Development and validation of a new dynamic in vitro model of the piglet colon (PigutIVM): application to the study of probiotics
- 798 Downloads
For ethical, technical, regulatory, and cost reasons, in vitro methods are increasingly used as an alternative to in vivo experimentations. The aim of the present study was to validate, according to in vivo data in living animals, a new in vitro model of the piglet colon, the PigutIVM, under both control conditions and antibiotic disturbance by the widely used colistin. The PigutIVM reproduces the main biotic and abiotic parameters of the piglet colon: temperature, pH, retention time, supply of ileal effluents, complex, and metabolically active microbiota and self-maintained anaerobiosis. Under both control and antibiotic-treated conditions, qPCR analyses showed that the main bacterial populations of piglet gut microbiota were similar in vitro and in vivo, with Pearson correlation coefficient higher than 0.9. During colistin administration, both in piglets and in the in vitro model, a significant decrease in Escherichia coli populations was observed together with changes in microbial composition of subdominant populations. SCFA concentrations were similar in vitro and in vivo and were not modified by colistin. Interestingly, the administration of the probiotic Saccharomyces cerevisiae var. boulardii CNCM I-1079 led in vitro to a decrease in E. coli levels, as previously observed when the antibiotic treatment was applied. This new in vitro model of the piglet colon provides a flexible, reproducible, and cost-effective tool for the screening of drugs or new dietary compounds, such as pre- or probiotics. It will be helpful for researchers, feed producers, or veterinarians when developing innovative non-antibiotic strategies.
KeywordsIn vitro model Piglet Gut microbiota Colistin Probiotic Saccharomyces cerevisiae var. boulardii
This study was supported by the Côtes d’Armor General Council and the Brittany and Auvergne regions.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Research involving animals
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
- Bian G, Ma S, Zhu Z, Su Y, Zoetendal EG, Mackie R, Liu J, Mu C, Huang R, Smidt H, Zhu W (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. Environ Microbiol 18:1566–1577. doi: 10.1111/1462-2920.13272 CrossRefPubMedGoogle Scholar
- Bindelle J, Pieper R, Montoya CA, Van Kessel AG, Leterme P (2011) Nonstarch polysaccharide-degrading enzymes alter the microbial community and the fermentation patterns of barley cultivars and wheat products in an in vitro model of the porcine gastrointestinal tract. FEMS Microbiol Ecol 76:553–563. doi: 10.1111/j.1574-6941.2011.01074.x CrossRefPubMedGoogle Scholar
- Blanquet-Diot S, Denis S, Chalancon S, Chaira F, Cardot JM, Alric M (2012) Use of artificial digestive systems to investigate the biopharmaceutical factors influencing the survival of probiotic yeast during gastrointestinal transit in humans. Pharm Res 29:1444–1453. doi: 10.1007/s11095-011-0620-5 CrossRefPubMedGoogle Scholar
- Brousseau JP, Talbot G, Beaudoin F, Lauzon K, Roy D, Lessard M (2015) Effects of probiotics Pediococcus acidilactici strain MA18/5 M and Saccharomyces cerevisiae subsp. boulardii strain SB-CNCM I-1079 on fecal and intestinal microbiota of nursing and weanling piglets. J Anim Sci 93:5313–5326. doi: 10.2527/jas.2015-9190 CrossRefPubMedGoogle Scholar
- Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, Fierer N, Pena AG, Goodrich JK, Gordon JI, Huttley CA, Kelley ST, Knights D, Koenig JE, Lozupone CA, McDonald D, Muegge BD, Pirrung M, Reeder J, Sevinsky JR, Turnbaugh PJ, Walters WA, Widmann J, Yatsunenko T, Zaneveld J, Knight R (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Methods 7:335–336. doi: 10.1038/nmeth.f.303 CrossRefPubMedPubMedCentralGoogle Scholar
- Collier CT, Carroll JA, Ballou MA, Starkey JD, Sparks JC (2011) Oral administration of Saccharomyces cerevisiae boulardii reduces mortality associated with immune and cortisol responses to Escherichia coli endotoxin in pigs. J Anim Sci 89:52–58. doi: 10.2527/jas.2010-2944 CrossRefPubMedGoogle Scholar
- R Core Team (2015) R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing. R Foundation for Statistical Computing Vienna, Austria.Google Scholar
- DeSantis TZ, Hugenholtz P, Larsen N, Rojas M, Brodie EL, Keller K, Huber T, Dalevi D, Hu P, Andersen GL (2006) Greengenes, a chimera-checked 16S rRNA gene database and workbench compatible with ARB. Appl Environ Microbiol 72:5069–5072. doi: 10.1128/AEM.03006-05 CrossRefPubMedPubMedCentralGoogle Scholar
- EU 2010 (2010) Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes. Official journal of the European Union L276/33–276/79.Google Scholar
- FAO/WHO (2002) Guidelines for the evaluation of probiotics in food. Report of a Joint FAO/WHO Working Group on Drafting Guidelines for the Evaluation of Probiotics in Food. London, OntarioGoogle Scholar
- Gérard-Champod M, Blanquet-Diot S, Cardot J-M, Bravo D, Alric M (2010) Development and validation of a continuous in vitro system reproducing some biotic and abiotic factors of the veal calf intestine. Appl Environ Microbiol 76:5592–5600. doi: 10.1128/AEM.00524-10 CrossRefPubMedPubMedCentralGoogle Scholar
- Joint FAO/WHO (2006) Evaluation of certain veterinary drug residues in food: 66th report of the joint FAO/WHO expert committee on food additives. WHO technical Report Series n° 939. Rome, ItalyGoogle Scholar
- Jonathan MC, van den Borne JJGC, van Wiechen P, Souza da Silva C, Schols HA, Gruppen H (2012) In vitro fermentation of 12 dietary fibres by faecal inoculum from pigs and humans. Food Chem 133:889–897. doi: 10.1016/j.foodchem.2012.01.110
- Kim HB, Borewicz K, White BA, Singer RS, Sreevatsan S, Tu ZJ, Isaacson RE (2012) Microbial shifts in the swine distal gut in response to the treatment with antimicrobial growth promoter, tylosin. Proc Natl Acad Sci 109:15485–15490. doi: 10.1073/pnas.1205147109 CrossRefPubMedPubMedCentralGoogle Scholar
- Kraler M, Schedle K, Schwarz C, Domig KJ, Pichler M, Oppeneder A, Wetscherek W, Prückler M, Pignitter M, Pirker KF, Somoza V, Heine D, Kneifel W (2015) Fermented and extruded wheat bran in piglet diets: impact on performance, intestinal morphology, microbial metabolites in chyme and blood lipid radicals. Arch Anim Nutr 69:378–398. doi: 10.1080/1745039X.2015.1075671 CrossRefPubMedGoogle Scholar
- Molist Gasa F, Ywazaki M, Gomez de Segura Ugalde A, Hermes RG, Gasa Gasó J, Pérez Hernández JF (2010) Administration of loperamide and addition of wheat bran to the diets of weaner pigs decrease the incidence of diarrhoea and enhance their gut maturation. Br J Nutr 103:879–885. doi: 10.1017/S0007114509992637
- Pivetta MR, Berto DA, Amorim AB, Saleh MAD, Pinheiro DF, Paulino M de LMV, Pinto JP de AN, Gonçalves HC (2014) Use of maltodextrin and a prebiotic in the feed of weaned piglets. Semina Ciênc Agrár 35:2129–2146. doi: 10.5433/1679-0359.2014v35n4p2129
- Saraoui T, Parayre S, Guernec G, Loux V, Montfort J, Le Cam A, Boudry G, Jan G, Falentin H (2013) A unique in vivo experimental approach reveals metabolic adaptation of the probiotic Propionibacterium freudenreichii to the colon environment. BMC Genomics 14:1. doi: 10.1186/1471-2164-14-911 CrossRefGoogle Scholar
- Tan CQ, Wei HK, Sun HQ, Long G, Ao JT, Jiang SW, Peng J (2015) Effects of supplementing sow diets during two gestations with konjac flour and Saccharomyces boulardii on constipation in peripartal period, lactation feed intake and piglet performance. Anim Feed Sci Technol 210:254–262. doi: 10.1016/j.anifeedsci.2015.10.013 CrossRefGoogle Scholar
- Tanner SA, Berner AZ, Rigozzi E, Grattepanche F, Chassard C, Lacroix C (2014) In vitro continuous fermentation model (PolyFermS) of the swine proximal colon for simultaneous testing on the same gut microbiota. PLoS One 9:e94123. doi: 10.1371/journal.pone.0094123 CrossRefPubMedPubMedCentralGoogle Scholar
- Tran TH, Boudry C, Everaert N, Théwis A, Portetelle D, Daube G, Nezer C, Taminiau B, Bindelle J (2016) Adding mucins to an in vitro batch fermentation model of the large intestine induces changes in microbial population isolated from porcine feces depending on the substrate. FEMS Microbiol Ecol 92(2). doi: 10.1093/femsec/fiv165
- Van den Abbeele P, Grootaert C, Marzorati M, Possemiers S, Verstraete W, Gérard P, Rabot S, Bruneau A, El Aidy S, Derrien M, Zoetendal E, Kleerebezem M, Smidt H, Van de Wiele T (2010) Microbial community development in a dynamic gut model is reproducible, colon region specific, and selective for Bacteroidetes and Clostridium cluster IX. Appl Environ Microbiol 76:5237–5246. doi: 10.1128/AEM.00759-10 CrossRefPubMedPubMedCentralGoogle Scholar
- Van den Abbeele P, Roos S, Eeckhaut V, MacKenzie DA, Derde M, Verstraete W, Marzorati M, Possemiers S, Vanhoecke B, Van Immerseel F, Van de Wiele T (2012) Incorporating a mucosal environment in a dynamic gut model results in a more representative colonization by lactobacilli. Microb Biotechnol 5:106–115. doi: 10.1111/j.1751-7915.2011.00308.x