Abstract
Weaning is a challenging period for gut health in piglets. Previous studies showed that dietary supplementations with either amino acids or polyphenols promote piglet growth and intestinal functions, when administered separately. Thus, we hypothesized that a combination of amino acids and polyphenols could facilitate the weaning transition. Piglets received during the first two weeks after weaning a diet supplemented or not with a mix of a low dose (0.1%) of functional amino acids (l-arginine, l-leucine, l-valine, l-isoleucine, l-cystine) and 100 ppm of a polyphenol-rich extract from grape seeds and skins. The mix of amino acids and polyphenols improved growth and feed efficiency. These beneficial effects were associated with a lower microbiota diversity and a bloom of Lactobacillaceae in the jejunum content while the abundance of Proteobacteria was reduced in the caecum content. The mix of amino acids and polyphenols also increased the production by the caecum microbiota of short-chain fatty acids (butyrate, propionate) and of metabolites derived from amino acids (branched-chain fatty acids, valerate, putrescine) and from polyphenols (3-phenylpropionate). Experiments in piglet jejunum organoids revealed that the mix of amino acids and polyphenols upregulated the gene expression of epithelial differentiation markers while it reduced the gene expression of proliferation and innate immunity markers. In conclusion, the supplementation of a mix of amino acids and polyphenols is a promising nutritional strategy to manage gut health in piglets through the modulation of the gut microbiota and of the epithelial barrier.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adamczyk B, Simon J, Kitunen V et al (2017) Tannins and their complex interaction with different organic nitrogen compounds and enzymes: old paradigms versus recent advances. ChemistryOpen 6:610–614. https://doi.org/10.1002/open.201700113
Allaire JM, Crowley SM, Law HT et al (2018) The intestinal epithelium: central coordinator of mucosal immunity. Trends Immunol 39:677–696. https://doi.org/10.1016/j.it.2018.04.002
Andersen-Civil AIS, Arora P, Williams AR (2021) Regulation of enteric infection and immunity by dietary proanthocyanidins. Front Immunol. https://doi.org/10.3389/fimmu.2021.637603
Beaumont M, Blachier F (2020) Amino acids in intestinal physiology and health. Adv Exp Med Biol 1265:1–20. https://doi.org/10.1007/978-3-030-45328-2_1
Beaumont M, Blanc F, Cherbuy C et al (2021a) Intestinal organoids in farm animals. Vet Res 52:33. https://doi.org/10.1186/s13567-021-00909-x
Beaumont M, Cauquil L, Bertide A et al (2021b) Gut microbiota-derived metabolite signature in suckling and weaned piglets. J Proteome Res 20:982–994. https://doi.org/10.1021/acs.jproteome.0c00745
Bian G, Ma S, Zhu Z et al (2016) Age, introduction of solid feed and weaning are more important determinants of gut bacterial succession in piglets than breed and nursing mother as revealed by a reciprocal cross-fostering model: gut bacterial succession in piglets. Environ Microbiol 18:1566–1577. https://doi.org/10.1111/1462-2920.13272
Bokulich NA, Subramanian S, Faith JJ et al (2013) Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing. Nat Methods 10:57–59. https://doi.org/10.1038/nmeth.2276
Bonetti A, Tugnoli B, Piva A, Grilli E (2021) Towards zero zinc oxide: feeding strategies to manage post-weaning diarrhea in piglets. Animals (basel). https://doi.org/10.3390/ani11030642
Boudry G, Jamin A, Chatelais L et al (2013) Dietary protein excess during neonatal life alters colonic microbiota and mucosal response to inflammatory mediators later in life in female pigs. J Nutr 143:1225–1232. https://doi.org/10.3945/jn.113.175828
Brenes A, Viveros A, Chamorro S, Arija I (2016) Use of polyphenol-rich grape by-products in monogastric nutrition. A review. Anim Feed Sci Technol 211:1–17. https://doi.org/10.1016/j.anifeedsci.2015.09.016
Burgueño JF, Abreu MT (2020) Epithelial toll-like receptors and their role in gut homeostasis and disease. Nat Rev Gastroenterol Hepatol. https://doi.org/10.1038/s41575-019-0261-4
Campbell JM, Crenshaw JD, Polo J (2013) The biological stress of early weaned piglets. J Anim Sci Biotechnol 4:19. https://doi.org/10.1186/2049-1891-4-19
Casanova-Martí À, González-Abuín N, Serrano J et al (2020) Long term exposure to a grape seed proanthocyanidin extract enhances l-cell differentiation in intestinal organoids. Mol Nutr Food Res 64:2000303. https://doi.org/10.1002/mnfr.202000303
Chalvon-Demersay T, Luise D, Le Floch N et al (2021) Functional amino acids in pigs and chickens: implication for gut health. Front Vet Sci. https://doi.org/10.3389/fvets.2021.663727
Choy YY, Quifer-Rada P, Holstege DM et al (2014) Phenolic metabolites and substantial microbiome changes in pig feces by ingesting grape seed proanthocyanidins. Food Funct 5:2298–2308. https://doi.org/10.1039/C4FO00325J
Cires MJ, Wong X, Carrasco-Pozo C, Gotteland M (2017) The gastrointestinal tract as a key target organ for the health-promoting effects of dietary proanthocyanidins. Front Nutr. https://doi.org/10.3389/fnut.2016.00057
Dowarah R, Verma AK, Agarwal N (2017) The use of Lactobacillus as an alternative of antibiotic growth promoters in pigs: a review. Anim Nutr 3:1–6. https://doi.org/10.1016/j.aninu.2016.11.002
Escudié F, Auer L, Bernard M et al (2018) FROGS: find, rapidly, OTUs with galaxy solution. Bioinformatics 34:1287–1294. https://doi.org/10.1093/bioinformatics/btx791
Gardiner GE, Metzler-Zebeli BU, Lawlor PG (2020) Impact of intestinal microbiota on growth and feed efficiency in pigs: a review. Microorganisms. https://doi.org/10.3390/microorganisms8121886
Ghosh S, Whitley CS, Haribabu B, Jala VR (2021) Regulation of intestinal barrier function by microbial metabolites. Cell Mol Gastroenterol Hepatol 11:1463–1482. https://doi.org/10.1016/j.jcmgh.2021.02.007
Giacomoni F, Le Corguillé G, Monsoor M et al (2015) Workflow4Metabolomics: a collaborative research infrastructure for computational metabolomics. Bioinformatics 31:1493–1495. https://doi.org/10.1093/bioinformatics/btu813
Girard M, Bee G (2020) Invited review: Tannins as a potential alternative to antibiotics to prevent coliform diarrhea in weaned pigs. Animal 14:95–107. https://doi.org/10.1017/S1751731119002143
Gresse R, Chaucheyras-Durand F, Fleury MA et al (2017) Gut microbiota dysbiosis in postweaning piglets: understanding the keys to health. Trends Microbiol 25:851–873. https://doi.org/10.1016/j.tim.2017.05.004
Grosu IA, Pistol GC, Taranu I, Marin DE (2019) The impact of dietary grape seed meal on healthy and aflatoxin B1 afflicted microbiota of pigs after weaning. Toxins 11:25. https://doi.org/10.3390/toxins11010025
Han M, Song P, Huang C et al (2016) Dietary grape seed proanthocyanidins (GSPs) improve weaned intestinal microbiota and mucosal barrier using a piglet model. Oncotarget 7:80313–80326. https://doi.org/10.18632/oncotarget.13450
Hao R, Li Q, Zhao J et al (2015) Effects of grape seed procyanidins on growth performance, immune function and antioxidant capacity in weaned piglets. Livest Sci 178:237–242. https://doi.org/10.1016/j.livsci.2015.06.004
Hou Q, Dong Y, Huang J et al (2020) Exogenous l-arginine increases intestinal stem cell function through CD90+ stromal cells producing mTORC1-induced Wnt2b. Commun Biol 3:1–16. https://doi.org/10.1038/s42003-020-01347-9
Kafantaris I, Stagos D, Kotsampasi B et al (2018) Grape pomace improves performance, antioxidant status, fecal microbiota and meat quality of piglets. Animal 12:246–255. https://doi.org/10.1017/S1751731117001604
Kamada N, Chen GY, Inohara N, Núñez G (2013) Control of pathogens and pathobionts by the gut microbiota. Nat Immunol 14:685–690. https://doi.org/10.1038/ni.2608
Kim CJ, Kovacs-Nolan J, Yang C et al (2009) l-cysteine supplementation attenuates local inflammation and restores gut homeostasis in a porcine model of colitis. Biochim Biophys Acta 1790:1161–1169. https://doi.org/10.1016/j.bbagen.2009.05.018
Lallès J-P, Bosi P, Smidt H, Stokes CR (2007) Weaning—a challenge to gut physiologists. Livest Sci 108:82–93. https://doi.org/10.1016/j.livsci.2007.01.091
Lan J, Dou X, Li J et al (2020) l-arginine ameliorates lipopolysaccharide-induced intestinal inflammation through inhibiting the TLR4/NF-κB and MAPK pathways and stimulating β-defensin expression in vivo and in vitro. J Agric Food Chem 68:2648–2663. https://doi.org/10.1021/acs.jafc.9b07611
Larrosa M, Luceri C, Vivoli E et al (2009) Polyphenol metabolites from colonic microbiota exert anti-inflammatory activity on different inflammation models. Mol Nutr Food Res 53:1044–1054. https://doi.org/10.1002/mnfr.200800446
Le Floch N, Wessels A, Corrent E et al (2018) The relevance of functional amino acids to support the health of growing pigs. Anim Feed Sci Technol 245:104–116. https://doi.org/10.1016/j.anifeedsci.2018.09.007
Leonard W, Zhang P, Ying D, Fang Z (2021) Hydroxycinnamic acids on gut microbiota and health. Compr Rev Food Sci Food Saf 20:710–737. https://doi.org/10.1111/1541-4337.12663
Mao X, Liu M, Tang J et al (2015) Dietary leucine supplementation improves the mucin production in the jejunal mucosa of the weaned pigs challenged by porcine rotavirus. PLoS ONE 10:e0137380. https://doi.org/10.1371/journal.pone.0137380
Marín L, Miguélez EM, Villar CJ, Lombó F (2015) Bioavailability of dietary polyphenols and gut microbiota metabolism: antimicrobial properties. Biomed Res Int 2015:e905215. https://doi.org/10.1155/2015/905215
McMurdie PJ, Holmes S (2013) phyloseq: an R package for reproducible interactive analysis and graphics of microbiome census data. PLoS ONE 8:e61217. https://doi.org/10.1371/journal.pone.0061217
Modina SC, Polito U, Rossi R et al (2019) Nutritional regulation of gut barrier integrity in weaning piglets. Animals 9:1045. https://doi.org/10.3390/ani9121045
Monagas M, Gómez-Cordovés C, Bartolomé B et al (2003) Monomeric, oligomeric, and polymeric flavan-3-ol composition of wines and grapes from Vitis vinifera L. Cv. Graciano, Tempranillo, and Cabernet Sauvignon. J Agric Food Chem 51:6475–6481. https://doi.org/10.1021/jf030325+
Nallathambi R, Poulev A, Zuk JB, Raskin I (2020) Proanthocyanidin-rich grape seed extract reduces inflammation and oxidative stress and restores tight junction barrier function in Caco-2 colon cells. Nutrients 12:1623. https://doi.org/10.3390/nu12061623
Noda T, Iwakiri R, Fujimoto K et al (2002) Exogenous cysteine and cystine promote cell proliferation in CaCo-2 cells. Cell Prolif 35:117–129. https://doi.org/10.1046/j.1365-2184.2002.00229.x
Odenwald MA, Turner JR (2017) The intestinal epithelial barrier: a therapeutic target? Nat Rev Gastroenterol Hepatol 14:9–21. https://doi.org/10.1038/nrgastro.2016.169
Portune KJ, Beaumont M, Davila A-M et al (2016) Gut microbiota role in dietary protein metabolism and health-related outcomes: the two sides of the coin. Trends Food Sci Technol 57:213–232. https://doi.org/10.1016/j.tifs.2016.08.011
Prates JAM, Freire JPB, de Almeida AM et al (2021) Influence of dietary supplementation with an amino acid mixture on inflammatory markers, immune status and serum proteome in LPS-challenged weaned piglets. Animals 11:1143. https://doi.org/10.3390/ani11041143
Quast C, Pruesse E, Yilmaz P et al (2013) The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res 41:D590-596. https://doi.org/10.1093/nar/gks1219
Ren W, Chen S, Yin J et al (2014) Dietary arginine supplementation of mice alters the microbial population and activates intestinal innate immunity. J Nutr 144:988–995. https://doi.org/10.3945/jn.114.192120
Ren M, Cai S, Zhou T et al (2019) Isoleucine attenuates infection induced by E. coli challenge through the modulation of intestinal endogenous antimicrobial peptide expression and the inhibition of the increase in plasma endotoxin and IL-6 in weaned pigs. Food Funct 10:3535–3542. https://doi.org/10.1039/c9fo00218a
Rohart F, Gautier B, Singh A, Cao K-AL (2017) mixOmics: an R package for ‘omics feature selection and multiple data integration. PLoS Comput Biol 13:e1005752. https://doi.org/10.1371/journal.pcbi.1005752
Song ZH, Tong G, Xiao K, et al (2016) L-cysteine protects intestinal integrity, attenuates intestinal inflammation and oxidant stress, and modulates NF-κB and Nrf2 pathways in weaned piglets after LPS challenge. Innate Immun 22:152–161. https://doi.org/10.1177/1753425916632303
Sun Y, Wu Z, Li W et al (2015) Dietary L-leucine supplementation enhances intestinal development in suckling piglets. Amino Acids 47:1517–1525. https://doi.org/10.1007/s00726-015-1985-2
Toden S, Ravindranathan P, Gu J et al (2018) Oligomeric proanthocyanidins (OPCs) target cancer stem-like cells and suppress tumor organoid formation in colorectal cancer. Sci Rep 8:3335. https://doi.org/10.1038/s41598-018-21478-8
Tsang C, Auger C, Mullen W et al (2005) The absorption, metabolism and excretion of flavan-3-ols and procyanidins following the ingestion of a grape seed extract by rats. Br J Nutr 94:170–181. https://doi.org/10.1079/bjn20051480
Valeriano VDV, Balolong MP, Kang D-K (2017) Probiotic roles of Lactobacillus sp. in swine: insights from gut microbiota. J Appl Microbiol 122:554–567. https://doi.org/10.1111/jam.13364
van der Hee B, Wells JM (2021) Microbial regulation of host physiology by short-chain fatty acids. Trends Microbiol. https://doi.org/10.1016/j.tim.2021.02.001
Verhelst R, Schroyen M, Buys N, Niewold T (2014) Dietary polyphenols reduce diarrhea in enterotoxigenic Escherichia coli (ETEC) infected post-weaning piglets. Livest Sci 160:138–140. https://doi.org/10.1016/j.livsci.2013.11.026
Verschuren LMG, Calus MPL, Jansman AJM et al (2018) Fecal microbial composition associated with variation in feed efficiency in pigs depends on diet and sex. J Anim Sci 96:1405–1418. https://doi.org/10.1093/jas/sky060
Vreugdenhil ACE, Dentener MA, Snoek AMP et al (1999) Lipopolysaccharide binding protein and serum amyloid a secretion by human intestinal epithelial cells during the acute phase response. J Immunol 163:2792–2798
Wu G (2010) Functional amino acids in growth, reproduction, and health. Adv Nutr 1:31–37. https://doi.org/10.3945/an.110.1008
Wu H, Luo T, Li YM et al (2018) Granny Smith apple procyanidin extract upregulates tight junction protein expression and modulates oxidative stress and inflammation in lipopolysaccharide-induced Caco-2 cells. Food Funct 9:3321–3329. https://doi.org/10.1039/C8FO00525G
Wu Y, Ma N, Song P et al (2019) Grape seed proanthocyanidin affects lipid metabolism via changing gut microflora and enhancing propionate production in weaned pigs. J Nutr 149:1523–1532. https://doi.org/10.1093/jn/nxz102
Yang Z, Liao SF (2019) Physiological effects of dietary amino acids on gut health and functions of swine. Front Vet Sci. https://doi.org/10.3389/fvets.2019.00169
Yang Z, Huang S, Zou D et al (2016) Metabolic shifts and structural changes in the gut microbiota upon branched-chain amino acid supplementation in middle-aged mice. Amino Acids 48:2731–2745. https://doi.org/10.1007/s00726-016-2308-y
Yang G, Bibi S, Du M et al (2017) Regulation of the intestinal tight junction by natural polyphenols: a mechanistic perspective. Crit Rev Food Sci Nutr 57:3830–3839. https://doi.org/10.1080/10408398.2016.1152230
Zhou H, Yu B, Gao J et al (2018) Regulation of intestinal health by branched-chain amino acids. Anim Sci J 89:3–11. https://doi.org/10.1111/asj.12937
Acknowledgements
The authors are grateful to the genotoul bioinformatics platform Toulouse Occitanie (Bioinfo Genotoul, https://doi.org/10.15454/1.5572369328961167E12) and Sigenae group for providing computing and storage resources to Galaxy instance (http://sigenae-workbench.toulouse.inra.fr). The authors acknowledge the metabolomics platform Axiom (MetaToul-MetaboHUB, Toulouse, France) and the BioToMyc research group (Toxalim, Université de Toulouse, INRAE, ENVT, INP-Purpan, UPS, Toulouse, France) for providing the pig jejunum sample used for organoid culture. We are grateful to Sylvie Combes (GenPhySE, Université de Toulouse, INRAE, ENVT, Castanet Tolosan, France) for careful revision of the manuscript.
Funding
The present work received the financial support from Ajinomoto Animal Nutrition Europe.
Author information
Authors and Affiliations
Contributions
MB, WL and TCD conceived the experiments. MB, CL, PO, LP and JVD performed the experiments. MB and TCD analyzed the data. MB and TCD wrote the manuscript. All authors approved the final version of the manuscript.
Corresponding author
Ethics declarations
Conflicts of interest
TCD and WL are employees of METEX NOOVISTAGO.
Ethical approval
For in vivo experiments, animals were handled in accordance with proper animal welfare guidelines from the Bangkok animal research center Co., Ltd (Bangkok, Thailand). Sampling of pig intestinal tissue for organoid culture was approved by a local ethical committee (N°TOXCOM/0163PP, Toulouse, France).
Additional information
Handling editor: G. Wu.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Beaumont, M., Lencina, C., Painteaux, L. et al. A mix of functional amino acids and grape polyphenols promotes the growth of piglets, modulates the gut microbiota in vivo and regulates epithelial homeostasis in intestinal organoids. Amino Acids 54, 1357–1369 (2022). https://doi.org/10.1007/s00726-021-03082-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00726-021-03082-9