Four groups of six 21 days-old piglets were used to evaluate the effect of a prebiotic or probiotic on the intestinal fermentative
activity. In each group, piglets received one of the following diets: basal diet (C); basal diet supplemented with xylooligosaccharide
(C-XOS); basal diet supplemented with Saccharomyces cerevisiae (C-SC); and basal diet supplemented with xylooligosaccharide
and S. cerevisiae (C-XOS+SC).
The short chain fatty acids in the colon of piglets were decreased with the inclusion of S. cerevisiae in the diet (Pb0.01). The
xylanolytic activity was higher (Pb0.05) in the small intestine of piglets fed C-XOS+SC diet, but no significant differences were
found in the caecum and colon. In the caecum contents, the cellulolytic activity was increased (Pb0.05) by the C-XOS and C-SC
diets, but remained similar when the diet was supplemented with the two additives combined

Cunha, L.F.

Freire, J.P.B.

Lordelo, M.M.

Marinho, M.C.

Livestock Science

English

Universidade Técnica de Lisboa

Microbial activity in the gut of piglets: effect of prebiotic and probiotic supplementation

UTL Repository

This article was originally published in a journal published byElsevier, and the attached copy is provided by Elsevier for theauthor’s benefit and for the benefit of the author’s institution, fornon-commercial research and educational use including withoutlimitation use in instruction at your institution, sending it to specificcolleagues that you know, and providing a copy to your institution’sadministrator.All other uses, reproduction and distribution, including withoutlimitation commercial reprints, selling or licensing copies or access,or posting on open internet sites, your personal or institution’swebsite or repository, are prohibited. For exceptions, permissionmay be sought for such use through Elsevier’s permissions site at:http://www.elsevier.com/locate/permissionusematerialAuthor's   personal   copyMicrobial activity in the gut of piglets: I. Effect of prebiotic andprobiotic supplementation☆M.C. Marinho a, M.M. Lordelo b, L.F. Cunha b, J.P.B. Freire b,⁎a Escola Superior Agrária de Castelo Branco, Apartado 119, 6000–909 Castelo Branco, Portugalb Universidade Técnica de Lisboa, Instituto Superior de Agronomia, Tapada da Ajuda 1349–017 Lisboa, PortugalAbstractFour groups of six 21 days-old piglets were used to evaluate the effect of a prebiotic or probiotic on the intestinal fermentativeactivity. In each group, piglets received one of the following diets: basal diet (C); basal diet supplemented with xylo-oligosaccharide (C-XOS); basal diet supplemented with Saccharomyces cerevisiae (C-SC); and basal diet supplemented with xylo-oligosaccharide and S. cerevisiae (C-XOS+SC).The short chain fatty acids in the colon of piglets were decreased with the inclusion of S. cerevisiae in the diet (Pb0.01). Thexylanolytic activity was higher (Pb0.05) in the small intestine of piglets fed C-XOS+SC diet, but no significant differences werefound in the caecum and colon. In the caecum contents, the cellulolytic activity was increased (Pb0.05) by the C-XOS and C-SCdiets, but remained similar when the diet was supplemented with the two additives combined.© 2007 Elsevier B.V. All rights reserved.Keywords: Piglets; Xylo-oligosaccharides; Saccharomyces cerevisiae; Intestinal fermentation1. IntroductionOne potential way to control the intestinal fermen-tation process is using prebiotics or probiotics as feedadditives. Prebiotics are non-digestible feed ingredientsthat beneficially affect the host by stimulating thegrowth and activity of specific bacteria groups mostly inthe colon. Xylo-oligosaccharides are considered pre-biotics presents in different by-products like corn cob orpeanuts shell (Maxwell et al., 2004).Oligosaccharides are hydrolysed to their monomericunits (glucose, galactose, xylose or arabinose) and thenfermented to short chain fatty acids (SCFA), mainly(C2), (C3) and (C4) by gut bacteria. They are animportant energy source and help to maintain a balancedintestinal ecosystem (Midtvedt, 1994). In fact, acetateand propionate are used by the pig as energy source(Montagne et al., 2003) and butyrate stimulates thedevelopment and growth of large and small intestine, byimproved epithelial cell proliferation (Sakata, 1987).The supplementation with Saccharomyces cerevisiaecan improve microbial balance in the gut of young pigs(Heugten et al., 2003), due to either the yeast cell wallcomponents (mannan-oligosaccharides) effect on theimmune modulation (Newman and Newman, 2001), ora direct effect to reduce pathogenic bacteria and toxicmetabolites (Anderson et al., 1999).Livestock Science 108 (2007) 236–239www.elsevier.com/locate/livsci☆ This paper is part of the special issue entitled “DigestivePhysiology in Pigs” guest edited by José Adalberto Fernández,Mette Skou Hedemann, Bent Borg Jensen, Henry Jørgensen, KnudErik Bach Knudsen and Helle Nygaard Lærke.⁎ Corresponding author. Tel.: +351 213653408; fax: +351213630734.E-mail address: jpfreire@isa.utl.pt (J.P.B. Freire).1871-1413/$ - see front matter © 2007 Elsevier B.V. All rights reserved.doi:10.1016/j.livsci.2007.01.081Author's   personal   copyIt is known that probiotics act mostly in the uppercompartments of the gastrointestinal tract; while pre-biotics are suppose to be used as substrate for potentiallybeneficial bacteria in the hindgut of monogastric animals.Therefore it is expected that the association of a specificprebiotic and probiotic may have a synergetic positiveeffect on intestinal microflora Nemcova et al. (1999).The objective of this work was to evaluate the effectof xylo-oligosaccharide and S. cerevisiae supplementa-tion, alone or combined, on the microbial activity in thegut of the piglet.2. Materials and methodsFour groups of six 21 days-old weaning piglets wereused. Piglets were individually housed in metabolismcages, during 4 weeks. In each group piglets receivedone of the following diets: (C) basal diet (wheat:600 g·kg−1; maize starch: 140 g·kg−1; soybean oil:50 g·kg−1; soluble fish protein concentrate: 150 g·kg−1),(C-XOS) basal diet supplemented with xylo-oligosac-charide (20 g·kg−1), (C-SC) basal diet supplementedwith S. cerevisiae (6×108 CFU·kg−1) and (C-XOS+SC)basal diet supplemented with xylo-oligosaccharide andS. cerevisiae. At the end of the experimental period thepiglets were slaughtered and pooled samples from thesmall intestine (SI), caecum and colon contents werecollected. The SCFA concentration (C2, C3 and C4) werequantified by gas chromatography following Jouany(1982). The microbial enzymes activity was analysed byspectrophotometry following the method described byFalcão-e-Cunha et al. (2004). Data were subjected toanalyses of variance using the GLM procedure of SAS(2001).Table 1Effect of xylo-oligosaccharide (XOS) or Saccharomyces cerevisiae (SC) supplementation on total concentration (mmol·l−1) and molar proportions ofSCFA in intestinal contentsC C-XOS C-SC C-XOS+SC XOS SC XOS×SC RSD (2)Small intestineC2+C3+C4(1) 22.8 20.7 23.0 15.1 0.1946 0.4742 0.4501 9.1C2:C3 3.44 3.30 3.99 3.93 0.7359 0.0617 0.8968 0.73C2:C4 5.05 4.10 5.20 4.91 0.1164 0.2163 0.3807 0.92C3:C4 1.45 1.33 1.32 1.29 0.5291 0.4860 0.6900 0.37CaecumC2+C3+C4 53.9 61.4 58.8 53.3 0.8837 0.8173 0.3580 17.0C2:C3 2.04 1.91 2.04 2.04 0.3828 0.3952 0.3588 0.17C2:C4 3.13 2.98 2.87 3.04 0.9600 0.5954 0.3937 0.44C3:C4 1.54 1.56 1.41 1.49 0.5471 0.2310 0.7049 0.20ColonC2+C3+C4 118.4a 81.6ab 55.4b 49.4b 0.1155 0.0016 0.2503 31.9C2:C3 2.96ab 3.50a 2.53b 2.71ab 0.1693 0.0285 0.4939 0.63C2:C4 9.49a 10.31a 4.72b 4.31b 0.8655 0.0002 0.6124 2.94C3:C4 3.14a 2.95a 1.89b 1.61b 0.4232 0.0003 0.8797 0.72(1)C2: Acetic, C3: Propionic, C4: Butyric. (2)For effects: Probability of significance of the F value. RSD: Residual standard deviation. In the samerow, values with different letters superscripts differ significantly (Pb0.05).Table 2Effect of xylo-oligosaccharide (XOS) or Saccharomyces cerevisiae (SC) supplementation on enzymatic(1) activities in intestinal contents of pigletsC C-XOS C-SC C-XOS+SC XOS SC XOS×SC RSD (2)Small intestineCellulolytic 132.9 58.6 156.0 52.6 0.0675 0.8540 0.7539 106.6Xylanolytic 46.1a 96.5a 56.3a 229.8b 0.0189 0.1173 0.1757 107.4CaecumCellulolytic 102.2a 354.6b 523.6c 125.0a 0.2246 0.1164 b0.0001 131.1Xylanolytic 288.1 329.2 273.5 253.2 0.8550 0.4291 0.5903 134.1ColonCellulolytic 74.2 92.4 92.9 76.7 0.9358 0.9069 0.1914 31.2Xylanolytic 387.5 636.8 507.0 404.2 0.4170 0.5296 0.0602 216.5(1)Enzymatic activities: mg hydrolysed substrate/g DM intestinal contents/h. (2)For effects: probability of significance of the F value. RSD: Residualstandard deviation. In the same row, values with different letters superscripts differ significantly (Pb0.05).237M.C. Marinho et al. / Livestock Science 108 (2007) 236–239Author's   personal   copy3. ResultsThe results of SCFA are showed in Table 1, and themicrobiological enzymatic activities data are presented inTable 2. The inclusion of xylo-oligosaccharide orS. cerevisiae in the piglet diet did not affect the levels ormolar proportions of SCFA in the SI and caecum.However, the level of total SCFA (mmol·l−1) in thecolon, decreased with the inclusion of S. cerevisiae on thediet (P=0.0016). Similarly, molar proportions of SCFAwere affected by S. cerevisiae supplementation only in thecolon contents. Molar proportions of acetic:butyric andpropionic:butyric acids were higher in piglets fed thecontrol diet, than on those fed diets supplemented withS. cerevisiae (P=0.0002; P=0.0003, respectively). Com-pared to the control diet, the cellulolytic activity in thecaecum contents was increased by the prebiotic orprobiotic supplementation, but remained similar whenthe diet was supplemented with the two additivescombined, thus a very significant interaction betweenthe effect of these additives (Pb0.0001). Furthermore,no significant differences were found in the SI andcolon contents for the cellulolytic activity. The xy-lanolytic activity was higher (P=0.0189) in the smallintestine of piglets fed xylo-oligosaccharides, but thedifference was statistically significant only in case ofthe C-XOS+SC diet. On the other hand, no sig-nificant differences were found in the caecum andcolon.4. DiscussionAs the xylo-oligosaccharides can be degrade by theintestinal microflora of the pig (Maxwell et al., 2004), itwas expected that the inclusion of this prebiotic in thediet could increase the levels of SCFA in the intestinalcontents. However, in our experiment, the xylo-oligosaccharides mixture was included at the level of20 g·kg−1, while Mikkelsen et al. (2003) have testedprebiotic mixtures (FOS, TOS) at levels of 40 g·kg−1.Therefore, considering the level of total non-digestibleoligosaccharides (NDO) of the control diet, the amountof additive we have used was probably too low to havean effect on the SCFA levels. The low level of totalSCFA in the colon of piglets receiving the S. cerevisiaesupplemented diets suggests a decrease in the microbialactivity in the hindgut. Previous results from Mathewet al. (1998) have showed no effects of live yeastsupplementation on SCFA concentrations along thegastrointestinal tract of the weaning piglets. It is likelythat S. cerevisiae could degrade dietary NDO in theileum, reducing the level of fermentable substratesreaching the hindgut and consequently the microbialpopulation in the colon. Nevertheless, the absence ofeffect of S. cerevisiae on the concentration of SCFA inthe SI does not confirm this hypothesis. The low SCFAconcentration found in the colon contents of pigletsreceiving the C-XOS+SC diet failed to demonstrate apositive effect, on the intestinal microbial population, ofcombining the two additives. According to Nemcovaet al. (1999) the benefit of associating prebiotics toprobiotics in the diet is a complex problem, and dependson the prebiotic composition and the specific live microorganisms used.The effect of the two tested additives on theenzymatic activities of intestinal contents must beoverlooked with caution, because of the high residualstandard deviation of these results. Nevertheless, thehigher xylanolytic activity in the SI of C-XOS+SC fedpiglets and the higher cellulolytic activity in the caecumof the piglets fed C-XOS or C-SC diets, suggest agreater microbial activity in these compartments,although these results are not confirmed by the SCFAconcentrations.In conclusion, the inclusion of S. cerevisiae in thediet has significantly decreased the SCFA concentrationand the molar proportion of butyric acid in the colon.Results on the effect of xylo-oligosaccharides orS. cerevisiae supplementation on the intestinal fermen-tative activity have show high variability. A positiveeffect of combining the two additives on microbialenzymes was found only in the case of xylanolyticactivity of the small intestine contents.AcknowledgmentsThe authors are grateful to the Commission of theEuropean Communities and the FEDER Funds forfinancial support of this work (Project POCTI/CVT/37545/2002).ReferencesAnderson, D.B., McCracken, R.T., Aminov, J.M., Simpson, R.,Mackie, M., Verstegen, M.W.A., Gaskins, H.R., 1999. Gutmicrobiology and growth-promoting antibiotics in swine. PigsNew Inf. 20, 115–122.Falcão-e-Cunha, L., Peres, H., Freire, J.P.B., Castro-Solla, L., 2004.Effects of alfalfa, wheat bran or beet pulp, with or withoutsunflower oil, on caecal fermentation and on digestibility in therabbit. Anim. Feed Sci. and Technol. 117, 131–149.Heugten, E. van., Funderburke, D.W., Dorton, K.L., 2003. Growthperformance, nutrient digestibility, and fecal microflora inweanling pigs fed live yeast. J. Anim. Sci. 81, 1004–1012.Jouany, J.P., 1982. Dosage des acides gras volatile (AGV) et desalcools dans les contenus digestifs, les jus d'ensilage, les cultures238 M.C. Marinho et al. / Livestock Science 108 (2007) 236–239Author's   personal   copybactérienne et les contenus de fermenteurs anaerobies. Sci.Aliments 2, 131–144.Mathew, A.G., Chattin, S.E., Robbins, C.M., Golden, D.A., 1998.Effects of a direct-fed yeast culture on enteric microbialpopulations, fermentation acids, and performance of weanlingpigs. J. Anim. Sci. 76, 2138–2145.Maxwell, F.J., Duncan, H.S., Hold, G., Stewart, C.S., 2004. Isolation,growth on prebiotics and prebiotic potential of novel bifidobacteriafrom pigs. Anaerobe 10, 33–39.Midtvedt, T., 1994. In vivo models for short-chain fatty acidproduction. In: Binder, H.J., Cummings, J., Soergel, K. (Eds.),Short-chain fatty acids. Falk Symposium, vol. 73. KluwerAcademic Publishers, Dordrecht, pp. 61–68.Mikkelsen, L.L., Jakobsen, M., Jensen, B.B., 2003. Effects of dietaryoligosaccharides on microbial diversity and fructo-oligosaccharidedegrading bacteria in faeces of piglets post-weaning. Anim. FeedSci. and Technol. 109, 133–150.Montagne, L., Pluske, J.R., Hampson, D.J., 2003. A review ofinteractions between dietary fibre and the intestinal mucosa, andtheir consequences on digestive health in young non-ruminantanimals. Anim. Feed Sci. and Technol. 108, 95–117.Nemcova, R., Bomba, A., Gancarcikova, S., Herich, R., Guba, P.,1999. Study of the effect of Lactobacillus paracasei and fructo-oligosaccharides on the faecal microflora in weanling piglets. Berl.Munch Tierdrzil. Wschr. 112, 225–228.Newman, K.E., Newman, M.C., 2001. Evaluation of manannoligosaccharides on the microflora and immunoglobulin status ofsows and piglet performance. J. Anim. Sci. 79 (Suppl.1), 189(Abstr.).Sakata, T., 1987. Stimulatory effect of short-chain fatty acids onepithelial cell proliferation in the intestine: a possible explanationfor trophic effects of fermentable fibre, gut microbes and luminaltrophic factors. Br. J. Nutr. 58, 95–103.SAS, 2001. SAS User's Guide: Statistics. Version 8.02 ed. SAS Inst.Inc., Cary, NC.239M.C. Marinho et al. / Livestock Science 108 (2007) 236–239

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Microbial activity in the gut of piglets: effect of prebiotic and probiotic supplementation

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