ObjectiveA large number of substrates after harvesting the fruiting bodies of Cordyceps militaris have been discarded or only used as feed and fertilizer. In this study, we aimed to investigate the potential utilization value of residual substrates under industrial cultivation of C. militaris.MethodsThe various bioactive substances in C. militaris fruiting body and substrate were measured and compared by colorimetry, HPLC, burning method, Kjeldahl legal nitrogen method, and GC-MS et al. The fruiting body was taken as a reference to better reflect the internal quality of substrate. Finally, a response surface experiment was conducted to investigate the effects of drying temperature, humidity, and time on the retention rate of C. militaris polysaccharide.ResultsIn the substrate, the content of polysaccharide was 4.68%, the cordycepic acid content was 7.95%, which were significantly higher than the content in the fruiting body. The content of cordycepin was as high as 1.13%, which was more than 4 times of the fruiting body. The crude fat content was also higher than that in fruiting body. In addition, 17 amino acids were detected in the substrate, including 7 essential amino acids. Moreover, several volatile components with drug properties were detected in the substrate. Finally, the optimum drying parameters of the substrates are as follows: drying temperature, 59°C; humidity, 16%; time, 24 h.ConclusionsThis study highlighted the available value of substrate is no less than the fruiting body, which is an ideal food supplement and pharmaceutical agent, which can be served as a good source of healthy substances potentially bioavailable for humans.

This study aimed to control mycotoxigenic fungi in repeatedly used marinades and marinated foods and to elucidate the cooperative mechanism of linalool and surfactin against Aspergillus parasiticus growth and aflatoxin B1 (AFB1) synthesis by analyzing the changes in mycelial growth, AFB1 content, ATP synthesis, and metabolites. Results showed that linalool inhibited A. parasiticus growth more effectively than surfactin, while their combination exhibited a strong cooperative effect, completely suppressing mycelial growth and AFB1 synthesis at high doses. The linalool-surfactin combination effectively disrupted cell membrane integrity, reduced ATP synthesis, increased malondialdehyde content to 18.86 nmol/g, and decreased ATPase activity by 80.64%. Metabolomic analysis showed that the linalool-surfactin combination disrupted glucose metabolism and the tricarboxylic acid cycle, leading to the abnormal accumulation of metabolites such as glucose-6-phosphate and malic acid. Moreover, the combination induced abnormal expression of key metabolites such as L-glutamic acid, 3-sulfinylpyruvic acid, and cyclic adenosine monophosphate, disrupting pathways including amino acid metabolism, oxidative phosphorylation, and the cell cycle, thereby inhibiting the growth and toxin synthesis of A. parasiticus at multiple levels. This multi-level inhibition via membrane damage, energy disruption, and metabolic dysregulation provides a mechanistic basis for natural preservation strategies to control AFB1 in marinade-based systems and marinated products.

This study investigated the impact of Bifidobacterium animalis ssp. lactis (B. lactis) IU100 or/and 1.5% resistant starch type III (RS3) on fermented milk during storage. The co-supplementation with enhanced texture, increasing hardness from 10.52 g (control) to 14.88 g and springiness from 1.18 mm to 3.03 mm, and promoted a denser gel network. Volatile profiling combined with OAV analysis revealed that the addition of B. lactis IU100 significantly increased the total content of alcohols (from 1231.77 μg/L to 2841.43 μg/L), particularly promoting the accumulation of compounds such as n-butanol and 1-octen-3-ol are known to contribute fruity and mushroom-like notes in dairy systems. The individual supplementation of 1.5% RS3 markedly elevated the total aldehyde content (from 3761.05 μg/L to 7026.82 μg/L), with compounds such as 2-octenal, (2e)- is associated with distinct fatty and nutty aromas in model systems. When B. lactis IU100 was combined with RS3, the level of 1-hexanol was further elevated, enhancing a fresh green note. Untargeted metabolomics further indicated that 300 significantly differential metabolites were identified in the co-supplemented group, among which key intermediates such as dephospho-CoA and adenosine diphosphate ribose were notably upregulated. These metabolites were mainly mapped to cofactor biosynthesis, purine metabolism, and pyrimidine metabolism, suggesting coordinated roles in the formation and interconversion of flavor precursors. In summary, the combined supplementation of B. lactis IU100 and RS3 effectively enhanced the overall quality and flavor complexity of fermented milk by modulating the volatile composition and core metabolic network.

This study investigated the flavor regulatory mechanism of mixed-strain fermentation in low-salt fish sauce from yellowfin tuna dark meat, based on physicochemical properties, microbial communities, and metabolite profiles. Pediococcus acidilactici and Staphylococcus xylosus at the optimal ratio of 1:2 synergistically increased the amino acid nitrogen content to above 0.7 g/100 mL. Amplicon sequencing revealed distinct microbial communities among the unfermented, single-strain, and co-fermentation groups. Eight core differential genera were identified between co-fermentation and unfermented groups, with Lactiplantibacillus and Pediococcus dominant in the co-fermentation group. Compared with the unfermented group, SPME-GC-MS and LC-MS identified 16 key differential volatile metabolites and 20 downregulated non-volatile metabolites in the co-fermentation group. These metabolites enhanced fruity, fatty, cheesy, and other flavors of fish sauce, and were enriched in ABC transporter, purine/nucleotide metabolism, and other pathways. Overall, co-fermentation provides theoretical and technical support for the process development and flavor regulation of low-salt fish sauce.

Ribosomes are central to protein synthesis and a frequent target for antibiotics. In fast-growing bacteria, the ribosome content is proportional to the growth rate; how ribosomes and protein synthesis are regulated during nutrient starvation remains poorly understood, particularly in single cells. To address this, we fluorescently labeled ribosomal proteins (RPs) in Salmonella and explored their variations and regulation in single cells. We show that the RP levels become heterogeneous during the transition to the stationary phase. Unexpectedly, cells with higher RP levels responded less to the induction of gene expression but accumulated more virulence gene products. Our work further reveals that adenosine 3',5'-monophosphate (cAMP) signaling increases the heterogeneity of the levels of RPs and other gene products. Fluorescence dilution assay and proteomic analysis indicate that cAMP signaling promotes gene expression heterogeneity by directing proteome-wide adaptation, which enables growth heterogeneity, hence differential dilution of gene products during nutrient depletion.

Coffee is an important dietary source of bioactive antioxidant compounds contributing to the antioxidant properties of coffee beverages. While brewing affects yield of total antioxidants, it is still not really clear which individual (phenolic) compounds contribute to the antioxidant activity the most. A method combining chromatographic separation and individual antioxidant evaluation might therefore be useful. This study aimed at evaluating the antioxidant potential of the compounds in coffee beverages using a high-performance liquid chromatography approach directly coupled to the well-known trolox equivalent antioxidative capacity (TEAC) assay (HPLC-onlineTEAC). The study further evaluated the influence of different brewing methods ('Americano', 'V60', 'French press', and 'cold brew') on the bioactive compound profile and antioxidant potential of Arabica coffee beverages. The brewing method significantly affected caffeine content, chlorogenic acid composition, total phenolic content (TPC), and antioxidant activity of the analyzed beverages (p < 0.05). Cold brew samples exhibited the highest total radical scavenging activity and concentrations of major caffeoylquinic acid isomers (3-CQA, 4-CQA, and 5-CQA). In contrast, "French-pressed" beverages were characterized by the highest TPC values, while V60 samples generally showed the lowest antioxidant-related parameters. Chlorogenic acids accounted for more than 84% of the total antioxidant potential of all analyzed beverages, whereas monocaffeoylquinic acids represented the dominant fraction responsible for radical-scavenging activity. The results indicate that prolonged low-temperature extraction favors the recovery and preservation of highly reactive chlorogenic acid isomers and contributes to the enhanced antioxidant potential of coffee beverages, beyond the effect of coffee dose alone.

Functional starter cultures were evaluated for their effects on purine compounds, uric acid (UA), and xanthine oxidase (XOD) activity during fermented lamb jerky processing. Five treatment groups, including CG, 37×-3, x3-2b, 3×-2B, and 2111 CB, were analyzed across pickling, fermentation, drying, and maturation stages. The results showed that starter cultures significantly affected the dynamic changes in individual purines, total purines, UA, and XOD activity. The increase in purine contents on a FW basis during late processing was closely related to moisture-loss-driven concentration effects. After DM correction, x3-2b maintained relatively low total purine and A + Hx levels, suggesting a more stable effect on reducing purine accumulation, indicating its potential as a candidate starter culture for low-purine fermented lamb jerky production. Although 37×-3 showed strong XOD-inhibitory potential and reduced UA levels, its residual total purine content remained relatively high. These findings provide a basis for using functional starter cultures to regulate purine accumulation and UA formation in fermented meat products.

Gracilaria bailinae polysaccharides have limited applications due to their high viscosity, large molecular weight, and poor solubility. Microbial fermentation emerges as an efficient strategy to enhance the bioactivity of plant polysaccharides. This study aimed to prepare fermented water-soluble crude polysaccharide (GBV) from G. bailinae via fermentation with Bacillus velezensis, using water-extracted polysaccharide (GBP) as a control. The physicochemical properties, immunomodulatory activity, and trained immunity effects of GBV were systematically investigated. A significant decrease in molecular weight, accompanied by increased sulfate and uronic acid content, was observed in G. bailinae polysaccharides following fermentation. Modifications to the monosaccharide composition ratio and microstructure also occurred, though the basic polysaccharide structure remained unaffected. In vitro experiments revealed that both GBV and GBP were non-toxic to RAW264.7 macrophages and promoted cell proliferation, phagocytosis, and the secretion of nitric oxide (NO) and pro-inflammatory cytokines (IL-1β, IL-6, TNF-α). Notably, GBV exhibited superior immunostimulatory activity at low concentrations (6.25-50 μg/mL) compared to GBP. Furthermore, at a concentration of 12.5 μg/mL, GBV was found to trigger trained immunity in RAW264.7 cells, as demonstrated by elevated levels of cytokines and NO following LPS restimulation. Untargeted metabolomics analysis indicated that the trained immunity effect was associated with metabolic reprogramming of purine, pyrimidine, and arginine-polyamine pathways, with significant downregulation of spermine potentially amplifying the M1 inflammatory phenotype. These findings demonstrate that B. velezensis fermentation improves the immunomodulatory activity of G. bailinae polysaccharides, and GBV holds great potential as a natural immune activator. These findings provide a theoretical foundation for the development and application of probiotic-fermented seaweed polysaccharides.

This study investigated the differences in meat quality, mineral composition and metabolomics between Kazakh and Lop sheep raised on grasslands and saline-alkali soils in Xinjiang. Significant variations were observed in protein content and shear force, as well as in the mineral content of potassium, calcium, sodium, magnesium, zinc and selenium. Metabolomic profiling revealed 250 metabolites with differential abundance. Among these, 41 key biomarkers were identified and validated using BorutaShap and ensemble learning, which effectively discriminated between Kazakh and Lop lamb samples. Pathway analysis highlighted glucose/galactose metabolism, amino acid metabolism, purine metabolism, glycerophospholipid metabolism, and tricarboxylic acid cycling. Histidine metabolism was found to be closely linked to protein content and tenderness. Further correlation analysis suggested that Gly-Tyr, Tyr-Gly and Ala-Arg could be potential tenderness indicators. These findings clarify the metabolic basis of quality differences between geographical indication breeds in different ecological environments, providing methodological insights into identifying molecular markers of lamb quality.

The intramuscular fat (IMF) content is an important indicator of meat quality, affecting the sensory properties of meat. IMF is a complex trait with polygenic nature. This research aimed to identify differentially expressed (DE) genes and key transcription factors (TFs) associated with IMF deposition in the liver of two rabbit lines divergently selected for IMF (high-IMF: H and low-IMF: L). We used 48 rabbits (24 H and 24 L) belonging to the 9th generation of selection to determine their liver gene expression levels using 3' RNA sequencing. We found 263 DE genes between H and L lines; 114 upregulated and 149 downregulated in the H line. Among them, over 30 genes were related to lipid metabolism, energy homeostasis, glutathione-purine metabolism, and/or molecule transport that could influence the IMF deposition. Of the 263 DE genes, 175 target genes were predicted for 11 DE TFs through the gene regulatory networks. Notably, three TFs, namely ETV1, NR4A1, and IKZF3 appear to modulate gene expression of several putative targets involved in lipid and energy metabolism (e.g., CREM, GDF15, and NR4A2). Functional analysis of DE genes revealed an overrepresentation of 37 enriched terms, including the PPAR signalling pathway (associated genes: CPT1B, CYP4A6, FABP4, and PLIN2) and other lipid metabolism bioprocesses. Collectively, our results enhance the understanding of the liver-muscle crosstalk that contributes to IMF deposition and improvement of rabbit meat quality.

Cellulose, a primary component of plant cell walls, is synthesized by cellulose synthase complexes (CSCs) at the plasma membrane. Targeting this process with cellulose biosynthesis inhibitors (CBIs) has significantly advanced our understanding of plant cell wall formation and provided valuable compounds for herbicide development. Here, we identified a fungal natural product, 8-methyldichlorodiaporthin (MDD), as a broad-spectrum plant CBI. Structure-activity relationship analyses demonstrate that methylation modifications on the isocoumarin ring and chlorination of the side chain are crucial for MDD-induced growth inhibition. A chemical forward genetic screen in Arabidopsis thaliana revealed two semidominant CESA1 mutations, causing A903T and H1024Y substitutions, that confer insensitivity to MDD. Both mutations locate to transmembrane domains of CESA1, and we show that MDD depletes CSCs from the plasma membrane and reduces cellulose content. Further genetic analyses indicate that the cesa1mddi1-1 A903T mutant also confers resistance to CBIs quinoxyphen and C17, but not to CBIs isoxaben, indaziflam, or ES20. Stacking additional point mutations conferring resistance to other CBIs, cesa3ixr1-1 G998D, and cesa6es20-r3 G935E into the cesa1mddi1-1 A903T background yields multiple-drug-resistant lines that maintain normal growth. These findings establish MDD, as a natural CBI that likely targets CESA1, thereby extending our understanding of CSC regulation and abilities to develop multidrug-resistant crop varieties. These findings offer unique perspectives for weed management and plant biotechnology.

This study aimed to clarify the molecular mechanisms of muscle growth/development in Liaoning cashmere goats and intestinal flora's regulatory role to improve meat production. Muscle tissues (intercostal, hindlimb biceps femoris, longissimus dorsi) and digestive contents (rumen, small intestine, cecum, feces) from goats at 6, 12, 18 months were analyzed via transcriptomics, proteomics, metabolomics, 16S rRNA sequencing, and multi-omics integration. Results: hindlimb biceps femoris optimally developed at 12 months (significantly associated with jejunal g_Lachnospiraceae_NK3A20_group, HOXC10, MYL3, cAMP pathway); longissimus dorsi actively grew at 12 months (closely related to ruminal/cecal g_Christensenellaceae_R-7_group, MEF2A, MYH6; COL1A1/COL1A2 via purine metabolism for fat deposition); intercostal muscle proliferated at 6 months (closely correlated with g_Aeriscardovia, Actin, L-malic acid, l-serine) and GDI was potentially related to its 18-month fat deposition. Multi-omics showed Goat muscle development is coordinately regulated by specific genes, proteins, metabolites, and flora, providing breeding targets for better meat production.