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43 pages, 8518 KiB  
Review
Cutting-Edge Sensor Technologies for Exosome Detection: Reviewing Role of Antibodies and Aptamers
by Sumedha Nitin Prabhu and Guozhen Liu
Biosensors 2025, 15(8), 511; https://doi.org/10.3390/bios15080511 (registering DOI) - 6 Aug 2025
Abstract
Exosomes are membranous vesicles that play a crucial role as intercellular messengers. Cells secrete exosomes, which can be found in a variety of bodily fluids such as amniotic fluid, semen, breast milk, tears, saliva, urine, blood, bile, ascites, and cerebrospinal fluid. Exosomes have [...] Read more.
Exosomes are membranous vesicles that play a crucial role as intercellular messengers. Cells secrete exosomes, which can be found in a variety of bodily fluids such as amniotic fluid, semen, breast milk, tears, saliva, urine, blood, bile, ascites, and cerebrospinal fluid. Exosomes have a distinct bilipid protein structure and can be as small as 30–150 nm in diameter. They may transport and exchange multiple cellular messenger cargoes across cells and are used as a non-invasive biomarker for various illnesses. Due to their unique features, exosomes are recognized as the most effective biomarkers for cancer and other disease detection. We give a review of the most current applications of exosomes derived from various sources in the prognosis and diagnosis of multiple diseases. This review also briefly examines the significance of exosomes and their applications in biomedical research, including the use of aptamers and antibody–antigen functionalized biosensors. Full article
(This article belongs to the Special Issue Material-Based Biosensors and Biosensing Strategies)
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13 pages, 917 KiB  
Article
Relationship Between Prostaglandin and Interleukin Concentrations in Seminal Fluid and Their Influence on the Rate of Fertilization in Men Undergoing ICSI
by Houda Amor, Fatina W. Dahadhah, Peter Michael Jankowski, Rami Al Nasser, Lisa Jung, Ingolf Juhasz-Böss, Erich Franz Solomayer and Mohamad Eid Hammadeh
Int. J. Mol. Sci. 2025, 26(15), 7627; https://doi.org/10.3390/ijms26157627 (registering DOI) - 6 Aug 2025
Abstract
Sperm count, motility, and morphology are semen parameters that directly affect male fertility. The presence of cytokines in seminal plasma negatively or positively influences these parameters. Interleukins and prostaglandins are proinflammatory cytokines present in human seminal plasma and play crucial roles in fertilization, [...] Read more.
Sperm count, motility, and morphology are semen parameters that directly affect male fertility. The presence of cytokines in seminal plasma negatively or positively influences these parameters. Interleukins and prostaglandins are proinflammatory cytokines present in human seminal plasma and play crucial roles in fertilization, in general and after intracytoplasmic sperm injection (ICSI) procedures. This study aimed to investigate the possible influence of interleukins IL-17 and IL-18, and prostaglandins PGE2 and PGF2α on male infertility. Semen samples were collected from 58 males who underwent the ICSI procedure. An enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of IL-17, IL-18, PGE2, and PGF2α, and these concentrations were then correlated with semen parameters and the rate of fertilization. Furthermore, the chromatin integrity of the sperm was evaluated with an Acridine Orange (AO) assay. The results showed an inversely proportional relationship between the AO binding intensity and fertilization rate (r = −0.394; p ≤ 0.002). Furthermore, a negative correlation was observed between the IL-18 concentration and positive AO (p ≤ 0.021). Moreover, the IL-18 concentration was positively correlated with the fertilization rate (p ≤ 0.05). In contrast, IL-17 did not significantly correlate with any semen parameters or with the fertilization rate. Seminal PGE2 levels were significantly correlated with embryo cleavage at 72 h (p ≤ 0.05). To conclude, this study revealed that denaturation of sperm nuclear deoxyribonucleic acid (DNA) contributes to low fertilization rates. In addition, this study proposed a potential role for IL-18 in fertilization. PGE2 likely influences embryo development, but further studies are needed to examine the impact of seminal PGE2 on the oocyte to fully elucidate its contribution to this complex biological process. Full article
(This article belongs to the Special Issue Advances in Spermatogenesis and Male Infertility)
11 pages, 1056 KiB  
Article
Optimization of Duck Semen Freezing Procedure and Regulation of Oxidative Stress
by Zhicheng Wang, Haotian Gu, Chunhong Zhu, Yifei Wang, Hongxiang Liu, Weitao Song, Zhiyun Tao, Wenjuan Xu, Shuangjie Zhang and Huifang Li
Animals 2025, 15(15), 2309; https://doi.org/10.3390/ani15152309 (registering DOI) - 6 Aug 2025
Abstract
Waterfowl semen cryopreservation technology is a key link in genetic resource conservation and artificial breeding, but poultry spermatozoa, due to their unique morphology and biochemical properties, are prone to oxidative stress during freezing, resulting in a significant decrease in vitality. In this study, [...] Read more.
Waterfowl semen cryopreservation technology is a key link in genetic resource conservation and artificial breeding, but poultry spermatozoa, due to their unique morphology and biochemical properties, are prone to oxidative stress during freezing, resulting in a significant decrease in vitality. In this study, we first used four different freezing procedures (P1–P4) to freeze duck semen and compared their effects on duck sperm quality. Then, the changes in antioxidant indexes in semen were monitored. The results showed that program P4 (initial 7 °C/min slow descent to −35 °C, followed by 60 °C/min rapid descent to −140 °C) was significantly better than the other programs (p < 0.05), and its post-freezing sperm vitality reached 71.41%, and the sperm motility was 51.73%. In the P1 and P3 groups, the sperm vitality was 65.56% and 53.41%, and the sperm motility was 46.99% and 31.76%, respectively. In terms of antioxidant indexes, compared with the fresh semen group (CK), the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-px) in the P2 group were significantly decreased (p < 0.05), while the activities of SOD and CAT in the P4 group showed no significant changes (p > 0.05) except that the activity of GSH-px was significantly decreased (p < 0.05). And the CAT and GSH-px activities in the P4 group were significantly higher than those in the P2 group (p < 0.05). The content of malondialdehyde (MDA) in the P2 group was significantly higher than that in the fresh semen group (p < 0.05), and there was no significant difference between the P2 group and the P4 group (p > 0.05). The total antioxidant capacity (T-AOC) content of the P2 and P4 groups was significantly lower than that of the fresh semen group (p < 0.05). The staged cooling strategy of P4 was effective in reducing the exposure time to the hypertonic environment by balancing intracellular dehydration and ice crystal inhibition, shortening the reactive oxygen species accumulation and alleviating oxidative stress injury. On the contrary, the multi-stage slow-down strategy of P2 exacerbated mitochondrial dysfunction and the oxidative stress cascade response due to prolonged cryogenic exposure time. The present study confirmed that the freezing procedure directly affects duck sperm quality by modulating the oxidative stress pathway and provides a theoretical basis for the standardization of duck semen cryopreservation technology. Full article
(This article belongs to the Section Poultry)
12 pages, 1169 KiB  
Article
Field-Compatible Cytometric Assessment of Epididymal Alpaca Sperm Viability and Acrosomal Integrity Using Fluorochrome
by Alexei Santiani, Miguel Cucho, Josselyn Delgado, Javier Juárez, Luis Ruiz and Shirley Evangelista-Vargas
Animals 2025, 15(15), 2282; https://doi.org/10.3390/ani15152282 - 5 Aug 2025
Abstract
In remote alpaca breeding regions, access to advanced sperm analysis laboratories is limited. This study validates a practical cytometric method for evaluating sperm viability and acrosomal integrity in epididymal alpaca sperm using early fluorochrome staining, formaldehyde fixation, and intermediate storage. Thirty-two testes were [...] Read more.
In remote alpaca breeding regions, access to advanced sperm analysis laboratories is limited. This study validates a practical cytometric method for evaluating sperm viability and acrosomal integrity in epididymal alpaca sperm using early fluorochrome staining, formaldehyde fixation, and intermediate storage. Thirty-two testes were transported at 5 °C, and spermatozoa were collected from the cauda epididymis. After morphometric screening, 26 samples were included. Aliquots were stained with Zombie Green (viability) and FITC–PSA (acrosomal integrity), at time zero. Each aliquot was divided for cytometric analysis at T0 (immediately), T24 (24 h after formaldehyde fixation) and T1w (1 week post-fixation). Fixed samples showed higher viability and acrosomal integrity values (T24: 70.75%, 97.24%; T1w: 71.80%, 97.21%) than T0 (67.63%, 95.89%). This may reflect fluorescence alterations associated with fixation. Strong correlations and Bland–Altman analysis confirmed consistency across time points. This method enables accurate sperm quality evaluation up to one week after collection, offering a useful tool for reproductive monitoring in field conditions without immediate analysis. Further research on ejaculated semen and field protocols is recommended. Full article
(This article belongs to the Special Issue Advances in Camelid Reproduction)
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18 pages, 1102 KiB  
Review
Exploring Human Sperm Metabolism and Male Infertility: A Systematic Review of Genomics, Proteomics, Metabolomics, and Imaging Techniques
by Achraf Zakaria, Idrissa Diawara, Amal Bouziyane and Noureddine Louanjli
Int. J. Mol. Sci. 2025, 26(15), 7544; https://doi.org/10.3390/ijms26157544 - 5 Aug 2025
Abstract
Male infertility is a multifactorial condition often associated with disruptions in sperm metabolism and mitochondrial function, yet traditional semen analysis provides limited insight into these molecular mechanisms. Understanding sperm bioenergetics and metabolic dysfunctions is crucial for improving the diagnosis and treatment of conditions [...] Read more.
Male infertility is a multifactorial condition often associated with disruptions in sperm metabolism and mitochondrial function, yet traditional semen analysis provides limited insight into these molecular mechanisms. Understanding sperm bioenergetics and metabolic dysfunctions is crucial for improving the diagnosis and treatment of conditions such as asthenozoospermia and azoospermia. This systematic review synthesizes recent literature, focusing on advanced tools and techniques—including omics technologies, advanced imaging, spectroscopy, and functional assays—that enable comprehensive molecular assessment of sperm metabolism and development. The reviewed studies highlight the effectiveness of metabolomics, proteomics, and transcriptomics in identifying metabolic biomarkers linked to male infertility. Non-invasive imaging modalities such as Raman and magnetic resonance spectroscopy offer real-time metabolic profiling, while the seminal microbiome is increasingly recognized for its role in modulating sperm metabolic health. Despite these advances, challenges remain in clinical validation and implementation of these techniques in routine infertility diagnostics. Integrating molecular metabolic assessments with conventional semen analysis promises enhanced diagnostic precision and personalized therapeutic approaches, ultimately improving reproductive outcomes. Continued research is needed to standardize biomarkers and validate clinical utility. Furthermore, these metabolic tools hold significant potential to elucidate the underlying causes of previously misunderstood and unexplained infertility cases, offering new avenues for diagnosis and treatment. Full article
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24 pages, 4384 KiB  
Article
Untargeted Metabolomic Identifies Potential Seasonal Biomarkers of Semen Quality in Duroc Boars
by Notsile H. Dlamini, Serge L. Kameni and Jean M. Feugang
Biology 2025, 14(8), 995; https://doi.org/10.3390/biology14080995 (registering DOI) - 4 Aug 2025
Abstract
High semen quality is vital for reproductive success in the swine industry; however, seasonal fluctuations often compromise this quality. The molecular mechanism underlying these seasonal effects on semen quality remains largely unclear. This study employed untargeted metabolomic profiling of boar seminal plasma (SP) [...] Read more.
High semen quality is vital for reproductive success in the swine industry; however, seasonal fluctuations often compromise this quality. The molecular mechanism underlying these seasonal effects on semen quality remains largely unclear. This study employed untargeted metabolomic profiling of boar seminal plasma (SP) to identify metabolites and metabolic pathways associated with semen quality during the summer and winter months. Semen samples were collected from mature Duroc boars at a commercial boar stud and classified as Passed or Failed based on motility and morphology. SP from five samples per group was analyzed using ultra-high-performance liquid chromatography–mass spectrometry (UHPLC-MS). In total, 373 metabolites were detected in positive ion mode and 478 in negative ion mode. Several differentially expressed metabolites (DEMs) were identified, including ergothioneine, indole-3-methyl acetate, and avocadyne in the summer, as well as LysoPC, dopamine, and betaine in the winter. These metabolites are associated with key sperm functions, including energy metabolism, antioxidant defense, and capacitation. KEGG pathway analysis indicated enrichment in starch and sucrose metabolism, pyrimidine metabolism, and amino acid metabolism across the seasons. Overall, the results reveal that SP metabolomic profiles vary with the season, thereby influencing semen quality. The identified metabolites may serve as potential biomarkers for assessing semen quality and enhancing reproductive efficiency in swine production. Full article
(This article belongs to the Special Issue Reproductive Physiology and Pathology in Livestock)
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16 pages, 24404 KiB  
Article
Oxidation of HfB2-HfO2-SiC Ceramics Modified with Ti2AlC Under Subsonic Dissociated Airflow
by Elizaveta P. Simonenko, Aleksey V. Chaplygin, Nikolay P. Simonenko, Ilya V. Lukomskii, Semen S. Galkin, Anton S. Lysenkov, Ilya A. Nagornov, Artem S. Mokrushin, Tatiana L. Simonenko, Anatoly F. Kolesnikov and Nikolay T. Kuznetsov
Corros. Mater. Degrad. 2025, 6(3), 35; https://doi.org/10.3390/cmd6030035 - 1 Aug 2025
Viewed by 168
Abstract
Ultrahigh-temperature ceramic composites based on hafnium diboride have a wide range of applications, including as components for high-speed aircraft and energy generation and storage devices. Consequently, developing methodologies for their fabrication and studying their properties are of paramount importance, in particular in using [...] Read more.
Ultrahigh-temperature ceramic composites based on hafnium diboride have a wide range of applications, including as components for high-speed aircraft and energy generation and storage devices. Consequently, developing methodologies for their fabrication and studying their properties are of paramount importance, in particular in using them as an electrode material for energy storage devices with increased oxidation resistance. This study investigates the behavior of ceramic composites based on the HfB2-HfO2-SiC system, obtained using 15 vol% Ti2AlC MAX-phase as a sintering component, under the influence of subsonic flow of dissociated air. It was determined that incorporating the modifying component (Ti2AlC) altered the composition of the silicate melt formed on the surface during ceramic oxidation. This modification led to the observation of a protective antioxidant function. Consequently, liquation was observed in the silicate melt layer, resulting in the formation of spherical phase inhomogeneities in its volume with increased content of titanium, aluminum, and hafnium. It is hypothesized that the increase in the high-temperature viscosity of this melt prevents it from being carried away in the form of drops, even at a surface temperature of ~1900–2000 °C. Despite the established temperature, there is no sharp increase in its values above 2400–2500 °C. This is due to the evaporation of silicate melt from the surface. In addition, the electrochemical behavior of the obtained material in a liquid electrolyte medium (KOH, 3 mol/L) was examined, and it was shown that according to the value of electrical conductivity and specific capacitance, it is a promising electrode material for supercapacitors. Full article
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14 pages, 939 KiB  
Review
Revisiting Male Fertility in Livestock: The Case of Bull Sperm RNA
by Rene A. Ramírez-Sosa, Francisco J. Jahuey-Martínez, Monserrath Felix-Portillo and José A. Martínez-Quintana
Biology 2025, 14(8), 969; https://doi.org/10.3390/biology14080969 (registering DOI) - 1 Aug 2025
Viewed by 282
Abstract
To achieve the goals of productivity and sustainability across diverse livestock systems, reproductive factors play a pivotal role. Historically, reproductive research has primarily focused on females, as they are responsible for maintaining pregnancy and delivering offspring following oocyte fertilization. However, since the early [...] Read more.
To achieve the goals of productivity and sustainability across diverse livestock systems, reproductive factors play a pivotal role. Historically, reproductive research has primarily focused on females, as they are responsible for maintaining pregnancy and delivering offspring following oocyte fertilization. However, since the early 2000s, the biological significance of sperm RNAs has been increasingly recognized in various livestock species. These RNAs contribute both genetically and epigenetically at the time of fertilization and during early embryonic development. Multiple types of sperm RNA have been identified in bovine, porcine, ovine, buffalo, and caprine spermatozoa. Notably, transcriptomic profiling has shown potential to differentiate between high- and low-fertility males, even when conventional semen quality values appear normal in both groups. This opens the possibility for more accurate identification of highly fertile sires. Nevertheless, a definitive marker or set of markers has yet to be established, likely due to the transcriptome’s sensitivity to environmental conditions and to the variability in evaluation methodologies. Therefore, global scientific efforts should aim to establish standardized, robust protocols, as sperm RNA represents a promising avenue for enhancing the sustainability of animal production systems. Full article
(This article belongs to the Special Issue The Biology of Animal Reproduction)
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10 pages, 603 KiB  
Article
The Impacts of Protein Supplementation and Semen Exposure on Uterine Cytokines in Beef Heifers
by Kyle J. McLean, Kiernan J. Brandt, Taylor B. Ault-Seay, Rebecca R. Payton, Liesel G. Schneider, J. Lannett Edwards, Phillip R. Myer and Justin D. Rhinehart
Agriculture 2025, 15(15), 1642; https://doi.org/10.3390/agriculture15151642 - 30 Jul 2025
Viewed by 243
Abstract
Reproductive efficiency is largely impacted during heifer development, which generally requires nutrient supplementation for proper maturation. Nutritional status can also influence inflammation within the reproductive tract. Therefore, we hypothesized that cytokine concentrations in uterine luminal fluid (ULF) will be impacted by protein supplementation [...] Read more.
Reproductive efficiency is largely impacted during heifer development, which generally requires nutrient supplementation for proper maturation. Nutritional status can also influence inflammation within the reproductive tract. Therefore, we hypothesized that cytokine concentrations in uterine luminal fluid (ULF) will be impacted by protein supplementation following exposure to semen via artificial insemination (AI). Commercial heifers (n = 60) were utilized to determine the effects of protein supplementation and AI on cytokine concentrations in ULF. Heifers were randomly assigned to one of three crude protein (CP) treatments (11%, 15%, and 19% overall CP) via supplementation: (1) CON (10% CP), (2) P20 (20% CP), and (3) P40 (40% CP). All heifers underwent estrus synchronization and ULF was collected 14 d after insemination. Cytokine profiles were constructed in MetaboAnalyst 5.0, and R Studio was used for individual cytokine analyses. Control heifers had increased (p = 0.05) MIP-1β concentrations (148.7 ± 123.9 pg) over P20 heifers (42.3 ± 123.9 pg), and P40 heifers (75.5 ± 123.9 pg) had intermediate concentrations. Semen exposure (1877 ± 550 pg) showed a trend (p = 0.06) to increase concentrations of IP-10 compared with heifers who were not inseminated (1556 ± 550 pg). In conclusion, although protein supplementation and semen exposure had minimal effects on overall cytokine profiles, MIP-1β, IP-10, and MIP-1α were identified as potential key regulators of uterine inflammation during early gestation. Full article
(This article belongs to the Section Farm Animal Production)
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12 pages, 1176 KiB  
Article
Effect of Different Extenders on the Oxidative Status and Fertility of Sarda Ram Liquid Semen Stored at 15 °C
by Pasciu Valeria, Charbel Nassif, Maria Dattena, Sara Succu, Francesca Daniela Sotgiu, Antonello Cannas, Ignazio Cossu, Elena Baralla, Fabrizio Chessa, Fiammetta Berlinguer and Laura Mara
Antioxidants 2025, 14(8), 932; https://doi.org/10.3390/antiox14080932 - 30 Jul 2025
Viewed by 249
Abstract
Liquid storage is an important tool used to prolong fresh semen shelf-life while protecting spermatozoa from damage, conserving their overall functionality, and ensuring better fertility than frozen semen from sheep. The increased production of reactive oxygen species (ROS) during sperm storage leads to [...] Read more.
Liquid storage is an important tool used to prolong fresh semen shelf-life while protecting spermatozoa from damage, conserving their overall functionality, and ensuring better fertility than frozen semen from sheep. The increased production of reactive oxygen species (ROS) during sperm storage leads to a decline in sperm quality, particularly with regard to sperm nuclear DNA damage and mitochondrial membrane potential (MMP). This study evaluated the effect of storing Sarda ram semen at 15 °C for 7 h on its redox status, motility, morphology, acrosome integrity, ATP content, mitochondrial potential membrane, and in vivo fertility after artificial insemination. Two different extenders were compared: a lab-made skimmed milk (SM)-based extender and a commercial extender (OviXcell®, IMV-Technologies, France). Lower ROS levels in the SM (p < 0.001) indicated that its oxidative status was better maintained compared to the commercial extender (CE). Antioxidant defenses (total antioxidant capacity, TEAC; superoxide dismutase, SOD; total thiols) were higher in the SM (p < 0.01) than in the CE. SM also had higher MMP (p < 0.05), acrosome integrity (p < 0.05), ATP content (p < 0.01), and in vivo fertilizing capacity (p < 0.05) compared to the CE, which indicated higher semen quality. In conclusion, the SM extender, while maintaining a better oxidative/antioxidant balance, ensured higher semen quality after 7 h of storage at 15 °C in vitro compared to the CE. Full article
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21 pages, 3547 KiB  
Article
Enzymatic Degumming of Soybean Oil for Raw Material Preparation in BioFuel Production
by Sviatoslav Polovkovych, Andriy Karkhut, Volodymyr Gunka, Yaroslav Blikharskyy, Roman Nebesnyi, Semen Khomyak, Jacek Selejdak and Zinoviy Blikharskyy
Appl. Sci. 2025, 15(15), 8371; https://doi.org/10.3390/app15158371 - 28 Jul 2025
Viewed by 191
Abstract
The paper investigates the process of degumming substandard soybean oil using an enzyme complex of phospholipases to prepare it as a feedstock for biodiesel production. Dehumidification is an important refining step aimed at reducing the phosphorus content, which exceeds the permissible limits according [...] Read more.
The paper investigates the process of degumming substandard soybean oil using an enzyme complex of phospholipases to prepare it as a feedstock for biodiesel production. Dehumidification is an important refining step aimed at reducing the phosphorus content, which exceeds the permissible limits according to ASTM, EN, and ISO standards, by re-moving phospholipids. The enzyme complex of phospholipases includes phospholipase C, which specifically targets phosphatidylinositol, and phospholipase A2, which catalyzes the hydrolysis of phospholipids into water-soluble phosphates and lysophospholipids. This process contributes to the efficient removal of phospholipids, increased neutral oil yield, and reduced residual oil in the humic phase. The use of an enzyme complex of phospholipases provides an innovative, cost-effective, and environmentally friendly method of oil purification. The results of the study demonstrate the high efficiency of using the phospholipase enzyme complex in the processing of substandard soybean oil, which allows reducing the content of total phosphorus to 0.001% by weight, turning it into a high-quality raw material for biodiesel production. The proposed approach contributes to increasing the profitability of agricultural raw materials and the introduction of environmentally friendly technologies in the field of renewable energy. Full article
(This article belongs to the Special Issue Biodiesel Production: Current Status and Perspectives)
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17 pages, 5549 KiB  
Article
The Effects of Limonin, Myo-Inositol, and L-Proline on the Cryopreservation of Debao Boar Semen
by Qianhui Feng, Yanyan Yang, Bing Zhang, Wen Shi, Yizhen Fang, Chunrong Xu, Zhuxin Deng, Wanyou Feng and Deshun Shi
Animals 2025, 15(15), 2204; https://doi.org/10.3390/ani15152204 - 27 Jul 2025
Viewed by 282
Abstract
Semen cryopreservation is associated with sperm vulnerability to oxidative stress and ice crystal-induced damage, adversely affecting in vitro fertilization (IVF) success. This study aimed to investigate the effects of freezing diluent supplemented with antioxidant limonin (Lim), myo-inositol (MYO), and the ice crystal formation [...] Read more.
Semen cryopreservation is associated with sperm vulnerability to oxidative stress and ice crystal-induced damage, adversely affecting in vitro fertilization (IVF) success. This study aimed to investigate the effects of freezing diluent supplemented with antioxidant limonin (Lim), myo-inositol (MYO), and the ice crystal formation inhibitor L-proline (LP) through sperm motility, morphological integrity, and antioxidant capacity. The Lim (150 mM), MYO (90 mM), and LP (100 mM) significantly ameliorated the quality of post-thaw sperm in Debao boar, and combined treatment of these agents significantly enhanced sperm motility, structural integrity, and antioxidant capacity compared with individual agents (p < 0.05). Notably, the combined use of these agents reduced glycerol concentration in the freezing diluent from 3% to 2%. Meanwhile, the integrity of the sperm plasma membrane, acrosome membrane, and mitochondrial membrane potential was significantly improved (p < 0.05), and the result of IVF revealed the total cell count of the blastocysts was also greater in the 2% glycerol group (p < 0.05). In conclusion, the newly developed freezing diluent for semen, by adding Lim (150 mM), MYO (90 mM), and LP (100 mM), can enhance the quality of frozen–thawed Debao boar sperm and reduce the concentration of glycerol from 3% to 2% as high concentrations of glycerol can impair the quality of thawed sperm and affect in vitro fertilization outcomes. In conclusion, the improved dilution solution formulated demonstrated efficacy in enhancing the quality of porcine spermatozoa following cryopreservation and subsequent thawing. Full article
(This article belongs to the Section Pigs)
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23 pages, 2699 KiB  
Article
Changes in L-Carnitine Metabolism Affect the Gut Microbiome and Influence Sexual Behavior Through the Gut–Testis Axis
by Polina Babenkova, Artem Gureev, Irina Sadovnikova, Inna Burakova, Yuliya Smirnova, Svetlana Pogorelova, Polina Morozova, Viktoria Gribovskaya, Dianna Adzhemian and Mikhail Syromyatnikov
Microorganisms 2025, 13(8), 1751; https://doi.org/10.3390/microorganisms13081751 - 26 Jul 2025
Viewed by 393
Abstract
L-carnitine and Mildronate are substances that can significantly rearrange the energy metabolism of cells. This can potentially cause changes in the bacterial composition of the gut microbiome and affect testis functionality and male sexual health. Mice of the C57Bl/6 line were used. Sexual [...] Read more.
L-carnitine and Mildronate are substances that can significantly rearrange the energy metabolism of cells. This can potentially cause changes in the bacterial composition of the gut microbiome and affect testis functionality and male sexual health. Mice of the C57Bl/6 line were used. Sexual behavior was assessed using physiological tests, and gene expression patterns were assessed by qPCR. High-throughput sequencing of mouse fecal microbiota was performed. We showed that long-term administration of Mildronate has no significant effect on the intestinal microbiome, and there was a compensatory increase in the expression of genes involved in fatty acid and leptin metabolism. No impairment of sexual motivation in male mice was observed. Prolonged L-carnitine supplementation caused a decrease in alpha diversity of bacteria and a decrease in some groups of microorganisms that are components of a healthy gut microflora. A correlation was observed between the level of bacteria from Firmicutes phylum, indicators of sexual motivation of mice, and the dynamics of body weight gain. Our results may indicate that metabolic modulators can have a significant impact on the structure of the bacterial community of the gut microbiome, which may influence male sexual health through the gut–semen axis. Full article
(This article belongs to the Section Gut Microbiota)
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18 pages, 3945 KiB  
Review
Association of Lycopene and Male Reproductive Health: Systematic Review and Meta-Analysis
by Isabel Viña and Juan R. Viña
Int. J. Mol. Sci. 2025, 26(15), 7224; https://doi.org/10.3390/ijms26157224 - 25 Jul 2025
Viewed by 613
Abstract
Lycopene, a carotenoid found in tomatoes and watermelon, has been investigated for its potential to improve male fertility through its antioxidant and anti-inflammatory mechanisms. However, evidence of its effectiveness remains inconsistent. We conducted a systematic review and meta-analysis of studies published until February [...] Read more.
Lycopene, a carotenoid found in tomatoes and watermelon, has been investigated for its potential to improve male fertility through its antioxidant and anti-inflammatory mechanisms. However, evidence of its effectiveness remains inconsistent. We conducted a systematic review and meta-analysis of studies published until February 2025 in the PubMed, Scopus, Web of Science, and Medline databases. Clinical studies evaluating lycopene supplementation in relation to male fertility outcomes were included in this review. Standardized mean differences (SMDs) were calculated for the key outcomes. Four clinical studies involving 151 participants were included. Lycopene supplementation significantly improved sperm concentration (SMD 0.33, 95% CI [0.02–0.65], p = 0.037) and nonprogressive motility (SMD 0.45, 95% CI [0.04–0.87], p = 0.032). No statistically significant effects were observed on total motility, progressive motility, normal or abnormal morphology, semen volume, or DNA damage. Sensitivity analyses showed that the findings were generally robust, although publication bias and methodological heterogeneity were noted. Lycopene supplementation may offer modest benefits in improving sperm concentration and nonprogressive motility in men. However, evidence for other fertility-related outcomes is inconclusive. Larger, high-quality randomized trials are needed to confirm these findings and clarify the role of lycopene in male reproductive health. Full article
(This article belongs to the Special Issue Advanced Research in Immunotoxicity and Oxidative Stress)
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20 pages, 792 KiB  
Article
Could Individual Variability in Resistance to Cryopreservation (“Freezability”) Serve as a Biomarker Reflecting Boar Fertility?
by Eduardo de Mercado, Helena Nieto-Cristóbal, Adrián Martín-San Juan, María José Martinez-Alborcia and Manuel Álvarez-Rodríguez
Animals 2025, 15(15), 2180; https://doi.org/10.3390/ani15152180 - 24 Jul 2025
Viewed by 431
Abstract
Subfertile boars often go undetected until they cause significant reproductive losses. Current semen quality assessments are limited in their ability to predict fertility, highlighting the need for complementary biomarkers. This study explored whether semen freezability could serve as an indirect indicator of boar [...] Read more.
Subfertile boars often go undetected until they cause significant reproductive losses. Current semen quality assessments are limited in their ability to predict fertility, highlighting the need for complementary biomarkers. This study explored whether semen freezability could serve as an indirect indicator of boar fertility. Eighteen boars were classified based on historical fertility records and semen freezability, assessed by post-thaw quality. Fresh and post-thaw semen samples were analyzed using the CASA system and fluorescence microscopy. High-fertility boars showed significantly better motility and functional sperm parameters in fresh semen compared to low-fertility boars. However, these differences were mostly lost after cryopreservation. Conversely, boars with good freezability had consistently better post-thaw semen quality, though this did not correlate directly with higher fertility outcomes. Notably, a combined analysis revealed that boars with both high fertility and poor freezability had the lowest post-thaw semen quality. This suggests that cryopreservation may expose hidden sperm defects not detectable in fresh semen. Total motility was the only parameter associated with both fertility and freezability. In conclusion, while freezability alone may not directly predict fertility, it may help identify low-performing males. The combined assessment of fresh semen motility and freezability could support more effective boar selection strategies. Full article
(This article belongs to the Section Animal Reproduction)
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