Search Results (2,201)

Akkermansia muciniphila, a next-generation probiotic in the human intestinal mucus layer, exhibits significant health-promoting properties. However, traditional static culture systems fail to replicate the dynamic peristaltic environment of the gastrointestinal tract, limiting understanding of its metabolic characteristics. This study employed an improved gastrointestinal bioreactor simulating intestinal peristalsis to investigate A. muciniphila growth dynamics and metabolomic profiles under dynamic conditions. Dynamic cultivation significantly enhanced bacterial growth. Biomass reached 1.32 ± 0.03 g/L in bovine heart infusion (BHI) medium and 2.03 ± 0.05 g/L in BHI supplemented with 2.5 g/L porcine mucin. These values represent increases of 45.05% and 123.08% relative to static BHI cultures, respectively. Dynamic conditions markedly elevated short-chain fatty acid production (acetic, propionic, isobutyric, isovaleric acids). Untargeted metabolomics identified 1463 metabolites with 1294 showing significant differential expression. Dynamic cultivation substantially altered amino acid biosynthesis, fatty acid, purine, and pyrimidine metabolism. These findings advance the understanding of A. muciniphila physiology and provide insights into its metabolic characteristics under simulated intestinal conditions. Full article

The gut microbiota is a central regulator of metabolic function, and its disruption by a high-fat diet (HFD) is strongly linked to obesity and metabolic impairment. This study evaluated the potential of heat-killed Lactiplantibacillus plantarum beLP1 (beLP1^®) in alleviating HFD-induced metabolic and microbial imbalances in mice. Male C57BL/6N mice were fed an HFD for 10 weeks, with or without daily oral supplementation of beLP1 (≥3 × 10¹⁰ cells). Compared with untreated HFD mice, beLP1 supplementation reduced serum triglycerides by 35% and lowered liver enzymes AST and ALT by 17% and 36%, respectively. Blood glucose levels remained similar to the HFD group throughout the study period. Shotgun metagenomic analysis revealed that beLP1 restored gut microbial diversity, increased beneficial taxa such as Akkermansia and Faecalibaculum high. and reduced pro-inflammatory species including Streptococcus sp., Mucispirillum schaedleri and Clostridium cocleatum. These microbial changes were associated with partial normalization of the Firmicutes/Bacteroidota ratio and improvements in antibiotic resistance gene (ARG) profiles. Specifically, in silico analysis of the short-chain fatty acid (SCFA) synthesis pathways indicated that the potential for acetate and propionate production was maximized in the beLP1 group, resulting in the highest relative abundance among all groups. This functional enhancement directly correlated with the enrichment of key SCFA-producing taxa, particularly Akkermansia muciniphila, confirming that increased bacterial abundance suggests an enhanced functional potential for SCFA production. Furthermore, beLP1^® induced a selective modulation of gut ARGs, significantly reducing specific subtypes such as tetracycline and multidrug efflux genes, despite a slight increase in vancomycin resistance markers. Overall, our findings suggest that beLP1^® attenuated the rate of body weight gain during the initial weeks of HFD exposure and significantly improved markers of hepatic stress and lipid metabolism. Full article

This study aims to evaluate the blanching process of wasted Undaria pinnatifida as a ruminant feed source by assessing chemical compositions, in vitro nutrient digestibility, rumen fermentation characteristics, greenhouse gas emissions, and rumen microbes. The blanching process was conducted at different temperatures (15 vs. 80 vs. 90 °C) and times (2 vs. 4 min) to assess the chemical and mineral contents. Supplementation levels of U. pinnatifida (0 vs. 0.5 vs. 1 vs. 2%) were observed with the blanching process (non-blanching (NBL) vs. blanching (LOS)). Increasing blanching temperature and time decreased (p < 0.05) dry matter, crude ash, and the mineral contents, including sodium, phosphorus, and arsenic. Moreover, LOS treatment increased (p < 0.01) in vitro dry matter and neutral detergent fiber digestibility, ruminal pH, and the acetate-to-propionate ratio, but reduced (p < 0.01) CH₄ (mL/g NDFD). Additionally, 2% of LOS treatment reduced the abundance of protozoa, fungi, fibrolytic microbes, methanogenic archaea, Methanobrevibacter ruminantium, Methanosarcina barkeri, and Methanosphaera stadtmanae (p < 0.01). Therefore, blanching at 80 °C for 2 min improved the nutritional profile by reducing antinutritional minerals. Subsequent in vitro fermentation suggested that supplementing the diet with 0.5–1% of LOS improved digestibility and altered fermentation, potentially reducing methane yield (per NDFD). Full article

Lignocellulose represents an abundant repository of renewable carbon. Derived from various plant sources, it holds tremendous potential as a renewable and sustainable feedstock for the production of valuable chemicals and fuels. However, its solid fermentable compounds, cellulose and hemicellulose, are embedded within complex lignin structures and are therefore poorly accessible to microbial conversion. This paper describes an artificial rumen reactor (ARR) that uses anaerobic microbes from the cattle rumen to increase the release of fermentable carbon from recalcitrant biomass. We outline the development of an ARR for the efficient conversion of lignocellulosic grass into volatile fatty acids (VFAs), which are valuable precursors for the production of a range of bioproducts, including biofuels, biomaterials, and biochemicals. The ARR, a 4-L bioreactor equipped with a ceramic filtration unit, has been optimised and was operated for extended periods of continuous VFA production. Across distinct short- and long-term observation periods, and independent of the cow from which the rumen microbes originated, the bioreactor demonstrated the ability to sustain VFA production, indicating robustness and stability. At an input of 60–80 g dry grass d⁻¹, the system produced approximately 6 mol VFA per kg of dry matter input (DMI). The decoupling of the Solid Retention Time (SRT; 10 days) and the Liquid Retention Time (LRT; 0.5 days) prevented inhibition of the VFA production. The VFA profile was dominated by acetic and propionic acids, comprising 68% and 19%, respectively, with butyric acid and minor VFAs accounting for the remainder. The application of low oxygen levels (<10%) in the reactor via limited aeration did not affect the VFA yield or its profile. Full article

Background/Objectives: Autism spectrum disorder (ASD) is a neurodevelopmental condition increasingly associated with dysregulated neuroimmune signaling and altered neurotrophic homeostasis. Tumor necrosis factor-alpha (TNF-α) has been implicated in ASD pathophysiology; however, the downstream effects of TNF-α blockade on cytokine–neurotrophin interactions during neurodevelopment remain insufficiently characterized. In this study, we evaluated the effects of infliximab (IFX), a monoclonal anti-TNF-α antibody, on behavioral performance, neuroinflammatory cytokine profiles, glial activation, and brain-derived neurotrophic factor (BDNF) signaling in a propionic acid (PPA)-induced experimental ASD rat model. Methods: Experimental ASD was induced by propionic acid administration in rats. Animals were divided into control and treatment groups. Behavioral performance was assessed using the Morris Water Maze, direct social interaction, and three-chamber sociability tests. Levels of TNF-α, interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and BDNF were measured in serum, hippocampal, and cerebellar tissues. Microglial and astrocytic activation were evaluated using CD11 and GFAP immunohistochemistry. Results: PPA administration resulted in pronounced impairments in learning, memory, and social behaviors, accompanied by elevated proinflammatory cytokine levels, increased BDNF expression, and marked glial activation in the hippocampus and cerebellum. Although IFX treatment significantly reduced TNF-α levels in central tissues, it did not improve behavioral deficits and was associated with persistently elevated IL-1β and IL-6 levels, sustained glial reactivity, and further alterations in BDNF levels. Conclusions: These findings suggest that TNF-α suppression alone does not normalize the disrupted cytokine–neurotrophin axis and may differentially modulate BDNF-related neuroplastic signaling during development. In conclusion, this study indicates that non-selective TNF-α blockade during neurodevelopment fails to confer behavioral benefit in experimental ASD and highlights the importance of considering cytokine–BDNF pathway interactions when designing immunomodulatory strategies for neurodevelopmental disorders. Full article

This meta-analysis and meta-regression evaluated how malic acid supplementation modulates rumen fermentation and its consequences for growth performance, nutrient digestibility, and carcass traits in lambs. Effect sizes (ES) were estimated using a random-effects model. Dietary composition was explored by meta-regression as a key source of heterogeneity, and subgroup analyses were used to compare free malic acid (FMA) and malate. Ruminal pH was not affected by malic acid supplementation. In contrast, total volatile fatty acid concentration increased with malic acid supplementation, particularly in studies using FMA. No effects were detected for propionate concentration, whereas acetate concentration increased (ES = 0.502; p = 0.036). A tendency toward a reduced ruminal acetate proportion was observed (ES = −0.683; p = 0.072). Malic acid supplementation tended (p = 0.057) to increase body weight gain (BWG; ES = 0.325) and final body weight (FBW; ES = 0.234). Malic acid supplementation did not affect carcass traits or overall nutrient digestibility. Meta-regression consistently identified fiber intake-related variables as major moderators of the effects of malic acid. Overall, the effects of malic acid supplementation on lamb performance appear to be primarily driven by its modulation of rumen fermentation and strongly conditioned by dietary context. Full article

This study investigated the interactive effects of corn silage and ramie silage on in vitro rumen fermentation characteristics, aiming to provide a scientific basis and empirical evidence for the rational incorporation of ramie into ruminant diets. Four binary substrate mixtures were formulated based on dry matter (DM) mass ratios of corn silage to ramie silage: 100:0 (CON), 60:40 (R40), 20:80 (R80), and 0:100 (R100). Rumen fluid was collected from three adult Liuyang black goats surgically fitted with permanent rumen cannulas, and a standardized 48 h in vitro batch culture assay was conducted. Results demonstrated that increasing the proportion of ramie silage significantly decreased (p < 0.05) the DM degradation rate, neutral detergent fiber (NDF) degradation rate, acid detergent fiber (ADF) degradation rate, and total gas production per gram of substrate DM. Specifically, CON and R40 exhibited significantly higher values for all four parameters than R80 and R100 (p < 0.05). Methane production was significantly reduced in all ramie-containing treatments relative to CON (p < 0.05), whereas hydrogen production increased progressively with ramie inclusion level, with CON yielding significantly less H₂ than both R80 and R100 (p < 0.05). Regarding fermentation parameters, increasing ramie proportion elevated (p < 0.05) both fermentation fluid pH and the acetate-to-propionate ratio, while total volatile fatty acid (TVFA) concentration declined linearly (p < 0.05). TVFA concentrations did not differ significantly between CON and R40, yet both were significantly greater than those in R80 and R100 (p < 0.05). Collectively, these findings indicate that ramie silage is a nutritionally valuable forage with potential as a high-quality partial replacement for conventional silages in ruminant feeding systems; however, its inclusion in corn–ramie mixed silages should not exceed 40% (on a DM basis) to maintain optimal fermentative efficiency and nutrient degradability. Full article

Rumen microbial fermentation plays a central role in nutrient utilization and milk production in dairy cows. This study evaluated the effects of supplementation with exogenous fibrolytic enzymes, alone or in combination with live yeast on rumen microbiota, fermentation characteristics, nitrogen-related metabolites, and production performance during the transition from outdoor grazing to indoor housing. Thirty Lithuanian Red dairy cows were assigned to control (CTR), enzyme (E), or enzyme plus yeast (YE) treatments across outdoor (OD) and transit (T) periods, while nine cows (three per group) were selected for rumen and microbiota analysis. Rumen bacterial communities were characterized using 16S rRNA gene sequencing, and functional parameters were evaluated using linear mixed-effects models. Supplementation resulted in selective changes in several bacterial genera, including Blautia spp., WPS-2, Ruminococcus spp., Erysipelotrichaceae UCG-009, Sharpea spp., uncultured Bacteroidales, and Prevotellaceae UCG-003, and was associated with alterations in fermentation patterns, particularly propionate concentration, and in nitrogen metabolism, including putrescine dynamics. The transition period significantly influenced microbial diversity and total bacterial abundance across treatments. Cows in the YE group maintained higher milk yield during the transition period. Overall, dietary supplementation modulated specific rumen metabolic responses and contributed to production stability without causing large-scale changes in overall microbial structure. Full article

Short-chain fatty acids (SCFAs) are key biomarkers of gut microbiota activity; however, routine quantification in fecal samples relies largely on chromatography, which is instrument-intensive and throughput-limited chromatography techniques. Herein, we present a rapid machine-learning-assisted electroanalysis platform for SCFAs profiling that integrates a disposable three-electrode planar gold chip with voltammetric fingerprinting and artificial neural network (ANN)-based signal decoupling. To generate orthogonal chemical information and improve the discrimination of structurally similar species, a dual pretreatment strategy combining acid-catalyzed esterification and alkaline dissociation was employed prior to electrochemical analyses. Differential pulse voltammetry (DPV) and cyclic voltammetry (CV) were employed to acquire high-dimensional fingerprints, from which current-, potential-, and area-based descriptors were extracted using a cross-information feature strategy. A hierarchical modeling framework improved total SCFAs prediction by incorporating ANN-predicted propionate and butyrate concentrations as auxiliary inputs. While linear calibration was achievable in standard mixtures, direct linear models performed poorly in real fecal matrices due to strong sample-dependent matrix interference. In contrast, the ANN captured nonlinear relationships among multifeature inputs and suppressed matrix effects. Validation against gas chromatography–mass spectrometry in an independent fecal test cohort (n = 30) demonstrated excellent agreement and low prediction errors, with mean absolute error/root mean square error values of 0.063/0.072 mM (propionic acid), 0.029/0.034 mM (butyric acid), and 0.135/0.202 mM (total SCFAs). The DPV/CV acquisition requires only minutes per sample, whereas pretreatment takes 1~3 h depending on the target route but can be performed in parallel for batch processing; thus, overall throughput is determined mainly by batch pretreatment rather than per-sample instrument time. This electrochemical–ANN workflow provides a portable, high-throughput alternative to chromatography for fecal SCFAs profiling in clinical screening and microbiome research. Full article

Water-soluble carbohydrate (WSC) is the key to producing quality forage silage and an important energy source for ruminants. The aim of this study was to investigate the effect of different silages used as roughage sources [whole-plant sugarcane silage (WSS) vs. elephant grass silage (EGS)] with varying levels of WSC on silage quality, buffalo growth performance, apparent digestibility, rumen fermentation, and microbial communities. Sixteen healthy male crossbred buffaloes were randomly divided into two treatment groups, with eight buffaloes/treatment. One group was fed whole-plant sugarcane silage, and the other group was fed elephant grass silage. Compared with EGS, WSS had higher WSC, lactic acid, and ethanol, but lower pH, ammonia nitrogen, propionic acid, and butyric acid (BA) contents (p < 0.05). Potential probiotics (e.g., Lactiplantibacillus and Hanseniaspora) were more abundant in WSS than in EGS (p < 0.05). Moreover, the feed conversion rate was higher in HWS (p < 0.05). However, rumen fermentation parameters were unaffected by diet (p > 0.05). Moreover, feeding WSS had lower dry matter digestibility (DMD), organic matter digestibility (OMD), and lower acid detergent fiber digestibility (ADFD) (p < 0.05). After WSS feeding, ruminal Treponema_2 was strongly associated with DMD, OMD, and ADFD (p < 0.05), and also showed positive correlations with BA and PA contents in WSS (p < 0.05). Additionally, rumen Ruminiclostridium_5 and Pseudozyma was associated with DMD and ADFD after being fed EGS (p > 0.05), respectively, but the Pseudozyma was associated with BA (p < 0.05) and Clostridium_sensu_stricto_11 (p > 0.05) in EGS. Our findings indicated that WSS exhibited superior fermentation quality and harbored potential beneficial microbes, whereas EGS showed higher apparent nutrient digestibility in buffalo but also contained undesirable bacteria (e.g., Clostridium_sensu_stricto_11). Future research should investigate the long-term effects of WSS feeding on buffalo health, immunity, and production performance, as well as its impact on rumen microbiota stability, to fully assess its potential as a safe and sustainable roughage source. Full article

To explore the appropriate supplementation rate of yeast culture (YC) in Kazakh sheep during fattening, the effects of different YC supplementation rates on rumen fermentation parameters and microbial community were studied through in vitro rumen fluid fermentation experiments. A 0.40 g high-concentrate diet was used as the fermentation substrate, and five groups were added with YC at 0% (CK), 1.25% (YC1), 2.5% (YC2), 3.75% (YC3) and 5% (YC4) of dietary dry matter, respectively. Anaerobic fermentation was carried out for 48 h in 60 mL fermentation broth. The results showed that the 48 h GP and microbial crude protein (MCP) concentration in all YC supplementation groups were significantly higher than those in the CK group (p < 0.05). The concentrations of total volatile fatty acids (TVFA) and propionate in the YC1 and YC2 groups were significantly increased and the A/P ratio in the two groups was significantly decreased (p < 0.05). The Multi-factor Comprehensive Evaluation Index (MFAEI) calculation indicated that 1.25% was appropriate. The YC1 and YC2 groups significantly increased the richness and diversity of rumen bacterial communities (Chao1 and Shannon indices, p < 0.05), and significantly increased the relative abundance of Bacteroidota and NK4A214_group (p < 0.05), while significantly decreasing the relative abundance of the potential pathogenic bacterium Campylobacter (p < 0.05). Ustilago abundance was significantly suppressed in all the YC-supplemented groups (p < 0.05). The most effective YC supplementation rate among the tested doses was 1.25% according to the MFAEI and key microbial indicators. The results suggest that dietary supplementation of 1.25% YC (dry matter basis) may beneficially modulate rumen fermentation parameters under in vitro conditions, providing a reference for further in vivo studies on its application in fattening Kazakh sheep. Full article

In this study, we examined whether dietary ADY improves growth, digestibility of feed nutrients, meat quality, and rumen microbial ecology in lambs. This experiment enrolled 90 healthy, similarly weighted (29.0 ± 0.5 kg) four-month-old Duhan lambs, which were randomly and evenly distributed into two treatment groups: a control group fed the basal diet and an ADY group fed the basal diet supplemented with 0.3 g/d per lamb of active dry yeast. The supplementation amount was adjusted weekly according to feed intake to maintain a constant daily dose. The results showed that, compared with the control group, ADY significantly increased the lambs’ average daily gain (ADG) and enhanced the apparent digestibility of neutral detergent fiber (NDF), crude protein (CP) (p < 0.05), and significantly reduced the feed conversion ratio (F/G) (p < 0.05). These improvements were accompanied by a shift in rumen fermentation toward propionate production, evidenced by higher NH₃-N, Total volatile fatty acids (TVFAs) and propionate proportion and a lower acetate proportion and acetate-to-propionate ratio (p < 0.05). ADY also altered the rumen microbiota, increasing Proteobacteria and Succinivibrionaceae_UCG-001 while decreasing norank_o_Clostridia_UCG-014 (p < 0.05). In muscle, ADY significantly increased the proportions of C14:0 and C18:3n-3 (p < 0.05). In addition, the proportion of C13:0, C18:0 and C18:2n-6t were significantly reduced (p < 0.05). In conclusion, dietary supplementation with ADY enhanced rumen fermentation, improved rumen microbial composition, and promoted nutrient utilization in lambs, thereby improving growth performance and meat quality. In addition, certain rumen microbial taxa may be associated with the formation of specific muscle fatty acids. Full article

Background/Objectives: Recently, a randomized clinical trial evaluated whether a six-month probiotic administration could reduce symptom severity in preschool children with Autism Spectrum Disorders (ASD), with (GI) or without (NGI) gastrointestinal symptoms. Significant positive changes were observed only in NGI children. A second explorative study on children prior to intervention identified a fecal metabolome fingerprint associated with ASD severity. Building on these findings, the present study aimed to assess whether metabolomics could monitor changes in ASD severity following probiotic administration using a subset of samples from the same trial. Second, this study aimed to identify fecal metabolites to be monitored in children to predict whether their autism severity may decrease after probiotic or placebo treatment. Methods: Evaluations of the fecal metabolome and microbiota could be completed on 57 children before and after a double-blind administration of a probiotic mixture or a placebo. Results: In NGI children the probiotic was found to influence the concentration of the amino acids aspartate, leucine, tryptophan, and valine, together with nicotinate and the short chain fatty acids acetate, butyrate, isobutyrate, and propionate. Lactobacilli and Sutterella showed significant changes in response to probiotic administration (p < 0.05). Acetate, 4-hydroxyphenyl, galactose, proline, and tyramine were identified as key fecal metabolites for prediction purposes. Conclusions: The present exploratory analysis, despite the small sample size, suggests that fecal metabolomics may provide a useful approach for monitoring and potentially for predicting changes in ASD severity following probiotics administration. Full article

Styela plicata, an edible ascidian rich in diverse bioactive constituents, represents a promising source of marine natural products for therapeutic discovery. Here, bioactive components from a 95% ethanol extract of S. plicata (ESP) were characterized by HPLC-MS/MS, showing that the major constituents were oxygenated small molecules dominated by fatty acyls and carboxylic acid derivatives. In a mouse model of alcohol-induced liver injury, H-ESP treatment (300 mg/kg) significantly reduced serum levels of AST, ALT, and TG (p < 0.01), while effectively ameliorating pathological changes in liver tissue, reducing lipid accumulation and inflammatory responses. Transcriptome sequencing (H-ESP vs. model group) identified 1097 differentially expressed genes (172 upregulated and 925 downregulated), and KEGG analysis highlighted significant enrichment of the Toll-like receptor signaling pathway. ESP modulated hepatic metabolite expression, suppressed inflammation via TLR-4/NF-κB pathway inhibition, and boosted antioxidant defenses by activating Nrf2/HO-1 signaling, which was further confirmed by RT-qPCR and immunohistochemistry. ESP increased intestinal SCFAs (acetate, propionate, isobutyrate; p < 0.05), improved α-diversity and the Firmicutes/Bacteroidetes ratio, reversed shifts in Lactobacillus and Bifidobacterium, and partly restored Odoribacter, supporting a gut–liver axis mechanism. Overall, these findings indicate that ESP exerts hepatoprotective effects by modulating the gut–liver axis, and they provide insights for developing natural therapeutics against alcoholic liver disease. Full article

Background: Gut microbiome-derived metabolites, particularly short-chain fatty acids (SCFA) and tryptophan derivatives, are central mediators of the gut–brain axis. This ex vivo study assessed how nutritional interventions impact such metabolites during early life, a critical period for neurodevelopment. Methods: The effects of galacto-oligosaccharides (GOS), nutrient blends (vitamins, minerals and amino acids) and their combinations were evaluated in the gut microbiomes of infants (2–4 months, n = 6) and young children (2–3 years old, n = 6) using the ex vivo SIFR^® technology. Results: Baseline microbiome composition was age-dependent, with infants displaying lower α-diversity and greater interpersonal variability. After ex vivo incubation, nutrient blends increased the propionate/butyrate ratio and branched-chain fatty acids in young children and elevated several B-vitamins and amino acid-derived metabolites, including indole-3-carboxaldehyde, imidazoleacetic acid and pipecolinic acid. Combining nutrient blends with GOS exhibited potential synergistic effects on propionate (infants) and 2-hydroxyisocaproic acid (HICA, both age groups). GOS strongly stimulated Bifidobacteriaceae and increased metabolites linked to bifidobacterial metabolism like acetate, HICA, N-acetylated amino acids, aromatic lactic acids and acetylagmatine; in young children, butyrate and γ-aminobutyric acid (GABA) also increased. Conclusions: Combinations of GOS with nutrient blends impacted microbiome-derived metabolites associated with the gut–brain axis, with potential synergistic increases of metabolites with emerging roles in neurodevelopment, including GABA, acetylagmatine and HICA. Despite shared bifidogenic effects, differences between age groups indicate that microbiome maturity may influence responses to nutritional intervention. Future clinical studies are needed to determine whether these metabolite changes translate into neurodevelopmental benefits in vivo. Full article