Early weaning predisposes the pig intestine to structural and functional alterations, due to the increase in E. coli populations. These bacteria use the lipopolysaccharide (LPS) derived from their cell wall as an important pathogenic factor. Little is known about the effects of LPS on the intestinal morphology. Such knowledge could be helpful in understanding the pathogenesis of post-weaning enteritis, which is needed to design therapeutic strategies. Objective: this study aimed to evaluate the effects of the oral intake of LPS on the morphology of intestinal villi and glands of weaned pigs. Methods: the study used 52 pigs weaned at 21 days. The animals were fed a basal diet added with four levels of LPS (0.0, 0.3, 0.5 and 1.0 µg/mg of food) for 10 days. Pigs were sequentially slaughtered on days 1, 5, 7 and 10 after weaning, and samples of small intestine were taken to evaluate morphological parameters by computerized image analysis. The statistical design used was randomized blocks in a 4x4 factorial arrangement. Results: results showed that LPS decreases the height and area of intestinal villi, and increases the width of the villi and the depth and width of the intestinal glands. These effects probably contribute to a decreased intestinal nutrient absorption and increase co-infection with other pathogens, thus leading to the post-weaning diarrhea syndrome. Conclusions: this study stresses the usefulness of computerized morphometric analysis to evaluate the effect of LPS on intestinal morphology, so it may be used in future studies to investigate the pathophysiology of the causative agents of enteritis and to evaluate therapeutic strategies.
Key words: histology, LPS lipopolysaccharide, morphometry, small intestine, weaned pigs.
Albin DM, Wubben JE, Rowlett JM, Tappenden KA, and Nowak RA. Changes in small intestinal nutrient transport and barrier function after lipopolysaccharide exposure in two pig breeds. J Anim Sci 2007; 85:2517-2523.
Amador P, Garcia-Herrera J, Marca MC, de la Osada J, Acin S, Navarro MA, Salvador MT, Lostao MP and Rodriguez-Yoldi MJ. Intestinal D-galactose transport in an endotoxemia model in the rabbit. J Membr Biol 2007; 2153:125-133.
Berkes, J, Viswanathan, VK, Savkovic, SD, and Hecht G. Intestinal epithelial responses to enteric pathogens: effects on tight junction barrier, ion transport, and inØ ammation. Gut 2003; 52:439-451.
Council for International Organizations of Medical Sciences (CIOMS). International Guiding Principles for Biomedical Research Involving Animals. 1985. Geneva . 28pp.
Cramwell PD. Development of neonatal gut and enzyme systems. In Varley MA Editor. The neonatal pig: development and survival. UK Cab International, 1995. p. 99-145.
Fan MZ. Growth and ontogeny of the gastrointestinal tract. In: Xu. RJ, Cranwell P editors. The neonatal pig. Gastrointestinal physiology. and nutrition. Nottingham University Press. 2002:31-60.
GarcÌa-Herrera J, Abad B, RodrÌguez-Yoldi MJ. Effect of lipopolysaccharide on D-fructose transport across rabbit jejunum. InØ amm Res 2003; 52:177-184.
Garcia-Herrera J, Marca MC, Brot-Laroche EGuillen, N, Acin S, Navarro MA, de la Osada J, Rodriguez-Yoldi MJ. Protein kinases, TNF- and proteasome contribute in the inhibition of fructose intestinal transport by sepsis in vivo. Am J Physiol Gastrointest Liver Physiol 2008; 294:G155ñG164.
GÛmez A. Weaning and the physiology of piglets. In: I International Seminar on sustainable production systems in small animals. Universidad del Cauca. Popay·n, 2006. 34p.
Gomez AS, Vergara D, Argote F. Effect of diet and age of weaning on piglet digestive physiology. Revista BiotecnologÌa en el Sector Agropecuario y Agronindustrial 2008; 6:32-41.
Guerra ACM, Gal·n OJA, MÈndez AJJ, Murillo AE. EvaluaciÛn del efecto del extracto de orÈgano (Oreganum vulgare) sobre algunos par·metros productivos de cerdos destetos. Revista Tumbaga 2008; 3:16-29.
Hedemann MS, Hojsgaard s, Jensen BB. Small intestine morphology and activity of intestinal peptidases in piglets around weaning. J Anim Physiol Anim Nutr 2003; 87:32-41.
Hedemann MS, Eskildsen M, LÊrke HN, Pedersen CJ, Lindberg E, Laurinen P, Bach KE. Fiber concentrations and Æ ber properties Intestinal morphology and enzymatic activity in newly weaned pigs fed contrasting. J Anim Sci 2006; 84:1375- 1386.
Jin L, Reynolds LP, Redmer DA, Caton JS, Crenshaw JD. Effects of dietary Æ ber on intestinal growth, cell proliferation, and morphology in growing pigs. J Anim Sci 1994; 72:2270- 2278.
Johnson GB, Brunn GJ, Samstein B. New insight into the pathogenesis of sepsis and the sepsis syndrome. Surgery 2005; 137:393ñ395
LallËs JPl, Boudry G, Favier C, Le Flocíh N, Luron I, Montagne L, Oswald IP, PiÈ S, Piel C and SËve B. Gut function and dysfunction in young pigs: physiology. Anim Res 2004a; 53:301-316.
LallËs JP, Konstantinov SR, Rothkˆtter HJ. Bases physiologiques, microbiologiques et immunitaires des troubles digestifs du sevrage chez le porcelet : don nÈes rÈcentes dans le contexte de la suppression des antibiotiques additifs alimentaires. JournÈes Recherche Porcine 2004b; 36:139-150.
Li BT, Van Kessel AG, Caine WR, Huang SX and Kirkwood RN. Small intestine morphology and bacterial populations in ileal digesta and feches of newly weanead pigs receiving a high dietary level of zinc oxide. Can J Anim 2001; 81:511-516.
McCracken BA, Spurlock ME, Roos MA, Zuckermann FA, Gaskins HR. Weaning anorexia may contribute to local inØ ammation in the piglet small intestine. J Nutr 1999; 129:613ñ619.
Makkink CA, Berntsen PJM, Op Den Kamp BML, Kemp B, Verstegen WA. Gastric protein breakdown and pancreatic enzyme activities in response to two different dietary protein sources in newly weaned pigs. J Anim Sci 1994; 72:2843-2850.
Marion J, Biernat M, Thomas F, Savary G, Le Breton Y, Zabielski R, Le HuÎrou-Luron I, Le Dividich J. Small intestine growth and morphometry in piglets weaned at 7 days of age. Effects of level of energy intake. Reprod. Nutr. Dev 2002; 42:339-354.
Nabuurs MJA, Hoogendoorn A, Van der Molen EJ, Van Osta LM. Villus height and crypt depth in weanead and unweanead pigs, reared under various circumstances in the Netherlands. Res Vet Sci 1993; 55:78-84.
National Research Council (NRC). Nutrient Requirements of Swine. 10th ed. National Academy Press. Washington: The Institute, 1998.
PiÈ S, LallËs JP, Blazy F, LafÆ tte J, SËve B, Oswald IP. Weaning Is Associated with an Upregulation of Expression of InØ ammatory Cytokines in the Intestine of Piglets. J Nutr 2004; 134:641-647.
Pitman RS, Blumberg RS. First line of defense: the role of the intestinal epithelium as an active component of the mucosal immune system. J Gastroenterol 2000; 35:805-814.
Pluske JR, Hampson DJ, Williams IH. Factors inØ uencing the structure and function of the small intestine in weaned pigs: a review. Livest Prod Sci 1997; 51:215-236.
Pluske JR, Williams IH, Aherne FX. Maintenance of villous height and crypt depth in the small intestine of weaned pigs. En: Manipuilating pig production III. Products with wheat gluten on digestibility of nutrients and growth performance in nursey pigs. J Anim Sci 1991; 72:151-159.
Reis STC , Guerrero CMJ, Aguilera BA, Mariscal LG.Effect of different cereals on intestinal morphology of weaned piglets. Tec Pecu Mex 2005; 43:309-321.
Reis STC, Aguilera, BAM, Aguilera BA, Mariscal LG, Guerrero CMJ. Morphology of the digestive tract of pigs fed diets containing isolated or concentrated soy protein. Arch Latinoam Prod Anim 2007b, 15:139-146.
Reis STC, Mariscal LG, Aguilera BA, Cervantes JGH. Digestibility of protein and energy in diets for piglets supplemented with three different types of dried whey. Vet Mex 2007; 38:141-151.
Ruemmele FM, Russo P, Beaulieu JF, Dionne S, Levy E, Lentze MJ, Seidman EG. Susceptibility to FAS-induced apoptosis in human nontumoral enterocytes: Role of costimulatory factors. J Cell Physiol 1999; 181:45-54.
Rodrigues MMA, Silva ODA, Taketomi, EA, HernandezBlazquez FJ. IgA production, coliforms analysis and intestinal mucosa morphology of piglets that received probiotics with viable or inactivated cells. Pesq Vet Bras 2007; 27:241-245.
SAS. SAS/STAT Userís Guide. Institute Inc. Statistical Analysis Systems Institute. Version 9.1th Ed. Cary, NC.: SAS Institute Inc. 2006.
Steel RG, Torrie JH. Principles and Procedures of Statistics: a biometrical approach. 2 Ed. New York, US: McGraw-Hill Book Co.; 1985.
Touchette KJ, Carroll JA, Allee GL, Matter RL, Dyer CJ, Beausang, LA, Zannelli ME. Effect of spray-dried plasma and lipopolysaccharide exposure on weaned pigs: I. Effects on the immune axis of weaned pigs. J Anim Sci 2002; 80:494-501.
Van Beers-Schreus, HMG, Nabuurs MJA, Vellenga L, Kalsbeek-van der Valk HJ, Wensing T, Breukink HJ. Weaning and the weanling diet inØ uence the villous height and cript 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.
Vente-Spreeuwenberg MAM, Verdonk AC, Gaskins HR, Verstegen MWA. Small intestine epithelial barrier function is compromised in pigs with low feed intake at weaning. J Nutr 2001; 131:1520-1527.
Vente-Spreeuwenberg MAM, Verdonk JMAJ, Bakker GCM, Beynen AC, Verstegen MWA. Effect of dietary protein source on feed intake and small intestine morphology in newly weaned piglets. Livest Prod Sci 2004a. 86:169-177.
Vente-Spreeuwenberg, MAM, Verdonk JMAJ, Koninkx,JFJG, Beynen AC, Verstegen MWA. Dietary protein hydrolysates vs the intact proteins do not enhance mucosal integrity and growth performance in weaned piglets. Livest Prod Sci 2004b; 85:151- 164.
Wang Fengjun, Graham WV, Wang Y, Witkowski ED, Schwarz BT, Turner J. Interferon- and Tumor Necrosis Factor- Synergize to Induce Intestinal Epithelial Barrier Dysfunction by Up-Regulating Myosin Light Chain Kinase Expression. Am J Pathol 2005; 166:409-419.
Willing BP, Van Kessel AG. Enterocyte proliferation and apoptosis in the caudal small intestine is inØ uenced by the composition of colonizing commensal bacteria in the neonatal gnotobiotic pig. J. Anim Sci. 2007; 85:3256-3266.
Yen JT. Anatomy of the Digestive System and Nutritional Physiology. In: Lewis A.J. and L.L. Southern (Ed.). Swine Nutrition 2nd ed. Washington, DC. USA. CRC Press; 2002. p 31.
Yoo D, W Lo, S Goodman, W Ali, C Semrad, M Field. Interferon-gamma downregulates ion transport in murine small intestine cultured in vitro. Am. J. Physiol 2000; 279:G1323ñ G1332.
Yuji H, Takashi A, Hiroshi H, Naotetsu K, Kenji M, Kenji K, Kazuhiko T, Kazuwa N. Alterations of Plasma Ghrelin Levels in Rats with Lipopolysaccharide-Induced Wasting Syndrome and Effects of Ghrelin Treatment on the Syndrome. Endocrinology. 2003; 144: 5365-5371.
Zhenfeng Z, Deyuan O, Xiangshu P, Sung WK, Yanhong L, Junjun W. Dietary Arginine Supplementation Affects Microvascular Development in the Small Intestine of EarlyWeaned. Pigs J.Nutr 2008; 138:1304-1309.