Species variation in the fecal metabolome gives insight into differential gastrointestinal function.

The metabolic composition of fecal extracts provides a window for elucidating the complex metabolic interplay between mammals and their intestinal ecosystems, and these metabolite profiles can yield information on a range of gut diseases. Here, the metabolites present in aqueous fecal extracts of humans, mice and rats were characterized using high-resolution (1)H NMR spectroscopy coupled with multivariate pattern recognition techniques. Additionally, the effects of sample storage and preparation methods were evaluated in order to assess the stability of fecal metabolite profiles, and to optimize information recovery from fecal samples. Finally, variations in metabolite profiles were investigated in healthy mice as a function of time. Interspecies variation was found to be greater than the variation due to either time or sample preparation. Although many fecal metabolites were common to the three species, such as short chain fatty acids and branched chain amino acids, each species generated a unique profile. Relatively higher levels of uracil, hypoxanthine, phenylacetic acid, glucose, glycine, and tyrosine amino acids were present in the rat, with beta-alanine being unique to the rat, and glycerol and malonate being unique to the human. Human fecal extracts showed a greater interindividual variation than the two rodent species, reflecting the natural genetic and environmental diversity in human populations. Fecal composition in healthy mice was found to change over time, which might be explained by altered gut microbial presence or activity. The systematic characterization of fecal composition across humans, mice, and rats, together with the evaluation of inherent variation, provides a benchmark for future studies seeking to determine fecal biomarkers of disease and/or response to dietary or therapeutic interventions.

[1]  D. Gauguier,et al.  Statistical total correlation spectroscopy: an exploratory approach for latent biomarker identification from metabolic 1H NMR data sets. , 2005, Analytical chemistry.

[2]  Catherine Juste.  Acides gras alimentaires, flore intestinale et cancer , 2005 .

[3]  E Holmes,et al.  Curve-fitting method for direct quantitation of compounds in complex biological mixtures using 1H NMR: application in metabonomic toxicology studies. , 2005, Analytical chemistry.

[4]  J. Trygg,et al.  Evaluation of the orthogonal projection on latent structure model limitations caused by chemical shift variability and improved visualization of biomarker changes in 1H NMR spectroscopic metabonomic studies. , 2005, Analytical chemistry.

[5]  M. Plebani,et al.  Enzymes in feces: useful markers of chronic inflammatory bowel disease. , 2007, Clinica chimica acta; international journal of clinical chemistry.

[6]  J. Lindon,et al.  Metabonomics: a platform for studying drug toxicity and gene function , 2002, Nature Reviews Drug Discovery.

[7]  T. West.  Pyrimidine base catabolism in Pseudomonas putida biotype B , 2001, Antonie van Leeuwenhoek.

[8]  C. Andrieux,et al.  Digestion/Absorption/Malabsorption Metabolic Characteristics of the Faecal Microflora in Humans from Three Age Groups , 2002, Scandinavian journal of gastroenterology.

[9]  I. Wilson,et al.  Gut microorganisms, mammalian metabolism and personalized health care , 2005, Nature Reviews Microbiology.

[10]  V. Mooser,et al.  Metagenomics: the role of the microbiome in cardiovascular diseases , 2006, Current opinion in lipidology.

[11]  T. Wolever,et al.  Serum acetate:propionate ratio is related to serum cholesterol in men but not women. , 1996, The Journal of nutrition.

[12]  A. Bax,et al.  1H and13C Assignments from Sensitivity-Enhanced Detection of Heteronuclear Multiple-Bond Connectivity by 2D Multiple Quantum NMR , 1986 .

[13]  P. Sprumont.  Ovarian follicles of normal NMRI mice and homozygous “Nude” mice , 1978, Cell and Tissue Research.

[14]  Ad Bax,et al.  MLEV-17-based two-dimensional homonuclear magnetization transfer spectroscopy , 1985 .

[15]  L. Eriksson.  Multi- and megavariate data analysis , 2006 .

[16]  I. Wilson,et al.  Rapid and noninvasive metabonomic characterization of inflammatory bowel disease. , 2007, Journal of proteome research.

[17]  C. Bauer,et al.  1H NMR analysis of novel sialylated and fucosylated lactose-based oligosaccharides having linear GlcNAc(β1–6)Gal and Neu5Ac(α2–6)GlcNAc sequences , 1997 .

[18]  G. Macfarlane,et al.  Variation in human intestinal microbiota with age. , 2002, Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver.

[19]  M. Tanner,et al.  The synthetic peroxide OZ78 is effective against Echinostoma caproni and Fasciola hepatica. , 2006, The Journal of antimicrobial chemotherapy.

[20]  G. Neupert,et al.  Colorectal cancer screening by non-invasive metabolic biomarker fecal tumor M2-PK. , 2006, World journal of gastroenterology.

[21]  S. Thibodeau,et al.  Colorectal cancer screening by detection of altered human DNA in stool: feasibility of a multitarget assay panel. , 2000, Gastroenterology.

[22]  J. Piškur,et al.  A gene duplication led to specialized γ‐aminobutyrate and β‐alanine aminotransferase in yeast , 2007, The FEBS journal.

[23]  C. Wasternack,et al.  Degradation of pyrimidines and pyrimidine analogs--pathways and mutual influences. , 1980, Pharmacology & therapeutics.

[24]  P. Turnbaugh,et al.  Microbial ecology: Human gut microbes associated with obesity , 2006, Nature.

[25]  Anne L McCartney,et al.  Differences between the gut microflora of children with autistic spectrum disorders and that of healthy children. , 2005, Journal of medical microbiology.

[26]  J. B. Carey,et al.  Isolation of deoxycholic acid from normal human feces. , 1955, The Journal of biological chemistry.

[27]  J. Trygg.  O2‐PLS for qualitative and quantitative analysis in multivariate calibration , 2002 .

[28]  F. Guarner,et al.  Gut flora in health and disease , 2003, The Lancet.

[29]  A. Bax,et al.  Complete proton and carbon-13 assignments of coenzyme B12 through the use of new two-dimensional NMR experiments , 1986 .

[30]  H. Ishiguro,et al.  Fecal pancreatic elastase: a reproducible marker for severe exocrine pancreatic insufficiency , 2006, Journal of Gastroenterology.

[31]  G. Bodenhausen,et al.  Natural abundance nitrogen-15 NMR by enhanced heteronuclear spectroscopy , 1980 .

[32]  T. Hofmann,et al.  Both wheat (Triticum aestivum) bran arabinoxylans and gut flora-mediated fermentation products protect human colon cells from genotoxic activities of 4-hydroxynonenal and hydrogen peroxide. , 2006, Journal of agricultural and food chemistry.

[33]  J. Lindon,et al.  'Metabonomics': understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data. , 1999, Xenobiotica; the fate of foreign compounds in biological systems.

[34]  E. Woodmansey,et al.  Intestinal bacteria and ageing , 2007, Journal of applied microbiology.

[35]  J. Lindon,et al.  NMR‐based metabonomic approaches for evaluating physiological influences on biofluid composition , 2005, NMR in biomedicine.

[36]  Ian D Wilson,et al.  Analytical strategies in metabonomics. , 2007, Journal of proteome research.

[37]  M. Tanner,et al.  Metabonomic investigations in mice infected with Schistosoma mansoni: an approach for biomarker identification. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[38]  John C. Lindon,et al.  Metabonomics Techniques and Applications to Pharmaceutical Research & Development , 2006, Pharmaceutical Research.

[39]  K. Itoh,et al.  Transformation of bile acids and sterols by clostridia (fusiform bacteria) in Wistar rats , 1999, Lipids.

[40]  C. Zancanaro,et al.  Proton Nuclear Magnetic Resonance Analysis of Meconium Composition in Newborns , 2003, Journal of pediatric gastroenterology and nutrition.

[41]  R. Bridges,et al.  Inhibition of the intestinal transport of uracil by hexoses and amino acids. , 1986, Biochimica et biophysica acta.

[42]  J. Nicholson,et al.  750 MHz 1H-NMR spectroscopy of human blood plasma. , 1993, Journal of pharmaceutical and biomedical analysis.