Search Results (246)

Background: Vibrio harveyi is a major bacterial pathogen threatening turbot aquaculture, necessitating the development of more effective vaccines. Bacterial ghosts (BGs), which are empty bacterial envelopes with preserved surface antigens, offer a promising alternative to traditional formaldehyde-killed vaccines that often suffer from reduced immunogenicity. Methods: We developed an optimized BGs vaccine for V. harveyi by combining the nonionic surfactant NP-40 with sodium hydroxide (NaOH). This NP-40/NaOH combination demonstrated a synergistic lytic effect, halving the minimum inhibitory concentration of NaOH required for complete inactivation. Results: The resulting BGs exhibited intact cellular morphology with transmembrane pores, efficient removal of cytoplasmic contents, and significantly better preservation of lipopolysaccharide structure compared to NaOH-alone treatment. Vaccination trials in turbot demonstrated that the NP-40/NaOH BGs provided the highest relative percent survival (RPS = 58.8%) upon challenge, outperforming both NaOH-alone BGs (RPS = 55.0%) and a traditional formaldehyde-killed vaccine (RPS = 34.8%). The superior protection was correlated with the induction of a more robust and sustained immune response, characterized by significantly higher levels of specific IgM antibodies, elevated lysozyme activity, and increased total serum protein. Conclusions: This study establishes the NP-40/NaOH protocol as an effective strategy for producing high-quality BGs with enhanced immunogenicity, presenting a potent vaccine candidate for controlling vibriosis in aquaculture. Full article

Due to the limitations of conventional antibiotics, antimicrobial peptides (AMPs) have emerged as promising therapeutic alternatives for the prevention and treatment of oral infections. This study systematically evaluated in vitro evidence regarding the antimicrobial and anti-biofilm activity of natural AMPs against oral pathogens. A systematic search using the PICOT strategy was conducted in PubMed, EMBASE, and Scopus, retrieving 7711 articles. After title and abstract screening, 109 studies were selected for full-text analysis, resulting in 26 articles that met the eligibility criteria. Among the AMPs evaluated, nisin (n = 15) and LL-37 (n = 5) were the most frequently investigated, while other peptides included lactoferrin, lactoferricin, melittin, lysozyme, histatin-5, cystatin C, chromogranin A, parasin-1, protamine, AmyI-1-18, and DCD-1L. Natural AMPs of human and animal origin demonstrated antimicrobial activity against bacteria associated with oral infections, particularly Streptococcus mutans and Enterococcus faecalis. These peptides were tested in different formulations, including solutions, incorporation into dental materials and polymers, and application in sonodynamic antimicrobial therapy. Overall, the findings indicate that natural AMPs represent a promising class of biomolecules for controlling oral biofilms; however, further clinical studies are required to validate their long-term efficacy and safety. Full article

In this study, the synergistic effects of dietary Bacillus velezensis AAHM-BV2301, silica nanoparticles (SiNPs), and chitosan (CS) on the growth performance, innate immunity, gut microbiota, and disease resistance of Asian seabass (Lates calcarifer) fingerlings were evaluated. A total of 400 fish (11.25 ± 2.12 g) were assigned to five dietary treatments for 30 days: control, BV (1 × 10⁸ CFU/kg feed), BVSiNP (1 × 10⁸ CFU/kg + 2 mg SiNP/kg), BVCS (1 × 10⁸ CFU/kg + 15 g CS/kg), and BVSiNPCS (combined additives at the same concentrations). The growth indices (WG, SGR, RGR, and FCR) significantly increased in the fish fed BVSiNPs, whereas the level of innate immunity increased across all the supplemented groups, with BVCS and BVSiNPCS having the strongest respiratory burst and lysozyme activities. The tissue-specific modulation of immune-related genes (α2M, HSP70, Mx, and C3) was most pronounced in BVSiNP-fed fish, particularly in the gills and liver. Gut microbiome profiling revealed enrichment of Cetobacterium somerae in response to BV-based treatments, whereas BVSiNPCS induced the greatest increase in microbial richness and network connectivity. Postchallenge survival against Vibrio vulnificus was significantly greater in the BV and BVSiNP groups (p < 0.05). Overall, SiNPs acted as functional enhancers of the B. velezensis probiotic, supporting improved growth, immune activation, and microbiota restructuring. These results highlight the potential of nanoparticle-integrated synbiotics for microbiome-targeted health management in aquaculture. Full article

Cutaneous lupus erythematosus (CLE) can occur independently of lupus erythematosus. SLE, and its responsiveness to treatment, does not necessarily align with that of coexisting SLE. Extracellular vesicles (EVs) allow communication between cells and rapid delivery throughout the body. We hypothesized that EVs may support disease-specific inflammation in CLE and SLE patients. Plasma EVs from healthy controls (n = 5), CLE (n = 6), and dermatomyositis (n = 17) were purified by ultracentrifugation and size-exclusion chromatography, phenotyped by flow cytometry, and profiled by LC-MS/MS. Circulating EVs were mainly platelet-, endothelial-, and antigen-presenting cell-derived examples. CLE EVs harbored four proteins absent in the controls—mimecan, IFI27, fibulin-2, and snRNP B/B′ (anti-Sm an-tigens)—and their cumulative number increased with SLEDAI. Relative to the controls, 18 proteins were upregulated and 15 downregulated in CLE EVs. The number of upregulated proteins showed a trend toward a correlation with SLEDAI (r = 0.79, p = 0.06) but not with CLASI (r = 0.21). Among upregulated proteins, lysozyme C and hyaluronan-binding protein 2 tracked with cutaneous activity (CLASI r = 0.74 and r = 0.86) but not with systemic activity (SLEDAI r = 0.52 and r = 0.31). CLE plasma EVs were enriched in antigen-presenting cell markers and disease-related cargo, including anti-Sm antigens and proinflammatory proteins. Although overall protein diversity correlated primarily with systemic disease activity, a subset of proteins appeared to reflect cutaneous activity. Full article

This study investigated the effects of dietary supplementation with Hylocereus polyrhizus juice by-product extract (HJBE) on growth performance, digestive enzyme activity, antioxidant status, immune response, and disease resistance against Edwardsiella tarda in juvenile Pagrus major. The HJBE, prepared from juice-processing residues, contained measurable levels of bioactive compounds, including vitamin C, total phenolics, and flavonoids, and exhibited noticeable antioxidant activity. Five experimental diets containing 0 (control), 0.5, 1.0, 2.0, and 3.0 g/kg HJBE were fed to juvenile red seabream (initial weight of 7.0 ± 0.01 g) for eight weeks. Fish fed the diet containing 1.0 g/kg HJBE (HJBE1) showed significantly higher final weight, weight gain, specific growth rate, feed efficiency, and protein efficiency ratio compared with other groups. Trypsin activity in intestine was also significantly enhanced in the HJBE1 group, suggesting improved feed utilization. Whole-body composition and hematological indices did not differ among treatments. Plasma antioxidant parameters showed a dose-dependent trend, with catalase and glutathione levels lower at inclusion levels of 0.5–1.0 g/kg. Serum lysozyme activity and immunoglobulin M concentrations were significantly higher in fish fed HJBE1 compared to the controls, corresponding to improved survival after E. tarda challenge. These findings suggest that even moderate levels of bioactive compounds in HJBE can beneficially influence antioxidant homeostasis, immune defense, and growth performance. In conclusion, dietary inclusion of 1.0 g/kg HJBE effectively improved growth, digestive physiology, antioxidant balance, and disease resistance in juvenile P. major. Full article

Background/Objectives: The growing demand for functional foods and alternative therapeutic strategies has intensified the search for novel probiotic strains from underexplored ecosystems. This study aimed to isolate and phenotypically characterize lactic acid bacteria (LAB) from spontaneously fermented fruits found in the Legal Amazon (Ananas comosus, Humiria balsamifera, Manilkara zapota, and Platonia insignis) and to perform genome-based analysis of the most promising isolate to evaluate its probiotic potential. Methods: The isolates were identified by MALDI-TOF-MS and screened for tolerance to low pH, bile salts, lysozyme, growth at 39 °C, and antimicrobial activity against five enteric pathogens. The most promising isolate was evaluated by coaggregation and biofilm assays, in silico proteome and CAZyme analysis, bacteriocin cluster mining, and in vivo efficacy testing using Tenebrio molitor larvae. Results: Three isolates from H. balsamifera were identified as Lactiplantibacillus plantarum (M1, M2, M4) by MALDI-TOF-MS. These isolates exhibited high resilience to all tested physiological stressors. Antimicrobial activity was contact-dependent, with no inhibition by cell-free supernatants. M2 showed the strongest pathogen exclusion, moderate biofilm formation, and high coaggregation with S. enterica and E. faecalis. Genome analysis of M2 revealed a 3.40 Mb chromosome, absence of acquired resistance or virulence genes, two plantaricin gene clusters, and 93 CAZymes, including GT families linked to exopolysaccharides biosynthesis. SignalP predicted secretion signals in 10 CAZymes. M2 significantly improved larval survival against E. coli and S. enterica, especially under prophylactic treatment. Conclusions: L. plantarum M2 combines safety, stress tolerance, genomic features, and in vivo efficacy, positioning it as a promising probiotic candidate adapted to tropical niches. These findings highlight H. balsamifera as a reservoir of novel probiotic strains. Full article

Lactobacillus acidophilus is a widely researched probiotic bacterium with broad applications in health and biotechnology; however, its protoplast formation has not been extensively investigated. This study aimed to optimize conditions for L. acidophilus protoplast formation. Freeze-dried cells were suspended in 20 mM HEPES buffer (pH 7) supplemented with sucrose (1.0 M, 1.5 M, and 2.0 M) to induce hyperosmotic conditions, yielding a final cell density of 10⁸ cells/mL. The suspensions were treated with 125 µg/mL lysozyme and incubated at 37 °C for 30 min, 1 h, or 2 h. Prior to enzymatic treatment, the buffer, lysozyme, and cell suspensions were equilibrated at either 22 °C (room temperature) or 37 °C. Phase contrast microscopy was used to evaluate protoplast formation across all treatment combinations, and a three-way ANOVA was conducted to assess the effects of buffer molarity, incubation time, and temperature. Protoplasts are valuable tools for genetic manipulation, cell fusion, and cell wall studies, yet optimized protocols for their generation in L. acidophilus are lacking. The highest protoplast yield with minimal lysis was observed under 2.0 M sucrose conditions after 2 h of incubation, particularly when all components were equilibrated at 37 °C. Prolonged lysozyme exposure increased lysis, especially at lower buffer molarities. Elevated buffer molarity conferred a protective effect by maintaining cell integrity during enzymatic digestion. These findings highlight the importance of osmotic strength and thermal equilibration in optimizing protoplast formation and provide a reproducible framework for controlled enzymatic treatments in L. acidophilus. Full article

With the aim to investigate the immunomodulatory potential of selenitetriglycerides (SeTG), a new lipophilic Se (IV) compound, 30 sheep (15 sheep/treatment) were used in a completely random design to receive the SeGT supplement as follows: (1) no SeGT supplement (Control) and (2) daily dosage of 2 mL of SeGT (equivalent to 1 mg Se/kg BW) during the first 7 days of the evaluation, which lasted 28 d. Individually, blood samples were collected on days 0, 14, and 28 to measure and assess parameters of innate cellular and humoral immunity, including respiratory burst activity (RBA) and potential killing activity (PKA) of monocytes and granulocytes, proliferative response of lymphocytes stimulated with ConA (Concovalin A) and LPS (lipopolisaccharidde), lysozyme activity, ceruloplasmin activity, and gamma globulin levels. From the 14th day, supplemental SeTG saw significant increases (p ≤ 0.001) in RBA and PKA parameters, as well as enhanced proliferative responses of lymphocytes compared with controls. Both innate humoral immunity (elevated lysozyme activity) and adaptive humoral immunity (increased gamma globulin levels) were positively influenced (p ≤ 0.01), whereas ceruloplasmin activity remained unchanged. Under the conditions in which the current experiment was carried out, SeGT showed good promise to modulate immunity in a short period (28 d). Further research should explore experiments with a greater number of animals over long-term periods of evaluation under production system conditions. Full article

Post-weaning diarrhea caused by Enterotoxigenic Escherichia coli (ETEC) is a major disease in piglets and leads to substantial economic losses in the swine industry. Compared to conventional lysozyme, yak stomach lysozyme (YSL) demonstrates distinctive resistance to pepsin, trypsin, high temperature, and acidic conditions. This study investigated the effects of dietary YSL supplementation on intestinal health in weaned piglets challenged with ETEC, utilizing metabolomics and proteomics. A total of 18 weaned piglets were randomly divided into three groups: control (C), diarrhea (D), and YSL treatment (YLT). Groups C and D were fed a basal diet, while the YLT group received the basal diet supplemented with YSL at a dosage of 100,000 U/kg following ETEC challenge. Following an acclimation period, piglets in groups D and YLT were orally challenged with ETEC, while group C received the same volume of sterile LB broth. The feeding trial lasted for 21 days before sample collection. The results demonstrated that dietary supplementation with YSL significantly reduced the diarrhea rate (p < 0.05). Compared with the D group, the YLT group exhibited significantly increased serum albumin levels (p < 0.05), along with a tendency toward greater villus height (p = 0.085) and higher serum glucose levels (p = 0.052), indicating an improvement in nutritional and metabolic status Metabolomic analysis identified 260 differentially abundant metabolites between the YLT and D groups (81 upregulated, 179 downregulated), which were predominantly enriched in pathways related to amino acid biosynthesis and metabolism, purine metabolism, and nucleic acid metabolism. Proteomic profiling revealed 571 differentially expressed proteins (237 upregulated, 334 downregulated). Upregulated proteins were mainly involved in arginine biosynthesis and base excision repair, while downregulated proteins were associated with the PPAR signaling pathway and Salmonella infection. In summary, dietary YSL supplementation alters the metabolic and proteomic profiles in the intestines of diarrheic piglets, potentially improving gut barrier function and nutrient utilization. This study offers novel insights into the potential of YSL as a promising feed additive for prevention of post-weaning diarrhea in pigs. Full article

Human serum albumin (HSA), an endogenous protein, was employed in the synthesis of nanoparticles. The treatment of an HSA solution with ethanol and glutaraldehyde resulted in the formation of human serum albumin nanoparticles (HSA NPs), which exhibited a weak fluorescence emission peak at 515 nm upon excitation at 360 nm. Importantly, these synthesized HSA NPs displayed a pronounced fluorescence polarization (FP) signal under identical excitation and emission conditions. Furthermore, incubation of the HSA NPs with specific DNA aptamers targeting lysozyme and thrombin led to a significant decrease in the FP values. This reduction in FP was effectively reversed upon the addition of lysozyme and thrombin. Based on these observations, a label-free fluorescence polarization-based detection platform for lysozyme and thrombin was developed utilizing HSA NPs and a DNA aptamer system. Full article

Cancer progression in skeletal muscle (SkM) is very rare, and mechanisms remain unclear. This study assessed the potential of SkM (myocyte)-derived EVs (C2C12-EVs) as anti-cancer agents. Using murine in vitro models, we showed that following treatment with C2C12-EVs, lung carcinoma cells failed to colonise SkM cells, and that C2C12-EVs selectively exerted apoptosis on cancer cells. Uptake of C2C12-EVs by carcinoma cells caused changes in lysosomal function and mitochondrial membrane properties inducing cell death with elevated caspase 3 and 9. The C2C12-EVs also inhibited cell proliferation, affecting cell cycle arrest at S phase and inhibited cell migration. Proteomic analysis of C2C12-EV cargoes highlighted functional enrichment pathways involved in lysozyme function, HIF-1 and PI3K-Akt signalling, regulation of actin cytoskeleton, pyruvate metabolism, platelet activation, and protein processing in ER. Decorin, a muscle cell-specific cytokine released from myocytes in response to stress, was significantly enriched in C2C12-EVs and may contribute to C2C12-EVs’ inhibitory activity on cancer cells. C2C12-EVs may suppress cancer and potentially be used as therapeutic agents for cancer metastasis. Full article

Imidacloprid (IMI), a neonicotinoid insecticide, is widely recognized for its environmental persistence and toxicity to non-target aquatic organisms. Extended photoperiod exposure (EPE), an emerging anthropogenic stressor, further disrupts aquatic ecosystems by altering physiological and biological processes. However, their combined impacts on aquatic species remain insufficiently explored. This study evaluates the synergistic effects of IMI and EPE on Procambarus clarkii, an ecologically and economically significant crayfish species. Crayfish were exposed to 25 µg/L IMI under normal photoperiod (1000 lx, L:D = 12:12 h) and additional intensified and extended photoperiod (5000 lx, L:D = 18:6 h) treatments over one month. Key parameters, including survival rate, growth performance, oxidative stress markers, immune enzyme activities, neuroendocrine hormone levels, and gene expression, were assessed. The results indicate that EPE significantly amplifies the adverse effects of IMI. EPE reduced survival rates and growth performance, particularly in the 5000 lx group. IMI combined with EPE markedly elevated oxidative stress, as evidenced by increased malondialdehyde (MDA) levels and altered activities of superoxide dismutase (SOD) and catalase (CAT). Immune functions were impaired, with significant reductions in lysozyme (LZM) and acid phosphatase (ACP). Neuroendocrine disruption was observed through suppressed melatonin (MT) levels under EPE. Gene-expression analysis revealed upregulation of oxidative stress and apoptotic pathways (Cu/Zn-SOD, CAT and caspase-3) and downregulation of anti-apoptotic genes (bcl-2) and molt-inhibiting hormone (MIH). This study demonstrates that EPE exacerbates IMI-induced physiological and biochemical disruptions in P. clarkii. The findings highlight the pressing need for integrated management strategies addressing chemical and light pollution to protect aquatic ecosystems and sustain economically important species like crayfish. Full article

This study evaluated the effects of feeding juvenile southern catfish (Silurus meridionalis) with one of six isonitrogenous and isoenergetic diets where fish meal (FM) was replaced by black soldier fly larval meal (BSFLM) at 0%, 10%, 20%, 30%, 40%, and 50% levels on growth, muscle quality, antioxidant capacity, immune response, and gut microbiota of juvenile southern catfish (Silurus meridionalis). A total of 1620 fish (9.20 ± 0.15 g) were fed one of six experimental diets for 8 weeks. Results demonstrated that a 50% replacement (H50 group) significantly improved weight gain rate, specific growth rate, and protein efficiency ratio (p < 0.001). Antioxidant capacity (T-AOC) was enhanced in groups H30 and H50, while immune markers lysozyme (LZM) and alkaline phosphatase (AKP) showed mixed responses. Muscle texture properties such as chewiness and adhesiveness were significantly altered across treatments. Gut villi remained structurally intact in all groups, and liver histology appeared normal. No significant differences were found in muscle amino acid or fatty acid profiles. Gut microbiota analysis revealed shifts in microbial composition, with increased abundance of Clostridia and Escherichia and functional enrichment in metabolic pathways at higher substitution levels. Interspecies network analysis indicated potential cooperation among beneficial microbes through metabolite exchange. It is concluded that 50% BSFLM substitution optimizes growth performance, muscle quality, and antioxidant capacity, while modulating gut microbiota, indicating its promise as a sustainable FM alternative and functional ingredient in aquafeeds. Full article

This study presents a novel approach to intensifying the anaerobic digestion of sewage sludge through enzymatic pretreatment using hydrolytic enzymes—cellulase and lysozyme. It aims to determine how enzymatic activation affects the efficiency of methane fermentation, defined as the degree of organic matter decomposition and yield and composition of biogas. An experiment was carried out under mesophilic conditions over 20 days, analyzing the physicochemical properties of sludge, biogas production, methane content, and sanitary parameters. The addition of cellulase and lysozyme significantly enhanced process efficiency, increasing both the rate of organic matter degradation and biogas yield. The highest biogas production values (0.73 L·g⁻¹ d.m. for cellulase and 0.72 L·g⁻¹ d.m. for lysozyme) were obtained at a 4% (w/w) enzyme concentration, with a corresponding increase in the degree of organic matter decomposition to 78.7% and 80.0%, respectively. The produced biogas contained 58–61% methane, exceeding the values observed in the control sample, which indicates a positive effect of enzymatic activation on methane selectivity. Enhanced biogas production was attributed to improved hydrolysis of complex organic compounds, resulting in greater substrate bioavailability for methanogenic microorganisms. Moreover, methane fermentation led to the complete elimination of E. coli from all supernatants, confirming the hygienization potential of the process. The results of this study indicate that enzymatic pretreatment may serve as a viable strategy to improve both the energy efficiency and hygienic safety of anaerobic digestion processes, with relevance for future optimization and full-scale wastewater treatment applications. Full article

Climate change and global warming are concerning issues impacting various industries. In the aquaculture industry, these issues are more important in coldwater species, like rainbow trout Oncorhynchus mykiss. Hence, strategies to control these negative effects are worthy of study. Herbal feed additives are reliable tools to increase fish growth and health, thereby mitigating the drawbacks of climate change on fish. In this study, three diets containing 100 (100EO), 200 (200EO), and 400 (400EO) mg/kg essential oil of Abies sibirica (SBF) along with a control diet (CTL; unsupplemented) were fed to triplicate groups of fish for 60 days. Then the fish were exposed to a 96 h heat stress (25 °C) to monitor their survival and biochemical responses. The results showed that growth performance, feed efficiency, heat stress resistance, intestinal activity of digestive enzymes, and skin mucus immunological parameters significantly (p < 0.05) increased in the SBF essential oil treatments, and the highest increases were observed in the 100EO treatment, followed by the 200EO group. Dietary supplementation with SBF essential oil significantly (p < 0.05) mitigated heat stress-induced increases in plasma cortisol and glucose. Moreover, dietary SBF essential oil significantly (p < 0.05) enhanced immunological parameters such as plasma and intestinal lysozyme and immunoglobulin levels, and improved hepatic antioxidant defenses (including catalase, glutathione peroxidase, total antioxidant capacity, and reduced glutathione), while reducing lipid peroxidation. These effects were most pronounced in the 100EO and 200EO treatments, with the highest performance being observed in the former group. In conclusion, dietary SBF essential oil at 100 mg/kg is capable of augmenting growth performance, immunity, and antioxidant capacity, and suppressing physiological stress, thereby augmenting fish resilience against heat stress. Full article