FNS  Vol.2 No.4 , June 2011
The Influence of Apple- or Red-Grape Pomace Enriched Piglet Diet on Blood Parameters, Bacterial Colonisation, and Marker Gene Expression in Piglet White Blood Cells
Abstract: Proanthocyanidins and flavanoids, both subfamilies of the polyphenols, are highly concentrated in different fruits and berries as well as in fruit pomace. They have shown to exhibit anti-cancer, anti-microbial, anti-oxidative, and im- mune-modulatory effects in vertebrates. Herein the effect of additional apple pomace or red-grape pomace in conven- tional piglet starter feeds were investigated in 36 young growing piglets. Immunological marker gene expression was quantified by quantitative real-time RT-PCR in white blood cells, and intestinal bacterial flora was investigated from weaning to three weeks post weaning. Polyphenol content in red-grape pomace, gut content and tissues were analyzed with HPLC. Flavan-3-ols (epicatechin and catechin) and proanthocyanidins (B1, B2 and C1) were identified in the gas- tro-intestinal tract content, whereas only traces could be detected in various piglet organs. The blood parameters, he- moglobin and hematocrit, were affected and down-regulated in all groups over testing period. In both pomace treated groups more thrombocytes were present compared to the standard feeding group. It turns out, that the pomace diets had greatest impact on the bacterial content in the colon. Results demonstrate that feeding apple pomace and red- grape pomace tended to increase the number of total colonic bacteria. Steptococci/Enterococci increased in the red- grape pomace. C. perfringens was not detectable at the second time point. The number of lactobacilli increased in both applied diets. The number of Clostridium perfringens decreased with the age of the piglets. Trends of mRNA expression changes were found in white blood cell (WBC) between different feeding regimens, since the expression variability in the groups was very high. Between the different time points there were significant differences within the apple pomace group, where TNF? (p = 0.033), NF?B (p = 0.024) and Caspase 3 (p = 0.019) mRNA expression increased signifi- cantly during treatment. We conclude that both polyphenol rich feedings have the potential to positively influence the intestinal flora, blood parameters, and WBC mRNA gene expression pattern of immunological marker genes.
Cite this paper: nullJ. Sehm, D. Treutter, H. Lindermayer, H. Meyer and M. Pfaffl, "The Influence of Apple- or Red-Grape Pomace Enriched Piglet Diet on Blood Parameters, Bacterial Colonisation, and Marker Gene Expression in Piglet White Blood Cells," Food and Nutrition Sciences, Vol. 2 No. 4, 2011, pp. 366-376. doi: 10.4236/fns.2011.24052.

[1]   E. Revilla and M. Ryan, “Analysis of Several Phenolic Compounds with Potential Antioxidant Properties in Grape Extracts and Wines by High-Performance Liquid Chromatography-Photodiode Array Detection without Sa- mple Preparation,” Journal of Chromatography A, Vol. 881, No. 1-2, 2000, pp. 461-469. doi:10.1016/S0021-9673(00)00269-7

[2]   K. Stoll, “Der Apfel,” Verlag Negiri, Zürich, 1997.

[3]   J. Boyer and R. H. Liu, “Apple Phytochemicals and Their Health Benefits,” Nutrition Journal, Vol. 12, No. 3, May 2004, pp. 3-5.

[4]   A. Schieber, F. C. Stintzing and R. Carle, “By Products of Plant Food Processing as a Source of Functional Compounts-Recent Developments,” Trends in Food Science & Technology, Vol. 12, No. 11, March 2001, pp. 401-413. doi:10.1016/S0924-2244(02)00012-2

[5]   M. Palma and L. T. Taylor, “Extraction of Polyphenolic Compounds from Grape Seeds with near Critical Carbon Dioxide,” Journal of Chromatography A, Vol. 849, No. 1, July 1999, pp. 117-124. doi:10.1016/S0021-9673(99)00569-5

[6]   J. M. Souquet, B. Labarbe, C. Le Guerneve, V. Cheynier and M. Moutounet, “Phenolic Composition of Grape Ste- ms,” Journal of Agricultural and Food Chemistry, Vol. 48, No. 4, April 2000, pp. 1076-1080. doi:10.1021/jf991171u

[7]   S. Sembries, G. Dongowski, K. Mehrlander, F. Will and H. Dietrich, “Dietary Fiber-Rich Colloids from Apple Pomace Extraction Juices Do not Affect Food Intake and Blood Serum Lipid Levels, but Enhance Fecal Excretion of Steroids in Rats,” The Journal of Nutritional Biochemistry, Vol. 15, No. 5, May 2004, pp. 296-302. doi:10.1016/j.jnutbio.2003.12.005

[8]   C. Santos-Buelga and A. Scalbert, “Proanthocyanidins and Tannin-Like Compounds: Nature, Occurence, Dietary Intake and Effects on Nutrition and Health,” Journal of the Science of Food and Agriculture, Vol. 80, No. 7, May 2000, pp. 1094-1117. doi:10.1002/(SICI)1097-0010(20000515)80:7<1094::AID-JSFA569>3.0.CO;2-1

[9]   F. M. Aarestrup, “Association between the Consumption of Antimicrobial Agents in Animal Husbandry and the Occurrence of Resistant Bacteria among Food Animals,” International Journal of Antimicrobial Agents, Vol. 12, No. 4, August 1999, pp. 279-285. doi:10.1016/S0924-8579(99)90059-6

[10]   K. E. Bach Knudsen, “Development of Antibiotic Resistance and Options to Replace Antimocrobials in Animals Diets,” Proceedings of the Nutrition Society, Vol. 60, No. 3, March 2001, pp. 291-299.

[11]   A. C. Francois, “Mode of Action of Antibiotics on Growth,” World Review of Nutrition and Dietetics, Vol. 3, March 1961, pp. 21-64.

[12]   W. J. Visek, “The Mode of Growth Promotion by Antibiotics,” Journal of Animal Science, Vol. 46. No. 5, August 1978, pp. 1447-1469.

[13]   D. B. Anderson, V. J. McCracken, R. I. Aminov, J. M. Simpson, R. I. Mackie, M. W. A. Verstegen and H. R. Gaskins, “Gut Microbiology and Growth-Promoting Antibiotics in Swine,” Pig News and Information, Vol. 20, No. 4, 1999, pp. 115-122.

[14]   D. Treutter, C. Santos-Buelga, M. Gutmann and H. Kolodziej, “Identification of Flavan-3-Olsand Procyanidins by High-Performance Liquid Chromatography and Che- mical Reaction Detection,” Journal of Chromatography A, Vol. 667, No. 1-2, April 1994, pp. 290-297. doi:10.1016/0021-9673(94)89078-1

[15]   M. Gutmann and W. Feucht, “A New Method for Selective Localization of Flavan-3-Ols in Plant Tissues Involving Glycolmethacrylate Embedding and Microwave Irradiation,” Histochemistry, Vol. 96, No. 1, 1991, pp. 83- 86. doi:10.1007/BF00266765

[16]   U. Mayr, D. Treutter, C. Santos-Buelga, H. Bauer and W. Feucht, “Developmental Changes in the Phenol Concentrations of ‘Golden Delicious’ Apple Fruits and Leaves,” Phytochemistry, Vol. 38, No. 5, March 1995, pp. 1151- 1155. doi:10.1016/0031-9422(94)00760-Q

[17]   C ?lschl?ger, J. Milde, H. Schempp and D. Treutter, “Polyphenols and Antioxidant Capacity of Sorbus Domestica L. Fruits,” Journal of Applied Botany and Food Quality, Vol. 78, No. 2, 2004, pp. 112-116.

[18]   T. D. Schmittgen, “Real Time Quantitative PCR,” Methods, Vol. 25, No. 4, December 2001, pp. 383-385. doi:10.1006/meth.2001.1260

[19]   K. J. Livak and T. D. Schmittgen, “Analysis of Relative gene Expression Data Using Real-Time Quantitative PCR and the 2-??CT Method,” Methods, Vol. 25, No. 4, December, pp. 402-408. doi:10.1006/meth.2001.1262

[20]   S. Bollmann, “Untersuchungen zur Wirkung Nichtantibiotischer Futterzus?tze auf die Darmflora Sowie den Verlauf Einer Experimentellen Escheria Coli-Bzw. Salmonella Derby-Infektion bei Schweinen,” Ph.D. Thesis, Tier?rztliche Hochschule Hannover, Hanover, 2002.

[21]   S. Stat, “User’s Manual,” Version 5.0, Jandel Scientific Software, Chicago, 1995.

[22]   J. Sehm, H. Lindermayer, H. H. D. Meyer and M. W. Pfaffl, “The Influence of Apple- and Red-Wine Pomace Rich Diet on mRNA Expression of Inflammatory and Apoptotic Markers in Different Piglet Organs,” Animal Science, Vol. 82, March 2006, pp. 877-887. doi:10.1017/ASC200699

[23]   J. Sehm, H. Lindermayer, C. Dummer, D. Treutter and M. W. Pfaffl, “The Influence of Apple Pomace or Red-Wine Pomace Rich Diet on the Gut Morphology in Weaning Piglets,” Journal of Animal Physiology and Animal Nutrition, Vol. 91, No. 7-8, August 2007, pp. 289-296. doi:10.1111/j.1439-0396.2006.00650.x

[24]   D. W. Funderburke and R. W. Seerley, “The Effects of Postweaning Stressors on Pig Weight Change, Blood, Liver and Digestive Tract Characteristics,” Journal of Animal Science, Vol. 68, No. 1, January 1990, pp. 155- 162.

[25]   A. Scalbert and G. Williamson, “Dietary Intake and Bioavailability of Polyphenols,” Journal of Nutrition, Vol. 130, Supplement 8S, January 2000, pp. 2073-2085.

[26]   A. Scalbert, C. Morand, C. Manach and C. Rémésy, “Absortption and Metabolism of Polyphenols in the Gut and Impact on Health,” Biomedicine & Pharmacotherapy, Vol. 56, No. 6, August 2002, pp. 276-282. doi:10.1016/S0753-3322(02)00205-6

[27]   H. Schneider, A. Schwiertz, M. D. Collins and M. Blaut, “Anaerobic Transformation of Quercetin-3-Glucoside by Bacteria from the Human Intestinal Tract,” Archive of Microbiology, Vol. 171, No. 2, January 1999, pp. 81-91. doi:10.1007/s002030050682

[28]   T. Mitsuoka, “Intestinal Flora and Aging,” Nutrition Reviews, Vol. 50, No. 12, December 1992, pp. 438-446. doi:10.1111/j.1753-4887.1992.tb02499.x

[29]   A. Bezkorovainy, “Probiotics: Determinants of Survival and Growth in the Gut,” American Journal of Clinical Nutrition, Vol. 73, No. 2, February 2001, pp. 399-405.

[30]   H. Hara, N. Orita, S. Hatano, H. Ichikawa, Y. Hara, N. Matsumoto, Y. Kimura, A. Terada and T. Mitsuoka, “Effect of Tea Polyphenols on Fecal Flora and Fecal Metabolic Products of Pigs,” The Journal of Veterinary Medical Science, Vol. 57, No. 1, February 1995, pp. 45-49.

[31]   E. Bone, A. Tamm and M. Hill, “The Production of Urinary Phenols by Gut Bacteria and Their Possible Role in Causation of Large Bowel Cancer,” The American Journal of Clinical Nutrition, Vol. 29, No. 12, December 1976, pp. 1448-1454.

[32]   A. Vince, P. F. Down, J. Murison, F. J. Twigg and O. M. Wrong, “Generation of Ammonia from Non-Urea Sources in a Fecal Incubation System,” Clinical Science and Molecular Medicine, Vol. 51, No. 3, September 1976, pp. 313-322.

[33]   V. Baud and M. Karin, “Signal Transduction by Tumor Necrosis Factor and Its Relatives,” Trends in Cell Biology, Vol. 11, No. 9, September 2001, pp. 372-377. doi:10.1016/S0962-8924(01)02064-5

[34]   H. Y. Chang and X. Yang, “Proteases for Cell Suicide Functions and Regulation of Caspases,” Microbiology and Molecular Biology Reviews, Vol. 64, No. 4, December 2000, pp. 821-846. doi:10.1128/MMBR.64.4.821-846.2000

[35]   E. Shaulian and M. Karin, “AP-1 in Cell Proliferation and Survival,” Oncogene, Vol. 20, No. 19, April 2001, pp. 2390-2400. doi:10.1038/sj.onc.1204383

[36]   M. Philpott and L. R. Ferguson, “Immunonutrition and cancer,” Mutation Research, 2001, Vol. 551, No. 1-2, July 2004, pp. 29-42.

[37]   C. Velik-Salchner, C. Schnurer, D. Fries, P. R. Mussigang, P. L. Moser, W. Streif, C. Kolbitsch and I. H. Lorenz, “Normal Values for Thrombelastography (ROTEM) and Selected Coagulation Parameters in Porcine Blood,” Thrombosis Research, Vol. 117, No. 5, May 2006, pp. 597-602. doi:10.1016/j.thromres.2005.05.015

[38]   G. G. Gomez, O. Phillips and R. A. Goforth, “Effect of Immunoglobulin Source on Survival, Growth, and Hematological and Immunological Variables in Pigs,” Journal of Animal Science, Vol. 76, No. 1, January 1998, pp. 1-7.

[39]   R. A. McCance and E. M. Widdowson, “The Effect of Colostrums on the Composition and Volume of the Plasma of Newborn Piglets,” The Journal of Physiologie, Vol. 145, No. 3, March 1959, pp. 547-550.

[40]   N. C. Jain, “Schlam′s Veterinary Hematology,” 4th Edition, Lea and Febiger, Philadelphia, 1986.

[41]   S. S. Bentivegna and K. M. Whitney, “Subchronic 3- -Month Oral Toxicity Study of Grape Seed and Grape Skin Extracts,” Food and Chemical Toxicology, Vol. 40, No. 12, December 2002, pp. 1731-1743. doi:10.1016/S0278-6915(02)00155-2

[42]   L. Vellenga, T. Wensing, H. J. Breukink and F. H. Hagens, “Effects of Irradiated Sow Colostrum on Some Biochemical and Haematological Measurements in Newborn Piglets,” Research in Veterinary Science, Vol. 41, No. 3, November 1986, pp. 316-318.

[43]   C. E. Thorn, “Normal Hematology of the Pig,” In: B. F. Feldman, J. G. Zinkl and N. C. Jain, Eds., Schalm’s Veterinary Hematology, 5th Edition, Lippincott Williams & Wilkins, Philadelphia, 2000, pp. 1089-1095.

[44]   E. Middleton, Jr., C. Kandaswami and T. C. Theoharides, “The Effects of Plant Flavonoids on Mammalian Cells: Implications for Inflammation, Heart Disease, and Cancer,” Pharmacological Reviews, Vol. 52, No. 4, December 2000, pp. 673-751.

[45]   J. D. Folts, “Potential Health Benefits from the Flavonoids in Grape Products on Vascular Disease,” Advances in Experimental Medicine and Biology, Vol. 505, April 2002, pp. 95-111.