1D <sup>1</sup>H NMR as a Tool for Fecal Metabolomics

Ganobis, Caroline M.; Al‐Abdul‐Wahid, M. Sameer; Renwick, Simone; Yen, Sandi; Carriero, Charley; Aucoin, Marc G.; Allen‐Vercoe, Emma

doi:10.1002/cpch.83

Cited by 10 publications

(12 citation statements)

References 24 publications

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“…Human fecal matter is a waste product of food digestion and mainly contains water, undigested food remnants, including nitrogen and protein matter, carbohydrates, lipids, and hundreds of small molecular metabolites, as well as bacteria [1]. Fecal matter is also a rich source of metabolic information and the noninvasive sampling has increased interest toward exploring the human fecal metabolome as a function of diet, diseases, and other lifestyle conditions [1,2]. Recent studies have shown close relationships between the dynamics of the fecal metabolome and gut microbiome functions [3,4].…”

Section: Introductionmentioning

confidence: 99%

Human Fecal Metabolome Reflects Differences in Body Mass Index, Physical Fitness, and Blood Lipoproteins in Healthy Older Adults

et al. 2021

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This study investigated how body mass index (BMI), physical fitness, and blood plasma lipoprotein levels are related to the fecal metabolome in older adults. The fecal metabolome data were acquired using proton nuclear magnetic resonance spectroscopy and gas chromatography–mass spectrometry on 163 healthy older adults (65–80 years old, 80 females and 83 males). Overweight and obese subjects (BMI ≥ 27) showed higher levels of fecal amino acids (AAs) (valine, alanine, and phenylalanine) compared to normal-weight subjects (BMI ≤ 23.5). Adults classified in the high-fitness group displayed slightly lower concentrations of fecal short-chain fatty acids, propionic acid, and AAs (methionine, leucine, glutamic acid, and threonine) compared to the low-fitness group. Subjects with lower levels of cholesterol in low-density lipoprotein particles (LDLchol, ≤2.6 mmol/L) displayed higher fecal levels of valine, glutamic acid, phenylalanine, and lactic acid, while subjects with a higher level of cholesterol in high-density lipoprotein particles (HDLchol, ≥2.1 mmol/L) showed lower fecal concentration of isovaleric acid. The results from this study suggest that the human fecal metabolome, which primarily represents undigested food waste and metabolites produced by the gut microbiome, carries important information about human health and should be closely integrated to other omics data for a better understanding of the role of the gut microbiome and diet on human health and metabolism.

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Section: Introductionmentioning

confidence: 99%

Human Fecal Metabolome Reflects Differences in Body Mass Index, Physical Fitness, and Blood Lipoproteins in Healthy Older Adults

et al. 2021

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“…Aliquots of cecal fluid were serially centrifuged, and the supernatant was passed through a 0.22 uM filter and stored at −80 °C for future metabolite analysis. Fecal material underwent the same process, with the exception of the collection and vessel establishment samples (0 h), which were first diluted at 25% wt/vol [ 22 ] with sterile phosphate-buffered saline (PBS) and vigorously mixed prior to centrifugation.…”

Section: Methodsmentioning

confidence: 99%

“…Filtered supernatant samples for metabolite analysis were standardized through the addition of a DSS-d6 Chenomx Internal Standard (Chenomx, EDM, CAN). Samples were read on a nuclear magnetic resonance (NMR) spectrometer with an operating field of ≥600 MHz using the METNOESY sequence [ 22 ] at the University of Guelph Advanced Analysis Center. Sample pH was measured using Cytiva Whatman wide-range pH indicator strips (Fisher Scientific, Canada).…”

Section: Methodsmentioning

confidence: 99%

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A Comparison of Methods to Maintain the Equine Cecal Microbial Environment In Vitro Utilizing Cecal and Fecal Material

MacNicol

Renwick

Ganobis

et al. 2022

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The equine gastrointestinal (GI) microbiota is intimately related to the horse. The objective of the current study was to evaluate the microbiome and metabolome of cecal inoculum maintained in an anaerobic chamber or chemostat batch fermenter, as well as the fecal slurry maintained in an anaerobic chamber over 48 h. Cecal and fecal content were collected from healthy adult horses immediately upon death. Cecal fluid was used to inoculate chemostat vessels (chemostat cecal, n = 11) and vessels containing cecal fluid (anaerobic cecal, n = 15) or 5% fecal slurry (anaerobic fecal, n = 6) were maintained in an anaerobic chamber. Sampling for microbiome and metabolome analysis was performed at vessel establishment (0 h), and after 24 h and 48 h of fermentation. Illumina sequencing was performed, and metabolites were identified via nuclear magnetic resonance (NMR). Alpha and beta diversity indices, as well as individual metabolite concentrations and metabolite regression equations, were analyzed and compared between groups and over time. No differences were evident between alpha or beta diversity in cecal fluid maintained in either an anaerobic chamber or chemostat. The microbiome of the fecal inoculum maintained anaerobically shifted over 48 h and was not comparable to that of the cecal inoculum. Metabolite concentrations were consistently highest in chemostat vessels and lowest in anaerobic fecal vessels. Interestingly, the rate of metabolite change in anaerobic cecal and chemostat cecal vessels was comparable. In conclusion, maintaining an equine cecal inoculum in either an anaerobic chamber or chemostat vessel for 48 h is comparable in terms of the microbiome. However, the microbiome and metabolome of fecal material is not comparable with a cecal inoculum. Future research is required to better understand the factors that influence the level of microbial activity in vitro, particularly when microbiome data identify analogous communities.

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“…Each sample was scanned at 298K in a Bruker Avance III 600 MHz spectrometer with a 5 mm TCI 600 cryoprobe (Bruker, Billerica, MA, USA) at the Advanced Analysis Centre in the University of Guelph, ON, Canada according to (8), except the number of scans was increased from 32 to 1536 to increase sensitivity for the low concentration fecal filtrate samples. Sample spectra were processed and analyzed at the Advanced Analysis Centre in the University of Guelph in Chenomx NMR suite 9.0 according to (8) with 128K zero filling and 0.2 Hz line broadening. Targeted metabolite concentrations were determined by the best fit for the peak regions within the Chenomx library of compounds.…”

Section: Supplementary Figures 12mentioning

confidence: 99%

Dietary Nitrate and Corresponding Gut Microbiota Prevent Cardiac Dysfunction in Obese Mice

et al. 2023

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Impaired heart function can develop in diabetic individuals in the absence of coronary artery disease or hypertension, suggesting mechanisms beyond hypertension/increased afterload contribute to diabetic cardiomyopathy. Identifying therapeutic approaches that improve glycemia and prevent cardiovascular disease are clearly required for clinical management of diabetes-related comorbidities. Since intestinal bacteria are important for metabolism of nitrate, we examined if dietary nitrate and fecal microbial transplantation (FMT) from nitrate-fed mice could prevent high-fat diet (HFD)-induced cardiac abnormalities. Male C57Bl/6N mice were fed an 8-week low-fat diet (LFD), HFD, or HFD+Nitrate (4mM sodium nitrate). HFD-fed mice presented with pathological left ventricular (LV) hypertrophy, reduced stroke volume and increased end diastolic pressure, in association with increased myocardial fibrosis, glucose intolerance, adipose inflammation, serum lipids, LV mitochondrial reactive oxygen species (ROS), and gut dysbiosis. In contrast, dietary nitrate attenuated these detriments. In HFD-fed mice, FMT from HFD+Nitrate donors did not influence serum nitrate, blood pressure, adipose inflammation, or myocardial fibrosis. However, microbiota from HFD+Nitrate mice decreased serum lipids, LV ROS, and similar to FMT from LFD donors, prevented glucose intolerance and cardiac morphology changes. Therefore, the cardioprotective effects of nitrate are not dependent on reducing blood pressure, but rather mitigating gut dysbiosis, highlighting a nitrate-gut-heart axis.

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1D ¹H NMR as a Tool for Fecal Metabolomics

Cited by 10 publications

References 24 publications

Human Fecal Metabolome Reflects Differences in Body Mass Index, Physical Fitness, and Blood Lipoproteins in Healthy Older Adults

Human Fecal Metabolome Reflects Differences in Body Mass Index, Physical Fitness, and Blood Lipoproteins in Healthy Older Adults

A Comparison of Methods to Maintain the Equine Cecal Microbial Environment In Vitro Utilizing Cecal and Fecal Material

Dietary Nitrate and Corresponding Gut Microbiota Prevent Cardiac Dysfunction in Obese Mice

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1D 1H NMR as a Tool for Fecal Metabolomics

Cited by 10 publications

References 24 publications

Human Fecal Metabolome Reflects Differences in Body Mass Index, Physical Fitness, and Blood Lipoproteins in Healthy Older Adults

Human Fecal Metabolome Reflects Differences in Body Mass Index, Physical Fitness, and Blood Lipoproteins in Healthy Older Adults

A Comparison of Methods to Maintain the Equine Cecal Microbial Environment In Vitro Utilizing Cecal and Fecal Material

Dietary Nitrate and Corresponding Gut Microbiota Prevent Cardiac Dysfunction in Obese Mice

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1D ¹H NMR as a Tool for Fecal Metabolomics