Fecal Metabolite Concentrations Research Articles

Exploring the effects of the dietary fiber compound mediated by a longevity dietary pattern on antioxidation, characteristic bacterial genera, and metabolites based on fecal metabolomics

BackgroundAge-related dysbiosis of the microbiota has been linked to various negative health outcomes. This study aims to investigate the effects of a newly discovered dietary fiber compound (DFC) on aging, intestinal microbiota, and related metabolic processes. The DFC was identified through in vitro fermentation screening experiments, and its dosage and composition were determined based on a longevity dietary pattern.MethodsAged SPF C57BL/6 J mice (65 weeks old) and young mice (8 weeks old) were divided into three groups: a subgroup without dietary fiber (NDF), a low DFC dose subgroup (LDF, 10% DFC), and a high DFC dose subgroup (HDF, 20% DFC). The total antioxidant capacity (T-AOC), total superoxide dismutase (T-SOD) activity, malondialdehyde (MDA) content, and glutathione peroxidase (GSH-Px) activity in liver and serum samples of the mice were measured according to the manufacturer’s protocol. The expression levels of characteristic bacterial genera and fecal metabolite concentrations in mice were determined using quantitative real-time PCR (qPCR) and nuclear magnetic resonance hydrogen spectroscopy (1H NMR). Metabolomics analysis was further conducted to identify biological functions and potential pathways related to aging.ResultsAfter an 8-weeks dietary intervention, DFC supplementation significantly attenuated age-related weight loss, organ degeneration, and oxidative stress. And promoted the growth of Lactobacillus and Bifidobacterium and inhibited the growth of Escherichia coli (E. coli) and Bacteroides (p < 0.05) in the intestinal tracts of aged mice. Metabolomic analysis identified glycolipid and amino acid metabolic pathway biomarkers associated with aging that were differentially regulated by DFC consumption. Correlation analysis between the identified microbial flora and the biomarkers revealed potential mechanistic links between altered microbial composition and metabolic activity with aging markers.ConclusionsIn conclusion, this study revealed an important mechanism by which DFC consumption impacts healthspan and longevity, shedding light on optimizing dietary fiber or developing fiber-based interventions to improve human health.

Patterns of infant fecal metabolite concentrations and social behavioral development in toddlers.

Gut-derived metabolites, products of microbial and host co-metabolism, may inform mechanisms underlying children's neurodevelopment. We investigated whether infant fecal metabolites were related to toddler social behavior. Stool samples collected from 6-week-olds (n = 86) and 1-year-olds (n = 209) in the New Hampshire Birth Cohort Study (NHBCS) were analyzed using nuclear magnetic resonance spectroscopy metabolomics. Autism-related behavior in 3-year-olds was assessed by caregivers using the Social Responsiveness Scale (SRS-2). To assess the association between metabolites and SRS-2 scores, we used a traditional single-metabolite approach, quantitative metabolite set enrichment (QEA), and self-organizing maps (SOMs). Using a single-metabolite approach and QEA, no individual fecal metabolite or metabolite set at either age was associated with SRS-2 scores. Using the SOM method, fecal metabolites of six-week-olds organized into four profiles, which were unrelated to SRS-2 scores. In 1-year-olds, one of twelve fecal metabolite profiles was associated with fewer autism-related behaviors, with SRS-2 scores 3.4 (95%CI: -7, 0.2) points lower than the referent group. This profile had higher concentrations of lactate and lower concentrations of short chain fatty acids than the reference. We uncovered metabolic profiles in infant stool associated with subsequent social behavior, highlighting one potential mechanism by which gut bacteria may influence neurobehavior. Differences in host and microbial metabolism may explain variability in neurobehavioral phenotypes, but prior studies do not have consistent results. We applied three statistical techniques to explore fecal metabolite differences related to social behavior, including self-organizing maps (SOMs), a novel machine learning algorithm. A 1-year-old fecal metabolite pattern characterized by high lactate and low short-chain fatty acid concentrations, identified using SOMs, was associated with social behavior less indicative of autism spectrum disorder. Our findings suggest that social behavior may be related to metabolite profiles and that future studies may uncover novel findings by applying the SOM algorithm.

Effects of Dietary Protein Levels on Sheep Gut Metabolite Profiles during the Lactating Stage.

Diet-associated characteristics such as dietary protein levels can modulate the gut's primary or secondary metabolites, leading to effects on the productive performance and overall health of animals. Whereas fecal metabolite changes are closely associated with gut metabolome, this study aimed to see changes in the rumen metabolite profile of lactating ewes fed different dietary protein levels. For this, eighteen lactating ewes (approximately 2 years old, averaging 38.52 ± 1.57 kg in their initial body weight) were divided into three groups (n = 6 ewes/group) by following the complete randomized design, and each group was assigned to one of three low-protein (D_I), medium-protein (D_m), and high-protein (D_h) diets containing 8.58%, 10.34%, and 13.93% crude protein contents on a dry basis, respectively. The fecal samples were subjected to untargeted metabolomics using ultra-performance liquid chromatography (UPLC). The metabolomes of the sheep fed to the high-protein-diet group were distinguished as per principal-component analysis from the medium- and low-protein diets. Fecal metabolite concentrations as well as their patterns were changed by feeding different dietary protein levels. The discriminating metabolites between groups of nursing sheep fed different protein levels were identified using partial least-squares discriminant analysis. The pathway enrichment revealed that dietary protein levels mainly influenced the metabolism-associated pathways (n = 63 and 39 in positive as well as negative ionic modes, respectively) followed by protein (n = 15 and 8 in positive as well as negative ionic modes, respectively) and amino-acid (n = 14 and 7 in positive as well as negative ionic modes, respectively) synthesis. Multivariate and univariate analyses showed comparative changes in the fecal concentrations of metabolites in both positive and negative ionic modes. Major changes were observed in protein metabolism, organic-acid biosynthesis, and fatty-acid oxidation. Pairwise analysis and PCA reveal a higher degree of aggregation within the D-h group than all other pairs. In both the PCA and PLS-DA plots, the comparative separation among the D_h/D_m, D_h/D_I, and D_m/D_I groups was superior in positive as well as negative ionic modes, which indicated that sheep fed higher protein levels had alterations in the levels of the metabolites. These metabolic findings provide insights into potentiated biomarker changes in the metabolism influenced by dietary protein levels. The target identification may further increase our knowledge of sheep gut metabolome, particularly regarding how dietary protein levels influence the molecular mechanisms of nutritional metabolism, growth performance, and milk synthesis of sheep.

Gut Microbiota Ecological and Functional Modulation in Post-Stroke Recovery Patients: An Italian Study.

Ischemic stroke (IS) can be caused by perturbations of the gut-brain axis. An imbalance in the gut microbiota (GM), or dysbiosis, may be linked to several IS risk factors and can influence the brain through the production of different metabolites, such as short-chain fatty acids (SCFAs), indole and derivatives. This study examines ecological changes in the GM and its metabolic activities after stroke. Fecal samples of 10 IS patients were compared to 21 healthy controls (CTRLs). GM ecological profiles were generated via 16S rRNA taxonomy as functional profiles using metabolomics analysis performed with a gas chromatograph coupled to a mass spectrometer (GC-MS). Additionally fecal zonulin, a marker of gut permeability, was measured using an enzyme-linked immuno assay (ELISA). Data were analyzed using univariate and multivariate statistical analyses and correlated with clinical features and biochemical variables using correlation and nonparametric tests. Metabolomic analyses, carried out on a subject subgroup, revealed a high concentration of fecal metabolites, such as SCFAs, in the GM of IS patients, which was corroborated by the enrichment of SCFA-producing bacterial genera such as Bacteroides, Christensellaceae, Alistipes and Akkermansia. Conversely, indole and 3-methyl indole (skatole) decreased compared to a subset of six CTRLs. This study illustrates how IS might affect the gut microbial milieu and may suggest potential microbial and metabolic biomarkers of IS. Expanded populations of Akkermansia and enrichment of acetic acid could be considered potential disease phenotype signatures.

Fecal metabolite profiling identifies liver transplant recipients at risk for postoperative infection

Metabolites produced by the intestinal microbiome modulate mucosal immune defenses and optimize epithelial barrier function. Intestinal dysbiosis, including loss of intestinal microbiome diversity and expansion of antibiotic-resistant pathobionts, is accompanied by changes in fecal metabolite concentrations and increased incidence of systemic infection. Laboratory tests that quantify intestinal dysbiosis, however, have yet to be incorporated into clinical practice. We quantified fecal metabolites in 107 patients undergoing liver transplantation (LT) and correlated these with fecal microbiome compositions, pathobiont expansion, and postoperative infections. Consistent with experimental studies implicating microbiome-derived metabolites with host-mediated antimicrobial defenses, reduced fecal concentrations of short- and branched-chain fatty acids, secondary bile acids, and tryptophan metabolites correlate with compositional microbiome dysbiosis in LT patients and the relative risk of postoperative infection. Our findings demonstrate that fecal metabolite profiling can identify LT patients at increased risk of postoperative infection and may provide guideposts for microbiome-targeted therapies.

PSIX-3 A Veterinary Gastrointestinal Low-Fat Diet Affects Fecal Characteristics, Metabolites, Bile Acids, and Microbiota Concentrations of Antibiotic-Treated Dogs

Abstract In a recent dog study, the antibiotic metronidazole proved to be an effective disruptor of the fecal microbiota and resulted in looser stools and altered fecal bile acid (BA) profiles. Recovery took several weeks and was aided by a dietary supplement. The objective of this study was to determine how a veterinary gastrointestinal low-fat diet affected the fecal characteristics and metabolite, BA, and microbiota concentrations of dogs recovering from metronidazole administration. Twenty-four healthy adult dogs were used in an 8-wk completely randomized design study. The study began with a 2-wk baseline phase where all dogs consumed a leading grocery brand dry kibble diet (GBD; wk 0 to wk 2). Over the next 2 wk, dogs continued to consume GBD and received metronidazole (20 mg/kg BW; wk 2 to wk 4). At wk 4, dogs were randomly allotted to one of two treatments [GBD; Blue Buffalo Gastrointestinal Support Low-Fat (BB)] for the remaining 4 wk (wk 5 to wk 8). Fecal scores were recorded daily, and fresh fecal samples were collected at wk 2, 4, 5, 6, 7, and 8 for measurement of pH, dry matter, and metabolite and bile acid concentrations. Fecal microbiota populations were also analyzed by 16S rRNA sequencing and qPCR, which was used to calculate dysbiosis index. All data were analyzed as repeated measures using the Mixed Models procedure of SAS 9.4, with treatment treated as a fixed effect, dog as a random effect, and significance set at P &amp;lt; 0.05. As anticipated, metronidazole administration increased (P &amp;lt; 0.0001) fecal scores (looser stools) and drastically altered (P &amp;lt; 0.05) fecal metabolite concentrations, reducing fecal short-chain fatty acid (SCFA), branched-chain fatty acid (BCFA), phenol, and indole concentrations, increasing primary BA concentrations, and decreasing secondary bile acid concentrations. Metronidazole dramatically impacted fecal microbiota populations by reducing alpha diversity, altering the abundance of multiple bacterial taxa, and increasing dysbiosis index. Fecal outcome variables recovered over the next 4 wk, with some being impacted by diet. The change in fecal SCFA and immunoglobulin A concentrations were greater (P &amp;lt; 0.01), while the change in fecal BCFA, phenol, and indole concentrations were decreased (P &amp;lt; 0.05) in dogs fed BB. Dysbiosis index and alpha diversity measures recovered quickly for all dogs and without dietary effects. Recovery of over 25 bacterial genera were impacted by diet (P &amp;lt; 0.05). While many bacterial taxa returned to baseline levels after 4 wk, others had still not fully recovered. Fecal BA concentrations recovered quickly for all dogs. The change in lithocholic acid concentration was less (P &amp;lt; 0.0001) in dogs fed BB, but other BA concentrations were not affected by diet. In conclusion, metronidazole drastically alters the fecal microbiota and metabolites of dogs. While most variables return to normal over time, a veterinary gastrointestinal diet may aid in the recovery.

195 Restricted Feeding and Weight Loss Change Fecal Metabolite Concentrations and Microbiota Populations of Overweight Cats

Abstract Feline obesity negatively influences many metabolic processes, health outcomes, and the individual well-being of cats, but little research evaluates the modification in gastrointestinal microbiota in an overweight state. The use of specially formulated diets and restricted feeding, resulting in subsequent body weight (BW) loss, may be used to improve health and modify gastrointestinal microbiota of overweight cats. This study aimed to identify and determine the effects of restricted feeding and BW loss on fecal metabolite concentrations and microbiota populations of overweight cats. All procedures were approved by the University of Illinois IACUC before experimentation. Twenty-two overweight adult spayed female and neutered male cats [BW = 5.70 ± 1.0 kg; BCS = 7.68±0.6; age = 4 ± 0.4 yr] were used in a 24-wk BW loss study. A control diet (OR) was fed during a 4-wk baseline to identify intake needed to maintain BW. After baseline (wk 0), cats were allotted to OR or a BW loss diet (FT) and fed to lose ~1.0% BW/wk. Fecal samples were collected at baseline (wk 0), and weeks 4, 8, 12, 16, 20, and 24 for microbiota and metabolite analyses. Fecal microbiota data were analyzed and evaluated using QIIME2 and Mixed Models procedure of SAS, with P &amp;lt; 0.05 considered significant. Change from baseline fecal acetate, propionate, butyrate, total short-chain fatty acid, isobutyrate, valerate, phenol, indole, and total phenol and indole concentrations were reduced (P &amp;lt; 0.05) with restricted feeding and BW loss. Change from baseline fecal scores and valerate concentrations were also affected by diet, with greater (P &amp;lt; 0.05) reductions occurring in cats fed OR than those fed FT. Change from baseline fecal pH was also greater (P = 0.05) in cats fed OR than those fed FT. Fecal bacterial alpha diversity was not affected by diet or restricted feeding and BW loss. However, fecal bacterial beta diversity, assessed by weighted and unweighted UniFrac distances and presented as principal coordinate plots, showed separation between dietary groups, with some significant clustering as a result of restricted feeding and weight loss. The relative abundances of a single fecal bacterial phylum and 5 fecal bacterial genera were impacted by restricted feeding and weight loss. Diet had a greater effect, with 3 fecal bacterial phyla and 19 fecal bacterial genera being different (P &amp;lt; 0.05) between dietary treatment groups. Our data demonstrate that restricted feeding of specially formulated diets, and subsequent BW loss, modifies the fecal metabolite profiles and microbiota populations of overweight cats. The mechanisms by which these changes may affect feline gastrointestinal health and metabolism requires further study.

Reduced immunomodulatory metabolite concentrations in peri-transplant fecal samples from heart allograft recipients

BackgroundEmerging evidence is revealing the impact of the gut microbiome on hematopoietic and solid organ transplantation. Prior studies postulate that this influence is mediated by bioactive metabolites produced by gut-dwelling commensal bacteria. However, gut microbial metabolite production has not previously been measured among heart transplant (HT) recipients.MethodsIn order to investigate the potential influence of the gut microbiome and its metabolites on HT, we analyzed the composition and metabolite production of the fecal microbiome among 48 HT recipients at the time of HT.ResultsCompared to 20 healthy donors, HT recipients have significantly reduced alpha, i.e. within-sample, microbiota diversity, with significantly lower abundances of key anaerobic commensal bacteria and higher abundances of potentially pathogenic taxa that have been correlated with adverse outcomes in other forms of transplantation. HT recipients have a wide range of microbiota-derived fecal metabolite concentrations, with significantly reduced levels of immune modulatory metabolites such as short chain fatty acids and secondary bile acids compared to healthy donors. These differences were likely due to disease severity and prior antibiotic exposures but were not explained by other demographic or clinical factors.ConclusionsKey potentially immune modulatory gut microbial metabolites are quantifiable and significantly reduced among HT recipients compared to healthy donors. Further study is needed to understand whether this wide range of gut microbial dysbiosis and metabolite alterations impact clinical outcomes and if they can be used as predictive biomarkers or manipulated to improve transplant outcomes.

Effects of commercial and traditional kefir supplementation on apparent total tract macronutrient digestibility and the fecal characteristics, metabolites, and microbiota of healthy adult dogs.

Kefir is a fermented dairy beverage that has been consumed by humans for centuries, but poorly studied in pets. The objective of this study was to determine the effects of commercial or traditional kefir supplementation on apparent total tract macronutrient digestibility (ATTD) and fecal characteristics, microbiota populations, and metabolite and immunoglobulin (Ig) A concentrations of healthy adult dogs. Twelve healthy adult dogs (5.67 ± 1.72 yr, 7.27 ± 1.15 kg) were used in a replicated 3 × 3 Latin square design (n = 12/group). All dogs were fed a commercial diet and allotted to 1 of 3 treatments (60 mL/d): 2% reduced-fat milk treated with lactase [CNTL; 4.57E + 03 lactic acid bacteria (LAB) colony-forming units (CFU)/mL], commercial kefir (C-Kefir; 6.95E + 04 LAB CFU/mL), or traditional kefir brewed daily from 2% reduced-fat milk and kefir grains (T-Kefir; 1.79E + 09 LAB CFU/mL). The experiment was composed of three 28-d periods, with each consisting of a 22-d transition phase, a 5-d fecal collection phase, and 1 d for blood collection. Fecal samples were collected for determination of ATTD and fecal pH, dry matter, microbiota, and metabolite, and IgA concentrations. Data were analyzed using the Mixed Models procedure of SAS 9.4. The main effects of treatment were tested, with significance set at P ≤ 0.05 and trends set at P ≤ 0.10. Kefir products differed in microbial density and profile, but fecal microbiota populations were weakly impacted. Bacterial alpha diversity tended to be greater (P = 0.10) in dogs fed T-Kefir than those fed CNTL. Bacterial beta diversity analysis identified a difference (P < 0.0004) between dogs-fed CNTL and those fed C-Kefir. Dogs-fed C-Kefir tended to have a greater (P = 0.06) relative abundance of Fusobacteriota than those fed CNTL or T-Kefir. Dogs-fed T-Kefir had a greater (P < 0.0001) relative abundance of Lactococcus than those fed CNTL or C-Kefir. Dogs-fed T-Kefir also tended to have a lower (P = 0.09) relative abundance of Escherichia Shigella and greater (P = 0.09) relative abundance of Candidatus stoquefichus than dogs-fed CNTL or C-Kefir. Dogs-fed C-Kefir tended to have lower (P = 0.08) fecal valerate concentrations than those fed CNTL or T-Kefir. All other measures were unaffected by kefir treatments. Our results suggest that kefir supplementation had minor effects on the fecal microbiota populations and fecal metabolite concentrations of healthy adult dogs without impacting ATTD, fecal characteristics, or fecal IgA concentrations.

Effects of hydrolyzed chicken liver on digestibility, fecal and urinary characteristics, and fecal metabolites of adult dogs.

High-protein low-carbohydrate diets have been widely used for adult maintenance dogs, as well as in specific weight loss diets and maintenance programs. However, increasing dietary protein may increase undigested protein in the hindgut, modifying intestinal fermentation and fecal metabolite concentrations. The aim of this study was to evaluate the effects of protein source and concentration on apparent total tract digestibility (ATTD) of nutrients, metabolizable energy (ME), fecal and urinary characteristics, and fecal metabolites of dogs. Twelve healthy adult dogs were distributed into six treatments (n = 6 per diet) in a balanced incomplete Latin square design consisting of three periods of 30 days each. Six diets, varying in protein source [poultry byproduct meal (PBPM) and hydrolyzed chicken liver powder (HCLP)] and concentration [24, 32, and 40% crude protein (CP) on dry matter (DM) basis] were tested: PBPM24 (PBPM based diet with 24% CP); PBPM32 (PBPM with 32% CP); PBPM40 (PBPM with 40% CP); HCLP24 (HCLP based diet with 24% CP); HCLP32 (HCLP with 32% CP); HCLP40 (HCLP with 40% CP). The ATTD of CP was greater in dogs-fed HCLP and higher protein concentrations diets (P < 0.05). However, dogs-fed HCLP diets had lower ATTD of fat and carbohydrates, and ME (P < 0.05). Similarly, high-protein diets reduced the ATTD of DM, OM, fat, carbohydrates, and energy (P < 0.05). High-protein diets increased the daily fecal output and moisture (P = 0.004 and P < 0.05, respectively), as well as the fecal score (P < 0.0001), verified as soft, moist stools, but still within the ideal range. Fecal valerate concentration was greater in dogs-fed PBPM at 32% CP (P = 0.007). Fecal isobutyrate tended to increase in dogs-fed PBPM and high-protein diets (P < 0.10). Also, dogs-fed PBPM and high-protein diets had greater fecal concentrations of isovalerate, branched-chain fatty acids (BCFA), and ammonia (P < 0.05). Finally, the fecal lactate concentration increased in dogs-fed HCLP and high-protein diets (P < 0.05). The HCLP increased the ATTD of CP, being a highly digestible protein. Although the inclusion of HCLP slightly increased fecal score and moisture, it decreased the amount of fecal metabolites of protein fermentation ammonia and BCFA, both of which are associated with proteolytic fermentation in the colon.

Dietary enrichment of resistant starches or fibers differentially alter the feline fecal microbiome and metabolite profile

BackgroundCats are strict carnivores but possess a complex gastrointestinal (GI) microbial community that actively ferments dietary substrates that are not digested and reach the colon. The GI microbiota responses to dietary inclusion of resistant starches versus fibers have not been tested in cats. Thus, our objective was to evaluate the effects of diets enriched in resistant starch or fibers on the fecal characteristics, microbiome, and metabolite profiles of cats. Twelve healthy adult domestic shorthair cats (age = 9.6 ± 4.0 year; body weight = 3.9 ± 1.0 kg) were used in a replicated 3 × 3 Latin square design to test diets that were enriched with: (1) resistant starch (ERS), (2) a fiber-prebiotic-probiotic blend (FPPB), or (3) a fiber-prebiotic-probiotic blend + immune-modulating ingredients (iFPPB). In each 28-day period, 22 days of diet adaptation was followed by fecal and blood sample collection. Fecal samples were used for shotgun metagenomic sequencing. In addition, fecal and blood metabolite measurements and white blood cell stimulation was performed to assess immune function.ResultsA total of 1690 bacterial species were identified, with 259 species differing between fiber-rich and ERS treatments. In comparison with fiber-rich treatments that increased diversity and promoted Firmicutes and Bacteroidetes populations, resistant starch reduced microbial diversity and fecal pH, led to a bloom in Actinobacteria, and modified Kyoto Encyclopedia of Genes and Genomes orthology (KO) terms pertaining to starch and sucrose metabolism, fatty acid biosynthesis and metabolism, epithelial cell signaling, among others. Resistant starch also differentially modified fecal metabolite concentrations with relevance to GI and overall host health (increased butyrate; decreased propionate and protein catabolites - branched-chain fatty acids; phenols and indoles; ammonia) and reduced blood cholesterol, which correlated strongly with microbial taxa and KO terms, and allowed for a high predictive efficiency of diet groups by random forest analysis.ConclusionEven though domestic cats and other carnivores evolved by eating low-carbohydrate diets rich in protein and fat, our results demonstrate that the feline microbiome and metabolite profiles are highly responsive to dietary change and in directions that are predictable.

Possible Effects of Early Maternal Separation on the Gut Microbiota of Captive Adult Giant Pandas.

Simple SummaryIn the process of ex-situ conservation, young giant pandas face a variety of unfavorable environmental impacts such as frequent maternal separation, parenting by non-parents, noise from tourists, and frequent replacement of animal houses, which may cause psychological and physiological stress. The gut microbiota is an important carrier of the interaction between the body and the environment, and recent studies revealed an association between stress and alterations of the intestinal microbiota. So, is the stress caused by the unfavorable parenting environment in the early life of captive giant pandas related to the gut microbiota? To answer this question, we use fecal metagenomics and LC-MS technology to study the effect of different parenting patterns on the structure, diversity, and metabolites of the intestinal microbial community of captive giant pandas. In order to evaluate the possible adverse effects of the traditional parenting mode on the gut microbiota of captive giant pandas in the early life, it can provide an important scientific basis for improving the welfare level of captive giant pandas.Maternal deprivation (MD) in early life induces dysbiosis in the host gut microbiota, which is a key determinant of abnormal behavior in stress model individuals. Compared with the early parenting environment of the wild, captive giant pandas face frequent and premature maternal separation. Will this lead to imbalance in intestinal flora and stress in captive giant pandas? The purpose of this research is to evaluate the possible adverse effects of the traditional parenting mode on the gut microbiota of captive giant pandas. The results showed that the frequent and premature maternal separation at early stages of the young did not change α and β diversity indices of the gut microbes, but it increased the relative abundance of s_Clostridium_tetani and s_Clostridium_sp_MSJ_8 (significantly positively correlated with the metabolism of propionic acid) and also the concentrations of fecal metabolites that are related to stress (N-acetyl-l-aspartic acid and corticosterone) in the intestinal tract of giant pandas in adulthood. Thereby, the function of protein digestion and absorption in the intestines of captive giant pandas was decreased, and the metabolism of short-chain fatty acids was disturbed. In conclusion, the parenting experience of early maternal separation could adversely affect the stress caused by the unfavorable parenting environment in the early life of captive giant pandas related to the gut microbiota of the captive giant pandas in adulthood.

Impacts of dietary exposure to pesticides on faecal microbiome metabolism in adult twins

BackgroundDietary habits have a profound influence on the metabolic activity of gut microorganisms and their influence on health. Concerns have been raised as to whether the consumption of foodstuffs contaminated with pesticides can contribute to the development of chronic disease by affecting the gut microbiome. We performed the first pesticide biomonitoring survey of the British population, and subsequently used the results to perform the first pesticide association study on gut microbiome composition and function from the TwinsUK registry.MethodsDietary exposure of 186 common insecticide, herbicide, or fungicide residues and the faecal microbiome in 65 twin pairs in the UK was investigated. We evaluated if dietary habits, geographic location, or the rural/urban environment, are associated with the excretion of pesticide residues. The composition and metabolic activity of faecal microbiota was evaluated using shotgun metagenomics and metabolomics respectively. We performed a targeted urine metabolomics analysis in order to evaluate whether pesticide urinary excretion was also associated with physiological changes.ResultsPyrethroid and/or organophosphorus insecticide residues were found in all urine samples, while the herbicide glyphosate was found in 53% of individuals. Food frequency questionnaires showed that residues from organophosphates were higher with increased consumption of fruit and vegetables. A total of 34 associations between pesticide residue concentrations and faecal metabolite concentrations were detected. Glyphosate excretion was positively associated with an overall increased bacterial species richness, as well as to fatty acid metabolites and phosphate levels. The insecticide metabolite Br2CA, reflecting deltamethrin exposure, was positively associated with the phytoestrogens enterodiol and enterolactone, and negatively associated with some N-methyl amino acids. Urine metabolomics performed on a subset of samples did not reveal associations with the excretion of pesticide residues.ConclusionsThe consumption of conventionally grown fruit and vegetables leads to higher ingestion of pesticides with unknown long-term health consequences. Our results highlight the need for future dietary intervention studies to understand effects of pesticide exposure on the gut microbiome and possible health consequences.

Chemical composition of selected insect meals and their effect on apparent total tract digestibility, fecal metabolites, and microbiota of adult cats fed insect-based retorted diets.

Insect meals are novel and potentially sustainable protein sources. The objectives of this study were to determine the chemical composition and standardized amino acid digestibility using the cecectomized rooster model of three selected insect meals (i.e., speckled cockroach [SC], Madagascar hissing cockroach [MC], and superworm [SW]) and to determine the effects of these insect meals on food intake, apparent total tract digestibility (ATTD) of macronutrients, fecal scores, and metabolites of adult cats fed insect- or chicken-based retorted diets. This study consisted of a complete randomized design, with 28 adult cats randomly assigned to one of the four experimental retorted diets: Control (chicken-based diet), SC diet, MC diet, or SW diet. All animal procedures were approved by the University of Illinois Institutional Animal Care and Use Committee. All diets were formulated to be complete and balanced and meet or exceed the nutritional requirements of adult cats. The experimental period was 28 d, with the first 7 d allotted for diet adaptation. The total fecal collection was completed during the last 4 d of the experimental period. On day 21, a fresh fecal sample from each cat was collected for the determination of fecal metabolites and microbiota. Food was offered twice daily to maintain body weight and body condition score. Among the three selected insect meals evaluated, oleic acid, palmitic acid, linoleic acid, and stearic acid were the most prevalent fatty acids. Branched-chain amino acids and arginine were the most preponderant indispensable amino acids in these insect meals. ATTD of dry matter, organic matter, acid-hydrolyzed fat, and crude protein did not differ among treatments (P > 0.05), and all diets were well digested by the cats. Similarly, fecal scores did not differ among the treatments and were within ideal range. No differences (P > 0.05) in fecal metabolite concentrations or microbiota diversity were observed among cats fed different experimental diets; only a few genera from Firmicutes and Bacteroidota phyla differ (P < 0.05) in cats fed SW diet in contrast to other dietary treatments. In conclusion, the selected insect meals evaluated herein are rich in linoleic acid, an essential fatty acid for cats. Insect-based retorted diets led to comparable results to those achieved with a chicken-based retorted diet, suggesting that these novel protein sources might be adequate alternative ingredients in feline diets.

Effects of dietary macronutrient profile on apparent total tract macronutrient digestibility and fecal microbiota, fermentative metabolites, and bile acids of female dogs after spay surgery

Obesity and estrogen reduction are known to affect the gut microbiota and gut microbial-derived metabolites in some species, but limited information is available in dogs. The aim of this study was to determine the effects of dietary macronutrient profile on apparent total tract macronutrient digestibility, fecal microbiota, and fecal metabolites of adult female dogs after spay surgery. Twenty-eight adult intact female beagles (age: 3.02 ± 0.71 yr, BW: 10.28 ± 0.77 kg; BCS: 4.98 ± 0.57) were used. After a 5-wk baseline phase (week 0), 24 dogs were spayed and randomly allotted to one of three experimental diets (n = 8 per group): 1) control (CO) containing moderate protein and fiber (COSP), 2) high-protein, high-fiber (HPHF), or 3) high-protein, high-fiber plus omega-3 and medium-chain fatty acids (HPHFO). Four dogs were sham-operated and fed CO (COSH). All dogs were fed to maintain BW for 12 wk after spay and then allowed to consume twice that amount for 12 wk. Fecal samples were collected at weeks 0, 12, and 24 for digestibility, microbiota, and metabolite analysis. All data were analyzed using repeated measures and linear mixed models procedure of SAS 9.4, with results reported as a change from baseline. Apparent organic matter and energy digestibilities had greater decreases in HPHF and HPHFO than COSH and COSP. Increases in fecal acetate, total short-chain fatty acids, and secondary bile acids were greater and decreases in primary bile acids were greater in HPHF and HPHFO. Principal coordinates analysis of weighted UniFrac distances revealed that HPHF and HPHFO clustered together and separated from COSH and COSP at weeks 12 and 24, with relative abundances of Faecalibacterium, Romboutsia, and Fusobacterium increasing to a greater extent and Catenibacterium, Bifidobacterium, Prevotella 9, Eubacterium, and Megamonas decreasing to a greater extent in HPHF or HPHFO. Our results suggest that high-protein, high-fiber diets alter nutrient and energy digestibilities, fecal metabolite concentrations, and fecal gut microbiota, but spay surgery had minor effects. Future research is needed to investigate how food intake, nutrient profile, and changes in hormone production influence gut microbiota and metabolites of dogs individually and how this knowledge may be used to manage spayed pets.

Influence of Age, Sex, and Diet on the Human Fecal Metabolome Investigated by 1H NMR Spectroscopy.

The human fecal metabolome is increasingly studied to explore the impact of diet and lifestyle on health and the gut microbiome. However, systematic differences and confounding factors related to age, sex, and diet remain largely unknown. In this study, absolute concentrations of fecal metabolites from 205 healthy Danes (105 males and 100 females, 49 ± 31 years old) were quantified using 1H NMR spectroscopy and the newly developed SigMa software. The largest systemic variation was found to be highly related to age. Fecal concentrations of short-chain fatty acids (SCFA) were higher in the 18 years old group, while amino acids (AA) were higher in the elderly. Sex-related metabolic differences were weak but significant and mainly related to changes in SCFA. The concentrations of butyric, valeric, propionic, and isovaleric acids were found to be higher in males compared to females. Sex differences were associated with a stronger, possibly masking, effect from differential intake of macronutrients. Dietary fat intake decreased levels of SCFA and AA of both sexes, while carbohydrate intake showed weak correlations with valeric and isovaleric acids in females. This study highlights some possible demographic confounders linked to diet, disease, lifestyle, and microbiota that have to be taken into account when analyzing fecal metabolome data.

Faecal microbial metabolites of proteolytic and saccharolytic fermentation in relation to degree of insulin resistance in adult individuals.

The gut microbiota may affect host metabolic health through microbial metabolites. The balance between the production of microbial metabolites by saccharolytic and proteolytic fermentation may be an important determinant of metabolic health. Amongst the best-studied saccharolytic microbial metabolites are the short-chain fatty acids acetate, propionate and butyrate. However, human data on the role of other microbial fermentation by-products in metabolic health are greatly lacking. Therefore, we compared in a cross-sectional study the faecal microbial metabolites (caproate, lactate, valerate, succinate, and the branched-chain fatty acids (BCFA) (isobutyrate, isovalerate)) between insulin sensitive (homeostatic model assessment of insulin resistance (HOMA-IR), HOMA-IR<1.85, IS) and insulin resistant (HOMA-IR>1.85, IR) individuals. Additionally, we assessed the relationships between faecal metabolites and markers of metabolic health including fasting glucose, insulin, free fatty acids, insulin resistance (HOMA-IR) and fasting substrate oxidation in 86 individuals with a wide range of body mass index. Faecal metabolite concentrations did not significantly differ between IS and IR. Furthermore, there were no associations between microbial metabolites and metabolic health markers, except for a slight positive association of isovalerate with carbohydrate oxidation (E%, std β 0.194, P=0.011) and fat oxidation (E%, std β -0.075, P=0.047), also after adjustment for age, sex and BMI. In summary, faecal caproate, lactate, valerate, succinate, and BCFA (isobutyrate, isovalerate) were not different between IR and IS individuals, nor was there any association between these faecal metabolites and parameters of metabolic health. Further human intervention studies are warranted to investigate the role of these microbially-derived fermentation products and their kinetics in metabolic health and insulin sensitivity.

PSXI-36 Evaluation of fecal microbiota of dogs fed extruded diets containing white and red sorghum as primary carbohydrate source

Abstract Sorghum (Sorghum bicolor) is a cereal grain consisting of either white, yellow, red, brown or black endosperms. In human and pet nutrition, sorghum has gained increased attention as a gluten-free and non-GMO whole-grain option. However, a paucity of information exists about the effect of this cereal grain on the GI microbiome and host health. Therefore, the aim of this study was to evaluate the effects of extruded diets containing corn (CON), white sorghum (WHS) or red sorghum (RES) as the main carbohydrate source on fecal microbiota and metabolites in dogs. Animal care protocol used in this study was approved by the Institutional Care and Use Committee at the University of Illinois. Three diets containing either 30% of CON, WHS, or RES were formulated to meet or exceed the AAFCO (2018) nutrient profile for adult dogs. Nine adult female beagles were randomly assigned to one of the 3 dietary treatments using a replicated 3x3 Latin square design. Each experiment period consisted of 14d (10d of diet adaption and 4d of fecal collection). Fresh fecal samples were collected and allocated for microbial analysis. Illumina 16S rRNA sequencing from V4 region was completed using MiSeq and analyzed using QIIME 2. Linear models were used to evaluate the main effect of treatment. Over 1.8 million sequences were generated. Samples were rarified to 42,160 reads for analysis of diversity and species richness. Beta-diversity did not differ among dogs fed CON, WHS, or RES diets (q- and p-values &amp;gt; 0.05). Similarly, microbial richness was also not affected by treatment. Overall, fecal metabolite concentrations were similar among treatments and did not have a strong correlated with microbial taxa. Our findings indicate that extruded diets using sorghum as a substitute for corn as the main dietary carbohydrate result in similar fecal metabolite profile and microbiota in adult dogs.

Changes in the Fecal Metabolome Are Associated with Feeding Fiber Not Health Status in Cats with Chronic Kidney Disease

The objective was to determine the effects of feeding different fiber sources to cats with chronic kidney disease (CKD) compared with healthy cats (both n = 10) on fecal metabolites. A cross-over within split-plot study design was performed using healthy and CKD cats (IRIS stage 1, 2, and 3). After cats were fed a complete and balanced dry food designed to aid in the management of renal disease for 14 days during a pre-trial period, they were randomly assigned to two fiber treatments for 4 weeks each. The treatment foods were formulated similar to pre-trial food and contained 0.500% betaine, 0.586% oat beta glucan, and either 0.407% short chain fructooligosaccharides (scFOS) fiber or 3.44% apple pomace. Both treatment foods had similar crude fiber (2.0 and 2.1% for scFOS and apple pomace, respectively) whereas soluble fiber was 0.8 and 1.6%, respectively. At baseline, CKD had very little impact on the fecal metabolome. After feeding both fiber sources, some fecal metabolite concentrations were significantly different compared with baseline. Many fecal uremic toxins decreased, although in healthy cats some increased; and some more so when feeding apple pomace compared with scFOS, e.g., hippurate, 4-hydroxyhippurate, and 4-methylcatechol sulfate; the latter was also increased in CKD cats. Changes in secondary bile acid concentrations were more numerous in healthy compared with CKD cats, and cats in both groups had greater increases in some secondary bile acids after consuming apple pomace compared with scFOS, e.g., tauroursodeoxycholate and hyocholate. Although changes associated with feeding fiber were more significant than changes associated with disease status, differential modulation of the gut-kidney axis using dietary fiber may benefit cats.

The Impact of Fresh Hass Avocado on the Fecal Metabolome Among Adults with Overweight and Obesity: A Randomized, Controlled Trial

Abstract Objectives Avocados are nutrient-rich fruits that have been recently linked to beneficial alterations to the gastrointestinal microbiota. However, previous research on shifts in the fecal metabolome with avocado intake has largely been conducted in in vitro or preclinical models and little is known about their metabolomic impact in human subjects. Methods Adult participants (n = 109) 25–45 years of age with BMI ≥ 25.0 kg/m2 were enrolled in an investigator-blinded, parallel arm, randomized, controlled trial. Participants consumed isocaloric meals with or without fresh Hass avocado once daily for 12-weeks and reported ≥ 80% meal consumption over the intervention period. Untargeted fecal metabolites were quantified in a subsample of participants (n = 48) using gas chromatography mass spectroscopy and were normalized by sample weight. Kruskal-Wallis tests and false discovery rate type I error correction were conducted and orthogonal partial least squares discriminant analysis (OPLS-DA) was used to predict treatment group by fecal metabolite concentrations (RStudio, version 3.6.2). Results A total of 292 metabolites were identified at intervention follow-up. Of these, three metabolites differed significantly between treatment groups. Fecal concentrations of lanosterol (P = 0.0004, q = 0.04) and the fatty alcohols hexadecanol (P = 0.001, q = 0.04) and octadecanol (P = 0.001, q = 0.04), were greater in the group consuming avocado as compared to control. Seventeen additional metabolites, including nine fecal lipids, two fat soluble vitamin derivatives, and three monosaccharides/disaccharides differed at P &amp;lt; 0.05 but did not meet the q &amp;lt; 0.05 threshold. Treatment group assignment was predicted correctly in 70% of cases (R2 = 72%, Q2 = 33%) using the trained OPLS-DA model. Conclusions Fresh Hass avocado intake increased fecal lipid and sterol concentrations among healthy adults with overweight and obesity, demonstrating diet-related modifications to the fecal metabolome. Funding Sources Support for this research was provided by the Hass Avocado Board, the USDA National Institute of Food and Agriculture, Hatch project 1009249, and the USDA National Institute of Food and Agriculture AFRI Predoctoral Fellowship, project 2018–07785.