Metabolic Cofactors Research Articles

MftG is crucial for ethanol metabolism of mycobacteria by linking mycofactocin oxidation to respiration

Mycofactocin is a redox cofactor essential for the alcohol metabolism of mycobacteria. While the biosynthesis of mycofactocin is well established, the gene mftG, which encodes an oxidoreductase of the glucose-methanol-choline superfamily, remained functionally uncharacterized. Here, we show that MftG enzymes are almost exclusively found in genomes containing mycofactocin biosynthetic genes and are present in 75% of organisms harboring these genes. Gene deletion experiments in Mycolicibacterium smegmatis demonstrated a growth defect of the ∆mftG mutant on ethanol as a carbon source, accompanied by an arrest of cell division reminiscent of mild starvation. Investigation of carbon and cofactor metabolism implied a defect in mycofactocin reoxidation. Cell-free enzyme assays and respirometry using isolated cell membranes indicated that MftG acts as a mycofactocin dehydrogenase shuttling electrons toward the respiratory chain. Transcriptomics studies also indicated remodeling of redox metabolism to compensate for a shortage of redox equivalents. In conclusion, this work closes an important knowledge gap concerning the mycofactocin system and adds a new pathway to the intricate web of redox reactions governing the metabolism of mycobacteria.

MftG is crucial for ethanol metabolism of mycobacteria by linking mycofactocin oxidation to respiration.

Mycofactocin is a redox cofactor essential for the alcohol metabolism of mycobacteria. While the biosynthesis of mycofactocin is well established, the gene mftG, which encodes an oxidoreductase of the glucose-methanol-choline superfamily, remained functionally uncharacterized. Here, we show that MftG enzymes are almost exclusively found in genomes containing mycofactocin biosynthetic genes and are present in 75% of organisms harboring these genes. Gene deletion experiments in Mycolicibacterium smegmatis demonstrated a growth defect of the ∆mftG mutant on ethanol as a carbon source, accompanied by an arrest of cell division reminiscent of mild starvation. Investigation of carbon and cofactor metabolism implied a defect in mycofactocin reoxidation. Cell-free enzyme assays and respirometry using isolated cell membranes indicated that MftG acts as a mycofactocin dehydrogenase shuttling electrons toward the respiratory chain. Transcriptomics studies also indicated remodeling of redox metabolism to compensate for a shortage of redox equivalents. In conclusion, this work closes an important knowledge gap concerning the mycofactocin system and adds a new pathway to the intricate web of redox reactions governing the metabolism of mycobacteria.

Could metabolic imaging and artificial intelligence provide a novel path to non-invasive aneuploidy assessments? A certain clinical need.

Pre-implantation genetic testing for aneuploidy (PGT-A) via embryo biopsy helps in embryo selection by assessing embryo ploidy. However, clinical practice needs to consider the invasive nature of embryo biopsy, potential mosaicism, and inaccurate representation of the entire embryo. This creates a significant clinical need for improved diagnostic practices that do not harm embryos or raise treatment costs. Consequently, there has been an increasing focus on developing non-invasive technologies to enhance embryo selection. Such innovations include non-invasive PGT-A, artificial intelligence (AI) algorithms, and non-invasive metabolic imaging. The latter measures cellular metabolism through autofluorescence of metabolic cofactors. Notably, hyperspectral microscopy and fluorescence lifetime imaging microscopy (FLIM) have revealed unique metabolic activity signatures in aneuploid embryos and human fibroblasts. These methods have demonstrated high accuracy in distinguishing between euploid and aneuploid embryos. Thus, this review discusses the clinical challenges associated with PGT-A and emphasizes the need for novel solutions such as metabolic imaging. Additionally, it explores how aneuploidy affects cell behaviour and metabolism, offering an opinion perspective on future research directions in this field of research.

Global metabolomic alterations associated with endocrine-disrupting chemicals among pregnant individuals and newborns

BackgroundGestational exposure to non-persistent endocrine-disrupting chemicals (EDCs) may be associated with adverse pregnancy outcomes. While many EDCs affect the endocrine system, their effects on endocrine-related metabolic pathways remain unclear. This study aims to explore the global metabolome changes associated with EDC biomarkers at delivery.MethodsThis study included 75 pregnant individuals who delivered at the University of Cincinnati Hospital from 2014 to 2017. We measured maternal urinary biomarkers of paraben/phenol (12), phthalate (13), and phthalate replacements (4) from the samples collected during the delivery visit. Global serum metabolome profiles were analyzed from maternal blood (n = 72) and newborn (n = 63) cord blood samples collected at delivery. Fifteen of the 29 urinary biomarkers were excluded due to low detection frequency or potential exposures during hospital stay. We assessed metabolome-wide associations between 14 maternal urinary biomarkers and maternal/newborn metabolome profiles. Additionally, performed enrichment analysis to identify potential alterations in metabolic pathways.ResultsWe observed metabolome-wide associations between maternal urinary concentrations of phthalate metabolites (mono-isobutyl phthalate), phthalate replacements (mono-2-ethyl-5-carboxypentyl terephthalate, mono-2-ethyl-5-hydroxyhexyl terephthalate) and phenols (bisphenol-A, bisphenol-S) and maternal serum metabolome, using q-value < 0.2 as a threshold. Additionally, associations of phthalate metabolites (mono-n-butyl phthalate, monobenzyl phthalate) and phenols (2,5-dichlorophenol, BPA) with the newborn metabolome were noted. Enrichment analyses revealed associations (p-gamma < 0.05) with amino acid, carbohydrate, lipid, glycan, vitamin, and other cofactor metabolism pathways.ConclusionMaternal paraben, phenol, phthalate, and phthalate replacement biomarker concentrations at delivery were associated with maternal and newborn serum global metabolome.

Failure to repair damaged NAD(P)H blocks de novo serine synthesis in human cells

BackgroundMetabolism is error prone. For instance, the reduced forms of the central metabolic cofactors nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate (NADPH), can be converted into redox-inactive products, NADHX and NADPHX, through enzymatically catalyzed or spontaneous hydration. The metabolite repair enzymes NAXD and NAXE convert these damaged compounds back to the functional NAD(P)H cofactors. Pathogenic loss-of-function variants in NAXE and NAXD lead to development of the neurometabolic disorders progressive, early-onset encephalopathy with brain edema and/or leukoencephalopathy (PEBEL)1 and PEBEL2, respectively.MethodsTo gain insights into the molecular disease mechanisms, we investigated the metabolic impact of NAXD deficiency in human cell models. Control and NAXD-deficient cells were cultivated under different conditions, followed by cell viability and mitochondrial function assays as well as metabolomic analyses without or with stable isotope labeling. Enzymatic assays with purified recombinant proteins were performed to confirm molecular mechanisms suggested by the cell culture experiments.ResultsHAP1 NAXD knockout (NAXDko) cells showed growth impairment specifically in a basal medium containing galactose instead of glucose. Surprisingly, the galactose-grown NAXDko cells displayed only subtle signs of mitochondrial impairment, whereas metabolomic analyses revealed a strong inhibition of the cytosolic, de novo serine synthesis pathway in those cells as well as in NAXD patient-derived fibroblasts. We identified inhibition of 3-phosphoglycerate dehydrogenase as the root cause for this metabolic perturbation. The NAD precursor nicotinamide riboside (NR) and inosine exerted beneficial effects on HAP1 cell viability under galactose stress, with more pronounced effects in NAXDko cells. Metabolomic profiling in supplemented cells indicated that NR and inosine act via different mechanisms that at least partially involve the serine synthesis pathway.ConclusionsTaken together, our study identifies a metabolic vulnerability in NAXD-deficient cells that can be targeted by small molecules such as NR or inosine, opening perspectives in the search for mechanism-based therapeutic interventions in PEBEL disorders.Graphical

The Metabolic Treatabolome and Inborn Errors of Metabolism Knowledgebase therapy tool: Do not miss the opportunity to treat!

Inborn errors of metabolism (IEMs) are rare genetic conditions with significant morbidity and mortality. Technological advances have increased therapeutic options, making it challenging to remain up to date. A centralized therapy knowledgebase is needed for early diagnosis and targeted treatment. This study aimed to identify all treatable IEMs through a scoping literature review, followed by data extraction and analysis according to the Treatabolome principles. Knowledge of treatable IEMs, therapeutic categories, efficacy, and evidence was integrated into the Inborn Errors of Metabolism Knowledgebase (IEMbase), an online database encompassing all IEMs. The study identified 275 treatable IEMs, 18% of all currently known 1564 IEMs, according to the International Classification of Inherited Metabolic Disorders. Disorders of fatty acid and ketone body metabolism had the highest treatability (67%), followed by disorders of vitamin and cofactor metabolism (60%), and disorders of lipoprotein metabolism (42%). The most common treatment strategies were pharmacological therapy (34%), nutritional therapy (34%), and vitamin and trace element supplementation (12%). Treatment effects were most commonly observed in nervous system abnormalities (34%), metabolism/homeostasis abnormalities (33%), and growth (7%). Predominant evidence sources included case reports with evidence levels 4 (48%) and 5 (12%), and individual cohort studies with evidence level 2b (12%). Our study generated the Metabolic Treatabolome 2024. IEMs are the largest group of monogenic disorders amenable to disease-modifying therapy. With drug repurposing efforts and advancements in gene therapies, this number will expand. IEMbase now provides up-to-date, comprehensive information on clinical and biochemical symptoms and therapeutic options, empowering patients, families, healthcare professionals, and researchers in improving patient outcomes.

Investigation of Intestinal Microbes of Five Zokor Species Based on 16S rRNA Sequences.

Zokor is a group of subterranean rodents that are adapted to underground life and feed on plant roots. Here, we investigated the intestinal microbes of five zokor species (Eospalax cansus, Eospalax rothschildi, Eospalax smithi, Myospalax aspalax, and Myospalax psilurus) using 16S amplicon technology combined with bioinformatics. Microbial composition analysis showed similar intestinal microbes but different proportions among five zokor species, and their dominant bacteria corresponded to those of herbivores. To visualize the relationships among samples, PCoA and PERMANOVA tests showed that the intestinal microbes of zokors are largely clustered by host species, but less so by genetics and geographical location. To find microbes that differ among species, LefSe analysis identified Lactobacillus, Muribaculaceae, Lachnospiraceae_NK4A136_group, unclassified_f_Christensenellaceae, and Desulfovibrio as biomarkers for E. cansus, E. rothschildi, E. smithi, M. aspalax, and M. psilurus, respectively. PICRUSt metagenome predictions revealed enriched microbial genes for carbohydrate and amino acid metabolism in E. cansus and E. smithi, and for cofactor and vitamin metabolism as well as glycan biosynthesis and metabolism in E. rothschildi, M. aspalax, and M. psilurus. Our results demonstrated differences in the microbial composition and functions among five zokor species, potentially related to host genetics, and host ecology including dietary habits and habitat environment. These works would provide new insight into understanding how subterranean zokors adapt to their habitats by regulating intestinal microbes.

Defense-Related Enzyme Activities and Metabolomic Analysis Reveal Differentially Accumulated Metabolites and Response Pathways for Sheath Blight Resistance in Rice.

Rice sheath blight (RSB), caused by the pathogenic fungus Rhizoctonia solani, poses a significant threat to global food security. The defense mechanisms employed by rice against RSB are not well understood. In our study, we analyzed the interactions between rice and R. solani by comparing the phenotypic changes, ROS content, and metabolite variations in both tolerant and susceptible rice varieties during the early stages of fungal infection. Notably, there were distinct phenotypic differences in the response to R. solani between the tolerant cultivar Zhengdao22 (ZD) and the susceptible cultivar Xinzhi No.1 (XZ). We observed that the activities of five defense-related enzymes in both tolerant and susceptible cultivars changed dynamically from 0 to 72 h post-infection with R. solani. In particular, the activities of superoxide dismutase and peroxidase were closely associated with resistance to RSB. Metabolomic analysis revealed 825 differentially accumulated metabolites (DAMs) between the tolerant and susceptible varieties, with 493 DAMs responding to R. solani infection. Among these, lipids and lipid-like molecules, organic oxygen compounds, phenylpropanoids and polyketides, organoheterocyclic compounds, and organic acids and their derivatives were the most significantly enriched. One DAM, P-coumaraldehyde, which responded to R. solani infection, was found to effectively inhibit the growth of R. solani, Magnaporthe grisea, and Ustilaginoidea virens. Additionally, multiple metabolic pathways, including amino acid metabolism, carbohydrate metabolism, metabolism of cofactors and vitamins, and metabolism of terpenoids and polyketides, are likely involved in RSB resistance. Our research provides valuable insights into the molecular mechanisms underlying the interaction between rice and R. solani.

Gold Nanoparticle Detection with Two-Photon Excitation Fluorescence Lifetime Imaging of NAD(P)H in Cancer Cells: An Analytical Approach to Separate Nanoparticle and NAD(P)H Signals.

Gold nanoparticles (AuNPs) have shown promise for applications in the diagnosis and treatment of different diseases, including cancer. Understanding the effect of AuNPs on metabolic reprogramming in cancer cells at the single cell level is of high importance for improving the efficacy and safety. Fluorescence lifetime imaging microscopy (FLIM) of nicotinamide adenine dinucleotide (phosphate) hydrogen (NAD(P)H) as a main metabolic cofactor and an indicator of metabolic reprogramming in cancer cells enables real-time monitoring of cancer cell metabolism in response to different treatments, including AuNPs. However, NPs such as AuNPs can be a potential source of signals themselves, which provides opportunities to measure the NP internalization, but it is also important to minimize confounding effects on metabolic measurements. In this study, we detected inherent photoluminescence (PL) from the AuNPs in treated prostate cancer cells (PC-3 cell line) as well as in solution at the NAD(P)H emission wavelength. We developed an analysis approach to minimize the confounding effect of the AuNPs' PL on metabolic measurements. On the other hand, we assessed the reliability of the intracellular AuNPs' PL as an estimator of AuNP uptake. To assess if intracellular AuNPs' PL may be dependent on the exposed cell type, we performed NAD(P)H FLIM imaging of AuNP-exposed SKBR-3 breast cancer cells, where we observed a similar AuNP PL but at a much lower level compared to PC-3 cells. We proposed that this difference can be attributed to the different levels of AuNP uptake or varying intracellular microenvironments.

Effects of Biological Additives on the Fermentation Quality and Microbial Community of High-Moisture Oat Silage

The primary objective of this study was to explore the effects of biological additives, including Streptococcus bovis (SB), Bacillus subtilis (BS), xylanase (XT), and their combined treatments, including SB + BS (SBBS), SB + XT (SBXT), and BS + XT (BSXT), on the chemical composition, fermentation characteristics, and microbial community of high-moisture oat silage. Compared with the CK group (control group without additives), SB and SBBS treatments increased the lactic acid content (p &lt; 0.05) and reduced the contents of acetic acid, propionic acid, butyric acid, and ammonia nitrogen in silage (p &lt; 0.05). XT, SBXT, and BSXT treatments decreased the neutral detergent fiber and acid detergent fiber contents (p &lt; 0.05), increasing the water-soluble carbohydrate content (p &lt; 0.05). The SB, SBBS, and SBXT treatments increased the abundance of Lactiplantibacillus (p &lt; 0.05) and significantly decreased microbial richness with diversity (p &lt; 0.05), improving the microbial community structure in silage. The addition of XT increased the relative abundance of Clostridium and Enterobacteriaceae, but its combination with SB and BS increased the abundance of Lactiplantibacillus and inhibited the development of undesirable bacteria. Moreover, different additives changed the metabolism of carbohydrates, amino acids, energy, cofactors and vitamins of bacterial communities during ensiling. In summary, the addition of SB and SBBS was more conducive to improving the fermentation characteristics of oat, while XT, SBXT, and BSXT performed better in degrading lignocellulose in plants.

Characteristics of gut microbiota of premature infants in the early postnatal period and their relationship with intraventricular hemorrhage

BackgroundStudies have shown correlations between gut microbiota and neurocognitive function, but little was known about the early postnatal gut microbiota and intraventricular hemorrhage (IVH). We aimed to explore the characteristics of gut microbiota in premature infants and their relationship with IVH, further exploring potential therapeutic targets.MethodsPremature infants delivered at Peking University First Hospital from February 2023 to August 2023 were recruited as a cohort. Feces samples were collected on postnatal days 1, 3, and 5. Premature infants were divided into normal, mild IVH, and severe IVH groups based on cranial ultrasound. 16S rRNA amplicon sequencing technology was used to determine the fecal microbiota, and the results were analyzed.ResultsThirty-eight premature infants were enrolled. There was a significant difference in alpha and beta diversity among the three groups. The relative abundance of E. coli and A. muciniphila was different among the three groups. Further random forest analysis indicated that S. lutetiensis, L. mirabilis, and N. macacae can effectively distinguish premature infants with IVH. Finally, the phylogenetic investigation of communities by reconstruction of unobserved states2 (PICRUSt2) functional gene analysis predicted significant differences in energy metabolism, carbohydrate metabolism, metabolism of cofactors and vitamins, and membrane transport between normal and severe IVH groups.ConclusionsThe gut microbiota in the early postnatal period of premature infants is closely associated with the IVH status. As age increases, the differences in gut microbiota of premature infants with different degrees of IVH continue to increase, and the trend of changes with severity of IVH becomes more and more obvious. E. coli, A. muciniphila, S. lutetiensis, L. mirabilis, N. macacae, G. haemolysans, and S. oralis can effectively distinguish between IVH infants and normal premature infants. The results indicate that gut microbiota is expected to provide effective therapeutic targets for the diagnosis and treatment of IVH.

Multiomics profiling of DNA methylation, microRNA, and mRNA in skeletal muscle from monozygotic twin pairs discordant for type 2 diabetes identifies dysregulated genes controlling metabolism

BackgroundA large proportion of skeletal muscle insulin resistance in type 2 diabetes (T2D) is caused by environmental factors.MethodsBy applying multiomics mRNA, microRNA (miRNA), and DNA methylation platforms in biopsies from 20 monozygotic twin pairs discordant for T2D, we aimed to delineate the epigenetic and transcriptional machinery underlying non-genetic muscle insulin resistance in T2D.ResultsUsing gene set enrichment analysis (GSEA), we found decreased mRNA expression of genes involved in extracellular matrix organization, branched-chain amino acid catabolism, metabolism of vitamins and cofactors, lipid metabolism, muscle contraction, signaling by receptor tyrosine kinases pathways, and translocation of glucose transporter 4 (GLUT4) to the plasma membrane in muscle from twins with T2D. Differential expression levels of one or more predicted target relevant miRNA(s) were identified for approximately 35% of the dysregulated GSEA pathways. These include miRNAs with a significant overrepresentation of targets involved in GLUT4 translocation (miR-4643 and miR-548z), signaling by receptor tyrosine kinases pathways (miR-607), and muscle contraction (miR-4658). Acquired DNA methylation changes in skeletal muscle were quantitatively small in twins with T2D compared with the co-twins without T2D. Key methylation and expression results were validated in muscle, myotubes, and/or myoblasts from unrelated subjects with T2D and controls. Finally, mimicking T2D-associated changes by overexpressing miR-548 and miR-607 in cultured myotubes decreased expression of target genes, GLUT4 and FGFR4, respectively, and impaired insulin-stimulated phosphorylation of Akt (Ser473) and TBC1D4.ConclusionsTogether, we show that T2D is associated with non- and epigenetically determined differential transcriptional regulation of pathways regulating skeletal muscle metabolism and contraction.

Anesthetics change the oral microbial composition of children and increase the abundance of the genus Haemophilus.

Oral microbiome homeostasis is important for children's health, and microbial community is affected by anesthetics. The application of anesthetics in children's oral therapy has become a relatively mature method. This study aims to investigate the effect of different anesthesia techniques on children's oral microbiota. Sixty children who visited the Department of Stomatology of the Jiangxi Provincial Children's Hospital were recruited. Subjects who did not receive anesthesia during the surgery were divided into non-anesthesia (Noa) group. Other children who accepted anesthesia during the surgery were grouped into lidocaine group, sevoflurane group, and intravenous injection-inhalation (intra-inhalation) group. Subsequently, their saliva samples were collected for 16S rDNA sequencing. A total of 1,316 operational taxonomic units (OTUs) were identified in overall samples, and 75,275 reads per sample were obtained on average. There were 137 genera were shared among the Noa, lidocaine, sevoflurane, and intra-inhalation groups. The genera Neisseria, Prevotella-7, Streptococcus, and Veillonella had a higher abundance in the four groups. Compared to the Noa group, anesthetics increased the abundance of the genus Haemophilus in the anesthesia groups. Alpha and beta diversity analyses revealed significant differences in the comparisons of Noa vs. sevoflurane and Noa vs. intra-inhalation. In contrast, the difference between the lidocaine and the Noa groups was slight. Linear discriminant analysis effect size (LEfSe) analyses identified 52, 16, and 37 differential microbes in the Noa vs. sevoflurane, Noa vs. lidocaine, and Noa vs. intra-inhalation comparisons, respectively. Notably, Haemophilus genus was significantly enriched in the sevoflurane group compared to the Noa group. When comparing the Noa group with the other three anesthesia groups, between-group pathway differences were found in amino acid metabolism, energy metabolism, biofilm formation, cofactor and vitamin metabolism, and antibiotic synthesis. This study elucidated oral microbiome characteristics in children under different anesthesia technology and found an enrichment of Haemophilus genus in the sevoflurane group compared to the Noa group. Our findings provide new insights into the effect of anesthetic on oral microbiota of children.

Individual and Combined Effects of Nanoplastics and Cadmium on the Rhizosphere Bacterial Community of Sedum alfredii Hance.

Nanoplastics (NPs) and cadmium (Cd) coexist in soil, but the combined effects of NPs and Cd on the rhizosphere bacterial community remain unknown. In this study, high-throughput sequencing and PICRUSt2 functional analysis were employed to explore the individual and combined effects of polystyrene (PS) NPs (low concentration [N1, 100 mg·kg-1] and high concentration [N2, 1000 mg·kg-1]) and Cd (low concentration [C1, 0.6 mg·kg-1] and high concentration [C2, 4 mg·kg-1]) on the diversity, structural composition, and function of the rhizosphere bacterial community associated with Sedum alfredii Hance. Individually, PS NPs and Cd significantly reduced the soil pH, while the combined treatments induced a more significant decrease in pH. In contrast, combined PS NPs and Cd significantly increased the diethylenetriaminepentaacetic acid-Cd (DTPA-Cd) and total Cd concentrations. Compared with individual treatments, C2N2 significantly increased DPTA-Cd by 4.08%. N1 had no significant effect on the Chao1, observed species, or Shannon indices, while N2 significantly reduced the richness and diversity of the rhizosphere bacteria and altered their community structure. Furthermore, adding PS NPs exacerbated the effect of Cd on rhizosphere bacterial communities. Compared with individual Cd treatments, C2N2 significantly reduced the relative abundances of Actinobacteriota, Bacteroidota, Crenarchaeota, and Myxococcota by 19.76%, 2.01%, 1.49%, and 2.00%, respectively, and significantly increased the relative abundance of Acidobacteriota by 16.05%. A cluster heat map showed that the combined treatments attenuated glycan biosynthesis and metabolic function and enhanced the metabolism of cofactors and vitamins. These findings illuminate rhizosphere processes under co-contamination with heavy metals and PS NPs, supporting the practical application of phytoremediation to alleviate combined Cd and PS NP pollution.

WITHDRAWN: 16S rRNA sequencing reveals alterations of gut bacteria in Hirschsprung-associated enterocolitis

Hirschsprung-associated enterocolitis (HAEC) stands as most common and serious complication of Hirschsprung disease. Variations in the microbiota composition may account for the differences observed between HAEC and healthy individuals, offering crucial insights into the disease's pathogenesis. Here, we performed a study to Changes in the gut microbiome using 16sRNA amplicon sequencing in a cohort of HAEC patients (n=16) and healthy controls (n=14). Our result revealed a significant disparity in beta diversity between the two groups. Following correction for false discovery rate (FDR), a rank-sum test at the genus level indicated a notable decrease in the relative abundance of Bifidobacterium, Lactobacillus, and Veillonella, whereas the Enterococcus genus exhibited a substantial increase in HAEC, a finding further supported by additional LEfSe analysis. Functional analysis showed that putative transport and catabolism, Digestive system, and metabolism of cofactors and vitamins were proved to be some abundant KOs in healthy group, while Infectious disease, Membrane transport, Carbohydrate metabolism were the 3 KOs with the higher abundance in the HAEC group. Our data increased our insight into the HAEC, which may shed further light on HAEC pathogenesis. Our study firstly demonstrated the difference between fecal microbiota of HAEC patients and healthy individuals, which made a step forward in the understanding of the pathophysiology of Hirschsprung-associated enterocolitis.

Microbiome analysis of serum extracellular vesicles in gestational diabetes patients.

Gestational Diabetes Mellitus (GDM) is among the most common complications during pregnancy, posing serious risks to both the patient's and offspring's health and well-being. Alterations in the maternal microbiome are closely associated with the pathogenesis of GDM, with Extracellular Vesicles (EVs) facilitating communication between microbiota and the host. However, little is known about the relationship between the microbial composition within EVs and the pathogenesis of GDM. Therefore, this study aims to characterize the microbiota within serum EVs of GDM Patients (GDM group) and to identify microbial communities that significantly differ from those in Women With Normal Pregnancies (NonGDM group). Blood samples were collected from both groups of patients, and EVs derived from serum were isolated via centrifugation. Identification and characterization of EVs were performed using transmission electron microscopy and nanoparticle flow cytometry. Microbiome analysis of serum EVs from both groups was conducted using 16S rRNA sequencing. Results indicated altered diversity in microbial communities within serum EVs of GDM patients. Further analysis at the phylum, family, genus, and species levels revealed that Proteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes were the dominant taxa in the EVs of both the NonGDM and GDM groups. Specifically, Actinobacteria and Firmicutes showed increased relative abundance in GDM group EVs compared to NonGDM, leading to a higher Firmicutes/Bacteroidetes ratio, while Proteobacteria and Bacteroidetes exhibited decreased relative abundance. Tax4Fun analysis revealed enrichment of microbial functions related to amino acid metabolism, carbohydrate metabolism, energy metabolism, and metabolism of cofactors and vitamins in both patient groups. In conclusion, this study reveals a potential correlation between changes in the microbial composition and diversity of serum EVs and the onset and development of GDM. Furthermore, changes in the relative abundance of Actinobacteria, Proteobacteria,Bacteroidetes, and Firmicutes may play an important role in the pathogenesis of GDM.

Inclusion of Hybrid Pennisetum and Probiotics Enhanced Anaerobic Fermentation Quality and Bacterial Diversity of Alfalfa Silage

This study aims to assess the impact of Bacillus subtilis (BS) and Lactobacillus buchneri (LB) on the fermentation quality, microbial communities, and predicted metabolic pathways in mixed silage made from alfalfa and hybrid Pennisetum. We prepared mixed silage from fresh alfalfa and hybrid Pennisetum in a 1:1 ratio and inoculated it with BS, LB, or a combination of both (BSLB) or left it untreated as a control. The silage was fermented for 30 and 60 days. The results showed that inoculation with BS, LB, or their combination increased the lactic acid and crude-protein content while reducing the fiber content compared to the control. Additionally, BS and LB inoculation raised (p &lt; 0.05) the acetic acid content, and the combination of both strains increased (p &lt; 0.05) the ratio of lactic acid to acetic acid. LB alone and the combined inoculation also increased the relative abundance of Lactobacillus during the pre-silage period. Functional analysis through the Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed considerable variations among the different probiotic treatments. The silage process reduced nucleotide metabolism but enhanced carbohydrate, amino acid, energy, cofactor, and vitamin nucleotide metabolism. High-throughput sequencing combined with KEGG functional prediction demonstrated significant differences in community composition and functional changes at 30 and 60 days of fermentation. These findings enhance our understanding of bacterial communities and functional changes in mixed silage of alfalfa and hybrid Pennisetum, offering valuable insights into the fermentation mechanisms of legume and grass silage and informing practices for producing high-quality mixed silage.

Comparative study of gut content microbiota in freshwater fish with different feeding habits: A case study of an urban lake.

The gut microbiota plays a crucial role in various physiological functions of the host and can be modulated by numerous factors, including feeding habit or trophic level. In this study, the impact of host feeding habits on the gut microbiota of freshwater fish was explored. Ten fish species, classified into four feeding habit categories (herbivorous, omnivorous, planktivorous, and carnivorous) were sampled from West Lake, a renowned urban scenic lake, and their gut content microbiota was analysed using 16S ribosomal RNA gene sequencing. A total of 2531 operational taxonomic units, belonging to 34 bacterial phyla, were identified, with 33.4% shared across all feeding habits. Firmicutes and Proteobacteria were the predominant phyla. However, at the family level, Peptostreptococcaceae and Clostridiaceae_1 were the most dominant. Microbiota composition diversity was highest in herbivorous fish, followed by omnivores, carnivores, and planktivores. Statistically significant differences in microbiota diversity were found between different feeding categories, except for the omnivores, which did not differ from the carnivores or planktivores. The most abundant predicted metabolic pathways across all feeding habits were similar, with amino acid metabolism, carbohydrate metabolism, metabolism of cofactors and vitamins, and metabolism of other amino acids being dominant. However, comparing the relative abundance of gene functions between different feeding habits revealed notable variations across most comparisons. Co-occurrence network analysis for each feeding habit revealed that all networks were dominated by the strong positive correlation among pairs of bacterial genera abundances, while the basic properties varied, implying differences in gut microbiota interactions based on the feeding habit. In conclusion, these results confirmed that the feeding habit could affect the structure and composition of the gut content microbiota but also changed their functions and interactions.

Effect of Oat Hay as a Substitute for Alfalfa Hay on the Gut Microbiome and Metabolites of Yak Calves.

To evaluate the impact of different roughages on the intestinal microbiota of yak calves, we fed them oat hay in substitution of alfalfa hay, in addition to milk replacer and starter powder. Twenty-one 45-day-old male yak calves were selected and randomly assigned to three groups: the milk replacer + starter + alfalfa hay group (AH), the milk replacer + starter + oat hay group (OH), and the milk replacer + starter + mixed hay group (AO), in which the alfalfa hay and oat hay were administered in a 1:1 ratio. All calves in the three groups were fed the same milk replacer and an equivalent amount of dry matter. The formal experiment commenced after a 21-day pre-test period and lasted for 120 days. Following the experiment, the contents of the jejunum and colon were collected to investigate the intestinal microbiota and metabolites using 16S rRNA sequencing and LC-MS metabolomics. The result showed that the AO group had greater final body weights overall than the AH group and OH group (p < 0.05). The AH group and OH group had considerably greater feed-to-gain ratios than the AO group (p < 0.05). At the phylum level, the OH group exhibited an increased relative abundance of Bacteroidota and Spirochaetota in the jejunum (p < 0.05). The relative abundance of Actinobacteriota in the colon was increased in the AO group (p < 0.05). At the genus level, the AO group exhibited a decreased abundance of Clostridium sensu_stricto_1 (p < 0.05), and the OH group showed an increased abundance UCG-005 and Alistipes in the jejunum. There were many differential metabolites in the OH group and AO group compared to the AH group, and the different metabolites of the OH group were associated with the metabolic pathways of the nervous system, sensory system, amino acid metabolism, and lipid metabolism in the jejunum and with lipid metabolism, amino acid metabolism, and the nervous system in the colon. In the AO group, these metabolites were associated with the digestive system and the translation and metabolism of cofactors in the jejunum and with the metabolism of cofactors and vitamins in the colon. In summary, it is feasible to replace alfalfa hay with oat hay based on milk replacer and starter. The combination of the two forages enhanced nutrient absorption, improved immune function, maintained the internal homeostasis of yak calves, and was more beneficial to their growth and development.

Metagenomics reveals differences in spore-forming bacterial diversity in raw milk in different regions and seasons in China

The spore-forming bacteria in the dairy industry are notable for their spores resilience and capacity to survive heating processes, allowing them to germinate and enter the vegetative stage, potentially leading to spoilage of the milk. Additionally, these spores can form biofilms, becoming a persistent source of contamination in processing environments. In this study, we collected a total of 165 raw milk from six different parts in China in spring, summer, autumn, and winter, respectively. Metagenomics sequencing method was used to explore and compare the differences in spore-forming bacterial composition and diversity in raw milk samples. Among these samples, four genera and 207 species of spore-forming bacteria were identified, with the genus Bacillus and the species Paenibacillus darwinianus dominant. Seasonal variations had a greater impact on the composition and abundance of spore-forming bacteria in raw milk than regional differences. Notable, raw milk samples collected during the spring and summer exhibited a higher number of unique spore-forming bacterial species compared to those collected in other seasons. Moreover, different regions and seasons have their own dominant bacteria. Metabolism of cofactors and vitamins, energy metabolism, carbohydrate metabolism, and amino acid metabolism were the main metabolic pathways. Hence, specific strategies need to be adopted to prevent and control spore-forming bacteria in raw milk in different regions and seasons.