Prebiotics and Bioactive Milk Fractions Affect Gut Development, Microbiota, and Neurotransmitter Expression in Piglets
Kirsten Berding
Division of Nutritional Sciences, University of Illinois, Urbana-Champaign
Search for more papers by this authorMei Wang
Department of Food Science and Human Nutrition, University of Illinois, Urbana-Champaign
Search for more papers by this authorMarcia H. Monaco
Department of Food Science and Human Nutrition, University of Illinois, Urbana-Champaign
Search for more papers by this authorLindsey S. Alexander
Piglet Nutrition and Cognition Laboratory, Department of Animal Sciences, University of Illinois, Urbana-Champaign
Search for more papers by this authorAustin T. Mudd
Piglet Nutrition and Cognition Laboratory, Department of Animal Sciences, University of Illinois, Urbana-Champaign
Neuroscience Program, University of Illinois, Urbana-Champaign
Search for more papers by this authorMaciej Chichlowski
Mead Johnson Pediatric Nutrition Institute, Evansville, IN
Search for more papers by this authorRosaline V. Waworuntu
Mead Johnson Pediatric Nutrition Institute, Evansville, IN
Search for more papers by this authorBrian M. Berg
Division of Nutritional Sciences, University of Illinois, Urbana-Champaign
Mead Johnson Pediatric Nutrition Institute, Evansville, IN
Search for more papers by this authorMichael J. Miller
Division of Nutritional Sciences, University of Illinois, Urbana-Champaign
Department of Food Science and Human Nutrition, University of Illinois, Urbana-Champaign
Search for more papers by this authorRyan N. Dilger
Division of Nutritional Sciences, University of Illinois, Urbana-Champaign
Piglet Nutrition and Cognition Laboratory, Department of Animal Sciences, University of Illinois, Urbana-Champaign
Neuroscience Program, University of Illinois, Urbana-Champaign
Search for more papers by this authorCorresponding Author
Sharon M. Donovan
Division of Nutritional Sciences, University of Illinois, Urbana-Champaign
Department of Food Science and Human Nutrition, University of Illinois, Urbana-Champaign
Address correspondence and reprint requests to Sharon M. Donovan, Department of Food Science and Human Nutrition, University of Illinois, 339 Bevier Hall, 905 South Goodwin Ave, Urbana, IL 61801 (e-mail: [email protected]).Search for more papers by this authorKirsten Berding
Division of Nutritional Sciences, University of Illinois, Urbana-Champaign
Search for more papers by this authorMei Wang
Department of Food Science and Human Nutrition, University of Illinois, Urbana-Champaign
Search for more papers by this authorMarcia H. Monaco
Department of Food Science and Human Nutrition, University of Illinois, Urbana-Champaign
Search for more papers by this authorLindsey S. Alexander
Piglet Nutrition and Cognition Laboratory, Department of Animal Sciences, University of Illinois, Urbana-Champaign
Search for more papers by this authorAustin T. Mudd
Piglet Nutrition and Cognition Laboratory, Department of Animal Sciences, University of Illinois, Urbana-Champaign
Neuroscience Program, University of Illinois, Urbana-Champaign
Search for more papers by this authorMaciej Chichlowski
Mead Johnson Pediatric Nutrition Institute, Evansville, IN
Search for more papers by this authorRosaline V. Waworuntu
Mead Johnson Pediatric Nutrition Institute, Evansville, IN
Search for more papers by this authorBrian M. Berg
Division of Nutritional Sciences, University of Illinois, Urbana-Champaign
Mead Johnson Pediatric Nutrition Institute, Evansville, IN
Search for more papers by this authorMichael J. Miller
Division of Nutritional Sciences, University of Illinois, Urbana-Champaign
Department of Food Science and Human Nutrition, University of Illinois, Urbana-Champaign
Search for more papers by this authorRyan N. Dilger
Division of Nutritional Sciences, University of Illinois, Urbana-Champaign
Piglet Nutrition and Cognition Laboratory, Department of Animal Sciences, University of Illinois, Urbana-Champaign
Neuroscience Program, University of Illinois, Urbana-Champaign
Search for more papers by this authorCorresponding Author
Sharon M. Donovan
Division of Nutritional Sciences, University of Illinois, Urbana-Champaign
Department of Food Science and Human Nutrition, University of Illinois, Urbana-Champaign
Address correspondence and reprint requests to Sharon M. Donovan, Department of Food Science and Human Nutrition, University of Illinois, 339 Bevier Hall, 905 South Goodwin Ave, Urbana, IL 61801 (e-mail: [email protected]).Search for more papers by this authorSupplemental digital content is available for this article. Direct URL citations appear in the printed text, and links to the digital files are provided in the HTML text of this article on the journal's Web site (www.jpgn.org).
This study was supported by a grant from Mead Johnson Nutrition (Evansville, IN).
S.D., M.M., and R.D. have received grant funding. S.D. has served on advisory boards, and S.D., R.D., M.W., M.M., L.A., and R.D. have consulted for Mead Johnson Nutrition. M.C., R.W., and B.B. are employees of Mead Johnson Nutrition. The remaining authors report no conflicts of interest.
ABSTRACT
Objective:
This study tested the hypothesis that the addition of prebiotics and 2 functional milk ingredients to infant formula would maintain normal growth and gut development, and modify microbiota composition and neurotransmitter gene expression in neonatal piglets.
Methods:
Two-day-old male piglets (n = 24) were fed formula (CONT) or formula with polydextrose (1.2 g/100 g diet), galactooligosaccharides (3.5 g/100 g diet), bovine lactoferrin (0.3 g/100 g diet), and milk fat globule membrane-10 (2.5 g/100 g diet) (TEST) for 30 days. On study day 31, intestinal samples, ileal and colonic contents, and feces were collected. Intestinal histomorphology, disaccharidase activity, serotonin (5'HT), vasoactive intestinal peptide (VIP), and tyrosine hydroxylase (TH) were measured. Gut microbiota composition was assessed by pyrosequencing of the V3–V5 regions of 16S rRNA and quantitative polymerase chain reaction.
Results:
Body weight of piglets on TEST was greater (P ⩽ 0.05) than CONT on days 17 to 30. Both groups displayed growth patterns within the range observed for sow-reared pigs. TEST piglets had greater jejunal lactase (P = 0.03) and higher (P = 0.003) ileal VIP expression. TEST piglets tended to have greater (P = 0.09) sucrase activity, longer (P = 0.08) ileal villi, and greater (P = 0.06) duodenal TH expression. Microbial communities of TEST piglets differed from CONT in ascending colon (AC, P = 0.001) and feces (P ⩽ 0.05). CONT piglets had greater relative abundances of Mogibacterium, Collinsella, Klebsiella, Escherichia/Shigella, Eubacterium, and Roseburia compared with TEST piglets in AC. In feces, CONT piglets harbored lower (P ⩽ 0.05) proportions of Parabacteroides, Clostridium IV, Lutispora, and Sutterella than TEST piglets.
Conclusions:
A mixture of bioactive ingredients improved weight gain and gut maturation, modulated colonic and fecal microbial composition, and reduced the proportions of opportunistic pathogens.
REFERENCES
- 1.Walker WA. Initial intestinal colonization in the human infant and immune homeostasis. Ann Nutr Metab 2013; 63 (suppl 2): 8–15.
- 2. American Academy of Pediatrics, Section on Breastfeeding. Policy statement: breastfeeding and the use of human milk. Pediatrics 2012; 129: e827–e841.
- 3.Reznikov EA, Comstock SS, Yi C, et al. Dietary bovine lactoferrin increases intestinal cell proliferation in neonatal piglets. J Nutr 2014; 144: 1401–1408.
- 4.Comstock SS, Reznikov EA, Contractor N, et al. Dietary bovine lactoferrin alters mucosal and systemic immune cell responses in neonatal piglets. J Nutr 2014; 144: 525–532.
- 5.Liu KY, Comstock SS, Shunk JM, et al. Natural killer cell populations and cytotoxic activity in pigs fed mother's milk, formula, or formula supplemented with bovine lactoferrin. Pediatr Res 2013; 74: 402–407.
- 6.Mastromarino P, Capobianco D, Campagna G, et al. Correlation between lactoferrin and beneficial microbiota in breast milk and infant's feces. Biometals 2014; 27: 1077–1086.
- 7.Dewettinck K, Rombaut R, Thienpont N, et al. Nutritional and technological aspects of milk fat globule membrane material. Int Dairy J 2008; 18: 436–457.
- 8.Billeaud C, Puccio G, Saliba E, et al. Safety and tolerance evaluation of milk fat globule membrane-enriched infant formulas: a randomized controlled multicenter non-inferiority trial in healthy term infants. Clin Med Insights Pediatr 2014; 8: 51–60.
- 9.Timby N, Domellöf E, Hernell O, et al. Neurodevelopment, nutrition, and growth until 12 mo of age in infants fed a low-energy, low-protein formula supplemented with bovine milk fat globule membranes: a randomized controlled trial. Am J Clin Nutr 2014; 99: 860–868.
- 10.Donovan SM, Wang M, Li M, et al. Host-microbe interactions in the neonatal intestine: role of human milk oligosaccharides. Adv Nutr 2012; 3: 450S–455S.
- 11.Oozeer R, van Limpt K, Ludwig T, et al. Intestinal microbiology in early life: specific prebiotics can have similar functionalities as human-milk oligosaccharides. Am J Clin Nutr 2013; 98: 561S–571S.
- 12.Scalabrin DM, Mitmesser SH, Welling GW, et al. New prebiotic blend of polydextrose and galacto-oligosaccharides has a bifidogenic effect in young infants. J Pediatr Gastroenterol Nutr 2012; 54: 343–352.
- 13.Gespach C, Chastre E, Emami S, et al. Vasoactive intestinal peptide receptor activity in human fetal enterocytes. FEBS Lett 1985; 180: 196–202.
- 14.Mawe GM, Hoffman JM. Serotonin signaling in the gastrointestinal tract: functions, dysfunctions, and therapeutic targets. Nat Rev Gastroenterol Hepatol 2013; 10: 473–486.
- 15.Heitkemper MM, Marotta SF. Development of neurotransmitter enzyme activity in the rat gastrointestinal tract. Am J Physiol 1983; 244: G58–G64.
- 16.Janjatović AK, Valpotić H, Kezić D, et al. Secretion of immunomodulating neuropeptides (VIP, SP) and nitric oxide synthase in porcine small intestine during postnatal development. Eur J Histochem 2012; 56: e30.
- 17.Pohl CS, Medland JE, Moeser AJ. Early-life stress origins of gastrointestinal disease: animal models, intestinal pathophysiology, and translational implications. Am J Physiol Gastrointest Liver Physiol 2015; 309: G927–G941.
- 18.Saulnier DM, Ringel Y, Heyman MB, et al. The intestinal microbiome, probiotics and prebiotics in neurogastroenterology. Gut Microbes 2013; 4: 17–27.
- 19.Mudd AT, Alexander LS, Berding K. Dietary prebiotics, milk fat globule membrane and lactoferrin affects structural neurodevelopment in the young piglet. Front Pediatr 2016; 4: 4.
- 20.Hester SN, Comstock SS, Thorum SC, et al. Intestinal and systemic immune development and response to vaccination are unaffected by dietary (1, 3/1, 6)-β-D-glucan supplementation in neonatal piglets. Clin Vaccine Immunol 2012; 19: 1499–1508.
- 21.Hartke JL, Monaco MH, Wheeler MB, et al. Effect of a short term fast on intestinal disaccharidase activity and villus morphology of piglets suckling insulin-like growth factor-I transgenic sows. J Anim Sci 2005; 83: 2404–2413.
- 22.Li M, Bauer LL, Chen X, et al. Microbial composition and in vitro fermentation patterns of human milk oligosaccharides differ between formula-fed and sow-reared piglets. J Nutr 2012; 142: 681–689.
- 23.Klindworth A, Pruesse E, Schweer T, et al. Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies. Nucleic Acids Res 2013; 41: e1.
- 24.Dowd SE, Callaway TR, Wolcott RD, et al. Evaluation of the bacterial diversity in the feces of cattle using 16S rDNA bacterial tag-encoded FLX amplicon pyrosequencing (bTEFAP). BMC Microbiol 2008; 8: 125.
- 25.Handl S, Dowd SE, Garcia-Mazcorro JF, et al. Massive parallel 16S rRNA gene pyrosequencing reveals highly diverse fecal bacterial and fungal communities in healthy dogs and cats. FEMS Microbiol Ecol 2011; 76: 301–310.
- 26.Edgar RC, Haas BJ, Clemente JC, et al. UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 2011; 27: 2194–2200.
- 27.Edgar RC. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 2004; 32: 1792–1797.
- 28.Price MN, Dehal PS, Arkin AP. FastTree 2--approximately maximum-likelihood trees for large alignments. PLoS One 2010; 5: e9490.
- 29.Lozupone C, Knight R. UniFrac: a new phylogenetic method for comparing microbial communities. Appl Environ Microbiol 2005; 71: 8228–8235.
- 30.Edgar RC. Search and clustering orders of magnitude faster than BLAST. Bioinformatics 2010; 26: 2460–2461.
- 31.Cole JR, Wang Q, Cardenas E, et al. The Ribosomal Database Project: improved alignments and new tools for rRNA analysis. Nucleic Acids Res 2009; 37 (Database issue): D141–D145.
- 32.Caporaso JG, Kuczynski J, Stombaugh J, et al. QIIME allows analysis of high-throughput community sequencing data. Nat Methods 2010; 7: 335–336.
- 33.Tong J, Liu C, Summanen P, et al. Application of quantitative real-time PCR for rapid identification of Bacteroides fragilis group and related organisms in human wound samples. Anaerobe 2011; 17: 64–68.
- 34.Oksanen J, Blanchet FG, Kindt R, et al. Vegan: Community Ecology Package. R package version 2.0-10, 2013. http://CRAN.R-project.org/package=vegan. Accessed February 29, 2016.
- 35.van Beers-Schreurs HM, Nabuurs MJ, Vellenga L, et al. Weaning and the weanling diet influence the villous height and crypt depth in the small intestine of pigs and alter the concentrations of short-chain fatty acids in the large intestine and blood. J Nutr 1998; 128: 947–953.
- 36.Montage L, Pulske JR, Hampson DJ. A review of interactions between dietary fibre and the intestinal mucosa, and their consequences on digestive health in young non-ruminant animals. Anim Feed Sci Technol 2003; 108: 95–117.
- 37.Shulman RJ, Henning SJ, Nichols BL. The miniature pig as an animal model for the study of intestinal enzyme development. Pediatr Res 1988; 23: 311–315.
- 38.Leforestier G, Blais A, Blachier F, et al. Effects of galacto-oligosaccharide ingestion on the mucosa-associated mucins and sucrase activity in the small intestine of mice. Euro J Nutr 2009; 48: 457–464.
- 39.Yang SC, Chen JY, Shang HF, et al. Effect of synbiotics on intestinal microflora and digestive enzyme activities in rats. World J Gastroenterol 2005; 11: 7413–7417.
- 40.Gershon MD. The second brain: a groundbreaking new understanding of nervous disorders of the stomach and intestine. New York: Harper Perennial; 1998.
- 41.Nezami BG, Srinivasan S. Enteric nervous system in the small intestine: pathophysiology and clinical implications. Curr Gastroenterol Rep 2010; 12: 358–365.
- 42.Li Z, Chalazonitis A, Huang YY, et al. Essential roles of enteric neuronal serotonin in gastrointestinal motility and the development/survival of enteric dopaminergic neurons. J Neurosci 2011; 31: 8998–9009.
- 43.Lychkova AE. Gradients of serotoninergic innervation of the large intestine. Bull Exp Biol Med 2005; 139: 550–553.
- 44.Ku SK, Lee HS, Lee JH. An immunohistochemical study of the gastrointestinal endocrine cells in the C57BL/6 mice. Anat Histol Embryol 2003; 32: 21–28.
- 45.Zhang H, Zhang T, Wang L, et al. Immunohistochemical location of serotonin and serotonin 2B receptor in the small intestine of pigs. Acta Histochem 2009; 111: 35–41.
- 46.Sayadi H, Harmon JW, Moody TW, et al. Autoradiographic distribution of vasoactive intestinal polypeptide receptors in rabbit and rat small intestine. Peptides 1988; 9: 23–30.
- 47.Sjögren YM, Tomicic S, Lundberg A, et al. Influence of early gut microbiota on the maturation of childhood mucosal and systemic immune responses. Clin Exp Allergy 2009; 39: 1842–1851.
- 48.Borre YE, Moloney RD, Clarke G, et al. The impact of microbiota on brain and behavior: mechanisms & therapeutic potential. Adv Exp Med Biol 2014; 817: 373–403.
- 49.Lopetuso LR, Scaldaferri F, Petito V, et al. Commensal clostridia: leading players in the maintenance of gut homeostasis. Gut Pathog 2013; 5: 23.
- 50.Nakano V, Ignacio A, Rodriguez Fernandez M, et al. Intestinal Bacteroides and Parabacteroides species producing antagonistic substances. Curr Trends Microbiol 2012; 8: 61–64.
- 51.Arshad M, Seed PC. Urinary tract infections in the infant. Clin Perinatol 2015; 42: 17–28.
- 52.Lanata CF, Fischer-Walker CL, Olascoaga AC, et al. Child Health Epidemiology Reference Group of the World Health Organization and UNICEF. Global causes of diarrheal disease mortality in children <5 years of age: a systematic review. PLoS One 2013; 8: e72788.
- 53.Podschun R, Ullmann U. Klebsiella spp. as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors. Clin Microbiol Rev 1998; 11: 589–603.
- 54.Hoeflinger JL, Kashtanov DO, Cox SB, et al. Characterization of the intestinal Lactobacilli community following galactooligosaccharides and polydextrose supplementation in the neonatal piglet. PLoS One 2015; 10: e0135494.
- 55.Holmgren J, Svennerholm A-M, Lindblad M, Goldman AS. Inhibition of bacterial adhesion and toxin binding by glycoconjugate and oligosaccharide receptor analogues in human milk. Human Lactation 3 ed. Berkeley, CA: Perseus Publishing; 1987. 251–259.
10.1007/978-1-4899-0837-7_28 Google Scholar
- 56.Ochoa TJ, Cleary TG. Effect of lactoferrin on enteric pathogens. Biochimie 2009; 9: 30–34.
10.1016/j.biochi.2008.04.006 Google Scholar
- 57.Tian H, Maddox IS, Ferguson LR, et al. Influence of bovine lactoferrin on selected probiotic bacteria and intestinal pathogens. Biometals 2010; 23: 593–596.
- 58.Hu W, Zhao J, Wang J, et al. Transgenic milk containing recombinant human lactoferrin modulates the intestinal flora in piglets. Biochem Cell Biol 2012; 90: 485–496.
Citing Literature
