Search Results (7,471)

Food-associated microorganisms represent a diverse reservoir of bacterial strains with functional traits potentially relevant to host–microbiome interactions, stress resilience, metabolic processes, and microbial competition. In this study, we performed a multi-omics characterization of food-origin bacterial isolates to identify a potential candidate strain for further studies on microbiome–microgreens interactions. Over one hundred bacterial isolates were initially recovered from various food products and subjected to molecular screening, followed by whole-genome sequencing, metabolic profiling, transcriptomic chcracterisation and morphological analysis of selected strains. Comparative characterization revealed significant differences among the strains. Among the tested isolates, strain B107/23, identified as Priestia megaterium (formerly Bacillus megaterium), had the highest number of genes associated with stress responses, nutrient uptake, plant growth-promoting traits, and colonization mechanisms. Phenotypic characterization later demonstrated high metabolic flexibility and a low substrate stress index, while transcriptomic profiling revealed the expression of genes associated with stress responses and metabolic activity. Overall, the integration of genomic, phenotypic, and transcriptomic results distinguished B107/23 as the most promising candidate among the tested isolates and revealed the molecular traits potentially significant to interactions between the microbe and the host. These findings provide a basis for the validation of the selected strain in microgreens systems, including studies addressing plant stress resilience and post-harvest quality. Full article

Plant patches are a common feature of dryland ecosystems, but how within-patch spatial position is associated with plant functional traits–and whether these patterns differ among organs—remains poorly understood. We sampled twigs, leaves, and flowers of the C₄ desert plant Salsola collina from the centers and edges of natural patches in arid Xinjiang, China, using five plots with three patches per plot. For each organ, we measured the concentrations of carbon (C), nitrogen (N), phosphorus (P), sucrose (SUC), fructose (FRU), total soluble sugars (SS), and starch (ST), and calculated non-structural carbohydrate (NSC) concentrations as the sum of SS and ST. Treating the plot as the experimental unit and accounting for paired center–edge observations within plots, we evaluated the effects of patch position, organ type, and their interaction using two-way ANOVA and distance-based multivariate analyses. Organ type accounted for the largest proportion of multivariate trait variation (R² = 0.90), whereas patch position and the Patch × Organ interaction each explained approximately 4% (all p < 0.001). Significant Patch × Organ interactions were detected for 14 of 17 traits, indicating that center–edge differences varied among organs for most traits. N concentrations were higher at patch edges in all three organs, whereas P concentrations were higher in twigs and flowers only; C showed no significant center–edge differences. NSC concentrations were higher at patch edges in all three organs, while SS was higher in edge twigs and leaves and ST was higher in edge flowers. These patterns were accompanied by contrasting organ-specific differences in carbohydrate composition and elemental stoichiometry. Because environmental variables were not measured concurrently, the observed patterns should be interpreted as spatial associations rather than demonstrated environmental mechanisms. These findings indicate that organ identity and within-patch position are jointly associated with variation in the carbon-, nutrient-, and carbohydrate-related traits of S. collina, highlighting the importance of considering organ-specific variation when evaluating fine-scale spatial heterogeneity in dryland plants. Full article

Conventional unprocessed black soldier fly larval meal often exerts inconsistent growth-promoting effects on broilers. This variability may arise from multiple factors, including dietary inclusion level, rearing substrate, processing method, defatting status, and nutritional composition; indigestible macromolecular proteins and chitin are among these contributing factors. Enzymatically hydrolyzed Hermetia illucens larval homogenate (EH-BSFLH) is rich in small peptides and lauric acid, which theoretically improve nutrient bioavailability and intestinal health, making it a promising novel functional feed additive. This study aimed to characterize the nutritional profile of a commercially available enzyme hydrolyzed Hermetia illucens larval homogenate (EH-BSFLH), and to explore the effects of 2% and 6% dietary EH-BSFLH supplementation on 1–21-day-old Arbor Acres (AA) broilers. A total of 126 one-day-old AA broilers were randomly assigned to three treatments in this 21-day starter feeding trial, with seven pens per treatment. Nutrient composition of EH-BSFLH and multiple phenotypic indicators of broilers were measured and statistically compared. EH-BSFLH utilized in this essay possessed unique nutritional characteristics, including high acid-soluble protein proportion (96.41% relative to total protein), abundant lauric acid (26.62% of total fatty acids), and a calcium-to-phosphorus (Ca/P) ratio of 3.10:1. The 6% EH-BSFLH supplementation increased (p < 0.02, FDR < 0.05) 21-d live weight, average daily feed intake (ADFI), average daily gain (ADG), eviscerated yield and semi-eviscerated yield relative to the control, while no significant difference in feed conversion ratio (FCR) was detected across treatments. Dietary EH-BSFLH at 2% and 6% increased (p < 0.02, FDR < 0.05) 24 h post-mortem pH of breast and thigh muscles, without affecting meat color or shear force. The 6% EH-BSFLH group elevated (p < 0.02, FDR < 0.05) 10 amino acids in thigh muscle, while maintaining stable essential amino acids/total amino acids (EAA/TAA) and essential amino acids/non-essential amino acids (EAA/NEAA) ratios. Moreover, 6% EH-BSFLH decreased duodenal crypt depth and increased Villus Height/Crypt Depth (VH/CD) ratios in duodenum and jejunum (p < 0.02, FDR < 0.05). Serum biochemical variables were generally comparable between groups, though differences were observed for glucose, albumin, alanine aminotransferase, aspartate aminotransferase, and total cholesterol (TCHO) (p < 0.02, FDR < 0.05). None of these values indicated obvious pathological alterations. This modified insect product exhibits promising application potential for young broiler feeding under the current trial conditions, and the observed shifts in serum metabolic indices may provide partial reference for balancing production performance and broiler physiological status in practical diet formulation. Full article

Sarcopenia is an independent predictor of adverse outcomes in hospitalized patients, associated with increased mortality, prolonged length of stay, and functional decline, yet it remains underdiagnosed owing to inconsistent diagnostic criteria and the practical challenges of assessing acutely ill patients. This narrative review summarizes current evidence on sarcopenia in hospitalized patients, with emphasis on diagnostic approaches, nutritional requirements, and optimal nutritional support strategies. Sarcopenia is particularly prevalent among critically ill patients and is driven by the convergence of systemic inflammation, immobility, and anabolic resistance. Diagnosis remains challenging: no ICU-specific framework exists, and the recent Global Leadership Initiative of Sarcopenia consensus established a unified, setting-independent conceptual definition as a foundation for future operational criteria, with ICU-specific frameworks yet to be developed. Bedside tools, including handgrip strength, bioelectrical impedance analysis, and ultrasound, retain strong prognostic value despite imperfect agreement with reference imaging. Nutritional intervention is a key component of management; however, large-scale trials have demonstrated that higher energy or protein delivery does not universally improve outcomes and may be harmful in selected critically ill patients. A gradual increase in energy and protein delivery according to the phase of illness, with protein targets of ≥1.2–1.5 g/kg/day, is recommended; leucine-enriched supplementation may help counteract anabolic resistance, although direct evidence in non-ICU hospitalized patients remains limited. Early mobilization is an essential complement, as nutrient provision without a mechanical stimulus cannot reverse immobility-induced muscle loss. A practical clinical algorithm integrating screening, nutritional target-setting, route selection, and reassessment is proposed to guide decision-making in this population. Future research should focus on improving the accuracy and feasibility of muscle assessment tools in critically ill patients to enable the timely diagnosis of sarcopenia and nutritional intervention, alongside the development of validated multi-marker panels and adequately powered trials examining the combined effect of nutrition and exercise in this population. Full article

Long-term fertilization can reshape soil microbial communities and alter soil ecosystem functioning. However, few studies have comprehensively investigated changes in microbial functional potential for carbon, nitrogen, phosphorus, and sulfur (CNPS) cycling in bulk and rhizosphere soils in response to long-term fertilization, or the relationships between microbial taxonomic shifts and functional responses. We combined 16S rRNA gene amplicon sequencing with high-throughput quantitative PCR targeting 71 CNPS cycling-related genes in a 32-year field experiment comprising an unfertilized control (CK), mineral fertilizer (NPK), manure (M), and NPK combined with manure (NPKM). The results showed that both mineral NPK fertilizer and manure increased soil nutrient availability but had contrasting effects on soil pH. Relative to CK, NPK, M, and NPKM increased rapeseed grain yield by 303.9%, 108.7%, and 397.8%, respectively, and biomass by 303.0%, 99.6%, and 410.5%, respectively. Fertilization also reshaped bacterial community composition, and the responses were similar between bulk and rhizosphere soils. In contrast, functional-gene abundances showed significant soil compartment-dependent responses to fertilization. NPK fertilization reduced most rhizosphere gene groups associated with C degradation, N transformation, P mobilization, and S cycling while increasing nitrification-related genes, whereas manure increased genes involved in carbohydrate degradation, methane metabolism, multiple N transformations, organic P mineralization, phosphate solubilization, and sulfate reduction. Network analysis showed that genes mediating similar processes clustered within common modules and were associated with shared bacterial taxa. Crop productivity was associated with variation in genes involved in N fixation and organic P mineralization in rhizosphere soil. Our results revealed associations between functional-gene abundances, soil nutrients, crop productivity, and bacterial community changes under long-term fertilization, highlighting the importance of quantitative functional-gene analysis as an indicator for evaluating soil multifunctionality. Full article

Hulless barley (Hordeum vulgare L. var. nudum) is a nutritious cereal but suffers from prolonged cooking and asynchronous gelatinization with rice. Pre-gelatinization technology can address these issues, yet optimal processing parameters are needed to preserve its bioactive components. This study optimized the infiltration and drying processes for pre-gelatinized hulless barley rice. Infiltration cycles and durations were evaluated using sensory scores, texture analysis, and gelatinization degree; drying temperatures (80–160 °C) and times were optimized based on quality and nutrient retention. Secondary infiltration for 4 h significantly outperformed traditional soaking, yielding higher retention of polyphenols (130.73 mg/100 g), flavonoids (50.01 mg/100 g), and β-glucan (2.96%). Drying at 80–100 °C for 90–120 min produced the best sensory and textural properties, maximal retention of polyphenols (128.46 mg/100 g), flavonoids (45 mg/100 g), β-glucan (3.1%), and reducing sugars (482.84 mg/100 g at 80 °C/120 min), and the highest DPPH and ABTS⁺ radical scavenging activities. Higher temperatures (≥120 °C) caused significant degradation of these functional components and reduced antioxidant capacity. The optimal combination—secondary infiltration for 4 h followed by drying at 80–100 °C for 90–120 min—effectively balances processing efficiency with nutritional and functional quality. This optimized process provides a practical technical solution for large-scale production of high-quality pre-gelatinized hulless barley rice. Full article

Carbon-based nanomaterials have emerged as promising tools for addressing major challenges in sustainable agriculture, including declining soil fertility, climate change, resource inefficiency, and increasing environmental pollution. Among these materials, carbon nanotubes (CNTs) and carbon quantum dots (CQDs) have attracted considerable attention because of their unique physicochemical properties and diverse interactions with plants and soil systems. This review provides a comprehensive comparative analysis of CNTs and CQDs, emphasizing how their structural characteristics govern environmental fate, plant uptake, physiological responses, and stress mitigation mechanisms. CNTs primarily function as one-dimensional nanostructures that improve soil properties, facilitate nutrient delivery, and immobilize environmental contaminants, whereas CQDs, owing to their ultrasmall size, excellent water dispersibility, and intrinsic fluorescence, actively regulate plant metabolism, photosynthesis, nutrient acquisition, and antioxidant defense. Their distinct transport pathways, rhizosphere interactions, and subcellular localization are critically evaluated alongside recent advances in synthesis, surface functionalization, and physicochemical modification. The review further summarizes current evidence regarding their roles in enhancing tolerance to heavy metal toxicity, salinity, and drought stress through modulation of reactive oxygen species scavenging, osmotic regulation, ion homeostasis, and stress-responsive signaling pathways. Potential phytotoxicity, environmental persistence, ecological risks, and green synthesis strategies are also discussed to provide a balanced assessment of their agricultural applications. Emerging opportunities for synergistic CNT–CQD composite systems are discussed, together with major knowledge gaps and future research priorities, including machine learning-assisted nanomaterial design, multi-omics characterization, long-term field validation, and life cycle-based risk assessment. This review establishes a conditional, context-dependent structure–behavior–function conceptual framework that provides theoretical guidance for the rational design and safe implementation of carbon nanomaterials in next-generation sustainable agriculture. Full article

Extrusion technology enables the development of nutrient-dense snacks, but most commercial extrudates rely on nutrient-poor cereal flours. This study evaluates the effect of the incorporation of mushroom-fermented flours (MFF) from Ganoderma sessile (GS) and Pleurotus ostreatus (PO), at increasing substitution levels, on the physicochemical, structural, nutritional, and functional (bioactive) properties of corn-based extrudates, relative to a corn control. Partial substitution (10–20%) significantly reduced expansion index and hardness, and reduced or maintained density, relative to the corn control, without causing structural collapse. At 100% substitution, the two species diverged sharply: GS100 retained an expansion index statistically indistinguishable from the corn control and a density numerically closer to, though still significantly different from, the corn control, while developing a markedly lower final pasting viscosity than the corn-containing formulations; PO100 produced the highest density and lowest expansion index of the entire data set, with an essentially flat pasting profile, despite both flours showing comparable shifts in protein, ash, and carbohydrate content. Incorporation of MFF markedly increased protein, ash, total phenols, triterpenes, and reducing power relative to the corn control, with the largest gains at 100% substitution; free radical-scavenging capacity increased significantly only in the 100% MFF extrudates. These results demonstrate that solid-state fermentation combined with extrusion is an effective strategy for producing nutritionally and functionally enhanced snacks. Full article

Culled beef cows may retain practical value for fattening after reproductive use, but the rumen and host physiological characteristics associated with this response remain incompletely characterized. This study compared culled beef cows (CBC; n = 9) with fattening bulls (CON; n = 9) using 18 healthy Simmental crossbred cattle fed the same experimental total mixed ration (TMR) under identical feedlot conditions. Growth performance, body condition score (BCS), apparent nutrient digestibility, economic outcomes, blood indices, rumen fermentation, ruminal enzyme activities, rumen metagenomes, and plasma untargeted metabolomes were assessed. Compared with CON, CBC had higher final body weight, average daily gain, feed intake, and apparent digestibility of dry matter, crude protein, and neutral detergent fiber, whereas the feed-to-gain ratio did not differ significantly between groups (p = 0.692). BCS increased significantly from day 0 to day 60 within both cattle categories. Because different, non-cross-calibrated BCS reference frameworks were used, no between-group comparison of BCS was made. Economic analysis indicated that the value attributable to actual live-weight gain was similar between groups when within-category prices were held constant, whereas the relative economic outcome varied with feed costs and changes in live-cattle market prices. CBC also showed more propionate-oriented fermentation, greater fibrolytic enzyme activities, distinct carbohydrate-active enzyme profiles, higher insulin and leptin, and lower non-esterified fatty acids and β-hydroxybutyrate. Because lean and fat mass were not directly measured, the tissue basis of the greater weight gain remains unresolved. Overall, CBC exhibited a stronger fattening response accompanied by coordinated differences in rumen function and host metabolic profiles. Full article

Efficient management of soil nitrogen and phosphorus availability is important for soil fertility and crop productivity, but the field responses of rhizosphere microbial communities to arbuscular mycorrhizal fungi (AMF) and clay-mineral amendments remain unclear. A wheat-field experiment compared four treatments: an untreated control (A), bentonite (B; 39 kg ${\mathrm{plot}}^{-1}$), sepiolite (C; 39 kg ${\mathrm{plot}}^{-1}$), and AMF inoculation (E; 2.25 kg inoculum ${\mathrm{plot}}^{-1}$; 120 spores ${\mathrm{g}}^{-1}$). Each treatment comprised six 4 × 7.5 m plots. Soil physicochemical properties were evaluated using all six plots, whereas rhizosphere microbial sequencing used three plots per treatment (plots 2, 4, and 6; n = 3). Treatment E showed higher mean alkali-hydrolyzable nitrogen (101.50 vs. 93.33 mg ${\mathrm{kg}}^{-1}$) and available phosphorus (56.00 vs. 20.98 mg ${\mathrm{kg}}^{-1}$) than the control, whereas treatment B showed a 32.8% higher mean available potassium. None of the bacterial or AMF alpha-diversity indices differed significantly among treatments. Taxonomic summaries showed descriptive variation among treatments, whereas principal component analysis (PCA) and analysis of similarities (ANOSIM) did not detect significant treatment-level separation of either the bacterial or AMF communities. Shotgun metagenomic annotation, rather than 16S-based functional inference, further showed descriptive differences in nitrogen- and phosphorus-related gene-abundance profiles and functional potential; Pseudomonadota and Actinomycetota accounted for substantial shares of several nutrient-cycling gene categories, while pstB showed comparatively pronounced numerical variation among treatments. These metagenomic profiles represent genetic functional potential and do not demonstrate transcript abundance or metabolic activity. Overall, AMF inoculation and clay-mineral amendments were associated with descriptive differences in nutrient availability, taxonomic profiles, and metagenomic functional potential in alkaline wheat farmland, but significant treatment-level separation of the overall bacterial and AMF communities was not detected. Full article

Active dry yeast (ADY) as a feed additive has been proven to have functions such as enhancing feed digestion and utilization rates, strengthening the body’s immunity, alleviating stress and improving rumen fermentation. This experiment mainly explores the effects of a new type of ADY on the production performance, rumen fermentation and nutrient digestibility of dairy cows. A total of 100 Holstein cows with similar milk yield (46.9 ± 8.5 kg/d), days in milk (66.7 ± 5.4 d), and parity (2.5 ± 1.0) were selected and randomly allocated into four groups (25 cows per group). The treatments were as follows: (1) Control group (CON), cows fed the basal diet only; (2) ADY I group (ADY I), basal diet supplemented with 5 g/d of common ADY (RF7, 1 × 10¹⁰ CFU/g) per cow; (3) Low ADY II group (LADY II), basal diet supplemented with 5 g/d of Actisaf Sc48 (viable yeast cell concentration is 1 × 10¹⁰ CFU/g) per cow; (4) High ADY II group (HADY II), basal diet supplemented with 10 g/d of Actisaf Sc48 per cow. The 10-week experimental period consisted of a 1-week adaptation period followed by a 9-week formal experimental period. The results showed that, compared with the CON group, mean dry matter intake (DMI) increased by 3.48%, 5.35%, and 6.96% in the ADY I, LADY II, and HADY II groups, respectively (p < 0.05), with a significant treatment × time interaction (p < 0.001). Compared with the CON and ADY I groups, the HADY II group showed increases in milk yield of 5.76% and 4.76% and in apparent crude protein (CP) digestibility of 5.22% and 3.17%, respectively (p < 0.05). Milk lactose concentration was 3.43% higher in the HADY II group than in the ADY I group (p < 0.05). Additionally, milk linolelaidic acid concentration was lower in LADY II and HADY II than in CON (p < 0.05). Compared with the CON group, the LADY II and HADY II groups showed increases in ruminal total volatile fatty acid (TVFA) concentrations of 42.11% and 44.72% and in acetate concentrations of 45.76% and 46.71%, respectively (p < 0.05). In this trial, the group receiving ADY II at 10 g/cow/day showed the best overall performance. However, this finding does not establish 10 g/cow/day as the optimal inclusion level for this product. Future studies should compare the two products at matched doses and integrate multi-omics analyses with functional validation and economic evaluation to clarify their mechanisms of action and guide practical supplementation. Full article

Optimizing fertilization strategies is key to increasing peanut yields and maintaining soil health, but it is still unclear how the proportion of basal fertilizer (applied before sowing) and topdressing (applied through drip irrigation during growth) systematically affects the “soil microbe plant” system under integrated drip irrigation and water–fertilizer management. This study set up four fertilization strategies—no fertilizer (F1), full basal fertilizer (F2: N–P₂O₅–K₂O = 343.9–123.0–120.0 kg·hm⁻² applied at sowing), drip irrigation with full water-soluble fertilizer (F3: total NPK input same as F2, applied via drip irrigation at flowering–pegging, pod-setting, and pod-filling stages in a 6:2:2 ratio), and 50% basal fertilizer + 50% water-soluble fertilizer (F4: basal applied at sowing, water-soluble applied via drip irrigation at the three growth stages in a 2:4:4 ratio)—to explore their effects on the soil nutrient availability, enzyme activity, bacterial community, and plant nutrient uptake throughout the entire growth stage of peanuts. The results showed that drip irrigation and topdressing strategies significantly improved soil-available nutrients and enzyme activity in the later stages of growth, with the F4 treatment showing the most significant advantage. This treatment not only significantly improved bacterial diversity and community functional redundancy and maintained a stable presence of multifunctional beneficial bacterial genera but also significantly increased nitrogen, phosphorus, and potassium uptake and fertilizer use efficiency. Partial least-squares structural equation modeling (PLS-SEM) reveals that fertilization strategies mainly indirectly drive yield increase by regulating microbial communities and enzyme activity. In summary, under the condition of equal nutrient input, the strategy of evenly matching nutrient supply with peanut demand was achieved, which not only ensured high yield and efficiency but also enhanced soil microecological stability. It is the optimal strategy for integrating water and fertilizer into the drip irrigation of summer peanuts under the conditions of this experiment. Full article

The field of neuroplacentology is a growing area that focuses on the influence of placental function on brain development. This field has expanded our understanding of the placenta’s crucial role in proper neurodevelopment as well as placental anomalies that increase the risk of adverse neurodevelopmental outcomes. Neuroplacentology studies have revealed distinct roles of placentally provided hormones and nutrients in neurodevelopment whose disruption by perinatal problems, including inflammation, can increase the risk of neurodevelopmental disorders such as autism spectrum disorder and ADHD. Placental insulin-like growth factor signaling significantly influences brain growth in animal model studies and is tightly linked with human neurodevelopmental outcomes. Insulin-like growth factor signaling is well-established as a regulator of placental function and is fundamental in fetal brain cell developmental processes and plays a role in inflammation. Placental insulin-like growth factor signaling underlies the placental production and delivery of hormones and nutrients that are necessary for fetal neural cell proliferation, differentiation, and growth. It is especially important to study this pathway in the placenta due to aberrant insulin-like growth factor signaling found in conditions such as preterm birth, fetal growth restriction, gestational diabetes, and obesity. All these conditions are linked to a greater risk for neurodevelopmental disorders, neuroinflammation, and other anomalies. This research subfield has also revealed sex differences in the insulin-like growth factor signaling pathway that will contribute to understanding the etiologies of these conditions. This area of study has and will continue to identify specific mechanisms, many currently in model systems, which could eventually be leveraged in preventative and interventive care to improve neurodevelopmental outcomes. Full article

Soils underpin terrestrial ecosystem functioning and exert strong control over farmland productivity, nutrient turnover, environmental quality, and agricultural sustainability. Reliable prediction of soil nutrients and pH is therefore important for precision fertilization, soil-quality assessment, and the sustainability of farmland management, particularly in fragile alpine agroecosystems. However, the comparative performance of candidate algorithms for estimating soil properties in alpine croplands of the Qinghai-Tibetan Plateau remains poorly characterized. Here, we assessed nine approaches, namely random forest (RF), generalized boosting regression (GBR), multiple linear regression (MLR), artificial neural network (ANN), generalized linear regression (GLR), conditional inference tree (CIT), extreme gradient boosting (eXGB), support vector machine (SVM), and recursive partitioning regression tree (RRT), for predicting soil carbon, nitrogen, phosphorus, potassium, and pH from annual mean temperature, annual precipitation, annual solar radiation, and maximum normalized difference vegetation index (NDVImax). Field observations were obtained from 103 sampling plots, with four individual soil samples collected per plot and depth, resulting in 412 observations for each depth layer. Clear contrasts in model performance emerged under alpine agricultural conditions. Across all soil variables and depths, eXGB and RF showed the best overall performance, with the lowest mean relative bias values of 1.35% and 1.55%, respectively, and the lowest mean RMSE values of 0.43 and 0.47. These results indicate that nonlinear ensemble algorithms better captured the heterogeneous relationships between environmental covariates and soil properties than linear models or single-tree approaches. Overall, RF and eXGB provide a practical methodological basis for precision fertilization, soil-quality assessment, and long-term sustainability in fragile alpine agroecosystems. Full article

Previous research has established distinctions between microbial communities that associate with either the solid or liquid rumen microenvironments utilizing 16S rRNA sequencing. Whole genome metagenomic sequencing (WGS) facilitates comprehensive analysis of a microorganism’s entire genome, producing strain level data and providing insights into microbial functionality. The objective of our study was to utilize WGS to characterize microbial species and enzyme profiles associated with the liquid versus solid rumen microenvironments in beef steers consuming forage. Four steers consuming forage were utilized in a 4 × 4 Latin Square experiment; on d 14 of each period, rumen contents were separated into solid and liquid samples and DNA was extracted then sequenced using WGS. Alpha and beta diversity measures revealed differences between solid and liquid microenvironments. There were also differences in enzyme profiles: carbohydrate-active enzyme classes were different across microenvironments, except glycosyl transferase. Further, the solid microenvironment was enriched in enzymes related to energy production/conversion and nutrient transport and metabolism, while the liquid microenvironment was enriched in coenzyme transport and metabolism enzymes. Species associated with cellulolytic activities were generally more abundant in the solid microenvironment while those associated with amylolytic activity were more abundant in the liquid. Genes related to methanogenesis were also different across microenvironments. Our findings offer valuable insights into the stratification of microorganisms across rumen microenvironments, expanding on existing literature with findings from next-generation sequencing and application to forage-based beef systems. Full article