Author details:
1 Auxiliar Professor. Universidad Autónoma de las Américas. Ph.D. student in Agricultural Sciences. Universidad Nacional de Colombia - Sede Medellín - Facultad de Ciencias Agrarias - Grupo de Investigación BIOGEM, A.A. 1779, Medellín, Colombia; 2 Associate Professor. Universidad Nacional de Colombia - Sede Medellín - Facultad de Ciencias Agrarias - Grupo de Investigación BIOGEM, A.A. 1779, Medellín, Colombia; 3 Assistant Professor. Universidad Nacional de Colombia - Sede Medellín - Facultad de Ciencias Agrarias - Grupo de Investigación BIOGEM, A.A. 1779, Medellín, Colombia.
In order to assess the effect of the addition of E. coli lipopolysaccharide (LPS) on the expression of tight junction proteins in the jejunum of weaned piglets (the animals were weaned at 21-days-old), 64 piglets were slaughtered at 1, 5, 7 and 10 days postweaning and complete extraction of jejunum was carried out in order to evaluate the molecular expression of Claudine 3 (C3), Claudine 4 (C4), and Zonula Ocludens-1 (ZO-1). To induce intestinal inflammation, animals were fed a basal diet supplemented with four levels of E. coli LPS (0, 0.3, 0.5, and 1.0 µg mg-1 of feed). The statistical design consisted of randomized blocks in a 4X4 factorial arrangement (four LPS concentrations and four post-weaning periods). A decrease was observed (P<0.01) in the expression of C3, C4, and ZO-1 in the animals that were fed the diet with the highest LPS-inclusion level. LPS contributes to the appearance of anatomical and intestinal functional disorders which are denoted by a decrease in the molecular expression of proteins from the intestinal barrier. This, in turn, is associated with an indiscriminate paracellular transport of molecules, microorganisms, and toxic compounds into the circulatory system. This results in post-weaning diarrhea and a decrease in the productive efficiency of piglets.
Key words: Diarrhea, fever, digestive functions, gastrointestinal pathophysiology.
Amador, P., J. García, M.C. Marca, J. de la Osada, S. Acín, M.A. Navarro, M.T. Salvador, M.P. Lostao and M.J. Rodríguez-Yoldi. 2007. Intestinal D-galactose transport in an endotoxemia model in the rabbit. The Journal of Membrane Biology 215(2-3): 125-133.
Berkes, J., V.K. Viswanathan, S.D. Savkovic and G. Hecht. 2003. Intestinal epithelial responses to enteric pathogens: effects on tight junction barrier, ion transport, and inflammation. Gut 52(3): 439–451.
Bruewer, M., A. Luegering, T. Kucharzik, C.A. Parkos, J.L. Madara, A.M. Hopkins and A. Nusrat. 2003. Proinflammatory cytokines disrupt epithelial barrier function by apoptosis-independent mechanisms. Journal of Immunology 171(11):6164-72.
Bustin, S.A. 2000. Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. Journal of Molecular Endocrinology 25(2): 169-193.
Chin, A.C., D.A. Teoh, K.G. Scott, J.B. Meddings, W.K. Macnaughton and A.G. Buret. 2002. Strain-dependent induction of enterocyte apoptosis by Giardia lamblia disrupts epithelial barrier function in a caspase-3-dependent manner. Infection and Immunity 70(7): 3673-3680.
Council for International Organizations of Medical Sciences. (CIOMS). 1985. International Guiding Principles for Biomedical Research Involving Animals. Geneva. 28 p.
Chow, J., S.M. Lee, Y. Shen, A. Khosravi and S.K. Mazmanian. 2010. Host-bacterial symbiosis in health and disease. Advances in Immunology 107: 243-274.
Davis, M.E., S.C. Sears, J.K. Apple, C.V. Maxwell and Z.B. Johnson. 2006. Effect of weaning age and commingling after the nursery phase of pigs in a wean-to-finish facility on growth, and humoral and behavioral indicators of well-being. Journal of Animal Science 84: 743-756.
Fan, M.Z. 2002. Growth and ontogeny of the gastrointestinal tract. pp. 31-60. In: Xu, R.J. and P. Cranwell (eds.). The neonatal pig: gastrointestinal physiology and nutrition. Nottingham University Press, Nottingham. 360 p.
Freeman, W.M., S.J. Walker and K.E. Vrana. 1999. Quantitative RT-PCR: pitfalls and potential. Biotechniques 26(1): 112-22.
García, J., B. Abad and M.J. Rodríguez-Yoldi. 2003. Effect of lipopolysaccharide on D-fructose transport across rabbit jejunum. Inflammation Research 52(4): 177–184.
García, J, M.C. Marca, E. Brot-Laroche, N. Guillén, S. Acin, M.A. Navarro, J. Osada and M.J. RodríguezYoldi. 2008. Protein kinases, TNF-α, and proteasome contribute in the inhibition of fructose intestinal transport by sepsis in vivo. American Journal of Physiology. Gastrointestinal and Liver Physiology 294(1): G155–G164.
Hataya, Y., T. Akamizu, H. Hosoda, N. Kanamoto, K. Moriyama, K. Kangawa, K. Takaya and K. Nakao. 2003. Alterations of plasma ghrelin levels in rats with lipopolysaccharide-induced wasting syndrome and effects of ghrelin treatment on the syndrome. Endocrinology 144: 5365-5371.
Huggett, J., K. Dheda, S. Bustin and A. Zumla. 2005. Real-time RT-PCR normalisation; strategies and considerations. Genes and Immunity 6(4): 279-84.
Kojima, C.J., J.A. Carroll, R.L. Matteri, K.J. Touchette and G.L. Allee. 2007. Effects of weaning and weaning weight on neuroendocrine regulators of feed intake in pigs. Journal of Animal Science 85(9): 2133–2139
Kubista, M., J.M. Andrade, M. Bengtsson, A. Forootan, J. Jonák, K. Lind, R, Sindelka, R. Sjöback, B. Sjögreen, L. Strömbom, A. Ståhlberg and N. Zoric. 2006. The realtime polymerase chain reaction. Molecular Aspects of Medicine 27(2-3): 95-125.
Lallès, J.P., G. Boudry, C. Favier, N. Le Floc’h, S. Pié, C. Piel and B. Sève, 2004. Gut function and dysfunction in young pigs: physiology. Animal Research 53: 301- 316.
Lamb-Rosteki, J., L. Kalischuk, G. Inglis and A. Buret. 2008. Epidermal growth factor inhibits Campylobacter jejuni-induced Claudin-4 disruption, loss of epithelial barrier funtion, and Escherichia coli translocation. Infection and Immunity 76(8): 3390-3398.
Makkink, C.A., P.J.M. Berntsen, B.M.L. op den Kamp, B. Kemp and W.A. Verstegen. 1994. Gastric protein breakdown and pancreatic enzyme activities in response to two different dietary protein sources in newly weaned pigs. Journal of Animal Science 72(11): 2843-2850.
Mariani, V., S. Palermo, S. Fiorentini, A. Lanubile and E. Giuffra. 2009. Gene expression study of two widely used pig intestinal epithelial cell lines: IPEC-J2 and IPI-2I. Veterinary Immunology and Immunopathology 131(3-4): 278-284.
Marion, J., M. Biernat, F. Thomas, G. Savary, Y. Le Breton, R. Zabielski, I. Le Huërou-Luron and J. Le Dividich, 2002. Small intestine growth and morphometry in piglets weaned at 7 days of age. Effects of level of energy intake. Reproduction, Nutrition, Development 42(4): 339–354.
McLaughlin, J., P. Padfield, J. Burt and C. O´Neil. 2004. Ochratoxin a increases permeability through tight juntions by removal of specific claudin isoforms. American Journal of Physiology. Cell physiology 287(5): C1412-C1417.
Muller, P.Y., H. Janovjak, A.R. Miserez and Z. Dobbie. 2002. Processing of gene expression data generated by quantitative real time RT-PCR. Biotechniques 32(6): 1372-1374, 1376, 1378-1379.
Musch, M.W., M.M. Walsh-Reitz and E.B. Chang. 2006. Roles of ZO-1, occludin, and actin in oxidant-induced barrier disruption. American Journal of Physiology and Gastrointestinal Liver Physiology 290: G222-G23.
National Research Council (NRC). 2012. The nutrient requirements of swine. Eleventh Revised Edition. The National Academy Press, Washington. 400 p.
Parra, S.J.E., T.J. Agudelo, P.D. Sanín, DJ. Forero, L.C. Muskus and H.A. López. 2013. Intestinal expression of pro-inðAammatory cytokines induced by oral intake of lipopolysaccharide (LPS) from E. coli in weaned pigs. Revista Colombiana de Ciencias Pecuarias 26: 108-118.
Paulin, S.M., A. Jagannathan, J. Campbell, T.S. Wallis and M.P. Stevens. 2007. Net replication of Salmonella enterica serovars Typhimurium and Choleraesuis in porcine intestinal mucosa and nodes is associated with their differential virulence. Infection and Immunity 75(8): 3950-3960.
Petersen, Y.M., D.G. Burrin and P.T. Sangild. 2001. GLP2 has differential effects on small intestine growth and function in fetal and neonatal pigs. American Journal of Physiology. Regulatory, Integrative and Comparative Physiology 281(6): 1986-1993.
Pié, S., S.P. Lallès, F. Blazy, J. Laffitte, B. Sève and I.P. Oswald. 2004. Weaning is associated with an upregulation of expression of inflammatory cytokines in the intestine of piglets. Journal of Nutrition 134: 641–647.
Pinton, P., J. Nougayréde, J. Del Rio, C. Moreno, D. Marin, L. Ferrier, A. Bracarence, M. Kolf-Clauw and I. Oswald. 2009. The food contaminant deoxynivalenol, decreases intestinal barrier permeability and reduces claudin expression. Toxicology and Applied Pharmacology 237(1): 41-48.
Pitman, R.S. and R.S. Blumberg. 2000. First line of defense: the role of the intestinal epithelium as an active component of the mucosal immune system. Journal of Gastroenterology 35(11): 805–814.
Pluske, J.R., D.E. Hopwood y D.J. Hampson. 2003. Relación entre la microbiótica intestinal, el pienso y la incidencia de diarreas, y su influencia sobre la salud del lechón tras el destete. pp. 94-108. En: XIX Curso de Especialización FEDNA, Madrid.
Reis, S.T.C., C.M.J. Guerrero, B.A. Aguilera y L.G. Mariscal. 2005. Efecto de diferentes cereales sobre la morfología intestinal de lechones recién destetados. Técnica Pecuaria en México 43(3): 309-321.
Reis, S.T.C., L.G. Mariscal, B.A. Aguilera y J.G.H. Cervantes. 2007. Digestibilidad de la proteína y energía en dietas para lechones complementadas con tres diferentes tipos de suero de leche deshidratado. Revista Veterinaria México 38(2): 141-151.
SASTM. 2006. SAS/STAT User’s Guide. Institute Inc. Statistical Analysis Systems Institute. Version 9.1th edition. Cary, NC: SAS Institute Inc.
Saitou, M., M. Furuse, H. Sasaki, J.D. Schulzke, M. Fromm, H. Takano, T. Noda and S. Tsukita. 2000. Complex phenotype of mice lacking occludin, a component of tight junction strands. Molecular Biology of the Cell 11(12): 4131–4142.
Segalés, J. y M. Domingo. 2003. La necropsia en el ganado porcino, diagnóstico anatomopatológico y toma de muestras. Boehringer Ingelheim, Madrid. 43 p.
Shen, L. and J.R. Turner. 2005. Actin depolymerization disrupts tight junctions via caveolae-mediated endocytosis. Molecular Biology of the Cell 16(9): 3919–3936.
Shifflett, D.E., D.R. Clayburgh, A. Koutsouris, J.R. Turner and G.A. Hecht. 2005. Enteropathogenic E. coli disrupts tight junction barrier function and structure in vivo. Laboratory Investigation; a Journal of Technical Methods and Pathology 85(10): 1308-1324.
Steel, R.G. and J.H. Torrie. 1980. Principles and procedures of statistics: a biometrical approach. Second edition. McGraw-Hill, New York. 633 p.
Tlaskalová-Hogenová, H., R. Stepánková, T. Hudcovic, L. Tucková, B. Cukrowska, R. Lodinová-Zádníková, H. Kozáková, P. Rossmann, J. Bártová, D. Sokol, D.P. Funda, D. Borovská, Z. Reháková, J. Sinkora, J. Hofman, P. Drastich and A. Kokesová. 2004. Commensal bacteria (normal microflora), mucosal immunity and chronic inflammatory and autoimmune diseases. Immunology Letters 93(2-3): 97–108.
Tsukita, S. and M. Furuse. 2002. Claudin-based barrier in simple and stratified cellular sheets. Current Opinion in Cell Biology 14(5):531-536
Vandesompele, J., K. De Preter, F. Pattyn, B. Poppe, N. Van Roy, A. De Paepe and F. Speleman. 2002. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biology 3(7): Research 0034.
Waseem, T., M. Duxbury, H. Ito and M. Robinson. 2008. Exogenous ghrelin modulates release of pro- and antiinflammatory cytokines in LPS-stimulated macrophages through distinct signaling pathways. Surgery 143: 334-342.
Xu, S., J. Lee and M. Miyake. 2011. Expression of ZO-1 and occludin at mRNA and protein level during preimplantation development of the pig parthenogenetic diploids. Zygote 20(2): 147-158.
Yang, E.V., and R. Glaser. 2000. Stress-induced immunomodulation: Impact on immune defenses against infectious disease. Biomedicine and Pharmacotherapy 54: 245-250.
Yu, D., A.M. Marchiando, C.R. Weber, D.R. Raleigh, Y. Wang, L. Shen and J.R. Turner. 2010. MLCK-dependent exchange and actin binding region-dependent anchoring of ZO-1 regulate tight junction barrier function. Proceedings of the National Academy of Sciences of the United States of America 107(18): 8237-8241.