Search Results (1,791)

Assisted reproductive technologies (ART), particularly in vitro fertilization (IVF), are characterized by substantial emotional burden, uncertainty, and repeated decision-making under probabilistic conditions. Although psychological distress is common among individuals and couples undergoing ART, psychosocial care remains inconsistently integrated into fertility pathways and is rarely tailored to individual needs. This narrative review synthesizes evidence concerning infertility-related distress, anxiety and depressive symptoms, couple functioning, treatment discontinuation, psychosocial screening, and psychological interventions across the IVF trajectory. Key phases of treatment, including pretreatment assessment, ovarian stimulation, embryo transfer, the waiting period, cycle outcome, and decisions regarding repetition or discontinuation, are described, and phase-specific psychological vulnerabilities are identified. On the basis of the available literature, we propose a conceptual stepped-care framework ranging from universal psychoeducation and routine monitoring to structured psychological interventions, couple-based care, and specialist psychiatric management. Practical screening thresholds, referral criteria, reassessment intervals, and a safety pathway for suicidal ideation are outlined. Particular attention is given to male-factor infertility, strategies for engaging male partners, and the ethical and data-governance requirements of digital mental health tools. Current evidence supports psychosocial interventions for improving emotional well-being and quality of life and suggests a potential role in treatment continuation, but their effects on pregnancy and live-birth outcomes remain uncertain. The proposed framework should therefore be regarded as a clinically informed conceptual model rather than a validated prediction algorithm. Prospective studies are required to assess its feasibility, predictive performance, cost-effectiveness, and impact on patient-centered outcomes. Full article

Chronic epididymitis and orchitis represent significant yet frequently under-recognized contributors to male infertility, particularly among men of reproductive age. These conditions arise from persistent inflammatory or immunological processes affecting the epididymis and testis, leading to impaired spermatogenesis, altered sperm maturation and possible obstruction of the male reproductive tract. Infectious aetiologies, especially those linked to sexually transmitted pathogens and uropathogens, remain predominant; however, non-infectious mechanisms, including autoimmune activation, post-vasectomy changes and idiopathic inflammation, also play critical roles. The persistent inflammatory milieu induces cytokine release, oxidative stress and structural tissue remodelling, ultimately compromising the functional and immune-privileged microenvironment necessary for optimal sperm production and transport. Diagnostic evaluation requires a multimodal approach incorporating clinical examination, microbiological testing, semen analysis and scrotal ultrasonography, with advanced imaging and molecular assays reserved for complex or equivocal cases. Management is individualized and may involve antimicrobial therapy, anti-inflammatory treatment, immunomodulation or microsurgical intervention in cases of ductal obstruction. Assisted reproductive technologies provide additional options when natural conception is not feasible. Despite increased recognition of their impact, chronic epididymitis and orchitis remain insufficiently studied, with gaps in standardized definitions, biomarker validation and long-term outcome data. This review provides a focused synthesis and phenotype-driven clinical framework for chronic epididymitis and orchitis through a fertility-preservation lens, bridging urological and andrological perspectives and integrating evidence on subclinical inflammation, contemporary diagnostic biomarkers and a staged therapeutic pathway. Full article

MYB transcription factors (TFs) play essential roles in diverse biological processes, including anther and pollen development, vegetative growth, seed development and germination, and stress responses. However, functional characterization of MYB TFs in maize (Zea mays) lags far behind that in Arabidopsis thaliana and Oryza sativa. In this study, we performed a genome-wide identification of 196 maize MYB TFs, along with phylogenetic analysis and Gene Ontology (GO) annotation. To bridge the knowledge gap, we established an integrated cross-species comparative workflow that systematically maps functionally characterized MYB TFs from Arabidopsis and rice to their maize orthologs. By coupling this homology-based approach with spatiotemporal expression profiling of developing anthers across multiple inbred lines, we prioritized candidate MYB TFs likely involved in anther and pollen development. This integrated strategy provides a useful reference for translating the rich functional knowledge accumulated in model plants to crops with less-characterized genomes. Our study not only establishes a solid foundation for the functional investigation of maize MYB TFs, but also offers promising targets for the mechanistic dissection and molecular breeding application of male sterility in maize. Full article

Background/Objectives: The Y chromosome plays a crucial role in male fertility, sex determination, and spermatogenesis, yet it remains poorly characterized in Mediterranean river buffalo (Bubalus bubalis, 2n = 50) because of its high repeat content, extensive heterochromatin, and complex palindromic structures. Although a chromosome-level Y assembly is available for swamp buffalo (2n = 48), no equivalent reference exists for the river type. Methods: To address this gap, Y chromosomes from 10 Mediterranean buffalo bulls were isolated by laser microdissection following peripheral blood culture and whole-chromosome amplification. Probe specificity was verified by FISH, and amplified Y chromosomes were sequenced using Illumina NovaSeq 6000 (Illumina, San Diego, CA, USA). Sequencing data were assembled and analysed through de novo assembly, repeat identification, sequence alignment, and variant detection. Comparative analyses included alignment to the swamp buffalo Y chromosome and annotation of Y-linked genes using the Bos taurus reference genome. Results: FISH confirmed the specificity of the isolated material, showing strong signals on the Y chromosome and on X/Y PAR and heterochromatic regions. Sequencing generated over 240 million paired-end reads, and de novo assembly produced 566,815 contigs. Repeat analysis identified 3.91% repetitive elements, mainly SINEs, while variant calling detected more than 23,000 variants. Comparative analyses mapped several contigs to the swamp buffalo Y chromosome and Y-linked genes. Annotation against the B. taurus genome identified 26 unique genes, including homologs shared with the X chromosome, and revealed MSY gene duplications, including 10 copies of TSPY and 3 of HSFY. Conclusions: These findings show that laser microdissection with NGS enables effective access to the buffalo Y chromosome, representing a milestone in the characterization of the river type genome, and providing a basis for studies on buffalo male fertility and breeding programs. Full article

Reactive oxygen species (ROS) and reactive nitrogen species (RNS) are critical modulators of male reproductive health, influencing sperm function, hormonal regulation, and overall fertility. While physiological levels of ROS and RNS are essential for processes such as sperm capacitation and acrosome reaction, their overproduction leads to oxidative and nitrosative stress, contributing to male infertility. Excessive ROS and RNS can damage sperm DNA, proteins, and lipids, impairing motility, viability, and fertilizing capacity. Moreover, these reactive species disrupt the hypothalamic-pituitary-gonadal (HPG) axis, leading to hormonal imbalances that further compromise reproductive function. Environmental factors, lifestyle choices, and underlying health conditions exacerbate the production of ROS and RNS, highlighting the need for preventive and therapeutic strategies. Clinically, ROS- and RNS-mediated redox imbalance has been implicated in several male reproductive disorders, including varicocele, genital tract infection and inflammation, obesity, diabetes and other metabolic disorders, and toxicant-related reproductive dysfunction. Antioxidant supplementation has shown promise in mitigating oxidative stress; however, its efficacy varies, and further research is necessary to establish standardized treatment protocols. These findings underscore the clinical relevance of integrating oxidative stress assessment with conventional semen analysis to improve risk stratification and guide targeted interventions in male infertility. This review synthesizes current knowledge on the molecular mechanisms by which ROS and RNS affect male reproduction and discusses potential clinical interventions to address oxidative and nitrosative stress in male infertility. Full article

Accelerating eutrophication of aquatic ecosystems worldwide has increased concern regarding cyanotoxin exposure as an emerging environmental and public health issue, with Microcystin-LR (MC-LR) among the most extensively studied congeners due to its widespread occurrence and high toxicity. Evidence from experimental animal and cellular studies indicates that MC-LR elicits pronounced toxic impacts on both the male and female reproductive systems. In males, MC-LR induces overt testicular injury, compromises the structural and functional integrity of the blood–testis barrier, and triggers severe disorders in reproductive hormone synthesis and secretion. In females, it precipitates ovarian dysfunction, impedes normal follicular maturation and development, and induces distinct embryotoxic effects. The underlying pathogenic mechanisms involve the synergistic interplay of multiple signaling pathways, primarily including oxidative stress induction, aberrant apoptosis activation, endocrine disruption, and epigenetic modifications. Of particular significance, emerging evidence suggests that parental exposure to MC-LR may induce intergenerational or potentially transgenerational reproductive effects through epigenetic modifications in germ cells, impairing fertility and developmental outcomes in subsequent offspring and thus posing a sustained, long-term threat to population-level health. This review systematically delineates the reproductive toxicity profiles and underlying molecular mechanisms of MC-LR, evaluates its transgenerational health hazards, and aims to furnish robust scientific evidence for the formulation of targeted environmental health policies and risk management strategies. Full article

Ring finger protein 126 (RNF126) is a RING-type E3 ubiquitin ligase that has recently emerged as a multifaceted regulator of cellular homeostasis, stress adaptation, and disease progression. Through its structurally distinct zinc-finger and catalytic RING domains, RNF126 orchestrates substrate recognition and ubiquitin transfer, generating diverse ubiquitin linkages with both proteolytic and nonproteolytic functions. Initially characterized as a component of the protein quality control (PQC) machinery, RNF126 cooperates with chaperones such as BAG6 and UBQLN1 to eliminate mislocalized and misfolded proteins, thereby maintaining proteostasis. Beyond PQC, RNF126 plays pivotal roles in DNA damage response pathways by regulating homologous recombination, non-homologous end joining, checkpoint signaling, and genome stability through substrates, including MRE11, Ku80, RNF168, and 14-3-3σ. Genetic studies have further demonstrated its importance in embryogenesis and male fertility, and accumulating evidence has identified RNF126 as a critical driver of malignancy in multiple cancers. RNF126 promotes tumor progression by degrading or modulating key regulators, such as p21, PTEN, p53, PDKs, and LKB1, thereby enhancing proliferation, metabolic reprogramming, anoikis resistance, metastasis, and chemo/radioresistance. Intriguingly, RNF126 exhibits context-dependent functions, acting as an oncogene or tumor suppressor depending on the tissue type and substrate selection. In addition to cancer, RNF126 has been implicated in neurodegeneration, cardiac pathology, antiviral immunity and adaptive immune regulation. This review summarizes the current knowledge of RNF126 structure, ubiquitin signaling mechanisms, physiological functions, and pathological roles, while discussing emerging therapeutic strategies and future challenges for targeting RNF126 in precision medicine. Full article

Background/Objectives: The “Land of Fires” (LF) in the Campania Region has attracted considerable attention due to massive environmental contamination deriving from decades of illegal disposal, burial, and burning of urban, industrial, and toxic waste. Cadmium (Cd) has been repeatedly proven to affect male reproductive function by a plethora of endocrine and non-endocrine mechanisms. The scientific literature is almost devoid of large studies addressing semen quality in this area, particularly by directly correlating seminal parameters to objectively measured pollutant burden in biological samples. Therefore, the aim of the current study was to comprehensively evaluate semen quality of males of reproductive age living in the LF, by correlating seminal parameters to cumulative local male reproductive tract Cd burden objectively quantified in whole semen samples. Methods: The current single-center, observational, cross-sectional study evaluated semen quality in 493 males aged 14–50 (29.07 ± 7.17) years living in three LF municipalities. Moreover, the association of semen quality with whole semen Cd (sCd) levels measured by inductively coupled plasma mass spectrometry (ICP-MS) was addressed in a subgroup of participants; semen samples suitable for semen Cd measurements were available from 383/493 (77.7%) participants of the total cohort, and all analyses involving semen Cd were performed within the measured subset. Results: In the total cohort, seminal parameters were as follows: semen pH 8.32 ± 0.3, semen volume 3.13 ± 1.67 mL, sperm concentration 37.58 ± 30.18 × 10⁶/mL, total count 111.2 ± 104 × 10⁶/ejaculate, total motility 56.83 ± 16.09%, progressive motility 50.22 ± 16.63%, in situ motility 6.72 ± 3.43%, immotile spermatozoa 43.07 ± 15.88%, normal morphology 7.97 ± 4.02%, and viability 64.75 ± 15.34%. Prevalence of normozoospermia and pathological seminal parameters was as follows: normozoospermia 66.5% (328/493), pathological seminal parameters 33.5% (165/493), specifically, oligozoospermia 14% (69/493), cryptozoospermia 0.8% (4/493), azoospermia 2.2% (11/493), asthenozoospermia 3% (15/493), teratozoospermia 0.6% (3/493), oligo-astheno-teratozoospermia 6.1% (30/493), necrozoospermia 5.7% (28/493), and different combined seminal parameters alterations 7.1% (35/493). Whole semen Cd was below (undetectable) or above (detectable) the limit of detection (LoD) (0.2 μg/L) in 66.6% (255/383) and 33.4% (128/383) whole semen samples, respectively. In samples with detectable sCd, sCd level was below or above the median value (0.76 μg/L; min–max 0.1–5.95 μg/L) in 23.4% (30/128) and 76.6% (98/128) whole semen samples, respectively. Participants with detectable sCd levels had a significantly reduced sperm total count (93.28 ± 84.88 × 10⁶/ejaculate vs. 113.2 ± 101.5 × 10⁶/ejaculate; p = 0.037), and normal morphology (7.29 ± 3.71% vs. 8.23 ± 3.91%; p = 0.034), and a significantly lower prevalence of normozoospermia (60.2% vs. 72.2%; p = 0.02) and significantly higher prevalence of pathological seminal parameters (39.8% vs. 27.8%; p = 0.02), specifically, a significantly higher prevalence of oligozoospermia (21.1% vs. 12.6%; p = 0.036) than those with undetectable sCd levels. Whole semen Cd levels were significantly higher in participants with pathological seminal parameters (1.08 ± 0.84 μg/L vs. 0.93 ± 0.74 μg/L; p = 0.037) than those with normozoospermia. Participants with sCd levels above the median value (N = 98) had a significantly reduced sperm concentration (29.12 ± 24.84 × 10⁶/mL vs. 43.62 ± 29.55 × 10⁶/mL; p = 0.015) and displayed a trend towards reduced sperm normal morphology (6.92 ± 3.38% vs. 8.55 ± 4.49%; p = 0.057) than those with sCd levels below the median value (N = 30). Moreover, participants with sCd levels above the median value (N = 98) had a significantly reduced sperm concentration (29.12 ± 24.84 × 10⁶/mL vs. 35.3 ± 26.29 × 10⁶/mL; p = 0.03), total count (85.77 ± 80.52 × 10⁶/ejaculate vs. 113.2 ± 101.5 × 10⁶/ejaculate; p = 0.008) and normal morphology (6.92 ± 3.38% vs. 8.23 ± 3.91%; p = 0.006), and a significantly lower prevalence of normozoospermia (57.1% vs. 72.2%; p = 0.008) and significantly higher prevalence of pathological seminal (42.9% vs. 27.8%; p = 0.008), specifically, a significantly higher prevalence of oligozoospermia (23.5% vs. 12.6%; p = 0.014) than those with undetectable sCd levels. Conclusions: The results of the current study demonstrate an association between the environmental Cd exposure and the impairment of seminal parameters, with a significantly poorer semen quality in participants with detectable sCd, and, more markedly, in those with sCd level above the median value, compared to participants with undetectable sCd, although subgroups comparisons highlighted a homogeneous profile in major confounders including age, BMI, and smoking habits among subgroups of participants with different sCd burden. Full article

Background: Male infertility affects millions of couples, accounting for 50 percent of cases. Despite such a major contribution of the male factor, it is not properly evaluated and is often overlooked in infertility assessments. Semen analysis, which is routinely performed to assess infertility status, is unable to assess the defects at the molecular level which are important to assess the fertilizing capacity of the sperm. This study aims to determine the utility of sperm function tests as biomarkers for male infertility in addition to standard semen analysis. Methods: Thirty-five men (aged 25–45 years) were recruited and divided into two groups: those with at least one altered semen parameter (infertile group) and those with normal semen parameters but unable to conceive after more than one year of unprotected intercourse (unexplained male infertility group). The DNA Fragmentation Index (DFI), Nuclear Chromatin Decondensation Test (NCDT) and Hypoosmotic Swelling Test (HOS) were used in diagnosing sperm dysfunction in both groups. The Mann–Whitney U testand Spearman’s rank correlation were used for analyzing the parameters of the groups. A p value < 0.05 was considered statistically significant. Results: While motility and vitality were nearly identical in both groups, the infertile group showed more morphological abnormalities. The DFI was higher in the unexplained male infertility group (UMI) (82%) than in the infertile group (36%). Poor decondensation capacity was present in 27% of the unexplained male infertility group and 60% of the infertile group. Both groups’ hypoosmotic swelling values fell within the usual range. Spearman correlation analysis revealed that the NCDT showed significant positive correlations with sperm vitality (r = 0.36; p = 0.02) and morphology (r = 0.53; p = 0.001). The DFI demonstrated significant negative correlations with vitality (r = −0.45; p = 0.006) and motility (r = −0.39; p = 0.01). HOS was significantly positively correlated with motility (r = 0.56; p = 0.0004) and vitality (r = 0.57; p = 0.0003). Additionally, the NCDT and DFI showed a significant inverse correlation (r = −0.33; p = 0.04). Conclusions: These findings highlight the potential of sperm function tests as valuable diagnostic tools alongside conventional semen analysis for a more comprehensive assessment of male fertility. Full article

Early flowering-related factors play pivotal roles in coordinating plant growth and reproductive development. In this study, we investigated the biological function of early flowering gene (ELF) in Arabidopsis thaliana using CRISPR/Cas9-mediated genome editing and construction of overexpression approaches. Two independent ELF overexpression (OE-ELF) and genome-edited (ge-elf) lines were generated and systemically analyzed. ELF overexpression significantly enhanced early seedling performance, increasing germination rate and seedling fresh weight by up to 8.7%, while genome-edited lines exhibited a marked reduction. Root growth was strongly promoted in OE-ELF plants, with root length increase of 85% and 75%, whereas ge-elf lines showed a reduction of up to 48%. At later developmental stages, OE-ELF plants displayed enhanced vegetative growth, including increased leaf length (32%), leaf area (91%), and accelerated flowering (21% earlier than wild type). In contrast, ge-elf delayed flowering by up to 25% and resulted in compact plant architecture. Reproductive development was severely compromised in ge-elf plants, which exhibited malformed inflorescences, reduced pollen germination, shortened silique (45%), and a drastic decrease in seed number per silique (70%). Conversely, OE-ELF plants showed increased silique number and seed per silique. Molecular analysis revealed that ELF positively regulates key flowering-related genes, including FLC, SOC1, AP1, and LFY, which correlated strongly with growth and reproductive traits. Our results demonstrate that ELF functions as a central regulator integrating vegetative growth, floral development, male fertility, and seed production in Arabidopsis thaliana. Full article

Low-temperature-induced fertility restoration in thermo-sensitive genic male sterile (TGMS) lines severely impairs hybrid seed purity, which is a major bottleneck for two-line hybrid rice production. Most commercial TGMS lines rely on the single tms5 locus, leading to high climatic vulnerability. In this study, we developed a dual-locus strategy by target genome editing of TMS5 and MS1 in indica rice GH89. Adenine base editing at the MS1 locus exhibited a high editing efficiency of 93.5%. Transgene-free homozygous single mutants (GH89-tms5 and GH89-MS1) and double mutant (GH89-tms5 + MS1) were generated for phenotypic analysis. The double mutant GH89-tms5 + MS1 remained completely sterile for 5 and 10 days under controlled low temperature (23.5 °C), with only minimal fertility restoration after 15 days. In the field, it maintained complete sterility for 84 consecutive days and was fully insensitive to short-term low temperature fluctuations, outperforming single mutants and commercial control Y58S. Moreover, the double mutant retained most key yield-related agronomic traits of the wild type with only minor variations. This dual mutation forms a “double-lock” fertility regulatory system, significantly increasing the low-temperature duration threshold for fertility restoration. The GH89-tms5 + MS1 line exhibits promising potential for future rice breeding applications. Full article

This study evaluated the effects of dietary supplementation with 25-hydroxyvitamin D₃ (25-OH-D3), canthaxanthin (Cx), copper (Cu), and gluco-oligosaccharides (GO) on performance, egg quality, fertility, and hatchability of broiler breeder hens during early production (30 to 41 weeks of age). A total of 210 breeder hens and 21 males were allocated to three dietary treatments. Birds were fed a standard broiler breeder diet (control) or the same diet in which 0.5 kg/MT of an additive premix replaced sand. This premix supplied 16.6–17.7 mg/kg canthaxanthin (Cx) and 3700–4700 IU/kg 25-OH-D3 in Treatment 1, and 9.2–11.1 mg/kg Cx and 4100–4700 IU/kg 25-OH-D3 in Treatment 2. Treatment 2 also included Cu and GO (≥44 mg/g within the additive), with Cu provided at industry-standard levels. Each treatment consisted of seven replicates, with birds housed in floor pens containing 10 females and 1 male per replicate. Hen performance was recorded weekly, while egg quality was assessed at 30, 35, and 41 weeks. Fertility and hatchability were evaluated at 30 and 41 weeks. Treatment 2 improved lay rate, albumen height, Haugh unit, and feed conversion ratio (FCR) at selected time points. Both supplemented diets consistently produced darker yolk color compared with the control. Supplementation consistently enhanced yolk color, whereas effects on albumen height, Haugh unit, shell thickness, and FCR were observed only at specific ages or weeks. Lay rate differed among treatments only at week 40, with a trend observed at week 41. Fertility and hatchability were not significantly affected by dietary treatment. Full article

Pyrethroid insecticides are widely used in agriculture, but their sublethal effects on male reproduction in Antheraea pernyi (Guérin-Méneville, 1855) (Saturniidae) remain largely unknown. Here, we evaluated the effects of sublethal β-cypermethrin exposure (LC₂₀ = 0.0074 mg/L) on gonadal development, testicular morphology, sperm quantity and function, mating behavior, and post-mating egg traits from the larval to adult stages and investigated the molecular mechanisms using transcriptome sequencing. Insecticide stress impaired testicular development and disrupted spermatogenesis, leading to a 25% reduction in eupyrene sperm bundles, a 10.72% decrease in acrosin activity, and a 16.75% decrease in the curvilinear velocity of apyrene sperm. Mating willingness decreased by 17.78 percentage points, and females mated with treated males produced 20.45% fewer eggs. Transcriptome analysis identified 1193 differentially expressed genes (|log₂FC| > 1, FDR < 0.01). Detoxification and stress response genes (e.g., CYP3A27, GSTD1, ABCB1, and Hsp70) were predominantly up-regulated, while reproduction-related genes (e.g., dnal1, tubb1, ATPsynbeta, far1, JHAMT, mei-41, and dna2) were down-regulated. This study demonstrates that sublethal β-cypermethrin exposure induces male reproductive toxicity in A. pernyi through a “dual-hit” mechanism: persistently activating the detoxification system (leading to energy depletion) while directly suppressing reproductive genes, ultimately resulting in impaired germ cell development, defective sperm function, and reduced fertility. Full article

Reproductive emergencies in male dromedary camels (Camelus dromedarius) threaten fertility, health, and welfare but remain poorly documented. This review consolidates knowledge on the diagnosis, management, and prognosis of acute reproductive pathologies, including traumatic injuries (testicular trauma, penile hematoma, and preputial laceration), obstructive conditions (urethral obstruction, phimosis, and paraphimosis), vascular emergencies (spermatic cord torsion, priapism), inflammatory diseases (orchitis, epididymitis, pizzle rot), congenital anomalies (persistent frenulum), iatrogenic complications, and pharmacologic-induced dysfunction. Systematic breeding soundness examination and ultrasonography are indispensable for assessing testicular perfusion and guiding intervention. Given the scarcity of camel-specific literature, this narrative clinical review integrates evidence from published camelid studies, relevant reports from other domestic species, and representative clinical observations documented by the authors to provide a practical framework for diagnosis and management. Field constraints, the stoic nature of camels, and harmful ethnoveterinary practices often delay diagnosis and compromise outcomes. Prognosis varies considerably depending on the condition and timeliness of intervention; early aggressive management can preserve reproductive function, while delayed treatment frequently results in permanent infertility or death. Given the economic and cultural importance of camels in arid regions, improving outcomes for reproductive emergencies is essential for sustaining breeding programs and enhancing animal welfare. This narrative clinical review integrates published literature with representative clinical observations and retrospective field cases to summarize the diagnosis, management, and prognosis of reproductive emergencies in male dromedary camels. It also provides a structured clinical framework to support veterinarians in managing these acute conditions and highlights critical gaps requiring further research. Full article

Morus alba L. fruit extract (MFE), which is rich in cyanidin 3-glucoside (C3G), demonstrates antioxidant properties and pharmacological effects, but its reproductive safety remains poorly understood. Polyphenols modulate steroidogenesis, spermatogenesis, and sperm acrosome integrity; therefore, toxicity assessment is needed for use. This study aimed to evaluate the antioxidant profiles and subchronic reproductive effects of MFE. MFE was standardized using high-performance liquid chromatography (HPLC), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) assays. Male rats were administered MFE (250 or 500 mg/kg BW) for 56 days. Assessments included computer-assisted sperm analysis (CASA), testosterone, seminal fructosamine, and testicular CYP11A1 and androgen receptor (AR) expression. Acrosomal status was determined using PNA lectin staining. The results showed that MFE contained C3G (119.42 mg/g), antioxidant capacity (DPPH IC₅₀: 0.101 mg/mL; FRAP: 465.01 µmol Fe (II)/g), and total contents (phenolics: 41.15 mg GAE/g; flavonoids: 3.15 mg CE/g; anthocyanidins: 11.04 mg C3G/g). MFE did not alter testicular histology and seminiferous stages VII-IX. High doses significantly increased sperm concentration, while both doses reduced sperm beat cross frequency. Testosterone, fructosamine, and CYP11A1/AR expressions showed increasing trends. Significantly, high doses induced a precocious acrosome reaction. In conclusion, MFE has no reproductive toxicity and pro-fertility effects on sperm quantity or androgenic markers, supporting safe subchronic use. Full article