Abstract
Introduction
Collecting feces is easy. It offers direct outcome to endogenous and microbial metabolites.
Objectives
In a context of lack of consensus about fecal sample preparation, especially in animal species, we developed a robust protocol allowing untargeted LC-HRMS fingerprinting.
Methods
The conditions of extraction (quantity, preparation, solvents, dilutions) were investigated in bovine feces.
Results
A rapid and simple protocol involving feces extraction with methanol (1/3, M/V) followed by centrifugation and a step filtration (10 kDa) was developed.
Conclusion
The workflow generated repeatable and informative fingerprints for robust metabolome characterization.
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References
Berendsen, B. J. A., Wegh, R. S., Memelink, J., Zuidema, T., & Stolker L. A. M. (2015). The analysis of animal faeces as a tool to monitor antibiotic usage. Talanta, 132, 258–268. doi:10.1016/j.talanta.2014.09.022.
Bligh, E. G. & Dyer, W. J. (1959). A rapid method of total lipid extraction and purification. Revue Canadienne de Biochimie et Physiologie, 37(8), 911–917.
Cao, H., Huang, H., Xu, W., Chen, D., Yu, J., Li, J., et al. (2011). Fecal metabolome profiling of liver cirrhosis and hepatocellular carcinoma patients by ultra-performance liquid chromatography-mass spectrometry. Analytica Chimica Acta, 691, 68–75. doi:10.1016/j.aca.2011.02.038.
Cesbron, N., Sydor, A., Penot, M., Prevost, S., Le Bizec, B., & Dervilly-Pinel, G. (2016). Analytical strategies to detect enobosarm administration in bovines. Food Additives and Contaminants: Part A, doi:10.1080/19440049.2016.1258122.
Deda, O., Chatziioannou, A. C., Fasoula, S., Palachanis, D., Raikos, N., & Theodoridis, G. A., et al. (2016). Sample preparation optimization in fecal metabolic profiling. Journal of Chromatography B. doi:10.1016/j.jchromb.2016.06.047.
Deda, O., Gika, H. G., Wilson, I. D., & Theodoridis, G. A. (2015). An overview of fecal sample preparation for global metabolic profiling. Journal of Pharmaceutical and Biomedical Analysis, 113, 137–150.
Dervilly-Pinel, G., Chereau, S., Cesbron, N., Monteau, F., & Le Bizec, B. (2015). LC-HRMS based metabolomics screening model to detect various β-agonists treatments in bovines. Metabolomics, 11(2), 403–411.
Gregory, K. E., Bird, S. S., Gross, V. S., Marur, V. R., Lazarev, A. V., Walker, W. A., et al. (2013). Method development for fecal lipidomics profiling. Analytical Chemistry, 85(2), 1114–1123. doi:10.1021/ac303011k.
Han, J., Antunes, L. C. M., Finlay, B. B., & Borchers, C. H. (2010). Metabolomics: towards understanding host-microbe interactions. Future Microbiology, 5(2), 153–161. doi:10.2217/FMB.09.132.
Hou, W., Zhong, D., Zhang, P., Li, Y., Lin, M., Liu, G., Yao, M., Liao, Q., & Xie, Z. (2016). A strategy to targeted metabolomics analysis of 11 gut microbiota-host co-metabolites in rat serum, urine and feces by ultra high performance liquid chromatography-tandem mass spectrometry. Journal of Chromatography A, 1429, 207–2017. doi:10.1016/j.chroma.2015.12.031.
Jacob, C. C., Dervilly-Pinel, G., Biancotto, G., & Le Bizec, B. (2014). Evaluation of specific gravity as normalization strategy for cattle urinary metabolome analysis. Metabolomics. doi:10.1007/s11306-013-0604-z.
Kouassi Nzoughet, J. J., Dervilly-Pinel, G., Chéreau, S., Biancotto, G., Monteau, F., Elliott, C. T.,& Le Bizec, B. (2015). First insights into serum metabolomics of trenbolone/estradiol implanted bovines; screening model to predict hormone-treated and control animals’ status. Metabolomics, 11, 1184. doi:10.1007/s11306-015-0775-x.
Matysik, S., Le Roy, C. I., Liebisch, G., Claus, S. P. (2016). Metabolomics of fecal samples: A practical consideration. Trends in Food Science and Technology, doi:10.1016/j.tifs.2016.05.011.
Ng, J. S. Y., Ryan, U., Trengove, R. D., & Maker, G. L. (2012). Development of an untargeted metabolomics method for the analysis of human faecal samples using Cryptosporidium-infected samples. Molecular and Biochemical Parasitology, 185, 145–150. doi:10.1016/j.molbiopara.2012.08.006.
Regal, P., Anizan, S., Antignac, J.-P., Le Bizec, B., Cepeda, A., & Fente, C. (2011). Metabolomic approach based on liquid chromatography coupled to high resolution mass spectrometry to screen for the illegal use of estradiol and progesterone in cattle. Analytica Chimica Acta, 700, 16–25. doi:10.1016/j.aca.2011.01.005.
Rhoades, J. R., Duffy, G., & Koutsoumanis, K. (2009). Prevalence and concentration of verotoxigenic Escherichia coli, Salmonella enterica and Listeria monocytogenes in the beef production chain: A review. Food Microbiology, 26, 357–376. doi:10.1016/j.fm.2008.10.012.
Rojas, D., Dervilly-Pinel, G., Cesbron, N., Penot, M., Sydor, A., Prevost, S., et al. (2016). Selective androgen receptor modulators: Comparative excretion study of bicalutamide in bovine urine and feces. Drug Testing and Analysis. doi:10.1002/dta.2113.
Royer, A.-L., Gallart-Ayala, H., Antignac, J.-P., Dervilly-Pinel, G., Monteau, F., Le Bizec, B. (2014). A versatile multi-analytical platform for serum metabolomics and lipidomics profiling. Conference paper, RFMF, 19–24 May, Lyon, France. https://colloque.inra.fr/9_js_rfmf_lille_2015/layout/set/print/Soumission/METABOLITES-IDENTIFICATION-IN-NON-TARGETED-METABOLOMICS-APPROACHES-A-MULTI-ANALYTICAL-MASS-SPECTROMETRY-PLATFORM-VIEW/(offset)/1.
Van Lieshout, L., & Roestenberg, M. (2015). Clinical consequences of new diagnostic tools for intestinal parasites. Clinical Microbiology and Infection, 21, 520–528. doi:10.1016/j.cmi2015.03.015.
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All animals involved in this study were hosted in the Clinic for Ruminants (Oniris (Veterinary College), Nantes, France). Their faeces were taken from the clinical examinations realised during their hospitalization.
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Cesbron, N., Royer, AL., Guitton, Y. et al. Optimization of fecal sample preparation for untargeted LC-HRMS based metabolomics. Metabolomics 13, 99 (2017). https://doi.org/10.1007/s11306-017-1233-8
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DOI: https://doi.org/10.1007/s11306-017-1233-8