Fecal Amino Acid Research Articles

Fecal proteolytic profiling of pediatric inflammatory bowel disease: A pilot study.

Colonoscopy is the gold standard for diagnosing inflammatory bowel disease (IBD). However, this invasive procedure has a high burden for pediatric patients. Previous research has shown elevated fecal amino acid concentrations in children with IBD versus controls. We hypothesized that this finding could result from increased proteolytic activity. Therefore, the aim of this study was to investigate whether fecal protease-based profiling was able to discriminate between IBD and controls. Protease activity was measured in fecal samples from patients with IBD (Crohn's disease (CD) n = 19; ulcerative colitis (UC) n = 19) and non-IBD controls (n = 19) using a fluorescence resonance energy transfer (FRET)-peptide library. Receiver operating characteristic (ROC) curve analysis was used to determine the diagnostic value of each FRET-peptide substrate. Screening the FRET-peptide library revealed an increased total proteolytic activity (TPA), as well as degradation of specific FRET-peptides specifically in fecal samples from IBD patients. Based on level of significance (p < .001) and ROC curve analysis (AUC > 0.85), the fluorogenic substrates W-W, A-A, a-a, F-h, and H-y showed diagnostic potential for CD. The substrates W-W, a-a, T-t, G-v, and H-y showed diagnostic potential for UC based on significance (p < .001) and ROC analysis (AUC > 0.90). None of the FRET-peptide substrates used was able to differentiate between protease activity in fecal samples from CD versus UC. This study showed an increased fecal proteolytic activity in children with newly diagnosed, treatment-naïve, IBD. This could lead to the development of novel, noninvasive biomarkers for screening and diagnostic purposes.

Defatted black soldier fly larvae meal in lactating sow and pre-weaning piglet diets: impacts on growth performance, fecal microbiota, and metabolic pathways

Abstract To address the underexplored use of black soldier fly larval (BSFL) meal in pig farming for sustainable development, this study examined the effects of substituting conventional protein with defatted BSFL meal in the pre-weaning diet of piglets farrowed from sows fed defatted BSFL diets (0%, 2.3%, and 4.6%). Fifty-four piglets (18 piglets in each treatment) were housed with sows until 28 days of age and fed a pre-weaning diet containing 0%, 12.5%, or 25% defatted BSFL meal based on their respective sows’ dietary treatments (0%, 2.3%, and 4.6% defatted BSFL diets) from 21 to 35 days of age. The growth performances and fecal microbiota of piglets were investigated. The results showed that sow diets and defatted BSFL meal-incorporated pre-weaning diets did not have a significant effect on the growth performance of piglets (). However, piglet growth tended to decrease with increasing defatted BSFL meals in the pre-weaning diets. Additionally, piglets nursed by sows fed defatted BSFL diets showed higher fecal short-chain fatty acid and lower ammonia levels at 21 days of age than those nursed by sows fed a control diet (). Furthermore, defatted BSFL meal-incorporated pre-starter diets decreased fecal amino acids and modified the gut microbial metabolic pathways associated with stress response. These findings suggest that the appropriate inclusion of defatted BSFL meal in the diets of lactating sows and piglets did not adversely affect the growth performance of piglets and may potentially improve the health of piglets at weaning; however, attention should be paid to the chitin content in BSFL diets, which may affect nutrient digestibility. Incorporating 2.3% defatted BSFL to sow and 12.5% to pre-weaning piglet diets may serve as a feeding strategy in pig farming. The results obtained will aid the application of BSFL meal in the swine industry and the study of its dietary effect on weaning stress in piglets.

Fermented soybean meal improved laying performance and egg quality of laying hens by modulating cecal microbiota, nutrient digestibility, intestinal health, antioxidant and immunological functions

Antinutritional factors in feedstuffs may limit their utilization in livestock production, but fermentation process can be used to improve feed quality; however, studies on fermented soybeans for laying hens remain limited. We investigated the effect of fermented soybean meal (FSBM) at various inclusion levels as a partial replacement for soybean meal (SBM) on egg production, egg quality, amino acid digestibility, gut morphology and microbiota, antioxidant capacity and immune response of young laying hens. A total of 360 Hy-line Brown laying hens aged 18 weeks were selected and divided into 5 groups of 6 replicates each and 12 birds per replicate. The control group received a basal diet while the trial group received the basal diet with FSBM included at 2.5%, 5.0%, 7.5% and 10.0%, respectively, for 12 weeks. Our findings revealed that the nutritional value of FSBM was higher compared to that of SBM in terms of reduced content of trypsin inhibitors and increased contents of crude protein, amino acids and minerals. FSBM enhanced egg production (P < 0.05), feed-to-egg ratio (P < 0.05), and albumen quality (albumen height and Haugh unit) (P < 0.05). Furthermore, FSBM improved apparent fecal amino acid digestibility (P < 0.05), gut morphology (increased villus height, villus width, villus height-to-crypt depth ratio and decreased crypt depth) (P < 0.05), antioxidant capacity (reduced malondialdehyde and increased catalase, total superoxide dismutase, glutathione peroxidase and total antioxidant capacity) (P < 0.05) and immune function (increased concentrations of IgG, IgA, and IgM; increased levels of transforming growth factor beta and Toll-like receptor 2; and reduced levels of interleukin 1β and tumor necrosis factor alpha) (P < 0.05). Further analysis showed that FSBM altered the composition of the gut microbiota favoring beneficial microbes. These findings suggest that probiotic fermentation improved the nutritional value of SBM. The inclusion of FSBM in the diets of laying hens at 2.5% or 5.0% improved amino acid digestibility, gut health, immune function, egg production and egg quality.

Multi-omic integration of microbiome data for identifying disease-associated modules

Multi-omic studies of the human gut microbiome are crucial for understanding its role in disease across multiple functional layers. Nevertheless, integrating and analyzing such complex datasets poses significant challenges. Most notably, current analysis methods often yield extensive lists of disease-associated features (e.g., species, pathways, or metabolites), without capturing the multi-layered structure of the data. Here, we address this challenge by introducing “MintTea”, an intermediate integration-based approach combining canonical correlation analysis extensions, consensus analysis, and an evaluation protocol. MintTea identifies “disease-associated multi-omic modules”, comprising features from multiple omics that shift in concord and that collectively associate with the disease. Applied to diverse cohorts, MintTea captures modules with high predictive power, significant cross-omic correlations, and alignment with known microbiome-disease associations. For example, analyzing samples from a metabolic syndrome study, MintTea identifies a module with serum glutamate- and TCA cycle-related metabolites, along with bacterial species linked to insulin resistance. In another dataset, MintTea identifies a module associated with late-stage colorectal cancer, including Peptostreptococcus and Gemella species and fecal amino acids, in line with these species’ metabolic activity and their coordinated gradual increase with cancer development. This work demonstrates the potential of advanced integration methods in generating systems-level, multifaceted hypotheses underlying microbiome-disease interactions.

Abstract 1471: A novel mouse model of upper tract urothelial carcinoma influenced by gut microbiota

Abstract Introduction &amp; Objective: Recently, it has been revealed that bladder cancer (BC) and upper tract urothelial carcinoma (UTUC) are biologically distinct diseases. Mouse models of BC carcinogenesis have been established using N-butyl-N-(4-hydro-oxybutyl) nitorosamine (BBN) for a long time, but there are few reports of UTUC carcinogenesis with high incidence. Therefore, the UTUC carcinogenesis mouse model is expected. The aim of this study is to establish a novel mouse model of UTUC carcinogenesis and to identify the characteristics of this model. Methods: We used two strains (C57BL/6 and BALB/c) of male and female mice and fed 0.05% BBN from 6 weeks old to the harvest day. Both sides of upper urinary tract from mice drinking BBN or tap water were subjected to whole exome sequencing and whole transcriptome sequencing (WTS) to assess its genetic similarity to human UTUC. Microbiome in gut were assessed by 16S ribosomal RNA gene sequencing (16S rRNA-seq). Metabolic differences in feces, serum and liver between UTUC mice and healthy mice were evaluated by comprehensive metabolomic analysis. Inflammatory mediators were assessed by quantitative polymerase chain reaction (qPCR). Results: Only BALB/c female showed higher incidence of UTUC than BC, while the other strains rarely had UTUC. This model showed SBS5-like mutational pattern. The frequent mutated gene list showed similarity to human UTUC. In terms of gene expression, this model belonged basal subtype. We detected 593 differentially expressed genes between cancerous side and non-cancerous side of upper urinary tract in this model, which could also classify human UTUC and adjacent normal samples. The cancerous side of upper urinary tract enriched several inflammation related pathways such as Nfkb and Tnf pathway. We validated the upregulation of inflammatory gene expression in upper urinary tract (Il6, Il1b, Tnfa, Relb) by qPCR (all: p &amp;lt; 0.05). 16S rRNA-seq revealed that Firmicutes was enriched in feces. Metabolomic analysis revealed that some amino acids in feces and FAD in serum were upregulated. Feeding with amino acid mediated diet changed gut environment. Firmicutes in feces and FAD in both feces and serum were downregulated consistently. This diet suppressed inflammatory gene expression in upper urinary tract (Il6, Il1b, Tnfa, Relb, Cd38, Tnfrsf1b and Kdm1a; all: p &amp;lt; 0.05). Finally, UTUC was disappeared but not BC in this model treated by BBN with amino acid mediated diet. Conclusion: We have established a novel mouse model developing UTUC with high incidence that genetically mimics human UTUC. In addition, amino acid mediated diet can change both microbiome and metabolome in gut of this model, downregulate host’s inflammatory reaction to BBN in upper urinary trat and suppress the UTUC carcinogenesis. Citation Format: Akinaru Yamamoto, Atsunari Kawashima, Kentaro Jingushi, Sassi Nesrine, Yuki Horibe, Akihiro Yoshimura, Masaru Tani, Liu Yutong, toshiki Oka, Yohei Okuda, toshihiro Uemura, Gaku Yamamichi, Yu Ishizuya, Yoshiyuki Yamamoto, Taigo Kato, Koji Hatano, Norio Nonomura. A novel mouse model of upper tract urothelial carcinoma influenced by gut microbiota [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1471.

Effects of chronic unpredictable mild stress on gut microbiota and fecal amino acid and short-chain fatty acid pathways in mice

Depression is a serious disease that has a significant impact on social functioning. However, the exact causes of depression are still not fully understood. Therefore, it is necessary to explore new pathways leading to depression. In this study, we used 16 S rDNA to examine changes in gut microbiota and predict related pathways in depression-like mice. Additionally, we employed liquid chromatography-mass spectrometry (LC-MS) to identify changes in amino acids and gas chromatography-mass spectrometry (GC-MS) to identify changes in short-chain fatty acids (SCFAs) in fecal samples. We conducted Pearson/Spearman correlation analysis to investigate the associations between changes in amino acids/SCFAs and behavioral outcomes. The 16 S rDNA sequencing revealed significant alterations in gut microbiota at the phylum and genus levels in mice subjected to chronic unpredictable mild stress (CUMS). The relative abundances of Bacteroidetes, Proteobacteria, Bacteroides, and Alloprevotella were increased, while Firmicutes, Verrucomicrobia, Actinobacteria, Lactobacillus, Akkermansia, Lachnospirillum, and Enterobacter were decreased in the CUMS mice. We used PICRUSt software to annotate the kyoto encyclopedia of genes and genomes (KEGG) pathway function related to depression-like behavior in mice. Our analysis identified sixty functional pathways associated with the gut microbiota of mice exhibiting depression-like behavior. In the amino acid concentration analysis, we observed decreased levels of hydroxyproline and tryptophan, and increased levels of alanine in CUMS mice. In the SCFAs concentration assay, we found decreased levels of butyric acid and valeric acid, and increased levels of acetic acid in CUMS mice. Some of these changes were significantly correlated with depressive-like behaviors. Our study contributes to the understanding of the mechanism of the gut-brain axis in the occurrence and development of depression.

Effects of fermented soybean meal and supplemental methionine and lysine on growth, digestibility, body composition, and amino acid composition of Beluga (Huso huso).

There is an increasing concern about using limited high-quality protein sources in the aquafeed industry. The alternative solution to this problem is cost-effective plant proteins such as soybean meal (SBM). However, it is better to improve plant protein-containing diets through processing and supplemental amino acids. This study aimed to examine the effects of fermented soybean meal (FSBM) and supplemental methionine and lysine (ML) in diets on growth, digestibility, and body and amino acid composition in Beluga (Huso huso). Two basal diets (490g FM Kg-1) of fish meal (FM) and FM with ML (FM + ML) were replaced by 40%, 60%, and 80% of FSBM and FSBM + ML (FM, FSBM40, FSBM60, FSBM80, FM + ML, FSBM40 + ML, FSBM60 + ML and FSBM80 + ML; 444g kg-1 crude protein and 19.77 MJ kg-1 gross energy). The diets were fed to triplicate groups of Beluga (394.6 ± 4.3g) three times daily for 56 d. The results showed that Beluga fed 80% FSBM and FSBM + ML had lower growth and feed utilization. Moreover, Beluga-fed FSBM + ML performed poorer than those fed FSBM (P < 0.05). No interactions between fermented SBM and ML were observed in apparent digestibility coefficients (ADC) and body proximate composition except for ADC of crude lipid. ADC in Beluga fed 80% were less than other diets. Substitution of FSBM and FSBM + ML in diets significantly reduced dry matter, ash and crude lipid; Crude lipid increased in groups fed FSBM + ML (P < 0.05). FSBM and FSBM + ML in 60% had higher and lower total amino acids in muscle and feces (P < 0.05), respectively. Also, excreted total amino acids in feces increased with ML (P < 0.05). Generally, in Beluga, FSBM-containing diets showed better effects on growth, feed utilization, whole-body lipid, and muscle and fecal amino acids than those with FSBM + ML. Fermented SBM could be replaced up to 60%.

Altered intestinal microbiota in children with bronchiolitis.

To investigate the correlation between the alteration of intestinal microbiota and disease in children with bronchiolitis. Fifty seven children diagnosed with bronchiolitis from January 2020 to January 2022 in our pediatric department were included as the case group, and another 36 normal children were included as the control group. Stool and blood were collected from both groups for high-throughput sequencing, untargeted metabolite detection and ELISA. A mouse model of RSV infection was established to validate the results of clinical case detection. Body weight, passive smoking, and a host of other factors were possible as acute bronchiolitis influencing factors in the onset of acute bronchiolitis. The alpha diversity Shannon, Simpson and Pielou's evenness indices were significantly lower in children with acute bronchiolitis than in healthy children with gated levels of Firmicutes, Bacteroidetes and genus levels of Clostridium and other short chain fatty acid-producing bacteria. The relative abundance of short-chain fatty acid (SCFAs)-producing bacteria decreased and the abundance of genus-level sphingolipid-producing bacteria Sphingomonas increased; the progression of acute bronchiolitis is likely to be associated with the abundance of Clostridium and Sphingomonas and higher fecal amino acid concentrations, including FF-MAS, L-aspartic acid, thioinosinic acid, picolinic acid; supplementation with Clostridium butyricum significantly alleviated RSV infection-induced lung inflammation. The progression of bronchiolitis may be associated with altered intestinal microbiota, decreased SCFAs and elevated sphingolipids metabolism in children. Some fecal bacteria and metabolites may predict the onset of bronchiolitis, and oral administration of Clostridium butyricum may alleviate RSV infection-induced pulmonary inflammation.

Fecal metabonomics combined with 16S rDNA sequencing to analyze the changes of gut microbiota in rats fed with different protein source diets.

When blended, animal and plant proteins can complement each other in terms of amino acid composition and release time. In this study, we investigated whether the blended protein diet has a better feeding effect than the single protein diet, and to reveal the differences in growth and intestinal microbiota composition caused by the blended protein diet. Forty Sprague Dawley (SD) rats received diets with different protein sources, including casein (C), whey protein (WP), black soybean protein (BSP), and black soybean-whey blended protein (BS-WP), for eight weeks. To investigate the effects of blended protein supplement on gut microbiota and metabolites, we performed a high throughput 16S rDNA sequencing and fecal metabolomics profiling. In addition, we determined growth and serum biochemical indices, and conducted intestinal morphology analyses. Compared to those in the BSP and WP groups, the daily body weight gain and feed conversion efficiency increased in the BS-WP group. Serum biochemical indices indicated that the protein utilization efficiency of the WP and BS-WP groups was relatively high, and the BS-WP blended protein diet improved the protein adoption rate. The BS-WP blended protein diet also improved intestinal tissue morphology and promoted intestinal villi development compared to the single protein diets. Furthermore, dietary protein altered the composition of gut microbiota, the gut microbial diversity of rats fed with the BS-WP diet was significantly (P < 0.05) higher than that of the other groups. The difference in dietary protein corresponded with an alteration of fecal amino acids and their metabolites, and tryptophan and tyrosine metabolism were the key mechanisms leading to the changes in fecal microbial composition. Dietary protein sources played an important role in the growth and development of rats by influencing intestinal metabolism and microbial composition. The BS-WP blended protein diet was more conducive to nutrient absorption than the single protein diet. Furthermore, blended protein increased the diversity of intestinal microbes and aided the establishment of intestinal barrier function.

An Overview on Fecal Profiles of Amino Acids and Related Amino-Derived Compounds in Children with Autism Spectrum Disorder in Tunisia

Autism spectrum disorder (ASD) is a neurodevelopmental pathology characterized by the impairment of social interaction, difficulties in communication, and repetitive behaviors. Alterations in the metabolism of amino acids have been reported. We performed a chromatographic analysis of fecal amino acids, ammonium, biogenic amines, and gamma aminobutyric acid (GABA) in Tunisian autistic children from 4 to 10 years, and results were compared with their siblings (SIB) and children from the general population (GP). ASD presented significantly higher levels of fecal amino acids than SIB and GP; differences being more pronounced in younger (4-7 years) than in older (8-10 years) individuals whereas no changes were found for the remaining compounds. Lower levels of histidine were the only difference related with severe symptoms of autism (CARS scale). A linear discriminant analysis (LDA) based on fecal amino acid profiles clearly separated ASD, SIB, and GP at 4 to 7 years but not at more advanced age (8-10 years), evidencing more pronounced alterations in younger children. The relationship of fecal amino acids with autism needs deeper research integrating blood analytical parameters, brain metabolism, and intestinal microbiota. Fecal amino acids could be targeted for designing personalized diets to prevent or minimize cognitive impairments associated with ASD.

The Differences in Intestinal Viscosity Produced by Barley and β-Glucanase Alter Digesta Enzyme Activities and Ileal Nutrient Digestibilities More in Broiler Chicks than in Cocks

The objectives of this study were to determine whether intestinal viscosity caused by mixed linked barley β-glucan depresses ileal nutrient digestibility and digestive enzyme activities and to determine the interaction of intestinal viscosity, digestive enzyme activities and ileal nutrient digestibility in different ages of poultry. In Experiments 1 and 2, 1-d-old broiler chicks and 1-y-old cocks, respectively, were fed diets with 60% corn, low and high viscosity barley with or without β-glucanase, for 3 wk. A 3 × 2 factorial design was used. Comparisons were made only within the same age group. In Experiment 3, 1-d-old broiler chicks were fed high viscosity barley with and without β-glucanase to measure fecal nutrient and ileal and fecal amino acid digestibility. Broiler chicks fed barley ate less and gained less weight than those fed corn; added β-glucanase resulted in increases in both food consumption and weight gain for the barley-fed chicks (P < 0.05). Relative pancreas weight was higher (P < 0.05) in chicks fed barley than in those fed corn, and lower with β-glucanase (P < 0.05). Digesta from barley-fed birds had the highest viscosities, which were decreased (P < 0.05) by β-glucanase. Amylase and lipase were lower in broiler chicks fed high viscosity barley compared with corn (P < 0.05), and β-glucanase increased both activities and that of trypsin as well (P < 0.05). In barley-fed broiler chicks, ileal starch, protein and fat digestibility, as well as fecal fat, protein and amino acid digestibility were lower (P < 0.05), but increased with β-glucanase (P < 0.05). Although barley increased intestinal viscosity and β-glucanase lowered it in poultry of both ages, barley-fed broiler chicks had decreased digestive enzyme activities and ileal nutrient digestibility that were increased with β-glucanase, indicating that viscosity is more limiting in broiler chicks than in cocks.

Development of a simultaneous quantification method for the gut microbiota-derived core nutrient metabolome in mice and its application in studying host-microbiota interaction

The gut microbiota interacts with the host via production of various metabolites of dietary nutrients. Herein, we proposed the concept of the gut microbiota-derived core nutrient metabolome, which covers 43 metabolites in carbohydrate metabolism, glycolysis, tricarboxylic acid cycle and amino acid metabolism, and established a quantitative UPLC-Q/TOF-MS method through 3-nitrophenylhydrazine derivatization to investigate the influence of obesity on the gut microbiota in mice. All metabolites could be simultaneously analyzed via separation on a BEH C18 column within 18 min. The lower limits of quantification of most analytes were less than 1 μM. Validation results demonstrated suitability for the analysis of mouse fecal samples. The method was then applied to detect the gut microbiota-derived nutrient metabolome in the feces of high-fat diet induced obese (DIO) and ob/ob (leptin-deficient) mice, as well as obesity-prone (OP) and obesity-resistant (OR) mice. Compared to the control groups, there were 13, 23 and 10 differentially abundant metabolites detected in ob/ob, DIO and OP groups, respectively. Among them, amino acids including leucine, isoleucine, glycine, methionine, tyrosine and glutamine were co-downregulated in the obese or OP mice and exhibited inverse association with body weight. 16S rDNA analysis revealed that the genera Lactobacillus and Dubosiella were also inversely associated with body weight and positively correlated with fecal amino acids. Collectively, our work provides an effective and simplified method for simultaneous quantifying the gut microbiota-derived core nutrient metabolome in mouse feces, which could assist various future studies on host-microbiota metabolic interaction.

Changes in Faecal and Plasma Amino Acid Profile in Dogs with Food-Responsive Enteropathy as Indicators of Gut Homeostasis Disruption: A Pilot Study

Dogs suffering from food-responsive enteropathy (FRE) respond to an elimination diet based on hydrolysed protein or novel protein; however, studies regarding the amino acid profile in FRE dogs are lacking. The aim of this pilot study was to evaluate whether the plasma and faecal amino acid profiles differed between control and FRE dogs and whether these could serve as indicators of severity of illness. Blood, faecal samples, body condition score, and severity of clinical signs based on the canine inflammatory bowel disease activity index were collected before starting the elimination diet. FRE dogs had lower proportions of plasma Asparagine, Histidine, Glycine, Cystine, Leucine, and branched-chain/aromatic amino acids; however, Phenylalanine increased. In faecal samples, Cystine was greater whereas Phenylalanine was lesser in sick dogs compared to control. Leucine correlated negatively with faecal humidity (r = -0.66), and Leucine and Phenylalanine with faecal fat (r = -0.57 and r = -0.62, respectively). Faecal Phenylalanine (r = 0.80), Isoleucine (r = 0.75), and Leucine (r = 0.92) also correlated positively with total short-chain fatty acids, whereas a negative correlation was found with Glycine (r = -0.85) and Cystine (r = -0.61). This study demonstrates the importance of Leucine and Phenylalanine amino acids as indicators of the disease severity in FRE dogs.

Dynamic changes in intestinal microbiota and metabolite composition of pre-weaned beef calves.

Gut microbes and their metabolites are essential for maintaining host health and production. The intestinal microflora of pre-weaned calves gradually tends to mature with growth and development and has high plasticity, but few studies have explored the dynamic changes of intestinal microbiota and metabolites in pre-weaned beef calves. In this study, we tracked the dynamics of faecal microbiota in 13 new-born calves by 16S rRNA gene sequencing and analysed changes in faecal amino acid levels using metabolomics. Calves were divided into the relatively high average daily gain group (HA) and the relatively low average daily gain group (LA) for comparison. The results demonstrated that the alpha diversity of the faecal microbiota increased with calf growth and development. The abundance of Porphyromonadaceae bacterium DJF B175 increased in the HA group, while that of Lactobacillus reuteri decreased. The results of the LEfSe analysis showed that the microbiota of faeces of HA calves at eight weeks of age was enriched with P. bacterium DJF B175, while Escherichia coli and L. reuteri were enriched in the microbiota of faeces of LA calves. Besides, the total amino acid concentration decreased significantly in the eighth week compared with that in the first week (P < 0.05). Overall, even under the same management conditions, microorganisms and their metabolites interact to play different dynamic regulatory roles. Our results provide new insights into changes in the gut microbiota and metabolites of pre-weaned calves.

Target Metabolites to Slow Down Progression of Amyotrophic Lateral Sclerosis in Mice.

Microbial metabolites affect the neuron system and muscle cell functions. Amyotrophic lateral sclerosis (ALS) is a multifactorial neuromuscular disease. Our previous study has demonstrated elevated intestinal inflammation and dysfunction of the microbiome in patients with ALS and an ALS mouse model (human-SOD1G93A transgenic mice). However, the metabolites in ALS progression are unknown. Using an unbiased global metabolomic measurement and targeted measurement, we investigated the longitudinal changes of fecal metabolites in SOD1G93A mice over the course of 13 weeks. We further compared the changes of metabolites and inflammatory response in age-matched wild-type (WT) and SOD1G93A mice treated with the bacterial product butyrate. We found changes in carbohydrate levels, amino acid metabolism, and the formation of gamma-glutamyl amino acids. Shifts in several microbially contributed catabolites of aromatic amino acids agree with butyrate-induced changes in the composition of the gut microbiome. Declines in gamma-glutamyl amino acids in feces may stem from differential expression of gamma-glutamyltransferase (GGT) in response to butyrate administration. Due to the signaling nature of amino acid-derived metabolites, these changes indicate changes in inflammation, e.g., histamine, and contribute to differences in systemic levels of neurotransmitters, e.g., γ-Aminobutyric acid (GABA) and glutamate. Butyrate treatment was able to restore some of the healthy metabolites in ALS mice. Moreover, microglia in the spinal cord were measured by IBA1 staining. Butyrate treatment significantly suppressed the IBA1 level in the SOD1G93A mice. Serum IL-17 and LPS were significantly reduced in the butyrate-treated SOD1G93A mice. We have demonstrated an inter-organ communications link among microbial metabolites, neuroactive metabolites from the gut, and inflammation in ALS progression. The study supports the potential to use metabolites as ALS hallmarks and for treatment.

Intestinal proline is a potential anti-allergy factor for allergy diagnosis and therapy.

Allergy has become a public health problem worldwide, but effective diagnostic and therapeutic approaches are limited currently. Amino acids are essential macronutrients that potentially participated in the allergy process. This work aimed to investigate whether amino acids can be applied as a mediator for allergy diagnosis and therapy. Two cohort studies were performed to investigate the correlation between fecal amino acids and allergy responses, and a spleen cell model was used to validate the role of amino acids in regulating allergy. In a cohort study with 193 volunteers, fecal proline was found to be negatively correlated with serum IgE, and detailed data analysis revealed that people with high-IgE-mediated allergy had decreased odds of high intestinal proline. In another cohort study with distinct allergic and non-allergic individuals, proline concentration was significantly lower in the allergic group. Daily diet and metagenomics analysis showed that the proline intake and microbiota amino acid metabolism were not significantly different, implying that the body's proline metabolism might be different between allergic and non-allergic individuals. Furthermore, the spleen cell model demonstrated that proline specifically targeted Th2 and Treg activity. Overall, this work revealed a tight correlation between gut proline and serum IgE, indicating proline as a promising biomarker and a potential therapeutic method for allergic diseases.

Enhancing egg production and quality by the supplementation of probiotic strains (Clostridium and Brevibacillus) via improved amino acid digestibility, intestinal health, immune response, and antioxidant activity.

This study focused on evaluating the influence of Clostridium butyricum and Brevibacillus strains on egg production, egg quality, immune response and antioxidant function, apparent fecal amino acid digestibility, and jejunal morphology when supplemented as probiotics in the diets of laying hens in the peak phase. A total of 288 healthy 30-week-old Hy-Line Brown laying hens were arbitrarily assigned to four dietary groups, which included control diet and control diet supplemented with 0.02% C. butyricum zlc-17, C. butyricum lwc-13, or Brevibacillus zlb-z1, for 84 days. The results showed that dietary C. butyricum and Brevibacillus sp. exerted a positively significant influence (P ≤ 0.05) compared to the control group on the performance, egg quality, and physiological response of the birds. The diets could reduce mortality rate and enhance (P ≤ 0.05) egg weight and egg mass, egg production rate, and feed efficiency. Further analysis suggested that the probiotic strains can enhance (P ≤ 0.05) eggshell quality, Haugh unit, thick albumen content, and albumen height. Also, probiotics enhanced (P ≤ 0.05) the antioxidant status via increased antioxidant enzymes and jejunal morphology as evidenced by increased villi surface area (VSA), the ratio of villi height to crypt depth, villi width, and villi height, and a significant reduction in crypt depth. Besides, nutrient absorption and retention were enhanced, as apparent fecal amino acid digestibility of key essential amino acids was substantially improved in the diet-based group. The concentrations of immunoglobulin M and A (IgM and IgA) increased significantly (P ≤ 0.05) in the probiotics group and the same effect was notable for complement proteins (C3) and immune organ (Spleen). Conclusively, the supplementation of Clostridium butyricum zlc-17 in comparison to Clostridium butyricum lwc-13 and Brevibacillus zlb-z1 strains significantly (P ≤ 0.05) promoted the antioxidant status, modulated the intestinal structure, enhanced amino acid digestibility, and regulated the immunity index of the laying hens, which finally improves the laying performance and egg quality of the laying hens.

Dietary Fructooligosaccharides Effectively Facilitate the Production of High-Quality Eggs via Improving the Physiological Status of Laying Hens.

The focus of this study was to investigate the influence of prebiotics, such as fructooligosaccharides (FOS), on laying performance, egg quality, apparent fecal amino acid digestibility, jejunal morphology, hematological indices, immunological response, and antioxidant capacity in laying hens. A total of 216 healthy Hy-Line Brown laying hens aged 30 weeks were randomly assigned to one of three dietary treatments: basal diet, basal diet supplemented with 0.3 percent FOS, or 0.6 percent FOS. For 84 days, each treatment was fed the corresponding experimental diet. According to the findings, dietary supplementation with FOS enhanced laying performance and egg mass while lowering mortality rate. Albumen height, thick albumen content, Haugh unit, and eggshell thickness were also improved by the prebiotics. Prebiotics also boosted antioxidant status by increasing the activity of antioxidant enzymes, improved morphological development of the jejunum as demonstrated by significant increases in villi height, villi width, ratio of villi height to crypt depth, and reduced crypt depth. The prebiotics group showed a considerable increase in immunoglobulin M, G, and A (IgM, IgG, and IgA) levels, as well as a similar effect on complement proteins (C3). Furthermore, the apparent fecal amino acid digestibility of most essential amino acids was significantly enhanced. Conclusively, fructooligosaccharides at inclusion level of 0.6% efficiently enhanced laying performance and production of high-quality eggs while positively modulating amino acid digestibility, jejunal morphology, antioxidant status, and immune functions of the laying hens.

Metabolic phenotyping reveals a potential link between elevated faecal amino acids, diet and symptom severity in individuals with severe mental illness

The brain-gut axis is increasingly recognized as an important contributing factor in the onset and progression of severe mental illnesses such as schizophrenia spectrum disorders and bipolar disorder. This study investigates associations between levels of faecal metabolites identified using 1H-NMR, clinical parameters, and dietary components of forty-two individuals diagnosed in a transdiagnostic approach to have severe mental illness. Faecal levels of the amino acids; alanine, leucine, and valine showed a significant positive correlation with psychiatric symptom severity as well as with dairy intake. Overall, this study proposes a diet-induced link between the brain-gut axis and the severity of psychiatric symptoms, which could be valuable in the design of novel dietary or therapeutic interventions to improve psychiatric symptoms.

Impact of probiotic Limosilactobacillus reuteri DSM 17938 on amino acid metabolism in the healthy newborn mouse.

We studied the effect of feeding a single probiotic Limosilactobacillus reuteri DSM 17938 (LR 17938) on the luminal and plasma levels of amino acids and their derivatives in the suckling newborn mouse, using gas chromatography and high-performance liquid chromatography. We found that LR 17938 increased the relative abundance of many amino acids and their derivatives in stool, while it simultaneously significantly reduced the plasma levels of three amino acids (serine, citrulline, and taurine). Many peptides and dipeptides were increased in stool and plasma, notably gamma-glutamyl derivatives of amino acids, following ingestion of the LR 17938. Gamma-glutamyl transformation of amino acids facilitates their absorption. LR 17938 significantly upregulated N-acetylated amino acids, the levels of which could be useful biomarkers in plasma and warrant further investigation. Specific fecal microbiota were associated with higher levels of fecal amino acids and their derivatives. Changes in luminal and circulating levels of amino acid derivatives, polyamines, and tryptophan metabolites may be mechanistically related to probiotic efficacy.