Microbial Metabolic Profiles Research Articles

Impact of high altitude on composition and functional profiling of oral microbiome in Indian male population

The oral cavity of human contains bacteria that are critical for maintaining the homeostasis of the body. External stressors such as high altitude (HA) and low oxygen affect the human gut, skin and oral microbiome. However, compared to the human gut and skin microbiome, studies demonstrating the impact of altitude on human oral microbiota are currently scarce. Alterations in the oral microbiome have been reported to be associated with various periodontal diseases. In light of the increased occurrence of HA oral health related problems, the effect of HA on the oral salivary microbiome was investigated. We conducted a pilot study in 16 male subjects at two different heights i.e., H1 (210 m) and H2 (4420 m). Total of 31 saliva samples,16 at H1 and 15 at H2 were analyzed by utilizing the 16S rRNA high-throughput sequencing, to explore the relationship between the HA environment and salivary microbiota. The preliminary results suggesting that, the most abundant microbiome at the phylum level are: Firmicutes, Bacteroidetes, Proteobacteria, and Actinobacteria. Interestingly, 11 genera were identified at the both heights with different relative abundances. In addition, the salivary microbiome was more diverse at H1 compared to H2 as demonstrated by decreased alpha diversity. Further, predicted functional results indicate that microbial metabolic profiles significantly decreased at H2 as compared to H1, including two major metabolic pathways involving carbohydrates, and amino acids. Our findings show that HA induces shifts in the composition and structure of human oral microbiota which can affect host health homeostasis.

Gut Microbial Succession Patterns and Metabolic Profiling during Pregnancy and Lactation in a Goat Model.

The maternal gut microbiome affects the duration of pregnancy, delivery, and lactation. It also coordinates the stability of maternal metabolism by regulating and modulating inflammatory cytokines and reproductive hormones. This has been shown in several species; however, the situation in ruminants remains a black box. Here, we aimed to elucidate the relationship between the hindgut microbiota, metabolism, and reproductive hormones in domestic goats (Capra hircus) during nonpregnancy, pregnancy, and lactation stages. The hindgut microbiota was altered during these three stages, with a drastic decrease in the abundance of Family_XIII_AD3011_group in the second and third trimesters of pregnancy. Additionally, a decline in the abundance of Christensenellaceae_R-7_group and Turicibacter was observed from the nonpregnancy stage to late gestation. Family_XIII_AD3011_group and Paeniclostridium were strongly correlated with decreased fecal estradiol and progesterone. Furthermore, we generated a metabolome atlas of the gut and serum from nonpregnancy to lactation to reveal the specific metabolic fingerprints of each physiological stage. Several specific gut metabolites, including carnitine C8:1, γ-aminobutyric acid, and indole-3-carboxylic acid, were negatively correlated with the fecal and serum estradiol concentrations. In contrast, 2'-deoxyinosine, deoxyadenosine, and 5'-deoxyadenosine were positively correlated with the fecal and serum estradiol concentrations. The levels of 2'-deoxyinosine, deoxyadenosine, and 5'-deoxyadenosine in fecal samples were positively correlated with Family_XIII_AD3011_group. Other serum metabolites, such as (±)12-HEPE (hydroxy eicosapentaenoic acid), (±)15-HEPE, (±)18-HEPE, cytidine, uracil, and 5-hydroxyindole-3-acetic acid, were negatively correlated with the serum concentrations of estradiol and progesterone. Finally, Corynebacterium and Clostridium_sensu_stricto_1 in the fecal samples were positively correlated with the abundance of 11,12-EET (epoxy-eicosatrienoic acid), (±)18-HEPE, (±)15-HEPE, and (±)12-HEPE in the serum. IMPORTANCE Our findings revealed that the activity of Family_XIII_AD3011_group and Corynebacterium is strongly correlated with the beneficial regulation of physiological hormones and metabolic changes during pregnancy and lactation. These findings are key for guiding targeted microbial therapeutic approaches to modulate microbiomes in gestating and lactating mammals.

Salivary microbiome and metabolome analysis of severe early childhood caries

BackgroundSevere early childhood caries (SECC) is an inflammatory disease with complex pathology. Although changes in the oral microbiota and metabolic profile of patients with SECC have been identified, the salivary metabolites and the relationship between oral bacteria and biochemical metabolism remains unclear. We aimed to analyse alterations in the salivary microbiome and metabolome of children with SECC as well as their correlations. Accordingly, we aimed to explore potential salivary biomarkers in order to gain further insight into the pathophysiology of dental caries.MethodsWe collected 120 saliva samples from 30 children with SECC and 30 children without caries. The microbial community was identified through 16S ribosomal RNA (rRNA) gene high-throughput sequencing. Additionally, we conducted non-targeted metabolomic analysis through ultra-high-performance liquid chromatography combined with quadrupole time-of-flight mass spectrometry to determine the relative metabolite levels and their correlation with the clinical caries status.ResultsThere was a significant between-group difference in 8 phyla and 32 genera in the microbiome. Further, metabolomic and enrichment analyses revealed significantly altered 32 salivary metabolites in children with dental caries, which involved pathways such as amino acid metabolism, pyrimidine metabolism, purine metabolism, ATP-binding cassette transporters, and cyclic adenosine monophosphate signalling pathway. Moreover, four in vivo differential metabolites (2-benzylmalate, epinephrine, 2-formaminobenzoylacetate, and 3-Indoleacrylic acid) might be jointly applied as biomarkers (area under the curve = 0.734). Furthermore, the caries status was correlated with microorganisms and metabolites. Additionally, Spearman's correlation analysis of differential microorganisms and metabolites revealed that Veillonella, Staphylococcus, Neisseria, and Porphyromonas were closely associated with differential metabolites.ConclusionThis study identified different microbial communities and metabolic profiles in saliva, which may be closely related to caries status. Our findings could inform future strategies for personalized caries prevention, detection, and treatment.

The Analysis of the Mycobiota in Plastic Polluted Soil Reveals a Reduction in Metabolic Ability.

Plastic pollution is a growing environmental issue that results in its accumulation and persistence in soil for many decades, with possible effects on soil quality and ecosystem services. Microorganisms, and especially fungi, are a keystone of soil biodiversity and soil metabolic capacity. The aim of this research was to study soil fungal biodiversity and soil microbial metabolic profiles in three different sites in northern Italy, where macro- and microplastic concentration in soil was measured. The metabolic analyses of soil microorganisms were performed by Biolog EcoPlates, while the ITS1 fragment of the 18S ribosomal cDNA was used as a target for the metabarcoding of fungal communities. The results showed an intense and significant decrease in soil microbial metabolic ability in the site with the highest concentration of microplastics. Moreover, the soil fungal community composition was significantly different in the most pristine site when compared with the other two sites. The metabarcoding of soil samples revealed a general dominance of Mortierellomycota followed by Ascomycota in all sampled soils. Moreover, a dominance of fungi involved in the degradation of plant residues was observed in all three sites. In conclusion, this study lays the foundation for further research into the effect of plastics on soil microbial communities and their activities.

Microbial Biofilms Colonizing Plastic Substrates in the Ross Sea (Antarctica)

Very few studies have investigated marine microbial colonization in polar regions, but climate-changing scenarios stress the importance of these investigations to protect life in such extremely vulnerable ecosystems. In two different coastal sites of the Ross Sea (Road and Tethys Bays, Antarctica) exposed to different stressors, the microbial biofilm colonizing the surface of plastic (polyvinyl chloride, PVC, and polyethylene, PE) panels left submerged in two experiments at different timescales (“short-term”: 3 months, and “long-term”: 9 and 12 months) was studied. The abundance and metabolic enzymatic activities [leucine aminopeptidase (LAP), beta-glucosidase (GLU) and alkaline phosphatase (AP)] of the prokaryotes and the microalgal abundance and species composition were analyzed, in parallel with the main environmental parameters. The prokaryotic community showed higher abundance and metabolic activities on PVC than on PE as opposed to microalgae. A peak in the microfouling prokaryotic abundance and metabolic functions was frequently recorded after 3 months of immersion, corresponding to the late austral summer period. LAP and AP were the most active enzymes, suggesting that microbial metabolic profiles were modulated by labile organic substrates. Our results suggest that the composition and function of microbial biofilm could be considered as sentinels of natural or anthropic-related disturbances.

Redox conditions and a moderate anthropogenic impairment of groundwater quality reflected on the microbial functional traits in a volcanic aquifer

Groundwater is an important freshwater resource and hosts specialized microbial assemblages providing fundamental ecosystem services. The current knowledge on the role of aquatic microorganisms in subsurface ecosystems is still limited. This work aimed to explore the links between groundwater hydrogeochemical properties and microbial community traits in a volcanic unconfined aquifer, moderately impacted by anthropic activities. The main physical and chemical parameters of groundwater samples were analyzed, along with microbial biomass (total cell counts, ATP-active biomass concentration), potential metabolic activity, and physiological profiles at the microbial community level (Biolog ™ EcoPlates). The results showed the coexistence of oxidizing and reducing groundwater conditions across the study area. We discriminated two groups of oxidizing/reducing groundwater samples (Ox and Red), each including two subgroups with different chemical conditions attributed to contrasting levels of anthropogenic impact for non-intensive agricultural practices and waste disposal activities (Ox − and Ox + ; Red − and Red +). Although the microbial biomass was likely not affected by changing redox, the microbial metabolic potential and functional diversity changed significantly. In the Ox samples, the community-level physiological profiles were different, mainly owing to the utilization of carboxylic acids (Ox − > Ox +) and carbohydrates (Ox + > Ox −). In the Red samples, a wider set of organic substrates were consumed by the microbial communities, including those less bioavailable (e.g., phenols). Significant differences were also found between Red − and Red + , mainly owing to the relative increase in the utilization of amino acids in Red − , polymers and amines in Red + , along with the active biomass. By reflecting the local redox conditions and moderate levels of anthropogenic impact, the applied approach highlighted changes of microbial metabolic potential and physiological profiles that imply direct repercussions on biogeochemical cycling and the ecosystem services provided by groundwater microorganisms.

Metagenomic Features Characterized with Microbial Iron Oxidoreduction and Mineral Interaction in Southwest Indian Ridge.

The Southwest Indian Ridge (SWIR) is one of the typical representatives of deep-sea ultraslow-spreading ridges, and has increasingly become a hot spot of studying subsurface geological activities and deep-sea mining management. However, the understanding of microbial activities is still limited on active hydrothermal vent chimneys in SWIR. In this study, samples from an active black smoker and a diffuse vent located in the Longqi hydrothermal region were collected for deep metagenomic sequencing, which yielded approximately 290 GB clean data and 295 mid-to-high-quality metagenome-assembled genomes (MAGs). Sulfur oxidation conducted by a variety of Gammaproteobacteria, Alphaproteobacteria, and Campylobacterota was presumed to be the major energy source for chemosynthesis in Longqi hydrothermal vents. Diverse iron-related microorganisms were recovered, including iron-oxidizing Zetaproteobacteria, iron-reducing Deferrisoma, and magnetotactic bacterium. Twenty-two bacterial MAGs from 12 uncultured phyla harbored iron oxidase Cyc2 homologs and enzymes for organic carbon degradation, indicated novel chemolithoheterotrophic iron-oxidizing bacteria that affected iron biogeochemistry in hydrothermal vents. Meanwhile, potential interactions between microbial communities and chimney minerals were emphasized as enriched metabolic potential of siderophore transportation, and extracellular electron transfer functioned by multi-heme proteins was discovered. Composition of chimney minerals probably affected microbial iron metabolic potential, as pyrrhotite might provide more available iron for microbial communities. Collectively, this study provides novel insights into microbial activities and potential mineral-microorganism interactions in hydrothermal vents. IMPORTANCE Microbial activities and interactions with minerals and venting fluid in active hydrothermal vents remain unclear in the ultraslow-spreading SWIR (Southwest Indian Ridge). Understanding about how minerals influence microbial metabolism is currently limited given the obstacles in cultivating microorganisms with sulfur or iron oxidoreduction functions. Here, comprehensive descriptions on microbial composition and metabolic profile on 2 hydrothermal vents in SWIR were obtained based on cultivation-free metagenome sequencing. In particular, autotrophic sulfur oxidation supported by minerals was presumed, emphasizing the role of chimney minerals in supporting chemosynthesis. Presence of novel heterotrophic iron-oxidizing bacteria was also indicated, suggesting overlooked biogeochemical pathways directed by microorganisms that connected sulfide mineral dissolution and organic carbon degradation in hydrothermal vents. Our findings offer novel insights into microbial function and biotic interactions on minerals in ultraslow-spreading ridges.

Deciphering Alternaria metabolic responses in microbial confrontation via an integrated mass spectrometric targeted and non-targeted strategy

Microbial confrontation is ubiquitously present in nature, such as mycoparasitism, competition and antibiosis between biocontrol agents and microbial pathogens. However, the internal metabolic responses of fungal pathogens under microbial interaction scenario have been scarcely investigated. In this study, we set up an integrated mycotoxin analysis and non-targeted metabolomics workflow to decipher metabolic changes of Alternaria pathogens when confronted with selected Trichoderma strains, as well as mycotoxin metabolization in the Trichoderma spp. Results demonstrated that Trichoderma spp. significantly influenced mycotoxin production and whole metabolome of Alternaria pathogens when in cocultivation, and one Trichoderma strain could metabolize alternariol into its hydroxylated form. These differential metabolites revealed fungal physiological alternations in various confrontation conditions. In all, a MS-based strategy was proposed to investigate microbial metabolic profiles under fungal/fungal and fungal/mycotoxin cocultivation, and this generic methodology would be significant for understanding the occurrence and change of food contaminants in microbial interactions.

A signature of Prevotella copri and Faecalibacterium prausnitzii depletion, and a link with bacterial glutamate degradation in the Kenyan colorectal cancer patients.

Colorectal cancer (CRC) is the fifth most diagnosed cancer in Sub-Saharan Africa. In Kenya, CRC incidence rates tripled from 1997 to 2017. In the Moi Teaching and Referral Hospital, Moi University, there has been an increase in CRC cases, notably for younger patients. A suggested pathobiology for this increase is gut microbiome dysbiosis. Since, for the Kenyan CRC patient population, microbiome studies are rare, there is a need for a better understanding of how microbiome dysbiosis influences CRC epidemiology in Kenya. In this single-center study, the focus was on profiling the gut microbiome of Kenyan CRC patients and healthy volunteers and evaluating associations between microbiome profiles and the age of CRC patients. The gut mucosa-associated microbiome of 18 CRC patients and 18 healthy controls were determined by 16S rRNA sequencing and analyzed for alpha and beta diversity, differential abundance, and microbial metabolic profiling. Alpha diversity metrics showed no significant differences, but beta diversity metrics showed dissimilarities in the microbial communities between CRC patients and healthy controls. The most underrepresented species in the CRC group were Prevotella copri (P. copri) and Faecalibacterium prausnitzii (F. prausnitzii), although Bacteroides fragilis (B. fragilis) and Prevotella nigrescens were overrepresented (linear discriminant analysis, LDA score >2, P<0.05). Also, for CRC patients, significant metagenomic functional alterations were evident in microbial glutamate metabolic pathways (L-glutamate degradation VIII was enriched, and L-glutamate and L-glutamine biosynthesis were diminished) (P<0.05, log2 Fold Change >1). Moreover, the microbiome composition was different for patients under 40 years of age compared to older patients (LDA score >2, P<0.05). Microbiome and microbial metabolic profiles of CRC patients are different from those of healthy individuals. CRC microbiome dysbiosis, particularly P. copri and F. prausnitzii depletion and glutamate metabolic alterations, are evident in Kenyan CRC patients.

Early hematopoietic injury triggered by benzene characterized with inhibition of erythrocyte differentiation involving the mollicutes_RF39-derived citrulline.

Benzene poisoning is a common adverse blood outcome in occupational workers, manifested by hematopoietic dysfunction. However, the specific phenotype and its mechanisms of early hematopoietic toxicity caused by benzene remain unclear. After 15 days of exposure, the WBC levels were not significantly altered in benzene-exposed mice. However, the level of red blood cells (RBC) showed a significant decrease, and it was significantly and negatively correlated with urinary S-phenylmercapturic acid (SPMA). Notably, 5mg/kg benzene exposure significantly inhibited the renewal capacity and the number of colony formation of hematopoietic stem progenitor cells in mice, especially erythrocyte differentiation. These results suggested that the early hematopoietic toxicity phenotype caused by benzene was dominated by inhibition of erythroid differentiation rather than WBC-related inflammation. To further understand the underlying mechanisms of benzene-induced early hematopoietic toxicity, 16S rRNA sequencing and plasma metabolites analysis were conducted to investigate the impact of benzene exposure for 15 days on microbial composition and metabolic profile of mice. We found that short-term benzene exposure induced disturbances in gut microbiota and metabolism. The relative abundance of Mollicutes_RF39 at order levels was significantly reduced in benzene-exposed mice and was strongly correlated with hematopoietic indicators and urinary benzene markers. Interestingly, Mollicutes_RF39 might disturb the levels of eight metabolites, whereas Citrulline was highly linked to Mollicutes_RF39 (r=0.862, P=0.000). Consequently, Mollicutes_RF39-derived Citrulline might be the key regulator of early hematopoietic injury induced by benzene exposure. These findings promote the understanding of early hematotoxicity phenotypes and provide new perspectives on the underlying mechanisms of benzene-induced hematotoxicity.

Gut microbiota and metabolic profile as affected by Maillard reaction products derived from bighead carp meat hydrolysates with galactose and galacto-oligosaccharides during in vitro pig fecal fermentation

Maillard reaction products (MRPs) play pivotal roles in gut health by affecting the microbiome-host interactions. This study aimed at investigating the effects of MRPs derived from bighead carp meat hydrolysates with galactose and galacto-oligosaccharides on intestinal microbial composition and metabolic profile by in vitro pig fecal fermentation. The pH decreased sharply in the first 12 h and the highest production of butyric acid was observed in GM (glycated BCH with galacto-oligosaccharide) treatment with 64.7 μmoL/10 mL (p < 0.05) at 48 h. Clostridium_sensu_stricto_1, Streptococcus, and Enterococcus were dominant in the GM treatment, while Escherichia-Shigella was predominant in LgM (glycated BCH with galactose) treatment at 12 h. The up-regulated metabolites indicated that GM and LgM might participate in the fatty acids synthesis and modulate lipid metabolism, respectively. Overall, GM will be more beneficial for gut health by promoting the production of butyric acid and fatty acids synthesis.

Effects of the increased protein level in small intestine on the colonic microbiota, inflammation and barrier function in growing pigs

BackgroundAn increased level of the dietary protein alters the colonic microbial community and metabolic profile of pigs, but it remains unclear whether this leads to colonic inflammation and impairs barrier function in growing pigs.ResultsSixteen pigs (35.2 ± 0.3 kg) were infused with sterile saline (control) or soy protein hydrolysate (SPH) (70 g/day) through a duodenal fistula twice daily during a 15-day experimental period. The SPH treatment did not affect their average daily feed intake and daily weight gain (P > 0.05), but reduced colon index and length (P < 0.05). Illumina MiSeq sequencing revealed that species richness was increased following SPH intervention (P < 0.05). Furthermore, SPH reduced the abundance of butyrate- and propionate-producing bacteria—such as Lachnospiraceae NK4A136 group, Lachnospiraceae_uncultured, Coprococcus 3, Lachnospiraceae UCG-002, and Anaerovibrio—and increased the abundance of potentially pathogenic bacteria and protein-fermenting bacteria, such as Escherichia-Shigella, Dialister, Veillonella, Prevotella, Candidatus Saccharimonas, Erysipelotrichaceae UCG-006, Prevotellaceae_uncultured, and Prevotellaceae UCG-003 (P < 0.05). In addition, a lower content of total short-chain fatty acids, propionate, and butyrate and a higher concentration of cadaverine, putrescine, total biogenic amines, ammonia, and isovalerate were observed following SPH infusion (P < 0.05). Further analysis revealed that SPH increased the concentration of tumour necrosis factor-α, interleukin (IL)-1β, IL-6, and IL-8 in the colonic mucosa (P < 0.05). Interestingly, SPH intervention increased the expression of occludin, zonula occludens (ZO)-1, and claudin-1 in colonic mucosa (P < 0.05). Correlation analysis showed that different genera were significantly related to the production of metabolites and the concentrations of pro-inflammatory cytokines.ConclusionAn increased soy protein level in the small intestine altered the colonic microbial composition and metabolic profile, which resulted in the secretion of colonic proinflammatory cytokines and the increased expression of tight junction proteins.

Responses of microbial communities and metabolic profiles to the rhizosphere of Tamarix ramosissima in soils contaminated by multiple heavy metals

Heavy metals (HMs) contamination around smelters poses serious stress to soil microbiome. However, the co-effect of multiple HMs and native vegetation rhizosphere on the soil ecosystem remains unclear. Herein, effects of high HMs level and the rhizosphere (Tamarix ramosissima) on soil bacterial community structure and metabolic profiles in sierozem were analyzed by coupling high-throughput sequencing and soil metabolomics. Plant roots alleviated the threat of HMs by absorbing and stabilizing them in soil. High HMs level decreased the richness and diversity of soil bacterial community and increased numbers of special bacteria. Plant roots changed the contribution of HMs species shaping the bacterial community. Cd and Zn were the main contributors to bacterial distribution in non-rhizosphere soil, however, Pb and Cu became the most important HMs in rhizosphere soil. HMs induced more dominant metal-tolerant bacteria in non-rhizosphere than rhizosphere soil. Meanwhile, critical metabolites varied by rhizosphere in co-occurrence networks. Moreover, the same HMs-tolerant bacteria were regulated by different metabolites, e.g. unclassified family AKYG1722 was promoted by Dodecanoic acid in non-rhizosphere soil, while promoted by Octadecane, 2-methyl- in rhizosphere soil. The study illustrated that high HMs level and rhizosphere affected soil properties and metabolites, by which soil microbial community structure was reshaped.

Antimicrobial PCMX facilitates the volatile fatty acids production during sludge anaerobic fermentation: Insights of the interactive principles, microbial metabolic profiles and adaptation mechanisms

Para-chloro-meta-xylenol (PCMX), as a massively used antimicrobial agent, exhibited dose-dependent effects on the sludge fermentation for volatile fatty acids (VFAs). The presence of PMCX at environmental levels (10–20 mg/g TSS) promoted the VFAs production by 2.5–3.0 folds but caused evident inhibition at high level (100 mg/g TSS). PCMX synchronously promoted the sludge solubilization, hydrolysis, and acidification but inhibited methanogenesis. Mechanistic investigations revealed that PCMX could interact with the extracellular polymeric substances (EPS) in sludge and cause the release of biodegradable substrates (especially proteins) by altering the WAS structures. Moreover, the functional fermentative bacteria (i.e., Firmicutes and Chloroflexi) were enriched while those VFAs consumers (i.e., Methanothrix and Methanomassiliicoccus) were reduced with low level of PCMX exposure. The microbial enzymatic activities (i.e., protease) and critical genes relevant to the substrates transport (i.e., gluC and lhpN) and metabolisms (especially the proteins, i.e., pckA), and VFAs biosynthesis (i.e., fabG) were all upregulated with the low PCMX stimulus. The microbial response and adaptation to PCMX toxicity were mainly attributed to the regulation of two-component and quorum sensing systems. Overall, PCMX contributed to the VFAs accumulation during sludge fermentation by mainly interacting and disrupting the sludge structures, and improving the microbial metabolic function traits. This work provided new insights into exogenous pollutants’ interactions, potential influences, and reaction mechanisms in the complicated sludge matrix.

Ablation of Carotenoid Cleavage Enzymes BCO1 & BCO2 Alters Gut Microbial and Plasma Metabolomic Profiles in High Refined Carbohydrate Diet-Fed Mice

Objectivesβ-Carotene-15,15′-oxygenase (BCO1) and β-carotene-9′,10′-oxygenase (BCO2) cleave carotenoids to form vitamin A or apo-carotenoid species. Beyond these roles, BCO1 and BCO2 have been implicated in modulating lipid, cholesterol, and oxidative stress-related processes, yet less is known about their combined influence on shaping gut microbial and plasma metabolic profiles. Current intake of refined carbohydrates and added sugars exceeds the recommendations within the Dietary Guidelines for Americans, which may contribute to the development of nonalcoholic fatty liver disease (NAFLD). In the present study, we aimed to uncover whether mice with systemic ablation of BCO1-/-/BCO2-/- (DKO) display distinctive phenotypes in response to a high refined carbohydrate diet (HRCD) model of NAFLD. MethodsMale, six-week-old C57BL/6J wild type mice (WT) and DKO mice were fed a chow diet or a HRCD (66.5% carbs including sucrose) for 24 weeks. We used fecal and plasma samples to respectively perform 16S rDNA sequencing, and both targeted and untargeted metabolomics using our UHPLC-QTOF-MS platform, a targeted Biocrates MxP Quant 500 kit, and a complimentary untargeted analysis of phospholipids from the METLIN database. Data were analyzed using QIIME2-DEseq2, MetaboAnalyst 5.0, MassHunter Qualitative Analysis, and MicrobiomeAnalyst. ResultsHRCD-fed WT and DKO mice developed similar levels of hepatic steatosis although DKO mice had significantly higher hepatic levels of malondialdehyde (MDA). Interestingly, in both chow-fed and HRCD-fed DKO mice, plasma adiponectin and hepatic bile acids were significantly lower compared to respective WT groups. Our metabolomics data revealed that DKO mice displayed significant alterations in ceramide, cholesterol ester, triglyceride, and phospholipid classes under both chow and HRCD conditions. Additionally, when compared to HRCD-fed WT mice, HRCD-fed DKO mice displayed significantly reduced alpha diversity and possessed significant compositional differences based on Bray Curtis dissimilarity measures. ConclusionsBCO1/BCO2 DKO mice display altered metabolomic and gut microbial profiles. Carotenoid cleavage enzymes affect diverse physiological processes, which involve lipid metabolism and the gut microbiome. Funding SourcesUSDA/ARS and NIFA/AFRI.

Short-chain fatty acids promote the effect of environmental signals on the gut microbiome and metabolome in mice

Gut microorganisms and the products of their metabolism thoroughly affect host brain development, function and behavior. Since alterations of brain plasticity and cognition have been demonstrated upon motor, sensorial and social enrichment of the housing conditions, we hypothesized that gut microbiota and metabolome could be altered by environmental stimuli, providing part of the missing link among environmental signals and brain effects. In this preliminary study, metagenomic and metabolomic analyses of mice housed in different environmental conditions, standard and enriched, identify environment-specific microbial communities and metabolic profiles. We show that mice housed in an enriched environment have distinctive microbiota composition with a reduction in gut bacterial richness and biodiversity and are characterized by a metabolomic fingerprint with the increase of formate and acetate and the decrease of bile salts. We demonstrate that mice treated with a mixture of formate and acetate recapitulate some of the brain plasticity effects modulated by environmental enrichment, such as hippocampal neurogenesis, neurotrophin production, short-term plasticity and cognitive behaviors, that can be further exploited to decipher the mechanisms involved in experience-dependent brain plasticity.

Synergy of Dietary Quercetin and Vitamin E Improves Cecal Microbiota and Its Metabolite Profile in Aged Breeder Hens.

In the present study, the synergistic effects of quercetin (Q) and vitamin E (E) on cecal microbiota composition and function, as well as the microbial metabolic profile in aged breeder hens were investigated. A total of 400 (65 weeks old) Tianfu breeder hens were randomly allotted to four experimental groups (four replicates per group). The birds were fed diets containing quercetin at 0.4 g/kg, vitamin E (0.2 g/kg), quercetin and vitamin E (QE; 0.4 g/kg and 0.2 g/kg), and a basal diet for a period of 10 wks. After the 10 week experimental period, the cecal contents of 8 aged breeder hens per group were sampled aseptically and subjected to high-throughput 16S rRNA gene sequencing and untargeted metabolomic analysis. The results showed that the relative abundances of phyla Bacteroidota, Firmicutes, and Actinobacteriota were the most prominent among all the dietary groups. Compared to the control group, the relative abundance of the families Bifidobacteriaceae, Lachnospiraceae, Tannerellaceae, Mathonobacteriaceae, Barnesiellaceae, and Prevotellaceae were enriched in the QE group; and Bacteroidaceae, Desulfovibrionaceae, Peptotostretococcaceae, and Fusobacteriaceae were enriched in the Q group, whereas those of Lactobacillaceae, Veillonellaceae, Ruminococcaceae, Akkermansiaceae, and Rikenellaceae were enriched in the E group compared to the control group. Untargeted metabolomics analyses revealed that Q, E, and QE modified the abundance of several metabolites in prominent pathways including ubiquinone and other terpenoid–quinone biosynthesis, regulation of actin cytoskeleton, insulin secretion, pancreatic secretion, nicotine addiction, and metabolism of xenobiotics by cytochrome P450. Furthermore, key cecal microbiota, significantly correlated with important metabolites, for example, (S)-equol positively correlated with Alistipes and Chlamydia in E_vs_C, and negatively correlated with Olsenella, Paraprevotella, and Mucispirillum but, a contrary trend was observed with Parabacteroides in QE_vs_C. This study establishes that the synergy of quercetin and vitamin E alters the cecal microbial composition and metabolite profile in aged breeder hens, which lays a foundation for chicken improvement programs.

Fermented Myriophyllum aquaticum and Lactobacillus plantarum Affect the Distribution of Intestinal Microbial Communities and Metabolic Profile in Mice

This research explores the effects of fermented Myriophyllum aquaticum (F) and Lactobacillus plantarum BW2013 (G) as new feed additives on the gut microbiota composition and metabolic profile of mice. Crude protein (p = 0.045), lipid (p = 0.000), and ash (p = 0.006) contents in Myriophyllum aquaticum (N) were improved, whereas raw fiber (p = 0.031) content was decreased after solid-state fermentation by G. Mice were fed with no additive control (CK), 10%N (N), 10%N + G (NG), 10%F (F), and 10%F + G (FG). High-throughput sequencing results showed that, compared with the CK group, Parabacteroides goldsteinii was increased in treatment groups and that Lactobacillus delbrueckii, Bacteroides vulgatus, and Bacteroides coprocola were increased in the F and FG groups. Bacteroides vulgatus and Bacteroides coprocola were increased in the F group compared with the N group. Metabolomic results showed that vitamin A, myricetin, gallic acid, and luteolin were increased in the F group compared with the N group. Reduction in LPG 18:1 concentration in the N and F groups could be attenuated or even abolished by supplementation with G. Furthermore, 9-oxo-ODA was upregulated in the FG group compared with the F group. Collectively, N, F, and G have beneficial effects on gut microbiota and metabolic profile in mice, especially intake of FG.

Lower abundance of Bacteroides and metabolic dysfunction are highly associated with the post-weaning diarrhea in piglets.

Growing evidences show a direct link between diarrhea and disorders of gut microbiota in pigs. However, whether there are microbial markers associated with post-weaning diarrhea remains unknown. In the current study, we compared the microbial community, functions and metabolites between healthy weaned piglets (group H, n=7) and piglets with post-weaning diarrhea (group D, n=7), in order to find out diarrhea associated microbial markers. Each of 7 fecal samples was collected from H and D piglets (weaned at 21 d and sampled at 26 d). The metagenomic and untargeted metabolomic analysis revealed that the microbial composition, function and metabolic profile in D pigs was considerably reshaped, including the reduced abundance and number of Bacteroides, which significantly correlated with the diarrhea status of host. The carbohydrate metabolism, biosynthesis and metabolism, lipid metabolism, amino acid metabolism, and the activity of glycan and carbohydrates digestion related enzymes showed extensively down-regulated in D pigs compared with H pigs. Diarrhea significantly changed the metabolic profiles of fecal microbiota, and most of the altered metabolites were negatively or positively correlated with the change in the abundance of Bacteroides. In conclusion, the lower abundance of Bacteroides and its associated metabolic dysfunction may be regarded as microbial markers of physiological post-weaning diarrhea in piglets.

Gut Microbiota-Mediated Elevated Production of Secondary Bile Acids in Chronic Unpredictable Mild Stress.

A growing body of evidence suggests that gut microbiota could participate in the progression of depression via the microbiota–gut–brain axis. However, the detailed microbial metabolic profile changes in the progression of depression is still not fully elucidated. In this study, a liquid chromatography coupled to mass spectrometry-based untargeted serum high-throughput metabolomics method was first performed to screen for potential biomarkers in a depressive-like state in a chronic unpredictable mild stress (CUMS)-induced mouse model. Our results identified that the bile acid and energy metabolism pathways were significantly affected in CUMS progression. The detailed bile acid profiles were subsequently quantified in the serum, liver, and feces. The results showed that CUMS significantly promoted the deconjugation of conjugated bile acid and secondary bile acid biosynthesis. Furthermore, 16S rRNA gene sequencing revealed that the increased secondary bile acid levels in the feces positively correlated with Ruminococcaceae_UCG-010, Ruminococcus, and Clostridia_UCG-014 abundance. Taken together, our study suggested that changes in family Ruminococcaceae abundance following chronic stress increased biosynthesis of deoxycholic acid (DCA), a unconjugated secondary bile acid in the intestine. Aberrant activation of secondary bile acid biosynthesis pathway thereby increased the hydrophobicity of the bile acid pool, which might, in turn, promoted metabolic disturbances and disease progression in CUMS mice.