Abstract
The probiotic bacteria are helpful for nutritional and therapeutic purposes, and they are commercially available in various forms, such as capsules or powders. Increasing pieces of evidence indicate that different growth conditions and variability in manufacturing processes can determine the properties of probiotic products. In recent years, the lipidomic approach has become a useful tool to evaluate the impact that probiotics induce in host physiology. In this work, two probiotic formulations with identical species composition, produced in two different sites, the USA and Italy, were utilized to feed Caenorhabditis elegans, strains and alterations in lipid composition in the host and bacteria were investigated. Indeed, the multicellular organism C. elegans is considered a simple model to study the in vivo effects of probiotics. Nematodes fat metabolism was assessed by gene expression analysis and by mass spectrometry–based lipidomics. Lipid droplet analysis revealed a high accumulation of lipid droplets in worms fed US-made products, correlating with an increased expression of genes involved in the fatty acid synthesis. We also evaluated the lifespan of worms defective in genes involved in the insulin/IGF-1-mediated pathway and monitored the nuclear translocation of DAF-16. These data demonstrated the involvement of the signaling in C. elegans responses to the two diets. Lipidomics analysis of the two formulations was also conducted, and the results indicated differences in phosphatidylglycerol (PG) and phosphatidylcholine (PC) contents that, in turn, could influence nematode host physiology. Results demonstrated that different manufacturing processes could influence probiotics and host properties in terms of lipid composition.
Key points
• Probiotic formulations impact on Caenorhabditis elegans lipid metabolism;
• Lipidomic analysis highlighted phospholipid abundance in the two products;
• Phosphocholines and phosphatidylglycerols were analyzed in worms fed the two probiotic formulations.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Amrit FRG, Ratnappan R, Keith SA, Ghazi A (2014) The C. elegans lifespan assay toolkit. Methods 68(3):465–475. https://doi.org/10.1016/j.ymeth.2014.04.002
Ashrafi K (2007) Obesity and the regulation of fat metabolism. In: WormBook: the online review of C. elegans biology, Pasadena. https://doi.org/10.1895/wormbook.1.130.1
Ashrafi K, Chang FY, Watts JL, Fraser AG, Kamath RS, Ahringer J, Ruvkun G (2003) Genome-wide RNAi analysis of Caenorhabditis elegans fat regulatory genes. Nature 421:268–272. https://doi.org/10.1038/nature01279
Baumeister R, Schaffitzel E, Hertweck M (2006) Endocrine signaling in Caenorhabditis elegans controls stress response and longevity. J Endocrinol 190:191–202. https://doi.org/10.1677/joe.1.06856
Biagioli M, Laghi L, Carino A, Cipriani S, Distrutti E, Marchianò S, Parolin C, Scarpelli P, Vitali B, Fiorucci S (2017) Metabolic variability of a multispecies probiotic preparation impacts on the anti-inflammatory activity. Front Pharmacol 8:1–10. https://doi.org/10.3389/fphar.2017.00505
Bianchi L, Laghi L, Correani V, Schifano E, Landi C, Uccelletti D, Mattei B (2020) A combined proteomics, metabolomics and in vivo analysis approach for the characterization of probiotics in large-scale production. Biomol 10(1):157. https://doi.org/10.3390/biom10010157
Brooks KK, Liang B, Watts JL (2009) The influence of bacterial diet on fat storage in C. elegans. PLoS One 4:e7545. https://doi.org/10.1371/journal.pone.0007545
Cabreiro F, Au C, Leung KY, Vergara-Irigaray N, Cochemé HM, Noori T, Weinkove D, Schuster E, Greene NDE, Gems D (2013) Metformin retards aging in C. elegans by altering microbial folate and methionine metabolism. Cell 153:228–239. https://doi.org/10.1016/j.cell.2013.02.035
Chung H-J, Sim J-H, Min T-S, Choi H-K (2018) Metabolomics and Lipidomics approaches in the science of probiotics: a review. J Med Food 21:1086–1095. https://doi.org/10.1089/jmf.2017.4175
Cinque B, La Torre C, Lombardi F, Palumbo P, Evtoski Z, Santini S, Falone S, Cimini A, Amicarelli F, Cifone MG (2017) VSL#3 probiotic differently influences IEC-6 intestinal epithelial cell status and function. J Cell Physiol 232:3530–3539. https://doi.org/10.1002/jcp.25814
Coleman RA, Lewin TM, Van Horn CG, Gonzalez-baro MR (2002) Do long-chain acyl-CoA synthetases regulate fatty acid entry into synthetic versus degradative pathways? J Nutr 132:2123–2126. https://doi.org/10.1093/jn/132.8.2123
Del Boccio P, Perrotti F, Rossi C, Cicalini I, Di Santo S, Zucchelli M, Sacchetta P, Genovesi D, Pieragostino D (2017) Serum lipidomic study reveals potential early biomarkers for predicting response to chemoradiation therapy in advanced rectal cancer: a pilot study. Adv Radiat Oncol 2:118–124. https://doi.org/10.1016/j.adro.2016.12.005
Douillard FP, Mora D, Eijlander RT, Wels M, De Vos WM (2018) Comparative genomic analysis of the multispecies probiotic-marketed product VSL#3. PLoS One 13:1–19. https://doi.org/10.1371/journal.pone.0192452
Ehmke M, Luthe K, Schnabel R, Döring F (2014) S-adenosyl methionine synthetase 1 limits fat storage in Caenorhabditis elegans. Genes Nutr 9:1–14. https://doi.org/10.1007/s12263-014-0386-6
Fijan S (2014) Microorganisms with claimed probiotic properties: an overview of recent literature. Int J Environ Res Public Health 11:4745–4767. https://doi.org/10.3390/ijerph110504745
Gami MS, Wolkow CA (2006) Studies of Caenorhabditis elegans DAF-2/insulin signaling reveal targets for pharmacological manipulation of lifespan. Aging Cell 5:31–37. https://doi.org/10.1111/j.1474-9726.2005.00188.x
Grześkowiak L, Isolauri E, Salminen S, Gueimonde M (2011) Manufacturing process influences properties of probiotic bacteria. Br J Nutr 105:887–894. https://doi.org/10.1017/S0007114510004496
Henderson ST, Johnson TE (2001) daf-16 integrates developmental and environmental inputs to mediate aging in the nematode Caenorhabditis elegans. Curr Biol 11:1975–1980. https://doi.org/10.1016/S0960-9822(01)00594-2
Horikawa M, Sakamoto K (2010) Polyunsaturated fatty acids are involved in regulatory mechanism of fatty acid homeostasis via daf-2/insulin signaling in Caenorhabditis elegans. Mol Cell Endocrinol 323:183–192. https://doi.org/10.1016/j.mce.2010.03.004
Kaletsky R, Murphy CT (2010) The role of insulin/IGF-like signaling in C. elegans longevity and aging. Dis Model Mech 3:415–419. https://doi.org/10.1242/dmm.001040
Kechagia M, Basoulis D, Konstantopoulou S, Dimitriadi D, Gyftopoulou K, Skarmoutsou N, Fakiri EM (2013) Health benefit of probiotic a review. ISRN Nutr 481651:1–7. https://doi.org/10.5402/2013/481651
Kenyon C, Jean C, Grensch E, Adam R, Ramon T (1993) A C.elegans mutant that twice as long as wild type. Nature 366:461–464. https://doi.org/10.1038/366461a0
Kimura T, Jennings W, Epand RM (2016) Roles of specific lipid species in the cell and their molecular mechanism. Prog Lipid Res 62:75–92. https://doi.org/10.1016/j.plipres.2016.02.001
Koch K, Havermann S, Büchter C, Wätjen W (2014) Caenorhabditis elegans as model system in pharmacology and toxicology: effects of flavonoids on redox-sensitive signalling pathways and ageing. Sci World J 2014:1–15. https://doi.org/10.1155/2014/920398
Kumar A, Baruah A, Tomioka M, Iino Y, Kalita MC, Khan M (2019) Caenorhabditis elegans: a model to understand host–microbe interactions. Cell Mol Life Sci 77:1–21. https://doi.org/10.1007/s00018-019-03319-7
Li Y, Na K, Lee HJ, Lee EY, Paik YK (2011) Contribution of sams-1 and pmt-1 to lipid homoeostasis in adult Caenorhabditis elegans. J Biochem 149:529–538. https://doi.org/10.1093/jb/mvr025
Liu Y, Alookaran JJ, Rhoads JM (2018) Probiotics in autoimmune and inflammatory disorders. Nutr 10:1537. https://doi.org/10.3390/nu10101537
Liu YJ, Janssens GE, McIntyre RL, Molenaars M, Kamble R, Gao AW, Jongejan A, Van Weeghel M, Macinnes AW, Houtkooper RH (2019) Glycine promotes longevity in Caenorhabditis elegans in a methionine cycle-dependent fashion. PLoS Genet 15:1–23. https://doi.org/10.1371/journal.pgen.1007633
Murphy CT, McCarroll SA, Lieb JD, Bargmann CI, Kamath RS, Fraser A, Ahringer J, Li H, Kenyon CJ (2003) Genes that act downstream of DAF-16 to influence C. elegans lifespan. Nature 424:277–283. https://doi.org/10.1038/nature01789
Nivoliez A, Camares O, Paquet-Gachinat M, Bornes S, Forestier C, Veisseire P (2012) Influence of manufacturing processes on in vitro properties of the probiotic strain Lactobacillus rhamnosus Lcr35®. J Biotechnol 160:236–241. https://doi.org/10.1016/j.jbiotec.2012.04.005
Nivoliez A, Veisseire P, Alaterre E, Dausset C, Baptiste F, Camarès O, Paquet-Gachinat M, Bonnet M, Forestier C, Bornes S (2014) Influence of manufacturing processes on cell surface properties of probiotic strain Lactobacillus rhamnosus Lcr35®. Appl Microbiol Biotechnol 99:399–411. https://doi.org/10.1007/s00253-014-6110-z
Nomura T, Horikawa M, Shimamura S, Hashimoto T, Sakamoto K (2010) Fat accumulation in Caenorhabditis elegans is mediated by SREBP homolog SBP-1. Genes Nutr 5:17–27. https://doi.org/10.1007/s12263-009-0157-y
Ogg S, Paradis S, Gottlieb S, Patterson GI, Lee L, Tissenbaum HA, Ruvkun G (1997) The Fork head transcription factor DAF-16 transduces insulin-like metabolic and longevity signals in C. elegans. Nature 389:994–999. https://doi.org/10.1038/40194
Palumbo P, Lombardi F, Cifone MG, Cinque B (2018) The epithelial barrier model shows that the properties of VSL#3 depend from where it is manufactured. Endocrine Metab Immune Disord Drug Targets 19:199–206. https://doi.org/10.2174/1871530318666181022164505
Perez CL, Van Gilst MR (2008) A 13C isotope labeling strategy reveals the influence of insulin signaling on lipogenesis in C. elegans. Cell Metab 8:266–274. https://doi.org/10.1016/j.cmet.2008.08.007
Pilon M, Svensk E (2013) PAQR-2 may be a regulator of membrane fluidity during cold adaptation. Worm 2:e27123. https://doi.org/10.4161/worm.27123
Roselli M, Schifano E, Guantario B, Zinno P, Uccelletti D, Devirgiliis C (2019) Caenorhabditis elegans and probiotics interactions from a prolongevity perspective. Int J Mol Sci 20(20):5020. https://doi.org/10.3390/ijms20205020
Schifano E, Marazzato M, Ammendolia MG, Zanni E, Ricci M, Comanducci A, Goldoni P, Conte MP, Uccelletti D, Longhi C (2019a) Virulence behavior of uropathogenic Escherichia coli strains in the host model Caenorhabditis elegans. Microbiologyopen 8:1–10. https://doi.org/10.1002/mbo3.756
Schifano E, Zinno P, Guantario B, Roselli M, Marcoccia S, Devirgiliis C, Uccelletti D (2019b) The foodborne strain Lactobacillus fermentum MBC2 triggers pept-1-dependent pro-longevity effects in Caenorhabditis elegans. Microorganisms 7:45. https://doi.org/10.3390/microorganisms7020045
Shi X, Li J, Zou X, Greggain J, Rødkær SV, Færgeman NJ, Liang B, Watts JL (2013) Regulation of lipid droplet size and phospholipid composition by stearoyl-CoA desaturase. J Lipid Res 54:2504–2514. https://doi.org/10.1194/jlr.m039669
Trinchieri V, Laghi L, Vitali B, Parolin C, Giusti I, Capobianco D, Mastromarino P, De Simone C (2017) Efficacy and safety of a multistrain probiotic formulation depends from manufacturing. Front Immunol 8:1–13. https://doi.org/10.3389/fimmu.2017.01474
Walker AK, Jacobs RL, Watts JL, Rottiers V, Jiang K, Finnegan DM, Shioda T, Hansen M, Yang F, Niebergall LJ, Vance DE, Tzoneva M, Hart AC, Näär AM (2011) A conserved SREBP-1/phosphatidylcholine feedback circuit regulates lipogenesis in metazoans. Cell 147:840–852. https://doi.org/10.1016/j.cell.2011.09.045
Walter J (2008) Ecological role of lactobacilli in the gastrointestinal tract: implications for fundamental and biomedical research. Appl Environ Microbiol 74:4985–4996. https://doi.org/10.1128/AEM.00753-08
Watts JL, Ristow M (2017) Lipid and carbohydrate metabolism in C. elegans. Genetics 207:413–446. https://doi.org/10.1534/genetics.117.300106
Witting M, Schmitt-Kopplin P (2016) The Caenorhabditis elegans lipidome: a primer for lipid analysis in Caenorhabditis elegans. Arch Biochem Biophys 589:27–37. https://doi.org/10.1016/j.abb.2015.06.003
Zanni E, Laudenzi C, Schifano E, Palleschi C, Perozzi G, Uccelletti D, Devirgiliis C (2015) Impact of a complex food microbiota on energy metabolism in the model organism Caenorhabditis elegans. Biomed Res Int 2015:621709–621712. https://doi.org/10.1155/2015/621709
Zanni E, Schifano E, Motta S, Sciubba F, Palleschi C, Mauri P, Perozzi G, Uccelletti D, Devirgiliis C, Miccheli A (2017) Combination of metabolomic and proteomic analysis revealed different features among Lactobacillus delbrueckii subspecies bulgaricus and lactis strains while in vivo testing in the model organism Caenorhabditis elegans highlighted probiotic properties. Front Microbiol 8:1–12. https://doi.org/10.3389/fmicb.2017.01206
Zhang Y, Zou X, Ding Y, Wang H, Wu X, Liang B (2013) Comparative genomics and functional study of lipid metabolic genes in Caenorhabditis elegans. BMC Genomics 14:64. https://doi.org/10.1186/1471-2164-14-164
Acknowledgments
The authors acknowledge the Caenorhabditis Genetics Center (University of Minnesota, Minneapolis) for the nematode and E. coli OP50 strains.
Author information
Authors and Affiliations
Contributions
DU designed the C. elegans experiments and wrote the paper.
ES performed C. elegans experiments and wrote C. elegans results.
IC executed formal lipidomics data analysis and wrote lipidomics results.
DP edited lipidomics results and data processing and PDB performed conceptualization of lipidomics experiments. DU, HJH, and PDB critically revised and edited the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Ethics approval
Not applicable
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
ESM 1
(PDF 85 kb)
Rights and permissions
About this article
Cite this article
Schifano, E., Cicalini, I., Pieragostino, D. et al. In vitro and in vivo lipidomics as a tool for probiotics evaluation. Appl Microbiol Biotechnol 104, 8937–8948 (2020). https://doi.org/10.1007/s00253-020-10864-w
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-020-10864-w