Search Results (266)

This study aimed to investigate the effects of supplementing with an essential oil blend and ascorbic acid on performance, semen characteristics, antioxidant status, gut microbiota, immunity, and gene expression in heat-stressed male rabbits. One hundred and forty male New Zealand White rabbits, aged 6 months, were randomly distributed into four dietary groups: the control group receiving basal feed (CON), the group receiving an essential oil blend (EOB, 200 mg/kg), the group receiving ascorbic acid (ASA, 1000 mg/kg), and the group receiving an essential oil blend and ascorbic acid (MAO). The experimental period lasted for eight weeks. The MAO mixture supported rabbits’ tolerance to heat stress by enhancing stress markers, as demonstrated by decreased glucose and HSP70 and increased triiodothyronine (T3). In addition, increased body weight, carcass weight, and nutrient digestibility, but reduced mortality rate, were observed in rabbits fed the MAO mixture. Additionally, semen density and volume, as well as sperm progressive motility and normality, were enhanced in rabbits fed the MAO mixture. Furthermore, MAO mixture supplementation decreases plasma cholesterol, triglycerides, AST, urea, and creatinine levels, while raising HDL and total protein levels. Adding MAO mixture contributed to an increase in plasma SOD and GPx levels, as well as seminal fluid TAC and GSH levels. Additionally, the incorporation of MAO increases IgA, IgG, and IL-10 levels while decreasing IL-6 and TNF-α levels. Adding the MAO mixture reduced C. perfringens and E. coli, as well as increased the expression of the MUC-2, CAT-1, and CLDN-1 genes. Combining an essential oil blend and ascorbic acid may contribute to improvements in performance, semen quality, immune response, antioxidant capacity, and gut health in heat-stressed male rabbits. Full article

Sperm quality influences bovine in vitro embryo production (IVEP). Linear regression is a statistical tool that models the relationship between a dependent variable and one or more independent variables. It can be used to predict outcomes, analyze trends, and understand the impact of variables. These models are useful for indicating which sperm variables most influence IVEP results, facilitating the selection of superior samples to enhance IVEP. Using early IVEP indicators, such as cleavage rate, can assist in scheduling recipient preparation. This work aimed to construct linear regression models to study the influence of a comprehensive set of sperm variables and cleavage rate on IVEP yields. A dataset comprising 51 semen batches from 23 Nellore bulls was compiled, including 26 sperm variables from computer-assisted sperm analysis (CASA) and flow cytometry per batch, with 184 IVEP procedures. The most robust predictive model had a coefficient of determination of 0.6358; furthermore, the BULL variable was the most influential predictor, yielding an independent coefficient of determination of 0.5218. Models that were exclusively founded on sperm analysis yielded meager coefficients of determination (<0.04). However, to predict the best batch from a bull, individual models achieve coefficients of determination ranging from 0.58 to 0.99. Contributions, impacts, and positive or negative correlations of various sperm variables with in vitro performance were influenced by the bull. We conclude that the BULL variable was the dominant predictor of in vitro performance, with cleavage rates serving as an early estimator of blastocyst rates. The predictive utility of analyzed sperm traits remains limited. Nonetheless, individualized models offer a valuable tool for selecting optimal batches for preferred bulls within IVEP laboratories, culminating in heightened blastocyst rates. Full article

Members of the genus Brucella are bacterial pathogens of global importance, and their increasing detection in marine mammals has raised concerns for wildlife conservation and public health. In this study, we evaluated the biological and pathogenic characteristics of two Brucella sp. sequence type 27 (ST27) isolates obtained from a dwarf sperm whale (Kogia sima). We compared them with terrestrial and marine Brucella reference strains. We assessed resistance to polymyxin B and human serum complement, intracellular infection dynamics in HeLa epithelial cells, persistence in a murine model, and associated hematological and histopathological changes, and analyzed lipopolysaccharide (LPS) profiles. The Kogia isolates exhibited resistance to polymyxin B and serum complement, comparable to that of B. abortus 2308W and marine mammal Brucella strains. In HeLa cells, the isolates displayed distinct, strain-specific intracellular infection dynamics. In the murine model, both isolates persisted in the spleen and induced granulomatous lesions. However, splenic bacterial loads and histopathological scores were generally lower than those observed with B. abortus 2308W, which exhibited the highest virulence among the strains evaluated. Hematological alterations associated with Kogia isolates were also less pronounced than those induced by B. abortus 2308W, indicating an intermediate and strain-dependent virulence phenotype without evidence of enhanced virulence relative to the terrestrial reference strain. Western blot analyses showed that Brucella sp. ST27 isolates were not recognized by anti-B. abortus or anti-O-antigen monoclonal antibodies, while exhibiting a distinct recognition pattern with anti-B. canis serum, indicating differences in surface antigen composition. Comparative whole-genome analysis identified a limited number of isolate-specific variants affecting coding and intergenic regions. Collectively, these findings highlight phenotypic and genetic features of Brucella sp. ST27 from Kogia sima, which distinguishes it from other marine and terrestrial Brucella strains and supports further investigation into its biological behavior and potential public health relevance. Full article

In this study, the ultrastructural characteristics of the mature spermatozoon of the cestode Gerbillitaenia psammomi, a parasite of the fat sand rat (Psammomys obesus) in the south of Tunisia, were examined by means of transmission electron microscopy. The spermatozoon is of Levron et al.’s type VI. This type of sperm cell is mainly characterized by the presence of a single axoneme of the 9 + ‘1’ pattern typical of the trepaxonematan Platyhelminthes, crest-like bodies, a periaxonemal sheath, twisted cortical microtubules, and a spiraled nucleus. The spermatozoon of G. psammomi possesses two crest-like bodies of different length. The results obtained in the present study suggest certain similarities with the remaining analyzed catenotaeniids; namely, Catenotaenia pusilla, Skrjabinotaenia (Meggittina) gerbilli, and Spasskijela lobata. The results are compared and discussed according to several characteristics found in the catenotaeniids and other studied cestodes, particularly those of the order Cyclophyllidea. Full article

Environmental hazard-induced male infertility has become a major public health issue. The concealment and accumulation of environmental hazards, and their interactions with the endogenous immune network, have long been underappreciated. As the central organ for sperm maturation and motility acquisition, the epididymis plays a vital role in male fertility, and the homeostasis of the epididymal immune microenvironment (EIM) is essential. Nevertheless, a systematic synthesis of common environmental hazards and their impact on EIM, which can lead to male infertility, remains lacking. This review comprehensively summarizes the composition, functionality, and key characteristics of the EIM and underscores its critical role in preserving male reproductive health. We further evaluate and delineate the disruption of EIM homeostasis resulting from major categories of environmental exposures—including chemical, physical, biological, and behavioral hazards—and discuss their shared pathophysiological mechanisms. By integrating evidence linking environmental insults, EIM dysregulation, and male infertility, this work aims to identify pivotal molecular mechanisms from an immunological perspective. The findings provide a mechanistic foundation for the development of targeted interventions and preventive strategies against environmental hazard-induced male infertility. Full article

Over the past few decades, the production and application of nanoparticle-contained products have been increasing. With increasing production, nanoparticles (NPs) entered water and terrestrial environments, posing a threat to living organisms because their ecotoxicological characteristics are not yet fully understood. Upon entering the marine environment, NPs are subjected to various factors that can alter their properties. This could lead to changes in their toxic effects on marine organisms. One potential physical factor that affects NPs is UV radiation. The toxicity of different concentrations of UV-pre-irradiated TiO₂ NPs on dollars Scaphechinus mirabilis sperm was studied, which allowed comparison of the effects of UV-activated and non-activated NPs. According to the resazurin and comet assays, a significant toxic effect is observed at lower concentrations for pre-irradiated TiO₂ NPs compared to non-irradiated ones. Additionally, high concentrations of pre-irradiated TiO₂ resulted in a significant increase in levels of malondialdehyde (MDA) compared to non-irradiated. Thus, it was demonstrated that the UV pre-irradiation NPs caused a more pronounced toxic effect than the non-irradiated TiO₂ NPs. Full article

Parassi mullet (Mugil incilis) is an ecologically and economically important species that supports small-scale artisanal fisheries. However, scarce knowledge of its reproductive biology limits the development of management and conservation strategies. This research describes key milt and sperm characteristics, including milt volume, sperm concentration, motility, and ultrastructural features. Males produced an average of 40.0 ± 20 µL of milt, with sperm concentrations between 6.00 and 20.37 × 10⁹ spermatozoa mL⁻¹. Sperm motility varied between 10% and 80%, with a mean duration of 14.13 ± 4.49 min. Mature spermatozoa measured 33.79 ± 0.67 µm and exhibited a subspherical head without an acrosome, a short midpiece, and a cylindrical flagellum. The nucleus contains electron-dense heterogeneous chromatin. The centriolar complex was positioned outside the nuclear fossa consistent with Type II spermiogenesis. The flagellum comprises a main piece and tapering end piece. The axoneme had 9 + 0 arrangement at the basal body region and the typical 9 + 2 configuration along its length. These results provide the first detailed description of sperm morphology in parassi mullet and contribute to an understanding of its reproductive biology, supporting future applications in taxonomy, toxicology, conservation and aquaculture programs. Full article

Background and Objectives: The association between different anesthesia modalities and spermiogram parameters and reproductive outcomes in patients undergoing microscopic subinguinal varicocelectomy (MSV) remains unclear. In this retrospective cohort study, we aimed to compare spermiogram parameters and pregnancy rates between patients receiving general anesthesia (GA) versus spinal anesthesia (SA) for MSV with 2-year follow-up data. Materials and Methods: Male patients aged between 18–50 years, with ASA physical scores between I–III, who underwent unilateral or bilateral primary MSV, were included in the study. To minimize selection bias and balance the baseline characteristics between the GA group and SA group, we employed a propensity score matching approach, matching all 38 SA patients with 380 GA patients selected from a larger pool. Patients with complete 24-month follow-up data were included in the final analysis. The primary outcome of our study was determined as evaluating sperm count changes. Secondary outcomes included other sperm parameters (motility, morphology and semen volume), natural pregnancy rates, perioperative complications and recovery parameters. Results: The final analysis included 418 patients who met all inclusion criteria and completed the follow-up period. The study population comprised 380 patients in the GA group and 38 in the SA group. No significant difference was found between the groups in terms of sperm count. Greater improvement in sperm motility was observed in the SA group starting from the third month onwards (p = 0.027). Natural pregnancy was achieved in 16/38 (42.1%) of SA patients versus 125/380 (32.9%) of GA patients (p = 0.031). In addition, better results were obtained in terms of recovery parameters in the SA group. Other results were comparable between the groups. Conclusions: Spinal anesthesia for MSV was associated with greater improvement in sperm motility and higher natural pregnancy rates compared to general anesthesia, despite comparable sperm count improvements. These associations warrant further investigation in prospective randomized trials. Full article

Artificial insemination (AI) is a key reproductive biotechnology for genetic improvement in sheep. However, its efficiency remains lower and more variable than in most other livestock species. This review critically synthesizes the historical foundations of sheep AI, including methodological principles established by the Soviet school, and evaluates how these concepts have been further developed and adapted to contemporary reproductive biology. Particular emphasis is placed on estrous synchronization protocols, semen processing and cryopreservation, and insemination techniques. We highlight how anatomical constraints of the ovine cervix, seasonal reproductive physiology, and species-specific characteristics of ram sperm collectively limit fertility outcomes, especially when frozen–thawed semen is used. Comparative analysis of cervical, transcervical, and laparoscopic insemination methods indicates that laparoscopic AI remains the most reliable approach, although recent advances in catheter design and semen handling have improved the feasibility of less invasive techniques. This review further discusses emerging approaches, including sperm sex-sorting, alternative recovery methods, and early-stage spermatogonial stem cell–based technologies, emphasizing both their potential applications and current limitations. Overall, the available evidence suggests that future progress in sheep AI will depend on the integrated optimization of hormonal synchronization, semen preservation, and insemination strategies, rather than on isolated technical innovations. Full article

In vitro manipulation of human spermatozoa during Assisted Reproductive Technology (ART) can induce several damages to sperm structure and functions. This study investigated the protective effects of Raphanus sativus seed extract and its active compounds on several sperm parameters during in vitro incubation and cryopreservation. Extracts from five seed-batches were characterized by HPLC-DAD-MS and ¹H-NMR, identifying sinapine and sinipic glycosides as the main characteristic compounds. Sperm DNA fragmentation (sDF) was detected by the Sperm Chromatin Dispersion test and LiveTUNEL. Excessive reactive oxygen species (ROS) production was detected by MitoSOX Red in viable spermatozoa. Caspase activity was detected by FLICA. Cryopreservation was conducted with two alternative freezing media. In vitro incubation with the extract protected against the loss of motility and reduced the induction of sDF, sperm ROS production, and caspase activity. Similarly, during cryopreservation, it allowed much better recoveries of sperm viability, motility, and DNA integrity by decreasing sperm ROS production with both freezing media. Sinapine and sinapic acid completely mimicked the protective effects of the whole extract during both in vitro incubation and cryopreservation, suggesting that they are included among the active principles. These findings support Raphanus sativus seed extract and its active compounds as candidates for inclusion in handling and freezing media for human spermatozoa in ART. Full article

The present study aimed to investigate the effect of Turnera diffusa supplementation on the quantitative and qualitative characteristics of semen in Duroc boars (n = 4). The experiment was divided into two periods, each corresponding to the duration of one spermatogenic cycle: a control period (40 days) (CP) and an experimental period (40 days) (EP). Nutrition and environmental conditions were kept constant throughout both periods. During the experimental period, each boar received a daily supplement of 7 g of Turnera diffusa extract. In each period, five ejaculates were collected from each boar included in the study. The ejaculates were evaluated for volume, sperm concentration, motility, agglutination, number of insemination doses obtained per ejaculate after dilution, and sperm viability after 24, 48, and 72 h of storage. The results of a two-way repeated measures ANOVA showed that the combined effect of boar × treatment significantly influenced ejaculate volume (p < 0.01) and viability after 48 h of storage (p < 0.05). The results of the two-way repeated measures ANOVA showed that treatment with the tested additive T. diffusa significantly affected sperm survival during storage for 24 h (p < 0.01), 48 h (p < 0.001), and 72 h (p < 0.05). Bonferroni post hoc analysis indicated that T. diffusa significantly affected only the parameters related to sperm viability, namely survival rates at 24 h (p < 0.001), 48 h (p < 0.01), and 72 h (p < 0.01). The findings of this study demonstrate that the application of the tested supplement, at the specified dose and duration, has a positive effect on semen quality in boars. Full article

Haloarchaea are moderate and extreme halophilic microorganisms inhabiting hypersaline environments characterised by high ionic and oxidative stress due to extremely high salt concentrations and high incidence of UV radiation (mainly in spring and summer). To be alive and metabolically active under these harsh conditions, haloarchaeal strains have developed molecular adaptations, like hyperpigmentation. Among the carotenoids produced by haloarchaeal species, the C₅₀ carotenoid called bacterioruberin (BR) and its derivatives, monoanhydrobacterioruberin and bisanhydrobacterioruberin, are the predominant natural pigments produced. This review aims to highlight the most significant characteristics of BR and their derivatives, as well as a description of the biological activities already reported that could provide benefits for human health, including antitumoral, immunomodulatory, antioxidant, skin protectant, antilipidemic, antiglycemic, and anti-atrophic effects, in addition to showing potential positive effects on sperm cells cryopreservation. Overall, C₅₀ carotenoids are fascinating natural biomolecules that could be utilised in processed food and nutraceuticals or as tools in the context of new strategies and/or pharmaceutical formulations to combat various human diseases or metabolic disorders. Full article

Background/Objectives: In digenic populations, all females produce males and females in their offspring. Monogenic populations are composed of gynogenic (female-producing) and androgenic (male-producing) females. A theoretical population genetic model for evolution of digenic to monogenic populations is presented here. Methods: A controlling gene was associated with each of the four processes that characterise monogenic populations: (1) oogenesis is conventional, whereas spermatogenesis is unusual and it is characterised by the exclusive formation of X-bearing sperm (gene (s)), i.e., the paternal chromosomes are eliminated so that only the maternal ones are transmitted to the next generation; (2) the X chromosome that is eliminated in the zygote is the one inherited from the father (gene r); (3) an imprinting process occurs in the mother (gene g), which protects the maternally inherited X chromosome from elimination in the zygote and the whole maternal chromosome complement in spermatogenesis; (4) a maternal factor is produced during oogenesis (gene e), which inactivates the elimination factor [r] in the zygote, thus controlling the elimination of the paternal X chromosome. The sequences of emergence of the genes (e s r g) that transform a digenic population into a monogenic one were analysed. Results: The following evolutionary sequences were found: (1) the sequence (r s e) under dominant conditions of gene (s) and recessive conditions of gene (r); and (2) the sequences (s r e), (r s e), and (e s r) under recessive conditions of gene (s) and gene (r). It was also found that the process of genomic imprinting is a necessary condition for the generation of a monogenic population. Furthermore, a quantitative change in the interaction between the elimination factor and its maternal inhibitor modifies the genotypic formula of the monogenic state. Conclusions: The number and types of evolutionary transitions of a digenic to a monogenic population depends on the dominant or recessive characteristic of the newly emerging genes. The imprinting process must already be present in the digenic population from which the monogenic one evolves; otherwise, the population cannot reach the monogenic state. Full article

Background/Objectives: Acephalic spermatozoa syndrome (ASS) is a rare subtype of male infertility characterized by headless sperm due to defective head–tail coupling. Genetic factors are recognized as the primary etiology of ASS; however, known pathogenic mutations only explain a subset of ASS cases. Further investigations are required to elucidate the underlying genetic pathogenesis of ASS and the implications of such genetic defects for assisted reproductive technology (ART) outcomes. This study aimed to identify a novel PMFBP1 mutation in an ASS patient; investigate the effects of the identified mutation on sperm ultrastructure and PMFBP1 protein expression/stability, and assess ART outcomes using the patient’s sperm. Methods: One 34-year-old infertile male with ASS was enrolled. Genetic screening was performed via whole-exome sequencing (WES), followed by Sanger sequencing for mutation validation. Sperm morphological characteristics were evaluated using Diff-Quik staining (for general morphology), transmission electron microscopy (TEM, for ultrastructural analysis), and peanut agglutinin (PNA) staining. Protein expression and stability were analyzed by Western blot and cycloheximide (CHX)/MG132 assays. ART outcomes were compared between the in vitro fertilization (IVF) cycles using the patient’s sperm and those using donor sperm. Results: In IVF cycles, donor sperm achieved normal fertilization (characterized by two pronuclei [2PN] formation), whereas the patient’s sperm failed to form 2PN and leading to embryo fragmentation. Genetic sequencing identified a novel nonsense mutation in PMFBP1 (c.2641C>T), which introduced a premature stop codon and resulted in a premature protein product (p.Arg881Ter). Morphologically, this mutation led to complete sperm head–tail detachment, and abnormalities in acrosome structure and sperm head–neck junction. The absence of PMFBP1 protein in the patient’s spermatozoa was observed. The in vitro assays showed the c.2641C>T mutation induced expression of the truncated PMFBP1 protein and significantly altered PMFBP1 protein stability. Conclusions: The PMFBP1 c.2641C>T mutation impairs sperm head–tail adhesion, thereby contributing to the pathogenesis of ASS. This study expands the clinical mutational spectrum of PMFBP1-associated male infertility and provides valuable insights for the genetic diagnosis of ASS patients. Additionally, these findings may lay a foundation for the choice of therapeutic strategies targeting PMFBP1-related ASS. Full article

Background/Objectives: Approximately 50% of infertility cases are attributable to male factors; yet conventional semen examination can not identify the molecular abnormalities that hinder sperm functionality. Extracellular vesicles (EVs) derived from sperm, such as testicular EVs, prostasomes, and epididymosomes, have become important modulators of oocyte activation, sperm maturation, capacitation, acrosome stability, motility, and early embryonic development. This study aimed to evaluate the potential diagnostic and translational uses of sperm-associated extracellular vesicles (EVs) in male infertility and assisted reproduction, while also consolidating recent insights on their origins, composition, and functional significance. Methods: A focused narrative search of PubMed (2000–2025) was conducted using backward and forward citation tracking. Studies that qualified included human clinical cohorts, functional sperm extracellular vesicle tests, and omics analyses using MISEV-aligned extracellular vesicle isolation and characterisation methodologies. When human mechanistic understanding was constrained, knowledge from animal research was selectively integrated. Results: The cargo signatures specific to the source identified in sperm-derived and seminal EVs encompass proteins, small RNAs, lipids, and enzymatic modules that govern sperm maturation, capacitation, acrosome reaction, redox balance, calcium signalling, zona binding, and DNA integrity. Density-resolved seminal extracellular vesicle subfractions (EV-H/EV-M/EV-L) have unique functional and proteomic characteristics linked to progesterone-induced hyperactivation, oxidative stress, and motility. Asthenozoospermia and oligoasthenoteratozoospermia are associated with changes in extracellular vesicle composition, reduced embryonic developmental potential, compromised oocyte activation (related to PLCζ), and increased sperm DNA fragmentation. Numerous EV-related miRNA and protein signatures may predict TESE results, identify functional sperm anomalies not recognised by conventional semen analysis, and differentiate between obstructive and non-obstructive azoospermia. Conclusions: The available findings indicate that sperm-derived extracellular vesicles are significant functional regulators of sperm physiology and may serve as valuable non-invasive indicators for male infertility. The standardisation of EV isolation, characterisation, and clinical validation is essential prior to widespread use; nonetheless, their integration into liquid biopsy methods and assisted reproductive technology processes represents a significant improvement. Full article