A Di-D-Fructose Dianhydride-Enriched Caramel Modulates Pig Fecal Microbiota Composition

Luis A. Rubio, María Jesús Peinado, Ana Echávarri, Raquel Ruiz, Elena Suárez-Pereira, Carmen Ortiz Mellet, José M. García Fernández

Departamento de Química Orgánica, Facultad de Química, Universidad de Sevilla, Sevilla, Spain.
Fisiología y Bioquímica de la Nutrición Animal (EEZ, CSIC), Granada, Spain.
Instituto de Investigaciones Químicas (IIQ), CSIC—Universidad de Sevilla, Sevilla, Spain.

DOI: 10.4236/aim.2014.45031                

Abstract

A correlation has been previously described between bifidobacteria counts before and after the use of a dietary additive in human studies. However, to our knowledge no information on this topic has yet been reported in animals, and no information exists either on similar possible correlations of bacterial groups other than bifidobacteria. The potential prebiotic effects of di-D-fructose dianhydride (DFA)-enriched caramels have been previously reported in laboratory animals, poultry and pigs. In the present work, twelve growing male castrated pigs (41.8 ± 1.9 kg mean BW) were fed in succession on a control (no additive) or DFA-enriched caramel (20 g/kg) containing diets. Another group of 10 pigs (38.0 ± 3.7 kg mean BW) fed on a control diet without any additive was used as negative control. Bacterial log₁₀ number of copies of the 16S rRNA gene was determined in fecal samples by using qPCR. Increased (P< 0.05) lactobacilli,Clostridium coccoides/Eu-bacterium rectaleand bacteroides group log₁₀ number of copies were determined in fecal samples of pigs fed on the caramel containing diet compared with non-caramel controls. In addition, for all bacterial groups studied microbiological values co-variated with initial counts and, except for enterobacteria, variations in the fecal bacterial numbers after caramel supplementation correlated (P< 0.05) with the fecal numbers before supplementation. In conclusion, the supplementation of pig diets with DFA-enriched caramels induced significant increases in the fecal number of copies of bacterial groups regarded as beneficial, and variations in the fecal number of copies correlated with the initial fecal number of copies.

Keywords

Di-D-Fructose Dianhydrides, Fecal Microbiota, Correlations, Pig, Prebiotics

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Greko, C. (2001) Safety Aspects on Non-Use of Antimicrobials as Growth Promoters. In: Piva, A., Bach Knudsen, K.E. and Lindberg, J.E., Eds., Gut Environment of Pigs, Nottingham University Press, Nottingham, 219-230.
[2]	Aarestrup, F.M. (2003) Effects of Termination of AGP Use on Antimicrobial Resistance in Food Animals. Working Papers for the WHO International Review Panels Evaluation, Document WHO/CDS/CPE/ZFK/2003.1a, World Health Organization, Geneva, 6-11.
[3]	Ortiz Mellet, C. and García Fernández, J.M. (2010) Difructose Dianhydrides (DFAs) and DFA Enriched Products as Functional Foods. Topics in Current Chemistry,294,49-77. http://dx.doi.org/10.1007/ 128_2010_50
[4]	Arribas, B., Suárez-Pereira, E., Ortiz Mellet, C., García Fernández, J.M., Buttersack, C., Rodríguez-Cabezas, M.E., Garrido-Mesa, N., Bailon, E., Guerra-Hernández, E., Zarzuelo A. and Gálvez, J. (2010) Di-D-Fructose-Enriched Caramels: Effect on Colon Microbiota, Inflammation, and Tissue Damage in Trinitrobenzenesulfonic Acid-Induced Colitic Rats. Journal of Agricultural and Food Chemistry, 58, 6476-6484. http://dx.doi.org/10.1021/jf100513j
[5]	Orban, J.I., Patterson, J.A., Sutton, A.L. and Richards, G.N. (1997) Effect of Sucrose Thermal Oligosaccharide Caramel, Dietary Vitamin-Mineral Level, and Brooding Temperature on Growth and Intestinal Bacterial Populations of Broiler Chickens. Poultry Science, 76, 482-490. http://dx.doi.org/10.1093/ps/76.3.482
[6]	Orban, J.I., Patterson, J.A., Sutton, A.L. and Richards, G.N. (1996) Growth Performance and Intestinal Microbial Populations of Growing Pigs Fed Diets Containing Sucrose Thermal Oligosaccharide Caramel. Journal of Animal Science, 74, 170-175.
[7]	Roberfroid, M.B. (2007) Prebiotics: The Concept Revisited. Journal of Nutrition, 137, 830S-837S.
[8]	Davis, L.M.G., Martínez, I., Walter, J. and Hutkins, W. (2010) A Dose Dependent Impact of Prebiotic Galactooligo-saccharides on the Intestinal Microbiota of Healthy Adults. International Journal of Food Microbiology, 144, 285-292.http://dx.doi.org/10.1016/ j.ijfoodmicro.2010.10.007
[9]	Metges, C.C. (2010) Classical and Post-Genomic Methods to Study GIT Function with Emphasis on the Pig. Livestock Science, 133, 10-19. http://dx.doi.org/10.1016/j.livsci.2010.06.014
[10]	Defaye, J. and García Fernández, J.M. (1994) Protonic and Thermal Activation of Sucrose and the Oligosaccharide Composition of Caramel. Carbohydrate Research, 256, C1-C4. http://dx.doi.org/10.1016/0008-6215(94)84219-1
[11]	Defaye, J. and García Fernández, J.M. (1995) The Oligosaccharide Components of Caramel. Zuckerindustrie, 120, 700-704.
[12]	Rubio, E.M., Gómez, M., Ortiz-Mellet, C., García-Fernández, J.M., Zarzuelo, A., Gálvez J.J. and Duval, R. (2007) Novel Caramels with a High Prebiotic Oligosaccharide Content. ES Patent WO2008107506.
[13]	Suárez-Pereira, E., Rubio, E.M., Pilard, S., Ortiz-Mellet, C. and García-Fernández, J.M. (2010) Di-D-Fructose Dian-hydrade (DFA)-Enriched Caramels by Acid Ion-Exchange Resin-Promoted Caramelization of D-Fructose: Chemical Analysis. Journal of Agricultural and Food Chemistry, 58, 1777-1787. http://dx.doi.org/10.1021/jf903354y
[14]	Ruiz, R. and Rubio, L.A. (2009) Lyophyllization Improves the Extraction of PCR-Quality Community DNA from Pig Fecal Samples. Journal of the Science of Food and Agriculture, 89, 723-727. http://dx.doi.org/10.1002/jsfa.3465
[15]	Candela, M., Maccaferri, S., Turoni, S., Carnevali, P. and Brigidi, P. (2010) Functional Intestinal Microbiome, New Frontiers in Prebiotic Design. International Journal of Food Microbiology, 140, 93-101. http://dx.doi.org/10.1016/j.ijfoodmicro.2010.04.017
[16]	Gaggìa, F., Mattarelli, P. and Biavati, B. (2009) Probiotics and Prebiotics in Animal Feeding for Safe Food Production. International Journal of Food Microbiology, 141, S15-S28. http://dx.doi.org/10.1016/j.ijfoodmicro.2010.02.031
[17]	Mikkelsen, L.L., Bendixen, C., Jakobsen, M. and Jensen, B.B. (2003) Enumeration of Bifidobacteria in Gastrointestinal Samples from Piglets. Applied and Environmental Microbiology, 69, 654-658. http://dx.doi.org/10.1128/AEM.69.1.654-658.2003
[18]	Kleessen, B., Hartman, L. and Blaut, M. (2001) Oligofructose and Long Chain Inulin Influence the Gut Microbial Ecology of Rats Associated with a Human Fecal Flora. British Journal of Nutrition, 86, 291-300.-http://dx.doi.org/10.1079/BJN2001403
[19]	Gibson, G.R., Probert, H.M., Van Loo, J., Rastall, R.A. and Roberfroid, M.B. (2004) Dietary Modulation of the Human Colonic Microbiota: Updating the Concept of Prebiotics. Nutrition Research Reviews, 17, 259-275.-http://dx.doi.org/10.1079/NRR200479
[20]	Roberfroid, M.B., Van Loo, J.A.E. and Gibson, G.R. (1998) The Bifidogenic Nature of Chicory Inulin and Its Hydrolysis Products. Journal of Nutrition, 128, 11-19.
[21]	Hidaka, H., Eida, T., Takizawa, T., Tokunaga, T. and Tashiro, Y. (1986) Effects of Fructo-Oligosaccharides on Intestinal Flora and Human Health. Bifidobacteria and Microflora, 5, 37-50.
[22]	Rycroft, C.E., Jones, M.R., Gibson, G.R. and Rastall, R.A. (2001) Fermentation Properties of Gentio-Oligosaccharides. Letters in Applied Microbiology, 32, 156-161. http://dx.doi.org/10.1046/j.1472-765x.2001.00875.x
[23]	Simpson, J.M., McCracken, V.J., Rex Gaskins, H. and Mackie, R.I. (2000) Denaturing Gradient Gel Electrophoresis Analysis of 16S Ribosomal DNA Amplicons to Monitor Changes in Fecal Bacterial Populations of Weaning Pigs after Introduction of Lactobacillus reuteri Strain MM53. Applied and Environmental Microbiology, 66, 4705-4714. -http://dx.doi.org/10.1128/AEM.66.11.4705-4714.2000
[24]	Janczyk, P., Pieper, R., Smidt, H. and Souffrant, B. (2010) Effect of Alginate and Inulin on Intestinal Microbial Ecology of Weanling Pigs Reared under Different Husbandry Conditions. FEMS Microbiology Ecology, 72, 132-142. http://dx.doi.org/10.1111/j.1574-6941.2009.00826.x
[25]	Guarner, F., Khan, A.G., Garisch, J., Eliakim, R., Gangl, A., Thomson, A., Krabshuis, J. and Lemair, T. (2011) Probiotics and Prebiotics. World Gastroenterology Organisation, Milwaukee.
[26]	Guo, X., Xia, X., Tang, R., Zhou, J., Zhao, H. and Wang, K. (2008) Development of a Real-Time PCR Method for Firmicutes and Bacteroidetes in Feces and Its Application to Quantify Intestinal Population of Obese and Lean Pigs. Letters in Applied Microbiology, 47, 367-373. http://dx.doi.org/10.1111/j.1472-765X.2008.02408.x
[27]	Delroisse, J.M., Bolvin, A.L., Parmentier, I., Dauphin, R.D., Vandenbol, M. and Portetelle, D. (2008) Quantification of Bifidobacterium spp. and Lactobacillus spp. in Rat Fecal Samples by Real-Time PCR. Microbiology Research,163,663-670. http://dx.doi.org/ 10.1016/j.micres.2006.09.004
[28]	Castillo, M., Martin-Orue, S.M., Manzanilla, E.G., Badiola, I., Martin, M. and Gasa, J. (2006) Quantification of Total Bacteria, Enterobacteria and Lactobacilli Populations in Pig Digesta by Real-Time PCR. Veterinary Microbiology, 144, 165-170. http://dx.doi.org/10.1016/j.vetmic.2005.11.055
[29]	Layton, A., McKay, L., Williams, D., Garrett, V., Gentry, R. and Sayler, G. (2006) Development of Bacteroides 16S rRNA Gene Taqman-Based Real-Time PCR Assays for Estimation of Total, Human and Bovine Fecal Pollution in Water. Applied and Environmental Microbiology, 72, 4214-4224. http://dx.doi.org/10.1128/AEM.01036-05
[30]	Matsuki, T., Watanabe, K., Fujimoto, J., Takada, T. and Tanka, R. (2004) Use of 16S rRNA Gene-Targeted Group-Specific Primers for Real-Time PCR Analysis of Predominant Bacteria in Human Feces. Applied and Environmental Microbiology, 70, 7220-7228. http://dx.doi.org/10.1128/AEM.70.12.7220-7228.2004
[31]	Malinen, E., Kassinen, A., Rinttilä, T. and Palva, A. (2003) Comparison of Real-Time PCR with SYBR Green I or 5’-Nuclease Assays and Dot-Blot Hybridization with rDNA-Targeted Oligonucleotide Probes in Quantification of Selected Fecal Bacteria. Microbiology, 149, 269-277. http://dx.doi.org/ 10.1099/mic.0.25975-0.