Search Results (5,188)

Semen quality and fecal microbial composition were compared between native Oula rams reared on the Qinghai–Tibet Plateau and Hu sheep rams introduced from lowland regions. Semen quality was analyzed in eight adult Oula rams and eight Hu rams, and fecal microbial composition was assessed via 16S rRNA sequencing. Results indicated that sperm acrosome integrity was significantly higher in Hu sheep than in Oula sheep (p < 0.001); other semen parameters showed no significant differences. Significant differences were also observed in fecal microbial communities between the two breeds. Compared with Hu sheep, Oula sheep exhibited higher microbial abundance and diversity at the phylum level, particularly Campylobacterota, Euryarchaeota, Planctomycetota, Verrucomicrobiota, Myxococcota, and Deferibacterota (p < 0.05). At the genus level, Oula sheep had significantly higher abundances of Treponema, Campylobacter, Methanobrevibacter, UCG-009, Family_XIII_AD3011_group, [Eubacterium]nodatum group, Candidatus Soleaferrea, Akkermansia, and unidentified_Ruminococcaceae (p < 0.05). Correlation analysis indicated associations between sheep semen quality and the top 30 abundant fecal microbial genera. Six genera showed significant positive correlations with acrosome integrity rate, and eight genera exhibited significant negative correlations (p < 0.05). Two genera were correlated positively with plasma membrane integrity rate (p < 0.05). Prevotellaceae_UCG-004 was positively correlated with sperm motility and Progressive Motility spermatozoa proportion (p < 0.05); Ruminococcus showed a significant positive correlation with sperm linear motility and a significant negative correlation with acrosome integrity rate (p < 0.05). These findings indicate that the microbial groups enriched in Oula sheep fecal samples and exhibiting negative correlations with acrosome integrity—including Ruminococcus, Treponema, Akkermansia, and Euryarchaeota—are associated with sperm quality through physiological adaptation mechanisms specific to high-altitude environments. Full article

The genus Parabacteroides comprises widespread gastrointestinal commensals, known to produce immunomodulatory molecules and extracellular vesicles, yet its full diversity is incompletely cataloged. This study describes strain ASD2025^T, isolated from healthy child feces, using a polyphasic taxonomic approach including phenotypic profiling, chemotaxonomy, and comparative genomics. Cells were non-motile, polymorphic rods that produced extracellular vesicles. Phylogenomic analysis placed ASD2025^T within the genus Parabacteroides within a species complex consisting of P. acidifaciens, P. hominis, “P. massiliensis”, P. merdae, and P. johnsonii, with average nucleotide identities to the type strains of 85.5–89.9%. The large genome (5.16 Mbp, 46.2% GC content) contained integrative conjugative elements harboring antibiotic resistance genes and hankyphage-related prophage. The strain produced succinate as the major metabolic end product, and its major fatty acids were anteiso-C_15:0, iso-C_17:0 3-OH, and C_15:0. Conditioned medium from ASD2025^T antagonized the interleukin-8 response caused by E. coli lipopolysaccharide in HT29 cells. The majority of related metagenome-assembled genomes originate from mouse microbiomes. Based on these distinct characteristics, strain ASD2025^T (=VKM B-3926^T = JCM 37967^T) represents a novel species of the genus Parabacteroides, for which the name Parabacteroides vesiculifaciens sp. nov. is proposed. Full article

Nanotechnology, based on nanoparticles, has become an emerging interdisciplinary tool in reproductive biotechnology, offering innovative opportunities to improve fertilization efficiency and reproductive performance in farm animals. The purpose of this review is to provide an updated synthesis of current research on nanoparticle-based approaches that enhance in vitro fertilization outcomes and other assisted reproductive technologies. The focus is on the biological mechanisms, potential benefits, and limitations of nanoparticle use in animal reproduction. Nanoparticles—including gold, silver, zinc oxide, selenium, and magnetic iron oxide—exhibit distinctive physicochemical properties that enable targeted interactions with gametes and reproductive cells. When used in semen extenders or culture media, nanoparticles improve sperm motility, acrosome and membrane integrity, and reduce oxidative stress and apoptosis. These effects contribute to enhanced fertilization rates and higher embryo developmental competence. In addition, nanoparticles can function as carriers for hormones, antioxidants, and growth factors, stabilizing reagents essential for oocyte maturation, sperm capacitation, and early embryo culture. The review also discusses nanopurification (selectively isolating and removing particles) and nanosorting (separating or organizing nanoscale objects) techniques that allow for non-invasive selection of viable gametes, and fluorescence- and magnet-assisted sorting systems that increase precision in sperm sexing. The mechanical aspects of nanoparticle–cell interactions are analyzed, emphasizing the influence of particle size, dose, and surface modification on both biological efficacy and cytotoxicity. Safety, toxicological concerns, and regulatory frameworks—including International Organization for Standardization (ISO) standards and European Commission recommendations—are critically reviewed to highlight the need for harmonized biocompatibility criteria. Although nanoparticle use in animal reproduction remains largely experimental, accumulated evidence demonstrates its potential to improve reproductive efficiency and reduce economic losses. Integrating nanoparticle-based systems with existing reproduction platforms may represent a transformative step toward sustainable and precision-driven livestock breeding. Full article

Background/Objectives/Methods: A bioassay-informed investigation of the Australian pasture soil-derived Streptomyces sp. S4S-00193A39 yielded the anthelmintic principals as three new spiroketal polyketide alkaloids, goondoxazoles A–C (1–3), with structures assigned by detailed spectroscopic analysis. Results: A structure–activity relationship based on the ability to inhibit the motility of Dirofilaria immitis microfilariae (mf) revealed a positive correlation for the benzoxazole moiety present in 2 and 3 (EC₅₀ 55–85 nM) versus the ring-opened aminobenzoic acid moiety evident in 1 (EC₅₀ 1.38 µM). This hypothesis was strengthened by extension of the SAR assessment to the known benzoxazole natural products A-33583 (12), UK-1 (13) and nataxazole (14), and the new analogue 5-hydroxynataxazole (15), which were isolated in our lab from three additional Australian pasture soil-derived Streptomyces spp. Of note, while the benzoxazole methyl esters 13–15 exhibited approximately 9- to 65-fold lower potency against D. immitis mf compared with 2 and 3, the carboxylic acid substituted benzoxazole 12 displayed comparable activity (EC₅₀ 72 nM) against D. immitis mf, and >5-fold improved potency against D. immitis L4 larvae (EC₅₀ 0.43 µM). Conclusions: These observations reveal the promising anthelmintic potential (against D. immitis) for the new structurally complex and chiral goondoxazoles (e.g., 2 and 3), and demonstrate that this effect can be replicated, even improved, by simpler, achiral benzoxazole microbial natural products (e.g., 12). Full article

Melanoma is a highly malignant neoplasm of the skin with early metastatic spread and increasing incidence worldwide. Although there are significant therapeutic advances in immunotherapy, especially with the checkpoint inhibitors targeting PD-1 and CTLA-4, challenges such as treatment-related toxicities, a heterogeneous response to therapy, and drug resistance continue to exist. There are unmet needs for novel therapeutic strategies and/or approaches to complement the existing treatment options. Potential targets for future melanoma treatment are the gap junction proteins, connexins, which show an altered pattern of regulation during melanoma progression. In this review, we highlight the regulation of gap junctions during melanoma progression and the characterization of gap junctions as tumor suppressors during early-stage tumor development and then the reversion to enhancers of tumor metastasis during late-stage melanoma progression. We provide a comprehensive overview of gap junctions in the skin and how the connexin proteins, which comprise gap junctions, are alternatively regulated in melanoma progression. Connexins are protein channels in the human body that consist of 21 isoforms. These isoforms form gap junctions that provide important intercellular signaling and permeability channels. Each connexin protein consists of four transmembrane domains and a C-terminal tail, which is an important part of its function and regulation. Permeants of gap junctions include signaling molecules such as cyclic AMP and inositol triphosphate which are linked to key cellular behaviors such as proliferation and migration, making them essential for several tumor-related processes. At least ten connexin isoforms are found in normal skin. Connexin 43 (Cx43) is classified as the most prevalent isoform while Connexin 26 (Cx26) has been reported to be more specialized with restricted expression patterns. Cx43 and Cx26 regulate the growth, differentiation, and repair of the epidermis after injury. Evidence suggests that connexins have a stage-related function in melanoma. Loss of connexin expression and gap junctional intercellular communication is linked to tumor suppression and loss of differentiation in early-stage melanoma, while re-expression or overexpression of specific connexins, notably Cx43, may promote metastasis through enhanced tumor–stromal interactions and increased motility in late-stage melanoma. Such opposing actions of connexins support their candidacy as biomarkers and therapeutic targets. Understanding the dual-stage related functions of connexins in melanoma development and progression may lead to less cytotoxic and more efficient future therapeutic approaches. Full article

Zelenci is a limnocrene spring in the South-Eastern Alps, attractive for its unique structure and known among researchers for its high diatom diversity. Our aim was to assess how abiotic factors influence the structure of diatom communities in different habitats and to compare the trophic status over 10 years of investigation. Four sampling sites were chosen: two for tychoplankton and two for epipelon. Achnanthidium minutissimum was the most abundant species in both habitats, while Navicula was the most diverse genus (17 species). Planktonic diatoms dominated plankton samples, while the motile ecological type dominated the epipelon. Of all diatom taxa, 23.5% had some endangerment status. Key factors influencing the Shannon–Wiener diversity index were water level, temperature, and concentration of NH₄⁺. The most important abiotic factors for the tychoplankton community were temperature and NH₄⁺, while the most important abiotic factors influencing the structure of epipelon communities were water level, NH₄⁺, pH, and dissolved oxygen concentration. Trophic index revealed increasing inflow of nutrients to the spring; 10 years ago, they were oligotrophic to oligo-mesotrophic, whereas in the present, they are predominantly eu-mesotrophic to eutrophic, indicating human pressure from the catchment area. Full article

Endocrine-disrupting chemical (EDC) exposure from plastics and everyday products is widespread and linked to infertility. We conducted a 3-month uncontrolled feasibility pilot study among five idiopathically infertile couples to assess whether an intensive lifestyle intervention was associated with within-person changes in urinary EDC biomarkers and exploratory changes in reproductive parameters. The intervention was embedded in a film project (“The Plastic Detox”) and integrated personalized education, product substitutions, at-home urine biomonitoring, sperm testing, and weekly coaching. Urine and semen samples were collected at baseline, 6 weeks, and 12 weeks. Linear mixed-effects models were used to estimate biomarker changes. BPA was designated a priori as the primary biomarker endpoint. Directional reductions were observed in urinary bisphenol A (BPA), mono-n-butyl phthalate (MBP), and monobenzyl phthalate (MBzP) over the intervention period. Within-person reductions in products containing ingredients of concern were associated with lower BPA levels. Descriptive upward trends of semen parameters were observed, with the majority of the subfertile men testing >40 million motile sperm/ejaculate after the intervention. Participants had increased environmental health literacy, were more motivated to reduce exposures, and reported improved wellness endpoints. Four couples achieved pregnancy and live birth during follow-up; given the uncontrolled design and small sample size, these outcomes are presented descriptively. Overall, this pilot study demonstrates feasibility and measurable biomarker change, supporting evaluation in larger, controlled trials. Full article

Listeria monocytogenes (LM) is a major foodborne pathogen causing illnesses ranging from gastroenteritis to severe systemic infections. The key virulence factors include bacterial motility, hemolysin and lecithinase production, and invasion of host tissues. This study investigated the anti-virulence effects of cannabidiol (CBD), the main non-psychoactive compound in Cannabis sativa, against LM. The minimum inhibitory concentration (MIC, 2289 μM; 719.8 µg/mL) and sub-inhibitory concentration (SIC, 11.92 μM; 3.75 µg/mL) of CBD were determined for LM strains Scott A and ATCC 19115. Cultures were treated with SIC, 6× SIC, 1/4× MIC, and MIC to assess effects on motility, hemolysin and lecithinase production, and adhesion and invasion of human intestinal (Caco-2) and brain endothelial (HBMEC) cells, alongside virulence gene expression by RT-qPCR. Cannabidiol’s efficacy was also determined using a Galleria mellonella larval infection model at SIC and 6× SIC. Cannabidiol at 6× SIC significantly reduced motility, toxin production, and host cell adhesion and invasion (p < 0.05). RT-qPCR revealed downregulation of key virulence genes, including prfA, hly, plcA, plcB, iap, motA, motB, actA, inlA, and inlB. In vivo, CBD enhanced larval survival in a dose-dependent manner and cytotoxicity was observed at concentrations above 33.75 µg/mL. These results indicate that CBD, at non-bactericidal levels, effectively suppresses multiple virulence mechanisms in LM, highlighting its potential as a novel anti-virulence agent for food safety and therapeutic applications. Full article

Castration is widely used in goat production to improve meat quality and manage reproduction, yet conventional surgical methods raise significant animal welfare concerns. Immunocastration targeting gonadotropin-releasing hormone (GnRH) offers a promising, welfare-friendly alternative, but its efficacy in slow-growing indigenous breeds remains poorly defined. In this study, we developed a novel tandem-repeat GnRH(30) recombinant vaccine and evaluated its effects on growth performance, reproductive function, and meat quality in male Leizhou goats. Eighteen six-month-old bucks were assigned to an immunocastration group (IM), surgical castration group (SC), or intact control group (IC). Vaccinated goats produced sustained anti-GnRH antibodies and exhibited significantly suppressed testosterone levels comparable to surgical castrates. Immunocastration induced marked testicular atrophy, disrupted spermatogenesis, reduced semen volume and sperm motility, and increased sperm abnormalities. Importantly, early growth performance and final body weight were not significantly affected. Compared with intact males, both IM and SC goats showed improved meat quality traits, including reduced drip loss and shear force, accompanied by increased intermuscular fat deposition, with overall patterns in the IM group closely resembling those of surgical castration. Overall, these findings indicate that the GnRH(30) vaccine can effectively suppress spermatogenesis and improve meat quality without affecting growth, providing an effective technical approach for castration management in indigenous goats. Full article

Listeria innocua is a bacterium frequently detected in food and food production plants (FPPs). Understanding the heterogeneity of L. innocua food isolates is essential for predicting potential food safety threats and developing preventive and control measures. This study aimed to characterize L. innocua isolated from raw drinking milk by investigating the genomic features related to virulence, antimicrobial resistance, and persistence using whole-genome sequencing (WGS), along with phenotypic antimicrobial susceptibility testing using the disk diffusion method. All ten isolates analyzed in this study belonged to sequence type (ST) 492 and were distantly related to the reference strain. A total of 80 virulence-associated genes were identified, including the complete Listeria Pathogenicity Islands-3 (LIPI-3) and LIPI-4 clusters typically found in virulent L. monocytogenes clones, as well as 66 additional genes involved in adhesion, invasion, motility, post-translational modification, regulation, immune modulation, and stress survival. Stress survival islet 2 (SSI-2) and genes encoding the Clp protease complex (clpC, clpE, clpP), which support both persistence and virulence, were also detected, whereas LIPI-1 and internalin genes were not detected. The antimicrobial resistance determinants included fosX, lin, norB, sul, and three multidrug efflux pumps (lde, mdrL and mdrM). Mobile genetic elements (plasmids, prophages, or transposons) were not detected. All isolates were phenotypically susceptible to benzylpenicillin, ampicillin, meropenem, erythromycin, and trimethoprim–sulfamethoxazole. These findings underscore the importance of ongoing genomic surveillance of L. innocua in food environments and highlight the need to assess the potential risk posed by specific lineages, such as ST492, to food safety. Full article

Listeria monocytogenes (Lm) is a foodborne pathogen whose virulence depends on the coordinated action of multiple virulence factors. Although deletion of either LIPI-4 or inlB reduces the virulence of Listeria monocytogenes, it remains unknown whether these two factors are functionally or regulatory connected. Therefore, we constructed an inlB deletion mutant and its complemented strain in the Lm928 and ΔLIPI-4 backgrounds. We assessed bacterial growth, biofilm formation, motility, host cell interactions (adhesion, invasion, intracellular proliferation), plaque formation, mouse organ colonization. Growth curve analysis showed no significant differences among strains. qPCR revealed that LIPI-4 modulates inlB expression in a cell-type-specific manner: inlB was downregulated in ΔLIPI-4 under culture and HTR-8 infection, but upregulated during hCMEC/D3 infection—yet functional defects persisted in all cases. Biofilm assays showed that ΔLIPI-4 and the double mutant exhibited enhanced biofilm formation, with the double mutant exceeding ΔLIPI-4, demonstrating synergistic enhancement. Motility assays indicated that LIPI-4 dominates bacterial movement, with ΔLIPI-4 and the double mutant showing identical severe defects. Plaque formation analysis showed that LIPI-4 is essential for cell-to-cell spread, while inlB deletion unexpectedly enhanced plaque formation—an effect completely abolished in the absence of LIPI-4. Host cell assays across Caco-2, HTR-8, and hCMEC/D3 models revealed that LIPI-4 is the core determinant of adhesion, invasion, and intracellular proliferation, whereas inlB contributes in the context of LIPI-4 and its effects vary with the specific cellular process examined. In mice, LIPI-4 was essential for systemic colonization of the liver and spleen, with inlB acting as a co-factor, whereas inlB unexpectedly promoted higher bacterial burdens in the brain, suggesting that inlB modulates LIPI-4-mediated neuroinvasion. Overall, our results establish LIPI-4 as the central determinant of Lm virulence, with inlB acting as a context-dependent co-factor that modulates LIPI-4-mediated pathogenesis in a cell type- and tissue-specific manner. Full article

Nano-graphene oxide (NGO) is a nanomaterial that has been frequently used in the fields of health and bioengineering in recent years. However, its potential use in semen cryopreservation is still in the exploratory phase. In this study, Angora bucks, a breed with low resistance to cold shock, were used. Sperm was collected from five different Angora bucks, pooled, diluted with a Tris-based egg yolk diluent, and frozen with the addition of NGO at two different sizes (50 and 500 nm) and doses (10 and 50 µg/mL). Nanoparticle characterization was performed using field emission scanning electron microscopy (FE-SEM), dynamic light scattering (DLS), and Fourier-transform infrared spectroscopy (FTIR). Post-thaw sperm analyses were evaluated based on motility and kinematic parameters, mitochondrial membrane potential (MMP), plasma membrane and acrosome integrity (PMAI), and DNA fragmentation. Applying 50 nm NGO at a dose of 50 µg/mL led to statistically significant improvements in motility and PMAI (p < 0.05). The same dose of 500 nm NGO, however, only showed a statistically significant improvement in the PMAI parameter (p < 0.05). No significant differences were observed between the groups for MMP and kinematic parameters (p > 0.05). Conversely, it was found that all sizes and doses of NGO significantly protected post-thaw sperm regarding DNA integrity (p < 0.05). These findings indicate that the NGO, at a size of 50 nm and a dose of 50 µg/mL, improves the post-thaw quality of Angora buck sperm and provides a cryoprotective effect that depends on size and dose. This study provides preliminary data on the potential effects of NGO; however, comprehensive mechanistic and in vivo validation studies are required to establish the biological and clinical validity of these findings. Full article

Energy homeostasis arises from a complex interplay between gut-derived hormones, the central nervous system, and pancreatic function. Beyond the classical incretin axis, a broad spectrum of gut peptides acts in concert to coordinate appetite regulation, nutrient sensing, gastric motility, and systemic bioenergetic balance. Perturbation of this network contributes to metabolic disorders such as obesity, type 2 diabetes, and cachexia, underscoring its pivotal role in physiological and pathological energy regulation. This review provides an integrated analysis of the mechanisms through which gut–brain–pancreas communication maintains metabolic homeostasis, with particular attention to the dynamic cross-talk between peripheral endocrine signals and central regulatory circuits. Alterations in these pathways are examined in relation to their impact on energy expenditure and substrate utilisation, alongside recent translational efforts exploiting multi-receptor peptide agonism and combinatorial hormonal modulation to restore metabolic equilibrium. Emerging therapeutic approaches increasingly aim to engage multiple bioenergetic pathways simultaneously, supported by advances in peptide engineering and molecular design. By conceptualising metabolic regulation as a coordinated network rather than a linear hormonal cascade, this article delineates a physiological and translational framework for next-generation interventions targeting bioenergetic dysfunction in human disease. Full article

Objective: To investigate the therapeutic effects and safety profile of Wanshi Shachong Xiaoji Pills (WSXPs) on a functional dyspepsia (FD) mouse model and to preliminarily explore its potential mechanism and impact on associated hepatic metabolism. Methods: An FD model was established in mice using L-arginine. Gastrointestinal motility was assessed by measuring gastric emptying and intestinal propulsion rates. Serum levels of gastrointestinal hormones (MTL, GAS, VIP, CCK) and gut microbiota composition were analyzed. A one-month repeated-dose toxicity study was conducted in normal mice to evaluate safety. The effects of WSXPs on lipid metabolism and inflammation were further examined in a hepatocyte steatosis model in vitro, and network pharmacology was employed to predict potential mechanisms. Results: WSXPs significantly alleviated FD symptoms by improving gastrointestinal motility, bidirectionally regulating gut hormone levels, and increasing the abundance of beneficial bacteria (Akkermansia muciniphila). Long-term administration showed no significant toxicity. In vitro, WSXPS reduced lipid accumulation and inflammation in hepatocytes. Network analysis identified the PI3K-Akt signaling pathway as a potentially central common target, providing a hypothesis for future mechanistic studies. Conclusions: WSXPs effectively improve FD symptoms, modulates gut microbiota, and exhibits potential benefits on hepatic lipid metabolism in vitro, possibly via the PI3K-Akt pathway. This hepatocyte-level finding, combined with its in vivo efficacy in FD, suggests a promising avenue for future research into its potential applications in metabolic-associated conditions. This study provides a scientific foundation for the further development and clinical application of WSXPs in treating FD. Full article

Melatonin (MLT) is a potent antioxidant that reduces oxidative stress (OS)-induced sperm damage. However, few studies have explored its effects in the field investigated here. This study aimed to evaluate the effects of MLT supplementation in extenders used for staining, sorting, and freezing on the quality of sorted Nili-Ravi buffalo sperm, embryo development after in vitro fertilization (IVF), and pregnancy outcomes following artificial insemination (AI). Computer-aided sperm analyzer (CASA) showed that progressive motility (PM) and velocity parameters of sorted, frozen–thawed sexed sperm were higher in the MLT-treated groups (p < 0.05). Fluorescence microscopy demonstrated that MLT significantly improved sperm plasma membrane integrity after staining and increased the proportion of frozen–thawed sex-sorted sperm with high mitochondrial membrane potential (HMMP) (p < 0.05). Raman spectroscopy further identified several distinctive Raman bands at 936, 1300, and 1651–1652 cm⁻¹, which may serve as biomarkers for assessing sperm quality. Additionally, cleavage rate, blastocyst formation, and pregnancy rates following IVF and AI were higher in the MLT-treated group (p < 0.05). In conclusion, MLT can serve as a valuable additive during sperm sex-sorting procedures to enhance sperm quality, thereby improving embryo developmental competence and pregnancy outcomes. Full article