Cell-free DNA (cfDNA) fragments represent emerging biomarkers of major interest in the assessment of male fertility and the management of couples undergoing assisted reproductive technology (ART). Easy to quantify and present in most biological fluids, seminal cfDNA could reflect apoptosis and necrosis occurring within the male reproductive system. In this study, we compared cfDNA concentrations, measured by means of qPCR targeting the RNase P gene, in the seminal plasma of men with severe sperm quality abnormalities, including very severe asthenozoospermia (n = 25), total azoospermia (n = 25), very severe oligozoospermia (n = 25), and very severe teratozoospermia (n = 25), with those of normozoospermic controls (n = 25). Mean cfDNA concentrations were particularly elevated in men with azoospermia (5.47 ± 1.17 µg/mL) and very severe teratozoospermia (3.25 ± 1.21 µg/mL), compared with controls (1.96 ± 0.27 µg/mL). After adjustment for multiple testing, only azoospermia and very severe teratozoospermia remained significantly associated with increased cfDNA levels (Holm–Bonferroni adjusted p < 0.003), whereas the other phenotypes did not maintain statistical significance. These results suggest that seminal cfDNA is markedly increased in the most severe forms of spermatogenic impairment, potentially reflecting enhanced germ cell apoptosis or testicular cell involvement. Despite some limitations, including the modest sample size and single-center design, our observations support the value of cfDNA as a promising non-invasive biomarker of sperm quality. Larger multicenter and longitudinal studies are needed to validate its clinical relevance and determine standardized reference thresholds for clinical application.

The SPINK2 protein, encoded by the SPINK2 gene, plays an essential role in the normal development of spermatozoa, and its deficiency is associated with spermatogenesis disorders ranging from aspermia to azoospermia. This study aimed to identify the most deleterious variants of the SPINK2 gene and to evaluate their effects on protein structure and function through an in silico approach. A total of 8,028 variants were identified, including 72 missense variants. Using 11 bioinformatics tools, six variants (P50L, T58I, C66Y, E62A, P42S, and P45L) were predicted to have deleterious effects. Protein-protein interaction analysis using the STRING database revealed strong functional associations between SPINK2, SPINK1, and ACR, and medium-confidence associations with SPINK4, SPINK13, PMPCA, KLK4, SPINK9, SPINK6, SPACA1, and NUDT8. Local structural analysis showed that variants such as T58I and C66Y gained additional hydrophobic interactions, whereas P50L and P42S lost key interactions, potentially impairing protein stability and function. Molecular dynamics simulations using GROMACS revealed that P50L enhances protein stability, reduces amino acid flexibility, and increases the overall dimensions of the protein. T58I had a mild effect on stability, whereas E62A and C66Y decreased stability and flexibility while increasing protein size. P42S and P45L induced slight stability alterations, reduced flexibility, and enlarged the protein. Overall, these structural and dynamic changes suggest functional impairment of SPINK2. To our knowledge, this is the first study to identify six deleterious SPINK2 variants with potential roles in the disruption of spermatogenesis, providing a foundation for future functional and clinical investigations.

Aquaculture has evolved as one of the most dynamic industries in food production, representing the fastest-growing activity in meeting global food demand. Nevertheless, its rapid expansion is accompanied by significant challenges, including water pollution and the proliferation of pathogens that induce stress in aquatic organisms, leading to disease outbreaks and high mortality rates. To mitigate these problems, antibiotics and chemical agents are widely used to control infections. However, their excessive application results in residual contamination and promotes the emergence of drug-resistant bacterial strains. As a sustainable and environmentally friendly alternative, probiotics, prebiotics, and phytobiotics have attracted growing interest as substitutes for conventional chemotherapy in aquaculture. These functional feed additives enhance disease resistance and exhibit diverse bioactivities, such as antibacterial, antiviral, antifungal, and antiparasitic effects. Additionally, they improve growth performance, strengthen immune responses in cultured species, and contribute to better water quality. This review synthesizes current findings on the role of probiotics, prebiotics and phytobiotics in advancing sustainable aquaculture practices worldwide, while critically discussing their limitations, such as species-specificity, dose and duration responses, and potential long-term risks, thereby providing valuable insights to guide future research and innovation toward environmentally responsible and health-promoting solutions in the aquaculture industry.

Exopolysaccharides produced by bacteria, particularly lactic acid bacteria, have emerged as versatile polysaccharides with significant structural diversity, functional properties, and broad applicability. This review explores the classification and biosynthesis of bacterial EPSs, alongside current methods used for screening EPS-producing strains, and the isolation, purification, and quantification of these carbohydrate polymers. It also highlights advanced analytical techniques, including GPC, GC-MS, XRD, FTIR, and NMR used to characterize their molecular architecture. Structural features such as monosaccharide composition, molecular weight, and glycosidic linkages are considered in relation to their potential influence on the biofunctional properties of EPS. These include prebiotic, antimicrobial, antioxidant, and immunomodulatory effects. Furthermore, EPSs have demonstrated considerable promise in therapeutic delivery systems. Their wide-ranging applications extend the food industry, cosmetic formulations, and agricultural practices. Compared to plant, animal, or algae-derived polysaccharides, bacterial EPSs offer the advantages of controlled, year-round production and high biocompatibility. Through this comprehensive review, we aim to highlight the potential of LAB-derived EPSs as sustainable and multifunctional biopolymers, paving the way for innovative solutions in health, food, and environmental biotechnology.

The intestinal microbiota of fish represents a valuable source of antimicrobial and health-promoting bacteria. In this context, we report for the first time the isolation of Bacillus siamensis strain AFSC23 from the intestine of Conger conger. The strain was comprehensively characterized using an integrated genomic and functional approach to assess its safety and probiotic potential in aquaculture. The strain exhibited resistance to lysozyme (100 mg/L), gastrointestinal conditions, bile salts (0.3

This study aims to identify antioxidants capable of protecting cells against oxidative stress. We evaluated Euphorbia paralias, a Mediterranean coastal plant rich in polyphenols, for its protective potential against hydrogen peroxide (H2O2)-induced oxidative stress in Tetrahymena pyriformis for the first time. A combined in vitro and in silico approach was employed. In vitro, the minimum effective concentration of the extract was determined, and its effects on growth, swimming speed, morphophysiological parameters, and key biochemical markers: catalase (CAT), superoxide dismutase (SOD), and malondialdehyde (MDA) were analyzed. In silico, molecular docking studies assessed the affinity of the main compounds for antioxidant enzymes (Keap1, SOD, and CAT) and their potential toxicity. Results demonstrated that the extract enhances cell growth and mobility, significantly activates antioxidant enzymes, and inhibits lipid peroxidation. Docking studies indicated a high affinity of the compounds for enzyme targets and low to negligible acute toxicity. These results suggest that Euphorbia paralias could be a promising source of antioxidants and highlight the value of an integrative approach combining in vitro and in silico studies to explore protective mechanisms against oxidative stress.

Carbapenem-resistant Gram-negative bacteria (CR-GNB) have experienced an alarming surge in prevalence in recent years, escalating into a critical global healthcare crisis. As carbapenems represent the last line of defense against such pathogens, infections caused by CR-GNB have become increasingly challenging to treat, given the restricted therapeutic options and heightened mortality risks. The discovery and development of alternative therapeutic strategies that present novel avenues against multi-drug-resistant organisms are gaining increased attention, presenting a pressing need for innovative solutions. Our comprehensive review delves into the multifaceted landscape of carbapenem resistance in Gram-negative bacteria in response to this urgent challenge. The scope of this review aims to provide an up-to-date and in-depth exploration regarding the mode of action of carbapenem and the resisting mechanisms of carbapenem in GNB. Additionally, it discusses the state of the art of some clinical therapies for the treatment of infections caused by CR-GNB. Moreover, it describes several combinational and alternative therapies to combat CR-GNB, including the computational approach of “molecular docking”. In light of the conclusions of this review, we call for the implementation of these strategies to develop comprehensive approaches to mitigate carbapenem resistance in Gram-negative bacteria.

Gluten-free baked goods exhibit reduced texture and taste characteristics compared to their gluten-containing counterparts. As a result, there is a renewed interest in the fermentation of gluten-free cereals with lactic acid bacteria, which is associated with an improvement in the final baked goods. Quinoa is garnering growing attention due to its different nutrients and bioactive substances, and it is notably employed to build gluten-free goods. In the present study, quinoa flour was fermented with Enterococcus strains (E. gallinarum SL2 and E. mundtii SL1), and further used in the manufacturing of gluten-free muffins. Several analyses were performed on the obtained sourdoughs and muffins, including a viscosity study, a textural and sensory analysis, and a polyphenol, organic acid, and carbohydrate content analysis. The results showed that the fermented quinoa flour exhibited enhanced nutritional value, with increased levels of organic acids such as lactic and acetic acid, as well as improved polyphenol content. The sensory and textural analyses revealed that both Enterococcus strains positively impacted the sensory characteristics and texture of the muffins. Notably, muffins prepared with E. mundtii SL1 demonstrated superior elasticity and overall taste. These results suggest that fermentation with these strains can significantly improve the nutritional profile and sensory quality of gluten-free baked goods, offering a promising approach for the development of healthier and more appealing gluten-free products.

Carbapenem resistance genes in Gram-negative bacteria (CR-GNB) are a major cause of critical infections and are considered an urgent public health concern. The present study aimed to describe the prevalence of CR-GNB and the dissemination of extended-spectrum beta-lactamase (ESBL) and carbapenemase genes in clinical isolates from Casablanca, Morocco. Firstly, the strains were collected and identified using phenotypic and biochemical methods, then the antibiotic susceptibility was evaluated by the disc diffusion assay to screen isolates resistant to carbapenems. Secondly, three traditional methods, the carbapenem inactivation method, the modified Hodge, and the in-house carba-NP, were performed to predict the carbapenemase production by the included strains. Finally, conventional PCR was utilized to validate and detect the carbapenemase- and ESBL-related genes. Concerning the results, out of the identified 122 strains, 48 were CR isolates, including 30 Klebsiella pneumoniae, 13 Escherichia coli, and 5 Pseudomonas aeruginosa. Furthermore, these strains presented a high level of resistance. Moreover, the prediction of carbapenemase production by the phenotypic methods showed variable results. Also, the PCR analysis revealed a high occurrence of β-lactamase (ESBL and carbapenemase) genes in the included clinical strains, and most strains harbored multiple resistance genes. Our findings suggest that the three existing methods have some limitations, and a validation study is still necessary for the carbapenemase diagnostics.

The production of β-lactamases is the main mechanism underlying carbapenem resistance. This study combined in silico and in vitro approaches to identify potential polyphenols as carbapenemase inhibitors. Molecular docking, molecular dynamics, and ADMET prediction were performed to assess the binding affinity, stability, and safety of quercetin, kaempferol, caffeic acid, and 3,4-dihydroxybenzoic acid against KPC-2, NDM-1, and OXA-48 carbapenemases. In vitro antibacterial assays and checkerboard analyses were conducted against Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeruginosa to assess antibacterial and synergistic effects. Then, the inhibition of the β-lactam hydrolytic activity was confirmed. In silico results showed that quercetin, kaempferol, and caffeic acid exhibited strong binding affinity and consistent stability towards the targets. Therefore, quercetin and kaempferol showed the strongest affinities (−8.0 kcal/mol) and stable interactions with key catalytic residues. ADMET profiles indicated good pharmacokinetic behavior and low acute toxicity. In vitro assays revealed that the polyphenols exhibited MIC values ranging from 12.5 to 25 mg/L and MBC values of 25–50 mg/L. Combined with cefotaxime, they enhanced bacterial susceptibility and inhibited β-lactam hydrolysis, with quercetin achieving complete inhibition at 200 mg/L. These findings highlight the potential of the four polyphenols as natural β-lactamase inhibitors. Further enzyme kinetics and in vivo studies are needed to confirm their therapeutic relevance.

Dysphania ambrosioides essential oil (EO) possesses significant antibacterial and antioxidant properties. However, its application as a food preservation agent is limited due to high volatility and instability. Given the industrial relevance of this EO, developing new products that incorporate microencapsulated D. ambrosioides EO is recommended. This study addresses these challenges by encapsulating the EO using inulin and gum arabic (IN/GA) biopolymers, known for their biocompatibility and biodegradability. We systematically evaluated the encapsulation efficiency and structural properties of the resulting microcapsules. Advanced characterization techniques, including FT-IR, SEM, and EDX, were used to analyze the chemical interactions and morphological characteristics of the microcapsules. The thermal stability of the microcapsules was assessed using TGA, while their stability and bioaccessibility were evaluated under simulated in vitro digestion conditions. The formulation (C1) used in this study demonstrated a high encapsulation efficiency (88 %). The IN/GA formulations successfully microencapsulated EO and alpha-terpinene, producing microcapsules with high stability (>80 %) and bioaccessibility (>40 %). These microcapsules showed controlled release during digestion and exhibited strong antibacterial activity against Staphylococcus aureus and Escherichia coli. These findings suggest that inulin and gum arabic are effective macromolecules for stabilizing this EO, offering valuable potential applications in the food industry.

This study aimed to explore the phytochemical analyses and antibacterial potential of three essential oils (EOs) using both in-vitro and in-silico experiments for drug discovery. The Eos of Rosmarinus officinalis, Lavandula angustifolia, and Salvia officinalis were extracted by hydrodistillation, and GC–MS determined the chemical composition. The antibacterial activities of each EO were studied by the disc diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of these EOs were determined by micro-dilution. In addition, the EOs were combined with ineffective antibiotics (Aztreonam et al.) against clinical bacteria (P. aeruginosa and K. pneumoniae) to determine this association's effect. The in-vitro toxicological study was based on the MTT cytotoxicity assay. Finally, in-silico methods were employed to estimate their possible antibacterial mechanisms. Molecular docking was performed to calculate the predictive binding affinities of five major volatile components to three proteins key in the bacterial cycle using the AutoDock Vina program; prediction of drug-likeness properties and Toxicity prediction were carried out respectively by SwissADME and ProToxII online server. The results showed that the three EOs possessed antibacterial activity against ATCC and clinical K. pneumoniae. We note that P. aeruginosa was resistant to lavender and sage EOs. Surprisingly, adding EOs to antibiotics ineffective against resistant bacteria showed a significant synergistic antibacterial effect. Cytotoxicity assay of essential oils showed different values ranging from 0.22 to 0.43 µL/mL. Finally, the molecular docking study revealed that both compounds (1,8-Cineol, 1-Dodecene, Linalool, cis-Thujone, and Camphor) from the three EOs have a significant potential to inhibit the protein target involved in bacteria resistance. Furthermore, the SwissADME prediction results showed that all five components satisfy the rule of five and exhibit acceptable drug-like characteristics. findings suggest that the three EOs have interesting antibacterial activity. The in-vitro and in-silico studies gave a great potentiation and may constitute a promising option to control the emergence of MDR P. aeruginosa and K. pneumoniae.

Squalene synthase (SQS) plays a crucial role in the cholesterol biosynthetic pathway. Its distinctive strategic position makes it a promising candidate for targeting and developing new anti-hypercholesterolemic agents. To uncover novel phytochemical scaffolds as potential inhibitors of SQS, we employed a structure-based virtual screening approach that involves screening 545 phytochemicals collected from Moroccan aromatic and medicinal plants and filtering them based on RMSD values and their affinity towards the target enzyme. Furthermore, we visualized the interacting amino acid residues to gain insight into the 2D and 3D interactions. The docking process was validated through the re-docking method with the reference co-crystallized complex (PDB id = 3v66, SQS-D3A). The screening resulted in the identification of two phytochemicals, Apigenin 7-O-rutinoside, and Apigenin 7-O-glucuronide, with high affinity for SQS binding sites. Both phytochemicals interact with functionally essential residues of SQS. The drug-likeness and toxicity of the phytochemicals were assessed through ADME-Tox analysis. Later, molecular dynamics simulations were performed by GROMACS software for 100 ns to evaluate the stability and fluctuations of protein-ligand complexes. By examining the trajectories produced by the MD simulations, we monitored complex stability, fluctuation, atomic gyration, H-bond, PCA, and FES. The simulations demonstrated that the interaction between the target and the compounds was stable and consistent. Binding free energy calculations indicated that Apigenin 7-O-rutinoside and Apigenin 7-O-glucuronide exhibited higher binding free energy than the co-crystallized inhibitor (D3A), providing a basis for further research in vitro and in vivo to develop potent SQS inhibitors for the treatment of hypercholesterolemia.

With the rapid expansion of aquaculture production, there has been a concomitant increase in fish diseases, generally combatted by the utilization of chemicals and antibiotics. However, their overuse leads to the emergence of drug-resistant microorganisms and the appearance of oxidative stress. Consequently, antibiotics contribute to a reduction in fish immune defense. In this work, eight marine bacteria were characterized using physiological, biochemical and microbiological tests, and were evaluated for their safety by studying their haemolytic and gelatinase activities. Then, we evaluated their ability to produce active biomolecules and tested their in vitro antibacterial activities against five bacterial pathogens that are highly prevalent in aquaculture. Moreover, we evaluated their antioxidant potential using DPPH, ABTS and ferric reducing power tests. The obtained results revealed that cell-free supernatants were effective, they exhibited good antioxidant and antibacterial activities. Their biomolecules showed good stability under different temperatures ranging from 60 to 121 degrees C and were found to be chemically composed of peptides, exopolysaccharides and polyphenols. Therefore, the secreted bioactive substances are of great biotechnological interest and constitute a promising eco-friendly alternative to antibiotics and chemical agents in the context of sustainable aquaculture.

Enterococci, known for their resilience, are commonly found in food, the environment, and the gastrointestinal tracts of humans and animals. In recent research, six strains of enterococcus were isolated from bat guano. These include Enterococcus mundtii SRBG1, Enterococcus gallinarum SRBG3, Enterococcus faecium SRBG2, Enterococcus casseliflavus EC1, and Enterococcus devriesei CAU 1344. Identification was done using 16S DNA analysis. Each strain underwent evaluation for its technological properties (such as tolerances to various NaCl concentrations and temperatures, as well as amylolytic, β-galactosidase, lipolytic, and proteolytic activities, and EPS production) and selected probiotic properties (including safety profile, resistance to 0.3 percent bile salts and gastric juice with a pH of 2.5, lysozyme tolerance, and antibacterial and antibiofilm activities against four foodborne pathogens). The results were analyzed using Principal Component Analysis. This analysis revealed that E. mundtii SRBG1 and E. gallinarum SRBG3, followed by E. faecium SRBG2, were most closely associated with a broad range of technological characteristics and were subsequently used for fermenting skimmed milk. The rheological properties of the samples indicated a shear-thinning or non-Newtonian behavior. Furthermore, during storage of the fermented milk at 4 °C over periods of 1, 7, 14, and 21 days, there were no significant changes in bacterial count (at around 7 log10 CFU/mL) and pH when fermented with the three evaluated strains.

Artificial insemination is a widely adopted method in livestock production for various reasons such as health security and genetic improvement. Although sperm motility is of paramount importance in this technique as it directly influences the sperm's ability to fertilize the oocyte. In previous research on human sperm, we observed that in vitro supplementation with Origanum Vulgare essential oil significantly improved sperm motility and antioxidant activities, all without negatively affecting the integrity of their DNA. Based on these promising results, we considered it crucial to explore the potential effects of supplementation with this essential oil on sperm of other species. In this study, we studied the effects of oregano essential oil supplementation on sperm motility of (bulls = 15) (dogs = 15) and (rabbits = 9) and the changes that in vitro incubation with this oil could induce on sub -motile sperm populations of different species. The results of the study showed that in vitro oregano essential oil supplementation had a significant impact on sperm motility in the three species studied. This improvement in sperm motility was accompanied by an increase in the proportion of subpopulations with high velocity and progressivity: an increase of (2.16%, 10% and 4.84%) for subpopulation 1, (6.50%, 5.5% and 3.17%) for subpopulation 4 in bulls, dogs and rabbits respectively. While the subpopulations representing low motile and non -progressive sperm have decreased. These results suggest that the use of oregano essential oil can be a beneficial approach to improve sperm motility in different species, which can have important implications for the success of artificial insemination.

Kisspeptin is a neuropeptide that plays a central role in fertility and neuroendocrine regulation of the hypothalamic-pituitary-gonadal axis. It has also been shown to act at the peripheral level in both men and women. Many studies have shown a correlation between kisspeptin blood levels and fertility in men. It is also involved in the maturation of sperm and even in the implantation of a pregnancy in women. In men, dysregulation of kisspeptin signaling can lead to hypogonadotropic hypogonadism. Recent studies have shown that Kisspeptin could constitute a new therapeutic target in the treatment of fertility disorders. Others have shown that the administration of exogenous Kisspeptin stimulates the release of gonadotropins in patients with fertility problems and even in healthy subjects. In addition, it plays an essential role in improving the quality of sperm in medically assisted procreation and even in the maturation of oocytes. In this literature review, we aim to examine the main functions of kisspeptin in male and female infertility.

The adaptive responses of the two yeasts Yarrowia lipolytica and Pichia pastoris to oxidative stress induced by the oxidants hydrogen peroxide (H2O2), sodium nitroprusside (SNP), and menadione (MD) have been studied. The ability of these compounds to inhibit the growth of the yeast cells under culture conditions was tested. The inhibitory concentrations of the compounds were evaluated. The cell viability, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity, and the lipids oxidation have also been investigated when exponential-phase cells are exposed to stress conditions. It was demonstrated that H2O2, SNP, and MD at 50%-inhibitory concentration affect the viability of both yeasts, probably by inhibiting the GAPDH and by causing other cell damages (lipid peroxidation), clearly augmented in the treated exponential-phase cells of Y. lipolytica and P. pastoris. On the other hand, enzymes involved in antioxidant defence systems such as catalase, glutathione peroxidase, and superoxide dismutase simultaneously significantly induced in the treated cells. The results showed that Y. lipolytica cells were more tolerant than P. pastoris cells to the presence of H2O2 and more sensitive in the presence of SNP and MD in the culture medium. Induction of these antioxidant enzymes with low doses of oxidants by pretreatment of yeast cells allowed them to tolerate higher doses of the compounds. Our data indicated that the increase of the cell viability agreed with the decrease of the lipid peroxidation and the increase of the GAPDH activity found after oxidative stress in pretreated cells.

Ichthyoplankton represent the first life stages of fish. The study of ichthyoplankton is crucial to understanding marine ecosystems and plays an important role in the management and durability of fisheries resources. During March and October of 2019, two oceanographic ichthyoplankton surveys were conducted in the Mediterranean Sea of Morocco from Tanger to Saadia by studying the horizontal structure of the ichthyoplankton species assemblage and its relation to environmental parameters. The average surface water temperature was (15.8°C in spring and 16.4°C in autumn). The fish eggs and larvae were more abundant in March than in October (21268 eggs/10m² and 14084 larvae/10m² in spring and 10094 eggs/10m² and 13796 larvae/10m²). In both seasons, fish eggs from the families Sternoptychidae and Sparidae were dominant (10101 eggs/10m² and 7527 eggs/10m² in spring and 4422 eggs/10m² and 3928 eggs/10m² in fall, respectively). However, Myctophidae larvae were the most abundant in the study area, reaching 7601 larvae/10m² in spring and 11021 larvae/10m² in autumn. The environmental parameters: temperature, salinity and chlorophyll-a (surface) seem to directly influence the spatial distribution of ichtyoplancton. On the other hand, it seems that predation by jellyfish (Pelagia noctiluca)was a very important factor that added to the factors that influenced the distribution of the species of fish eggs and larvae. This work represents the first survey conducted in the southwestern Alboran Sea, which studies the horizontal structure of the ichthyoplankton species assemblage and its relation to environmental factors in the spring and autumn of 2019.