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Dietary l-threonine supplementation attenuates lipopolysaccharide-induced inflammatory responses and intestinal barrier damage of broiler chickens at an early age

  • Yueping Chen (a1), Hao Zhang (a1), Yefei Cheng (a1), Yue Li (a1), Chao Wen (a1) and Yanmin Zhou (a1)...


This study was conducted to investigate the protective effects of l-threonine (l-Thr) supplementation on growth performance, inflammatory responses and intestinal barrier function of young broilers challenged with lipopolysaccharide (LPS). A total of 144 1-d-old male chicks were allocated to one of three treatments: non-challenged broilers fed a basal diet (control group), LPS-challenged broilers fed a basal diet without l-Thr supplementation and LPS-challenged broilers fed a basal diet supplemented with 3·0 g/kg l-Thr. LPS challenge was performed intraperitoneally at 17, 19 and 21 d of age, whereas the control group received physiological saline injection. Compared with the control group, LPS challenge impaired growth performance of broilers, and l-Thr administration reversed LPS-induced increase in feed/gain ratio. LPS challenge elevated blood cell counts related to inflammation, and pro-inflammatory cytokine concentrations in serum (IL-1β and TNF-α), spleen (IL-1β and TNF-α) and intestinal mucosa (jejunal interferon-γ (IFN-γ) and ileal IL-1β). The concentrations of intestinal cytokines in LPS-challenged broilers were reduced by l-Thr supplementation. LPS administration increased circulating d-lactic acid concentration, whereas it reduced villus height, the ratio between villus height and crypt depth and goblet density in both jejunum and ileum. LPS-induced decreases in jejunal villus height, intestinal villus height:crypt depth ratio and ileal goblet cell density were reversed with l-Thr supplementation. Similarly, LPS-induced alterations in the intestinal mRNA abundances of genes related to intestinal inflammation and barrier function (jejunal toll-like receptor 4, IFN- γ and claudin-3, and ileal IL-1 β and zonula occludens-1) were normalised with l-Thr administration. It can be concluded that l-Thr supplementation could attenuate LPS-induced inflammatory responses and intestinal barrier damage of young broilers.


Corresponding author

* Corresponding author: Y. Zhou, fax +86 25 84395314; email


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1. Alexander, C & Rietschel, ET (2001) Invited review: bacterial lipopolysaccharides and innate immunity. J Endotoxin Res 7, 167202.
2. Lu, YC, Yeh, WC & Ohashi, PS (2008) LPS/TLR4 signal transduction pathway. Cytokine 42, 145151.
3. Tan, J, Liu, S, Guo, Y, et al. (2014) Dietary l-arginine supplementation attenuates lipopolysaccharide-induced inflammatory response in broiler chickens. Br J Nutr 111, 13941404.
4. Li, Y, Zhang, H, Chen, YP, et al. (2015) Bacillus amyloliquefaciens supplementation alleviates immunological stress and intestinal damage in lipopolysaccharide-challenged broilers. Anim Feed Sci Technol 208, 119131.
5. Li, Y, Zhang, H, Chen, YP, et al. (2015) Bacillus amyloliquefaciens supplementation alleviates immunological stress in lipopolysaccharide-challenged broilers at early age. Poult Sci 94, 15041511.
6. Liu, L, Shen, J, Zhao, C, et al. (2015) Dietary Astragalus polysaccharide alleviated immunological stress in broilers exposed to lipopolysaccharide. Int J Biol Macromol 72, 624632.
7. Zhang, X, Zhao, L, Cao, F, et al. (2013) Effects of feeding fermented Ginkgo biloba leaves on small intestinal morphology, absorption, and immunomodulation of early lipopolysaccharide-challenged chicks. Poult Sci 92, 119130.
8. Kaiser, MG, Block, SS, Ciraci, C, et al. (2012) Effects of dietary vitamin E type and level on lipopolysaccharide-induced cytokine mRNA expression in broiler chicks. Poult Sci 91, 18931898.
9. Gilani, S, Howarth, GS, Kitessa, SM, et al. (2016) New biomarkers for intestinal permeability induced by lipopolysaccharide in chickens. Anim Product Sci 56, 19841997.
10. Groschwitz, KR & Hogan, SP (2009) Intestinal barrier function: molecular regulation and disease pathogenesis. J Allergy Clin Immunol 124, 320.
11. Wu, QJ, Zhou, YM, Wu, YN, et al. (2013) The effects of natural and modified clinoptilolite on intestinal barrier function and immune response to LPS in broiler chickens. Vet Immunol Immunopathol 153, 7076.
12. Tenenhouse, HS & Deutsch, HF (1966) Some physical-chemical properties of chicken γ-globulins and their pepsin and papain digestion products. Immunochemistry 3, 1120.
13. Schaart, MW, Schierbeek, H, van der Schoor, SRD, et al. (2005) Threonine utilization is high in the intestine of piglets. J Nutr 135, 765770.
14. van der Schoor, SRD, Wattimena, DL, Huijmans, J, et al. (2007) The gut takes nearly all: threonine kinetics in infants. Am J Clin Nutr 86, 11321138.
15. Floc’h, NL & Sève, B (2007) Catabolism through the threonine dehydrogenase pathway does not account for the high first-pass extraction rate of dietary threonine by the portal drained viscera in pigs. Br J Nutr 93, 447456.
16. van der Sluis, M, Schaart, MW, de Koning, BA, et al. (2009) Threonine metabolism in the intestine of mice: loss of mucin 2 induces the threonine catabolic pathway. J Pediatr Gastroenterol Nutr 49, 99107.
17. Wang, W, Zeng, X, Mao, X, et al. (2010) Optimal dietary true ileal digestible threonine for supporting the mucosal barrier in small intestine of weanling pigs. J Nutr 140, 981986.
18. Law, GK, Bertolo, RF, Adjiri-Awere, A, et al. (2007) Adequate oral threonine is critical for mucin production and gut function in neonatal piglets. Am J Physiol Gastrointest Liver Physiol 292, G1293G1301.
19. Hamard, A, Mazurais, D, Boudry, G, et al. (2010) A moderate threonine deficiency affects gene expression profile, paracellular permeability and glucose absorption capacity in the ileum of piglets. J Nutr Biochem 21, 914921.
20. Chee, SH, Iji, PA, Choct, M, et al. (2010) Functional interactions of manno-oligosaccharides with dietary threonine in chicken gastrointestinal tract. I. Growth performance and mucin dynamics. Br Poult Sci 51, 658666.
21. Chee, SH, Iji, PA, Choct, M, et al. (2010) Functional interactions of manno-oligosaccharides with dietary threonine in chicken gastrointestinal tract. II. Mucosal development, mucin dynamics and nutrient utilisation. Br Poult Sci 51, 667676.
22. Zhang, Q, Chen, X, Eicher, SD, et al. (2016) Effect of threonine deficiency on intestinal integrity and immune response to feed withdrawal combined with coccidial vaccine challenge in broiler chicks. Br J Nutr 116, 20302043.
23. Wils-Plotz, EL, Jenkins, MC & Dilger, RN (2013) Modulation of the intestinal environment, innate immune response, and barrier function by dietary threonine and purified fiber during a coccidiosis challenge in broiler chicks. Poult Sci 92, 735745.
24. National Research Council (1994) Nutrient Requirements of Poultry, 9th rev. ed. Washington, DC: National Academies Press.
25. Corzo, A, Kidd, MT, Dozier, IWA, et al. (2007) Dietary threonine needs for growth and immunity of broilers raised under different litter conditions. J Appl Poult Res 16, 574582.
26. Star, L, Rovers, M, Corrent, E, et al. (2012) Threonine requirement of broiler chickens during subclinical intestinal Clostridium infection. Poult Sci 91, 643652.
27. Taghinejad-Roudbaneh, M, Babaee, MJ, Afrooziyeh, M, et al. (2013) Estimation of dietary threonine requirement for growth and immune responses of broilers. J Appl Anim Res 41, 474483.
28. Min, YN, Liu, SG, Qu, ZX, et al. (2017) Effects of dietary threonine levels on growth performance, serum biochemical indexes, antioxidant capacities, and gut morphology in broiler chickens. Poult Sci 96, 12901297.
29. Chen, YP, Cheng, YF, Li, XH, et al. (2017) Effects of threonine supplementation on the growth performance, immunity, oxidative status, intestinal integrity, and barrier function of broilers at the early age. Poult Sci 96, 405413.
30. Zhang, Q, Chen, X, Eicher, SD, et al. (2017) Effect of threonine on secretory immune system using a chicken intestinal ex vivo model with lipopolysaccharide challenge. Poult Sci 96, 30433051.
31. Gonzalez, D, Mustacich, DJ, Traber, MG, et al. (2011) Early feeding and dietary lipids affect broiler tissue fatty acids, vitamin E status, and cyclooxygenase-2 protein expression upon lipopolysaccharide challenge. Poult Sci 90, 27902800.
32. Wang, X, Li, Y, Shen, J, et al. (2015) Effect of Astragalus polysaccharide and its sulfated derivative on growth performance and immune condition of lipopolysaccharide-treated broilers. Int J Biol Macromol 76, 188194.
33. Hosoda, N, Nishi, M, Nakagawa, M, et al. (1989) Structural and functional alterations in the gut of parenterally or enterally fed rats. J Surg Res 47, 129133.
34. Horn, NL, Donkin, SS, Applegate, TJ, et al. (2009) Intestinal mucin dynamics: response of broiler chicks and White Pekin ducklings to dietary threonine. Poult Sci 88, 19061914.
35. Livak, KJ & Schmittgen, TD (2001) Analysis of relative gene expression data using real-time quantitative pcr and the 2−ΔΔCT Method. Methods 25, 402408.
36. Zheng, XC, Wu, QJ, Song, ZH, et al. (2016) Effects of Oridonin on growth performance and oxidative stress in broilers challenged with lipopolysaccharide. Poult Sci 95, 22812289.
37. Zhang, X, Zhong, X, Zhou, Y, et al. (2010) Dietary RRR-α-tocopherol succinate attenuates lipopolysaccharide-induced inflammatory cytokines secretion in broiler chicks. Br J Nutr 104, 17961805.
38. Vichaya, EG, Hunt, SC & Dantzer, R (2014) Lipopolysaccharide reduces incentive motivation while boosting preference for high reward in mice. Neuropsychopharmacology 39, 28842890.
39. Spurlock, ME (1997) Regulation of metabolism and growth during immune challenge: an overview of cytokine function. J Anim Sci 75, 17731783.
40. Dhabhar, FS, Miller, AH, McEwen, BS, et al. (1995) Effects of stress on immune cell distribution. Dynamics and hormonal mechanisms. J Immunol 154, 5511.
41. Shen, YB, Piao, XS, Kim, SW, et al. (2010) The effects of berberine on the magnitude of the acute inflammatory response induced by Escherichia coli lipopolysaccharide in broiler chickens. Poult Sci 89, 1319.
42. Wang, X, Shen, J, Li, S, et al. (2014) Sulfated Astragalus polysaccharide regulates the inflammatory reaction in LPS-infected broiler chicks. Int J Biol Macromol 69, 146150.
43. Beutler, B (2000) Tlr4: central component of the sole mammalian LPS sensor. Curr Opin Immunol 12, 2026.
44. Miller, SI, Ernst, RK & Bader, MW (2005) LPS, TLR4 and infectious disease diversity. Nat Rev Microbiol 3, 3646.
45. Shan, M, Gentile, M, Yeiser, JR, et al. (2013) Mucus enhances gut homeostasis and oral tolerance by delivering immunoregulatory signals. Science 342, 447453.
46. Nichols, NL & Bertolo, RF (2008) Luminal Threonine concentration acutely affects intestinal mucosal protein and mucin synthesis in piglets. J Nutr 138, 12981303.
47. Faure, M, Choné, F, Mettraux, C, et al. (2007) Threonine utilization for synthesis of acute phase proteins, intestinal proteins, and mucins is increased during sepsis in rats. J Nutr 137, 18021807.
48. Rémond, D, Buffière, C, Godin, JP, et al. (2009) Intestinal inflammation increases gastrointestinal threonine uptake and mucin synthesis in enterally fed minipigs. J Nutr 139, 720726.
49. Li, P, Yin, YL, Li, D, et al. (2007) Amino acids and immune function. Br J Nutr 98, 237252.
50. Wang, X, Qiao, SY, Liu, M, et al. (2006) Effects of graded levels of true ileal digestible threonine on performance, serum parameters and immune function of 10–25kg pigs. Anim Feed Sci Technol 129, 264278.
51. Defa, L, Changting, X, Shiyan, Q, et al. (1999) Effects of dietary threonine on performance, plasma parameters and immune function of growing pigs. Anim Feed Sci Technol 78, 179188.
52. Ren, M, Liu, XT, Wang, X, et al. (2014) Increased levels of standardized ileal digestible threonine attenuate intestinal damage and immune responses in Escherichia coli K88+ challenged weaned piglets. Anim Feed Sci Technol 195, 6775.
53. Sun, XQ, Fu, XB, Rong, Z, et al. (2001) Relationship between plasma D(-)-lactate and intestinal damage after severe injuries in rats. World J Gastroenterol 7, 555558.
54. Guo, S, Al-Sadi, R, Said, HM, et al. (2013) Lipopolysaccharide causes an increase in intestinal tight junction permeability in vitro and in vivo by inducing enterocyte membrane expression and localization of TLR-4 and CD14. Am J Pathol 182, 375387.
55. Capaldo, CT & Nusrat, A (2009) Cytokine regulation of tight junctions. Biochim Biophys Acta 1788, 864871.
56. Al-Sadi, R, Ye, D, Dokladny, K, et al. (2008) Mechanism of IL-1β-induced increase in intestinal epithelial tight junction permeability. J Immunol 180, 56535661.
57. Turner, JR (2009) Intestinal mucosal barrier function in health and disease. Nat Rev Immunol 9, 799809.
58. Bein, A, Zilbershtein, A, Golosovsky, M, et al. (2017) LPS induces hyper-permeability of intestinal epithelial cells. J Cell Physiol 232, 381390.
59. Smirnova, MG, Guo, L, Birchall, JP, et al. (2003) LPS up-regulates mucin and cytokine mRNA expression and stimulates mucin and cytokine secretion in goblet cells. Cell Immunol 221, 4249.
60. Andrianifahanana, M, Moniaux, N & Batra, SK (2006) Regulation of mucin expression: mechanistic aspects and implications for cancer and inflammatory diseases. Biochim Biophys Acta 1765, 189222.
61. Garg, P, Ravi, A, Patel, NR, et al. (2007) Matrix metalloproteinase-9 regulates muc-2 expression through its effect on goblet cell differentiation. Gastroenterology 132, 18771889.
62. Sodhi, CP, Neal, MD, Siggers, R, et al. (2012) Intestinal epithelial toll-like receptor 4 regulates goblet cell development and is required for necrotizing enterocolitis in mice. Gastroenterology 143, 708718.e705.
63. Kim, YS & Ho, SB (2010) Intestinal goblet cells and mucins in health and disease: recent insights and progress. Curr Gastroenterol Rep 12, 319330.



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