to assess blood flow in the left and right distal supratesticular artery (DsTA) and their marginal branches (marginal arteries—MA). Peak systolic velocity (PSV), end diastolic velocity (EDV), pulsatility index (PI) and resistive index (RI) were evaluated in both blood vessels, and TAMV (time‐averaged mean velocity), TABF (total arterial blood flow) and TABF rate (TABF‐R) were calculated for MA. The mean diameter of MA was greater (p < 0.05; 0.24 ± 0.05 vs. 0.19 ± 0.05 cm; mean ± SD) but TABF‐R was less (p < 0.05; 0.004 ± 0.004 vs. 0.02 ± 0.01 mL/s/cm3) in sexually mature donkeys compared with prepubertal animals. Apart from RI values for the right testicle of prepubertal donkeys, PI and RI were consistently greater (p < 0.05) in DsTA compared with MA. Significant correlations were found among select biometric and haemodynamic attributes of the testes (height, width and length, TV, TTV and PSV‐ST) and ejaculate characteristics (volume, sperm defects‐total, head and midpiece) in sexually mature donkeys (n = 8). The present results highlight the importance of scrotal ultrasonography for the reproductive assessment of jacks and provide reference values, based on the available subpopulation of Miranda donkeys that can be used in their clinical and reproductive management and research, or conservation programmes. [ABSTRACT FROM AUTHOR]

Sperm cryopreservation in small ruminant is an efficient strategy to distribute spermatozoa for reproductive programmes, but this process reduces the fertility potential of frozen–thawed spermatozoa. The aim of the current research was to evaluate the impact of different concentrations of cysteamine (CYS) in soybean lecithin (SL)‐based medium on postthawed buck semen quality and fertility potential. Semen samples were collected from five bucks, twice a week, then diluted in the SL‐based extender containing different concentrations of CYS as follows: extender containing 0 mM (control, C0), 1 mM (C1), 2 mM (C2), 4 mM (C4) and 8 mM (C8) CYS. Motility characteristics, membrane integrity, abnormal morphology, mitochondrial activity, acrosome integrity, viability, apoptotic‐like changes, lipid peroxidation, DNA fragmentation, ROS concentration, pregnancy rate and kidding rate were evaluated after freeze–thaw process. In results, C1 resulted in greater (p ≤ 0.05) total motility, progressive motility, average path velocity, membrane integrity, mitochondrial activity, acrosome integrity, viability, pregnancy rate and kidding rate compared to the other groups. Furthermore, supplementation of freezing medium with 1 mM of CYS presented lower (p ≤ 0.05) apoptotic‐like changes, lipid peroxidation, DNA fragmentation and ROS concentration compared to the other groups. On the other hand, C8 presented the least (p ≤ 0.05) total motility, progressive motility, average path velocity, membrane integrity, mitochondrial activity, acrosome integrity and viability as well as the highest (p ≤ 0.05) apoptotic‐like changes, lipid peroxidation, DNA fragmentation and ROS concentration compared to the other groups. Therefore, supplementation of freezing medium with 1 mM CYS could be a helpful strategy to protect buck's spermatozoa quality and fertility potential during cryopreservation process. [ABSTRACT FROM AUTHOR]

Seminal extracellular vesicles (SemEVs) are repositories of biomolecules, including metabolites involved in the regulation of sperm function. The correlation between the metabolite profile of SemEVs and semen parameters, along with their role in regulating sperm function, is an unexplored area. This preliminary study evaluated the metabolomic content of SemEVs. Semen samples were obtained from 18 healthy men, and SemEVs were extracted from seminal plasma using the size exclusion chromatography qEV Gen 2–35 nm column coupled with an automatic fraction collector. The physical characterization of SemEVs was carried out with the ZetaView PMX-430-Z QUATT laser system. EV protein markers were detected using Western blot. In addition, these SemEVs were used for metabolomic profiling and functional bioinformatic analysis. The mean concentration of isolated SemEVs was 1.7 ± 1.1 × 1011/mL of seminal plasma, whereas SemEVs size and zeta potential were 129.5 ± 5.5 nm and −40.03 ± 3.99 mV, respectively. Western blot analysis confirmed the presence of EV specific markers such as CD81, ALIX, and TSG101. A total of 107 metabolites were identified using this untargeted metabolomic approach in SemEVs. Bioinformatics analysis further revealed that metabolites associated with tyrosine metabolism were highly enriched in these SemEVs. Ingenuity Pathway Analysis (IPA) also indicated that these metabolites present in SemEVs were involved in the regulation of the free radical scavenging pathway. Furthermore, our metabolomic results suggest that these SemEV-associated metabolites may play a pivotal role in the maintenance of seminal plasma redox homeostasis. [ABSTRACT FROM AUTHOR]

Simple Summary: Artificial insemination (AI) is a reproductive technique routinely used in livestock to deliver sperm directly to the cervix or uterus to achieve pregnancy. In swine, AI is performed with chill-stored sperm displaying minimum criteria evaluated using traditional procedures; however, variegated fertility appeals for additional tools for sperm evaluation. To enhance sperm evaluation, we assess the potential of near-infrared (NIR) and Raman spectroscopy in addition to traditional techniques to monitor boar sperm quality during 10-day chill-storage. Sperm samples showed differential responses to storage, with sperm quality nearly maintained in some samples, whereas it sharply deteriorated in others. Better than NIR, Raman spectral profiles obtained from freshly extended samples efficiently predict sperm survival to storage, making the technique a promising tool for in-depth assessment of sperm samples. Artificial insemination (AI) plays a critical role in livestock reproduction, with semen quality being essential. In swine, AI primarily uses cool-stored semen adhering to industry standards assessed through routine analysis, yet fertility inconsistencies highlight the need for enhanced semen evaluation. Over 10-day storage at 17 °C, boar semen samples were analyzed for motility, morphology, sperm membrane integrity, apoptosis, and oxidative stress indicators. Additionally, machine learning tools were employed to explore the potential of Raman and near-infrared (NIR) spectroscopy in enhancing semen sample evaluation. Sperm motility and morphology gradually decreased during storage, with distinct groups categorized as "Good" or "Poor" survival semen according to motility on Day 7 of storage. Initially similar on Day 0 of semen collection, "Poor" samples revealed significantly lower total motility (21.69 ± 4.64% vs. 80.19 ± 1.42%), progressive motility (4.74 ± 1.71% vs. 39.73 ± 2.57%), and normal morphology (66.43 ± 2.60% vs. 87.91 ± 1.92%) than their "Good" counterparts by Day 7, using a computer-assisted sperm analyzer. Furthermore, "Poor" samples had higher levels of apoptotic cells, membrane damage, and intracellular reactive oxygen species on Day 0. Conversely, "Good" samples maintained higher total antioxidant capacity. Raman spectroscopy outperformed NIR, providing distinctive spectral profiles aligned with semen biochemical changes and enabling the prediction of semen survival during storage. Overall, the spectral profiles coupled with machine learning tools might assist in enhancing semen evaluation and prognosis. [ABSTRACT FROM AUTHOR]

A rapid decline in wild animal biodiversity, as well as in the number of local livestock breeds of lower economical values, makes species and breeds protection and creation of genetic reserves a necessity and a challenge of the 21st century. For birds, semen freezing is still the best method of gene preservation ex situ in vitro. The aim of the presented study was to assess the susceptibility to freezing process and to establish the semen gene bank of twelve goose breeds covered by the Polish genetic resources conservation program. Sexually mature ganders were randomly selected from four northern local breeds: Kartuska, Pomorska, Rypinska, Suwalska, four southern local breeds: Garbonosa, Kielecka, Lubelska, Podkarpacka, and four foreign goose breeds: Kuban, Landes, Roman and Slovakian. Each breed was represented by 12 males, twelve repetitions were carried out. Semen samples were frozen in a computer-controlled cryogenic chamber using a method developed for White Koluda goose. The basic evaluation criterion was sperm morphology in fresh and freeze-thawed semen assessed in nigrosine-eosin stained histological smears. The percentage of live in total and live normal sperm in the fresh semen differed significantly (P<0.05) between breeds. Semen of Kartuska and Suwalska ganders (northern breeds), contained the highest percent of live sperm – 94.4 and 94.1 respectively, including 52.5% and 54.2% of live normal sperm, while the lowest number of these forms was observed for Slovakian goose – 86.4% and 29.2%, respectively. The freezing process resulted in a significant (P<0.05) reduction in the total number of live and live normal sperm and an increase in dead sperm in all groups. Cryopreservation process was the most effective in case of Suwalska goose semen (28.4% of live normal cells in thawed semen) and the less profitable for Landes (7.4% of live normal cells). The analyses showed significant differences between breeds in sperm morphology in fresh and thawed semen. Moreover, the high proportion of live spermatozoa in the fresh semen did not guarantee a high proportion of spermatozoa after thawing, indicating the differences in semen susceptibility to cryopreservation process depending on goose breed. [ABSTRACT FROM AUTHOR]

Background: Transgender and non-binary individuals face unique challenges when it comes to fertility preservation (FP). Objective: Despite the growing prevalence of gender dysphoria (GD) and gender transitioning, there is a lack of clear guidelines and consensus on the management of these patients in the FP setting. Clinicians and institutions providing FP services should ensure that they are aware of the needs and circumstances of this underrepresented group of patients and offer them accurate and evidence-based information when counseling and tailoring their FP treatment. Materials and methods: For this scoping review, three major search engines were used. Including Embase, Epistemonikos, Google Scholar, MEDLINE and PubMed. Sources of grey literature were also explored (ResearchGate and Web of Science). The combination of only two keywords [transgender] AND [fertility preservation] was used up to May 2023. Results: The available evidence on clinical management and FP outcomes in transgender patients is limited and mainly originates from case reports or small case series. The main limitation of current FP services for transgender and non-binary individuals is the lack of scientific evidence regarding their care. Discussion: Overall, FP in transgender patients requires individualized and realistic plans, and psychological counseling should be offered. This review aims to provide the latest evidence coming from original studies to facilitate proper counseling and fertility management for these individuals. Conclusions: Inclusive health systems that provide comprehensive reproductive health care to transgender individuals can help them make informed decisions about FP and improve their quality of life. Future research is needed to establish more robust evidence-based guidelines for the management of transgender and non-binary individuals in the FP setting. [ABSTRACT FROM AUTHOR]

Purpose: The impact of air pollution on semen quality has been confirmed, yet the joint effect remains unclear. We evaluate the individual and joint associations of particulate (PM2.5 and PM10) and gaseous pollutants (NO2, SO2, O3 and CO) with semen quality. Methods: We included 5,114 men in this study from 2014 to 2022. The individual and joint associations were measured by multiple linear regression models. Results: Sperm motility and semen volume were inversely associated with pollutant concentrations during every stage of sperm development, especially at lag days 0–9 and 10–14 (all P < 0.05). Stratified analyses showed that the study pollutants (except CO) had a positive effect on semen concentration during the stage of sperm development, especially in spring and autumn, while a decreased total sperm number was associated with CO (all P < 0.05). However, joint associations of particulate and gaseous pollutants with semen quality parameters were not statistically significant (all P > 0.05). Conclusions: During all stages of sperm development, particulate and gaseous pollutants had individual negative impacts on sperm motility and semen volume, and these impacts were less pronounced in spring and autumn. Our findings highlight the importance and necessity of reducing the exposure to pollutants especially in the critical stage of sperm development to improve semen quality. [ABSTRACT FROM AUTHOR]

Simple Summary: Improving the genetic traits influencing rooster semen quality and quantity is crucial for optimizing poultry production. Breeders can identify heritable traits that significantly influence semen characteristics by evaluating genetic parameters, ensuring that only the best roosters are selected for breeding purposes. The development of appropriate genetic models aids in accurately predicting these traits, thereby enhancing selection efficiency. Selection based on these models boosts fertility and hatchability and promotes the propagation of desirable traits across generations. This approach leads to healthier, more productive flocks, increased economic efficiency, and more sustainable poultry farming practices. Improving reproductive traits, particularly semen quality and quantity, is crucial for optimizing poultry production and addressing the current limitations in native chicken reproduction. The aim of this study was to develop a genetic model to estimate genetic parameters guiding the selection of individual Thai native roosters. Using data collected from 3475 records of 242 Thai native grandparent roosters aged 1–4 years, we evaluated semen traits (mass movement, semen volume, and sperm concentration) over 54 weeks. A random regression test–day model incorporating five covariance functions, including a linear spline function with four, five, six, and eight knots (SP4, SP5, SP6, and SP8) and second-order Legendre polynomial function (LG2), was used to estimate genetic parameters. The results showed that the SP8 model consistently outperformed the other models across all traits, with the lowest mean square error, highest coefficient of determination, and superior predictive ability. Heritability estimates for mass movement, semen volume, and sperm concentration ranged from 0.10 to 0.25, 0.22 to 0.25, and 0.11 to 0.24, respectively, indicating moderate genetic influence on these traits. Genetic correlations between semen volume and sperm concentration were highest in the SP8 model, highlighting a strong genetic association between these traits. The SP8 model also revealed a high genetic correlation between mass movement and semen volume, supporting the potential for selecting mass movement as a predictor of semen volume. In conclusion, this study highlights the effectiveness of random regression models with linear spline functions to evaluate the genetic parameters of semen traits in native Thai roosters. The SP8 model is a robust tool for breeders to enhance the reproductive performance of native Thai chickens, contributing to sustainable poultry production systems. [ABSTRACT FROM AUTHOR]

Semen preservation quality affects the artificial insemination success rate, and seminal exosomes are rich in various proteins that are transferable to sperm and conducive to sperm-function preservation during storage. However, the specific effects of these proteins remain unclear. In this study, the specific effects of these proteins on semen preservation quality and fertilization capacity were investigated through a proteomic analysis of seminal exosomes from boars with high conception rates (HCRs) and low conception rates (LCRs). The results revealed significant differences in the expression of 161 proteins between the two groups, with the GPX5 level being significantly higher in the HCR group (p < 0.05). The role of GPX5 was further investigated by constructing engineered exosomes enriched with GPX5 (Exo-GPX5), which could successfully transfer GPX5 to sperm. Compared to the control group, Exo-GPX5 could significantly improve sperm motility on storage days 4 and 5 and enhance the acrosome integrity on day 5 (p < 0.05). Additionally, Exo-GPX5 increased the total antioxidant capacity (T-AOC) of sperm, reduced the malondialdehyde (MDA) level, and decreased the expression of antioxidant proteins SOD1 and CAT (p < 0.05). In simulated fertilization experiments, Exo-GPX5-treated sperm exhibited higher capacitation ability and a significant increase in the acrosome reaction rate (p < 0.05). Overall, Exo-GPX5 can improve boar semen quality under 17 °C storage conditions and enhance sperm fertilization capacity. [ABSTRACT FROM AUTHOR]

In this study, we evaluated sheep sperm quality after using Tris‐citrate‐fructose‐based extender with and without egg yolk, a Tris‐citrate without fructose and with egg yolk and the commercial extender Biladyl®, preserving diluted semen at 15 and 23°C for different times (4, 24, 48 and 72 h). The results showed that the diluents with fructose and egg yolk gave the best results of seminal quality. Moreover, the production of ROS was higher for the temperature of 23°C compared to the temperature of 15°C (control). In addition, VCL and the percentage of spermatozoa with intact acrosome decreased with temperatures of 23°C. Finally, a drastic decrease in sperm quality was observed after 24 hours of preservation for most of the parameters evaluated. [ABSTRACT FROM AUTHOR]

The aim of this experiment was to assess the effect of media viscosity on ram sperm motility, kinematics and rheotaxis in vitro by using methylcellulose as a media thickener. Frozen–thawed semen of three rams was thawed and diluted in Tyrode's albumin lactate pyruvate (TALP) media supplemented with 0%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6% and 0.7% w/v of methylcellulose. Sperm motility and kinematic characteristics were analysed using computer‐assisted sperm analysis (CASA). The rheotactic behaviour was assessed in a microfluidic channel, and the number of spermatozoa that passed the 10 mm point of a microfluidic channel over a 2min period against a flow rate of 30 μm/sec was assessed. The use of media with higher viscosity (higher levels of methylcellulose) resulted in significantly lower (p <.05) sperm motility and kinematic parameters. Moreover, higher levels of methylcellulose reduced (p <.05) the number of spermatozoa that exhibited positive rheotaxis. In conclusion, viscosity affected the kinematic properties and rheotactic behaviour of ram sperm. [ABSTRACT FROM AUTHOR]

The present study compares two protocols for the cryopreservation of chicken semen. Both protocols had an initial low cooling rate in the first step, followed by higher cooling rates around ice nucleation (Protocol 1) or following the dissipation of the latent heat of fusion (Protocol 2) in the second step. Semen ejaculates obtained from 12 roosters were diluted with Rootex with 6% dimethylformamide and frozen following either Protocol 1 (from +5°C to −10°C at 5°C/min and from −10°C to −130°C at 60°C/min) or Protocol 2 (from +5°C to −35°C at 7°C/min and from −35°C to −140°C at 60°C/min). Compared with fresh semen, following both protocols, cryopreservation resulted in reduced post‐thaw sperm quality (p <.001). Post‐thaw percentage of sperm with an intact plasma membrane was greater using Protocol 2 than Protocol 1 (p <.05). The results suggest that high cooling rates around the time of ice nucleation are not recommendable. [ABSTRACT FROM AUTHOR]

Goat production is affected by reproductive seasonality. In vitro embryo production (IVEP) could overcome this effect. This study aimed to evaluate the impact of the season of semen collection/freezing on IVEP of prepubertal goat oocytes and on sperm quality and functionality concerning capacitation. Semen from six fertile bucks was collected, pooled and cryopreserved in spring and autumn and used for IVEP of oocytes recovered during the breeding season. Oocytes were IVM in TCM‐199 with hormones, EGF and cysteamine; fertilized and cultured in BO‐IVF and BO‐IVC media (IVF Bioscience, UK). Semen samples were assessed at 0 and 3 h after culture in capacitating (BO‐IVF, CAP) and non‐capacitating conditions for sperm plasma membrane and acrosome integrity, mitochondrial membrane potential (MMP), intracellular calcium and plasma membrane lipid disorder. Blastocyst production was higher with spring sperm compared to autumn (12.0% vs. 2.1%, respectively; p <.05). After CAP, acrosome reaction and intracellular calcium were higher (p <.05) in spring than autumn sperm. No differences were found in other sperm parameters. In conclusion, seasonal variations in the IVEP of prepubertal goats could be linked to differences in sperm ability to undergo in vitro capacitation. [ABSTRACT FROM AUTHOR]

A protocol for conventional in vitro fertilization (IVF) in horses using fresh semen has been described, using a prolonged incubation in FERT‐TALP medium (22 h) at 38.2°C in the presence of penicillamine, hypotaurine and epinephrine (PHE). Our work aimed to develop a protocol that maintains quality parameters in frozen–thawed equine spermatozoa incubated for 22 h in the presence of PHE using different media (FERT‐TALP and INRA96) and incubation temperatures (30 and 38.2°C). Twelve frozen ejaculates from four stallions were thawed and then incubated in either FERT‐TALP or INRA96 with PHE at 30 or 38.2°C for 22 h. Following incubation, total motility (TM), progressive motility (PM), viability and acrosome integrity were evaluated. The results showed that TM was significantly higher (p <.001) at 30°C in both media, while PM was higher for INRA96 at 30°C compared to 38°C (p <.05). Moreover, INRA96 at 30°C exhibited higher sperm viability and acrosome integrity (p <.001) compared to the other experimental groups. These preliminary results suggest that incubating thawed equine spermatozoa at 30°C with PHE in INRA96 successfully maintains motility, viability and acrosome integrity in equine spermatozoa, indicating its potential use for conventional equine IVF. [ABSTRACT FROM AUTHOR]