Associations of Lifestyle Factors with Cadmium and Nickel in Seminal Fluid of Potential Sperm Donors.

Spermatogenesis is worsened by reactive oxygen species (ROS) and antioxidants could reduce ROS induced sperm damage. This prospective cohort study evaluated the potential therapeutic effects of a combination of vitamins, minerals and antioxidants on the sperm quality parameters of infertile men with idiopathic low spermatogenesis. Seminal fluid analysis tests were performed before treatment, 3 and 6 months after treatment. The treatment resulted in a significant improvement in the rate (%) of sperm motility from 16.95±6.93 to 23.11±8.87, after 3 months and reached 23.68±8.73 after 6 months (p=0.0006) whereas a non-significant increase in sperm count (from 13.05±8.1 to 15.79±7.9 after 3 months and 15.26±10.3 million/ml after 6 months (p=0.1650). Morphology and agglutination showed little changes. A positive correlation between sperm count and motility was observed after 3 months of treatment (r = 0.594; p=0.007). The combination of vitamins and antioxidants improved the sperm motility significantly and to a lesser extent the sperm count, however, sperm morphology and agglutination remained relatively unchanged. Using antioxidants is safe and can improve semen parameters.

Exosomal ncRNAs in seminal fluid: Unraveling their regulatory roles in male infertility.

The impact of extracellular vesicles on spermatozoa: Navigating toward fertilization.

Male fertility investigation is currently limited to semen analysis. However, the origins of abnormal sperm parameters are not well-understood, and normal sperm do not assure fertility in men. Improved pathophysiological and prognostic insight might be achieved utilising additional measures of male reproductive tract function. Cytokine and chemokine levels in seminal plasma (SP) may be relevant, but evidence on their clinical significance is unclear. The utility of measuring SP cytokines remains uncertain, and no consensus exists on which cytokines are most informative. To perform a systematic review and meta-analysis on the association between fertility status and concentration of seminal plasma cytokines in men. The sixth edition of the WHO laboratory manual for the examination and processing of human semen raises the prospect of evaluating cytokines in SP as part of an extended examination. We performed a systematic search and meta-analysis to assess whether the current literature is sufficient to identify cytokines present in human SP that exhibit a relationship with fertility status in men. We searched PubMed, Web of Science, Scopus, and Embase from inception until April 2025, using keywords pertaining to seminal fluid and cytokines, restricted to humans and the English language. Original data with values reported as concentration of cytokines in SP of men clearly defined as infertile, compared to a discernible population of fertile/normozoospermic healthy control men, were included. A total of 5737 studies were identified, with 2737 duplicates removed. Title and abstract screening were performed for 3000 studies, then 291 studies underwent full-text screening, and 68 studies progressed to quality assessment using the NHLBI-NIH quality assessment tool and 52 studies underwent data extraction. We identified 52 research articles published from 1993 to 2025 that quantified at least one cytokine in the seminal plasma of 8153 men, after 19 studies of poor quality and/or containing serious flaws were excluded. Data on 30 cytokines in the SP of healthy control and infertile men were extracted and included in narrative synthesis. Compared to fertile controls, infertile men had elevated concentrations of IL6 (SMD 0.39, 95% CI; 0.14-0.64, I2 = 80.7%), TNFA (SMD 0.13, 95% CI; 0.00-0.25, I2 = 4.3%), and CXCL8 (SMD 0.24, 95% CI; 0.06-0.43, I2 = 0.0%) in seminal plasma. IL6 reported a high degree of heterogeneity between studies, whilst CXCL8 and TNFA reported low heterogeneity. No significant moderator effects due to study quality or composition of the control cohort were identified. With one in six couples experiencing infertility worldwide and a male factor identified as a primary or contributing cause in up to 50% of cases, there is a strong imperative to develop a better understanding of the pathophysiology of male infertility. This analysis identifies an association between infertility and elevated SP IL6, CXCL8, and TNFA, but whether these pro-inflammatory cytokines reflect events affecting fertility or are simply markers of transient inflammation and/or cofactors of fertility status remains to be determined. Greater precision in quantifying seminal plasma cytokines will be attained, and additional informative cytokines may be identified, by utilising standardised technical approaches in future studies. CRD42023398438.

Comparison of seminal fluid analysis parameters before and after COVID-19 infection

Seminal fluid proteins as regulation factors for optimizing reproduction: a modeling approach

In the male body, zinc accumulates most abundantly in prostatic cells, where it plays a key role in producing high amounts of citrate in seminal fluid. Intraprostatic accumulation of Zn increases during the development of benign prostatic hyperplasia (BPH), one of the most common diseases in men over 50 years of age. Continuing our investigations on intraprostatic androgens, in this study, we analyzed the mineral content (Zn, Ca, Cu, K, Mg, Mn, and Na) in the transitional zone (TZ) of the prostate using inductively coupled plasma optical emission spectrometry (ICP-OES). The concentrations of testosterone (T) and dihydrotestosterone (DHT) were determined by liquid chromatography-mass spectrometry (LC-MS). Group-wise and correlation analyses demonstrated a descriptive trend toward a volume-dependent increase in Zn concentrations within TZ tissue, whereas other elements exhibited heterogeneous covariance patterns; intraprostatic hormone levels, although elevated in larger prostates, showed no consistent linear correlations with elemental concentrations. Given the observational design of the present study, the reported tissue Zn profiles cannot be interpreted as evidence supporting supplementation in BPH, and any potential clinical implications warrant evaluation in rigorously designed interventional studies.

Presence of sperm-bound antisperm antibodies and their association with semen quality in dogs.

Prostate-specific antigen (PSA) or gamma-selenoprotein or kallikrein-3 (KLK3) is a glycoprotein enzyme secreted by the epithelial cells of the prostate glands. It plays a crucial role in male fertility and is commonly used as a marker of prostate cancer. Antibodies to PSA anti-gen might play a role in male immune infertility. To date, the tests available in the market pro-vide information only about the presence or absence of these antibodies in body fluids, which is further confirmed by the Western blot test. There are no tests available to quantify the amount of anti-PSA antibodies in human body fluid. Hence, the present patent relates to in vitro immuno-assay for detecting and quantifying anti-prostate specific antigen (anti-PSA) antibodies in a hu-man body fluid sample. In particular, the immunoassay is an indirect ELISA. The assay has demonstrated high sensitivity and specificity, capable of detecting anti-PSA antibodies in body fluids within a range of 4.61 ng/mL to 431.37 ng/mL. To further validate the assay's specificity, additional experiments have been conducted using various samples, including chicken seminal fluid, and the serum and semen of azoospermic individuals. These samples, standardized to the same volume, have been incubated with a fixed amount of human PSA-coated antigen. Similar experiments have been performed with anti-human, anti-rabbit, anti-mouse, anti-horse, and anti-goat antibodies, further confirming the assay's specificity.

Prostate cancer (PCa) remains a major global health burden, yet current screening tools often lead to overdiagnosis due to low specificity, highlighting the urgent need for more precise diagnostic approaches. Prostatic fluid (PSF) represents a promising but underexplored biofluid with exceptional diagnostic potential due to its direct contact with the PCa microenvironment. Here, we employed molecule-level interpretable surface-enhanced Raman spectroscopy (SERS) to comprehensively investigate PCa-associated alterations in two PSF components including metabolites and small extracellular vesicles (sEVs) and explored their potential interrelations via correlation analysis. Through molecule-resolvable SERS spectral set (MORE SERSome) technique, we identified ergothioneine and deoxyguanosine as differential metabolites between PCa and benign prostatic hyperplasia patients. We further constructed a fusion diagnostic model by integrating metabolites and sEVs information. The fusion model significantly outperformed the diagnostic accuracy by applying any single component, suggesting diagnostic complementarity between PSF metabolites and sEVs. Integration with clinical variables such as age and plasma prostate-specific antigen concentration further enhanced performance with the area under the curve as high as 0.93 for PCa diagnosis, substantially surpassing existing screening methods. These findings strengthen the importance of in-depth analysis of specific PSF components and further promise the potential of SERS-based PSF profiling as a noninvasive strategy for PCa diagnosis and biopsy guidance.

Semen from mature, healthy bulls contains commensal microbes. Potential internal sources of seminal microbes have not been extensively evaluated. Objectives were to 1) assess whether the testes of the bull contain commensal microbes and to compare the composition of the putative testicular microbiome to that of semen and rumen fluid and 2) determine whether other organs of the bull reproductive system contain microbial populations. Here, we demonstrate that the testes of the bull contain a low biomass, yet diverse, microbiome. Interestingly, the microbial composition of rumen fluid, semen and testicular tissue were each dissimilar from one another, indicating that each source contains a unique microbiota. Only four core amplicon sequence variants, Acinetobacter, Enterobacteriaceae (E. coli), Jeotgalicoccus, and Kiritimatiellae WCHB1-41, were shared between semen and the testes. Along with the testes, microbial populations were also present within the penile and pelvic urethra, seminal vesicles and epididymis of mature bulls. Of these anatomic niches, the microbial populations within the penile and pelvic urethra were the most diverse and shared the greatest number of core taxa (n = 72). The microbiota of the seminal vesicles, epididymis and the testes were significantly dissimilar from each other. Only one core taxa, Mycoplasma, was shared between the testes and epididymis. Collectively, our results demonstrate that semen and organs of the bull reproductive tract contain unique microbial populations. Further research is necessary to determine whether the microbial composition of organs of the bull reproductive system, such as the testes and epididymis, influence sperm viability and bull fertility.

Sperm undergo a last step of maturation known as capacitation in the female reproductive system, which prepares themselves to fertilize the ovum. Mechanisms of capacitation include changes in membrane composition, electrolyte concentration, pH levels, cellular and molecular pathways, signaling cascades, sperm protein phosphorylation, and sperm-specific channel activity. Capacitation is a strictly regulated process since it must occur at the appropriate time and place. In this study, Google Scholar, Web of Science Core Collection, PubMed/MEDLINE, and Scopus were used to search and retrieve data. The 2 main consequences of capacitation are modifications to the membrane composition and changes in sperm motility type. Decapacitation factors, which are primarily located in seminal fluid, should prevent ejaculated sperm from undergoing early capacitation. The activities of decapacitation factors diminish as seminal fluid is removed and sperm enter the female genital system, whereas the capacitation factors cause sperm to become capacitated. The proteins that either stimulate or hinder the process of capacitation have been investigated in this study, with a focus on expressing their role in human capacitation as accurately as possible. The role of suitable proteins for human assisted reproductive technology and in vitro fertilization, would likely be better understood if studies could concentrate on the human route after investigating the capacitation process in various animals.

The relevance of studying the microbiome of bull semen is due to the high importance of reproduction in animal husbandry, the problems of semen contamination with pathogens, and the need to improve the efficiency of artificial insemination. This review provides an overview of the taxonomic groups of microorganisms present in the seminal fluid of bulls. The review highlights the reasons for the limited research on the microbiota of bull semen and focuses on the advantages of modern molecular technologies, such as 16S rRNA sequencing, which allow for a more accurate determination of the microbial spectrum and their role in the reproductive system. The microbiota is diverse, including the phyla Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria , and Fusobacteria , which are likely to have a variety of effects on seed quality and fertility. It has been proven that despite the established influence of the reproductive system microbiome on fertility and sperm quality, which has been confirmed by studies on humans and other species of productive animals, there are currently no specific scientific publications that examine in detail the role of the seminal microbiome in bulls and its impact on relevant reproductive indicators. This review explores the external factors that influence the composition of the microbiome, such as feeding, season, and animal management practices. It was concluded that the use of probiotics in the feeding of bulls is a promising and environmentally friendly way to optimize the microbiota and improve reproductive function, increasing the quality of semen and fertility and reducing the need for antibiotics.

In the forensic examination of sexual assault cases, the Acid Phosphatase (AP) test is commonly used as a rapid presumptive screening method, while the PSA immunoassay is applied for confirmatory purposes due to its superior sensitivity. Immunochromatographic PSA (p30) tests are capable of detecting minute quantities of seminal material and often produce weak or trace positive results at dilutions where the AP test has already become negative. Because AP relies on an enzyme-based color reaction, its visible response diminishes at higher dilutions, limiting its practical detection threshold. PSA, a glycoprotein secreted by the epithelial cells of the prostate and present in high concentrations in seminal plasma, serves as a dependable biomarker for semen identification, particularly in cases where spermatozoa are absent due to azoospermia, vasectomy, or sample degradation. Comparative findings from both tests indicate that the PSA assay exhibits markedly higher sensitivity, detecting seminal fluid at greater dilutions (up to 1:2048 or more), whereas AP activity typically declines and becomes undetectable around 1:512–1:1024.Overall, while both AP and PSA tests are useful for identifying seminal stains, the PSA test offers a more sensitive and reliable means of detection at extreme dilutions where AP reactions become weak or inconclusive. Consequently, in rape investigations, PSA testing plays a critical role by enabling the sensitive detection of seminal fluid, reinforcing biological evidence, corroborating victim statements, facilitating DNA analysis, and providing robust scientific support within the criminal justice system and courts of law.

Physical and sexual violence remains among the most prevalent crimes in contemporary society, with women and children comprising the majority of victims. In such cases, biological evidence—such as blood, semen, and saliva—plays a critical role in the investigation and resolution of criminal incidents. However, this evidence may be compromised, either unintentionally due to environmental exposure or intentionally by the perpetrator or victim, with the aim of concealing the crime. This study investigates the potential for DNA recovery from blood and semen stains that have been deliberately exposed to diluted sodium hypochlorite (bleach), a common household chemical used in attempts to destroy forensic evidence. Biological stains were deposited on various fabric types and subsequently subjected to washing with bleach and detergent at 90°C. The samples were then examined using ultraviolet (UV) light and luminol for preliminary detection, followed by DNA isolation and quantification procedures. The findings demonstrate that DNA recovery is possible even after exposure to bleach, thereby affirming the viability of forensic analysis under compromised conditions. This study underscores the potential for retrieving genetic material in scenarios where biological evidence has been subjected to intentional destruction, thereby contributing valuable insights to forensic science and criminal investigations.

Declining reproductive health and fertility are global public health issues affecting an estimated 15% of reproductive-aged couples worldwide. The reasons for declining fertility are complex. However, a male contribution is thought to occur in ∼50% of infertile couples. Deficits in sperm number and/or function are undeniably a major cause of infertility, but compelling evidence suggests that additional factors in the male ejaculate also play an influential and underappreciated role. In this review, we focus specifically on extracellular vesicles within human seminal plasma and explore their emerging roles in reproduction and fertility. These seminal extracellular vesicles (SEVs) are nano-sized membrane structures secreted by various cell lineages in virtually all regions of the male reproductive tract and exert key roles in intercellular communication. Consideration is given to the well-characterized effects of SEVs in supporting sperm as they transit through the female reproductive tract and their ability to modulate the immune environment within the female reproductive tract. Building on these important roles, we also detail the emerging links between dysregulated SEV production and male fertility status, and highlight the potential utility of leveraging these vesicles to improve fertility and reproductive outcomes in infertile couples. Altogether, this review highlights how expanding knowledge of SEVs provides a new perspective on the complexity of seminal fluid physiology and the underlying aetiology of male infertility.

The extracellular calcium concentration is important for sperm function, maturation, and survival, however, studies exploring the link between seminal calcium and reproductive function have reported inconsistent findings. To investigate the association between seminal calcium levels and semen quality in both healthy and infertile men. A total of 301 young men from the general Danish population who participated in a study of semen quality, and 228 infertile men who were supplemented with calcium (500mg) + cholecalciferol (300,000 IU bolus followed by 1400 IU daily), or placebo in a randomized controlled trial were included. Each man underwent a physical examination, routine semen analysis, analysis of calcium concentrations in the seminal fluid, and blood sampling for assessment of PTH, testosterone, estradiol, inhibin B, and AMH. In young men from the general population, sperm concentration was higher in those within the highest seminal calcium quartile (Q4: 9.0-19.2mmol/L) compared with the lowest (Q1: 1.3-4.7mmol/L; 53.0 million/mL vs. 31.0 million/mL, p = 0.010). Accordingly, infertile men in Q1 had the lowest sperm concentration (e.g., 6.6 vs. 22.5 million/mL in Q3), but also fewer progressively motile spermatozoa (p = 0.030). Supplementation with calcium and cholecalciferol did not affect seminal calcium concentration (p = 0.33), however, infertile men who experienced an increase in their seminal calcium concentrations > 30% at the end of intervention had a higher sperm concentration (p = 0.048) and total sperm count (p < 0.001) compared with baseline. There was a weak positive correlation between longitudinal changes of seminal calcium and serum estradiol (r = 0.17, p = 0.016). Lower seminal calcium concentration is associated with poorer semen quality but was not influenced by calcium supplementation. Future research should focus on uncovering possible regulators, for instance, estrogens. ClinicalTrials.gov no. NCT01304927 (registered February 28, 2011).

The essential outcome of a successful mating is the transfer of genetic material from males to females in sexually reproducing animals from insects to mammals. In males, this culminates in ejaculation, a precisely timed sequence of organ contractions driven by the concerted activity of interneurons, sensory neurons, and motor neurons. Although central command circuits that trigger copulation have been mapped, the motor architecture and the chemical logic that couple specific neuronal subclasses to organ-specific contractility, seminal fluid secretion, and sperm emission remain largely uncharted. This gap in knowledge limits our ability to explain how neural circuits adapt to varying contexts and how their failure contributes to infertility. Here, we present an in-depth anatomical and functional analysis of the motor neurons that innervate the internal male reproductive tract of Drosophila melanogaster. We identify two classes of multi-transmitter motor neurons based on neurotransmitter usage, namely octopamine and glutamate neurons (OGNs) and serotonin and glutamate neurons (SGNs), each with a biased pattern of innervation: SGNs predominate in the accessory glands, OGNs in the ejaculatory duct, with equal contributions of each to the seminal vesicles. Both classes co-express vesicular transporters for glutamate (vGlut) and amines (vMAT), confirming their dual chemical identity. Their target organs differentially express receptors for glutamate, octopamine, and serotonin, suggesting combinatorial neuromodulation of contractility. Functional manipulations show that SGNs are essential for male fertility but OGNs are dispensable. Glutamatergic transmission from both classes is also dispensable for fertility. These findings provide the first high-resolution map linking multi-transmitter motor neurons to specific reproductive organs, reveal an unexpected division of labor between serotonergic and octopaminergic signaling pathways, and establish a framework for dissecting conserved neural principles that govern ejaculation and male fertility.

The essential outcome of a successful mating is the transfer of genetic material from males to females in sexually reproducing animals from insects to mammals. In males, this culminates in ejaculation, a precisely timed sequence of organ contractions driven by the concerted activity of interneurons, sensory neurons, and motor neurons. Although central command circuits that trigger copulation have been mapped, the motor architecture and the chemical logic that couple specific neuronal subclasses to organ-specific contractility, seminal fluid secretion, and sperm emission remain largely uncharted. This gap in knowledge limits our ability to explain how neural circuits adapt to varying contexts and how their failure contributes to infertility. Here, we present an in-depth anatomical and functional analysis of the motor neurons that innervate the internal male reproductive tract of Drosophila melanogaster. We identify two classes of multi-transmitter motor neurons based on neurotransmitter usage, namely octopamine and glutamate neurons (OGNs) and serotonin and glutamate neurons (SGNs), each with a biased pattern of innervation: SGNs predominate in the accessory glands, OGNs in the ejaculatory duct, with equal contributions of each to the seminal vesicles. Both classes co-express vesicular transporters for glutamate (vGlut) and amines (vMAT), confirming their dual chemical identity. Their target organs differentially express receptors for glutamate, octopamine, and serotonin, suggesting combinatorial neuromodulation of contractility. Functional manipulations show that SGNs are essential for male fertility but OGNs are dispensable. Glutamatergic transmission from both classes is also dispensable for fertility. These findings provide the first high-resolution map linking multi-transmitter motor neurons to specific reproductive organs, reveal an unexpected division of labor between serotonergic and octopaminergic signaling pathways, and establish a framework for dissecting conserved neural principles that govern ejaculation and male fertility.