Comparative Clinical Pathology

, Volume 26, Issue 2, pp 343–349 | Cite as

Identification of some plasma biomarkers associated with early weaning stress in crossbred piglets

  • U. K. De
  • S. Nandi
  • Reena Mukherjee
  • G. K. Gaur
  • M. R. Verma
Original Article


The present study was undertaken to examine the physiological effects of early weaning of crossbred piglets and identify a panel of plasma-based biomarkers. Eighteen crossbred (Landrace × indigenous) suckling piglets were assigned to one of the two treatments on the day of weaning: (1) control (n = 9), piglets were loose-housed with their dam and (2) weaned (n = 9), piglets were abruptly selected from their dam and kept in a separate pen. Ad libitum water was supplied to all pens for drinking and all the piglets received same concentrate diet with free access. The blood was collected from anterior vena cava on d −3, 1, 3, 7, and 15 relative to weaning (day 1) and measured for oxidative stress indicators (total antioxidant; TA and nitric oxide; NOx), acute phase proteins (haptoglobin; Hp and ceruloplasmin; Cp), high mobility group box 1 protein (HMGB1), inflammatory cytokines (interferon-gamma;IFN-γ, tumor necrosis factor- alpha;TNF-α, and interleukin-1beta; IL-1β), and chemokines (intercellular adhesion molecule-1; ICAM-1, monocyte chemoattractant protein-1;MCP-1 and transforming growth factor beta 1; TGF-β1). It is concluded that MCP-1, Hp, and HMGB1 protein are the potential biomarkers besides NOx and IL-1β which are the sensitive indicator of physiological stress in early weaned piglets. In the future, the determination of these markers would definitely help in identifying the weaned piglets with substantial stress condition which in turn help to find out the strategies for effective and efficient management of piglets to make the pig husbandry an economically profitable, environmentally sustainable, and ecologically viable industry.


HMGB1 ICAM-1 IFN-γ Il-1β MCP-1 TNF-α Stress Weaning piglet 


  1. Alexander TJ, Thornton K, Boon G, Lysons RJ, Gush AF (1980) Medicated early weaning to obtain pigs free from pathogens endemic in the herd of origin. Vet Rec 106:114–119CrossRefPubMedGoogle Scholar
  2. Al-Sadi RM, Thomas YM (2007) IL-1β causes an increase in intestinal epithelial tight junction permeability. J Immunol 178:4641–4649CrossRefPubMedPubMedCentralGoogle Scholar
  3. Al-Sadi R, Boivin M, Ma T (2009) Mechanism of cytokine modulation of epithelial tight junction barrier. Front Biosci 14:2765–2778CrossRefGoogle Scholar
  4. Arthington JD, Eicher SD, Kunkle WE, Martin FG (2003) Effect of transportation and commingling on the acute-phase protein response, growth, and feed intake of newly weaned beef calves. J Anim Sci 81:1120–1125CrossRefPubMedGoogle Scholar
  5. Bonnette ED, Kornegay ET, Lindemann MD, Hammerberg C (1990) Humoral and cell-mediated immune response and performance of weaned pigs fed supplemental vitamin E levels and housed at two nursery temperatures. J Anim Sci 68:1337–1345CrossRefPubMedGoogle Scholar
  6. Boudry G, Peron V, Le Huerou-Luron I, Lalles JP, Sève B (2004) Weaning induces both transient and long-lasting modifications of absorptive, secretory, and barrier properties of piglet intestine. J Nutr 134:2256–2262PubMedGoogle Scholar
  7. Brockmeier SL, Halbur PG, Thacker EL (2002) Porcine respiratory disease complex. In: Brogden KA, Guthmiller JM (eds) Polymicrobial diseases. ASM Press, Washington (DC)Google Scholar
  8. Campbell JM, Crenshaw JD, Polo J (2013) The biological stress of early weaned piglets. J Anim Sci Biotechnol 4:19CrossRefPubMedPubMedCentralGoogle Scholar
  9. Chen G, Li J, Ochani M, Rendon-Mitchell B, Qiang X, Susarla S, Ulloa L, Yang H, Fan S, Goyert SM, Wang P, Tracey KJ, Sama AE, Wang H (2004) Bacterial endotoxin stimulates macrophages to release HMGB1 partly through CD14 and TNF-dependent mechanisms. J Leukoc Biol 76:994–1001CrossRefPubMedGoogle Scholar
  10. Francisco CJ, Shryock TR, Bane DP, Unverzagt L (1996) Serum haptoglobin concentration in growing swine after intranasal challenge with Bordetella bronchiseptica and toxigenic Pasteurella multocida type D. Can J Vet Res 60:222–227PubMedPubMedCentralGoogle Scholar
  11. Gruys E, Toussaint MJM, Niewold TA, Koopmans SJ (2005) Acute phase reaction and acute phase proteins. J Zhejiang Univ Sci B 6:1045–1056CrossRefPubMedPubMedCentralGoogle Scholar
  12. Hohenshell LM, Cunnick JE, Ford SP, Kattesh HG, Zimmerman DR, Wilson ME, Matteri RL, Carroll JA, Lay DC Jr (2000) Few differences found between early- and late-weaned pigs raised in the same environment. J Anim Sci 78:38–49CrossRefPubMedGoogle Scholar
  13. Howe KL, Reardon C, Wang A, Nazli A, McKay DM (2005) Transforming growth factor-beta regulation of epithelial tight junction proteins enhances barrier function and blocks en- terohemorrhagic Escherichia coli O157:H7-induced increased permeability. Am J Pathol 167:1587–1597CrossRefPubMedPubMedCentralGoogle Scholar
  14. Hu CH, Xiao K, Luan ZS, Song J (2013) Early weaning increases intestinal permeability, alters expression of cytokine and tight junction proteins, and activates mitogen-activated protein kinases in pigs. J Anim Sci 91:1094–1101CrossRefPubMedGoogle Scholar
  15. Huang LF, Yao YM, Sheng ZY (2012) Novel insights for high mobility group box 1 protein-mediated cellular immune response in sepsis: a systemic review. World J Emerg Med 3:165–171CrossRefPubMedPubMedCentralGoogle Scholar
  16. Jiang R, Chang X, Stoll B, Fan MZ, Arthington J, Weaver E, Campbell J, Burrin DG (2000) Dietary plasma protein reduces small intestinal growth and lamina propria cell density in early weaned pigs. J Nutr 130:21–26PubMedGoogle Scholar
  17. Kalinina N, Agrotis A, Antropova Y, DiVitto G, Kanellakis P, Kostolias G, Ilyinskaya O, Tararak E, Bobik A (2004) Increased expression of the DNA-binding cytokine HMGB1 in human atherosclerotic lesions. Role of activated macrophages and cytokines. Arterioscler Thromb Vasc Biol 24:2320–2325CrossRefPubMedGoogle Scholar
  18. Kato S, Hussein MH, Kakita H, Goto T, Daoud GA, Kato T, Sugiura T, Nobata M, Nakajima Y, Endo T, Mizuno K, Ito T, Kato I, Suzuki S, Togari H (2009) Edaravone, a novel free radical scavenger, reduces high-mobility group box 1and prolongs survival in a neonatal sepsis model. Shock 32:586–592CrossRefPubMedGoogle Scholar
  19. Kim MH, Yang JY, Upadhaya SD, Lee HJ, Yun CH, Jong KH (2011) The stress of weaning influences serum levels of acute-phase proteins, iron-binding proteins, inflammatory cytokines, cortisol, and leukocyte subsets in Holstein calves. J Vet Sci 12:151–157CrossRefPubMedPubMedCentralGoogle Scholar
  20. Kojima CJ, Kattesth HG, Roberts MP, Sun T (2008) Physiological and immunological responses to weaning and transport in the young pig: modulation by administration of porcine somatotropin. J Anim Sci 86:2913–2919CrossRefPubMedGoogle Scholar
  21. Laine TM, Lyytikäinen T, Yliaho M, Anttila M (2008) Risk factors for post-weaning diarrhoea on piglet producing farms in Finland. Acta Vet Scand 50:21CrossRefPubMedPubMedCentralGoogle Scholar
  22. Liu Y (2015) Fatty acids, inflammation and intestinal health in pigs. J Anim Sci Biotechnol 6:41CrossRefPubMedPubMedCentralGoogle Scholar
  23. Luo Z, Zhu W, Guo Q, Luo W, Zhang J, Xu W, Xu J (2016) Weaning induced hepatic oxidative stress, apoptosis, and aminotransferases through MAPK signaling pathways in piglets. Oxid Med Cell Longev 2016. doi:10.1155/2016/4768541
  24. Madsen K, Lewis SA, Tavernini MM, Hibbard J, Fedorak RN (1997) Interleukin 10 prevents cytokine-induced disruption of T84 monolayer barrier integrity and limits chloride secretion. Gastroenterol 113:151–159CrossRefGoogle Scholar
  25. McCracken BA, Spurlock ME, Roos MA, Zuckermann FA, Gaskins HR (1999) Weaning anorexia may contribute to local inflammation in the piglet small intestine. J Nutr 129:613–619PubMedGoogle Scholar
  26. Mei J, Xu RJ (2005) Transient changes of transforming growth factor-b expression in the small intestine of the pig in association with weaning. Br J Nutr 93:37–45CrossRefPubMedGoogle Scholar
  27. Moeser AJ, Ryan KA, Nighot PK, Blikslager AT (2007) Gastrointestinal dysfunction induced by early weaning is attenuated by delayed weaning and mast cell blockade in pigs. Am J Physiol Gastrointest Liver Physiol 293:G413–G421CrossRefPubMedGoogle Scholar
  28. National Research Council (1998) Nutrient requirements of swine, 10th edn. National Academic Press, USA, Washington, DCGoogle Scholar
  29. Nofrarías M, Manzanilla EG, Pujols J, Gibert X, Majo N, Segales J, Gasa J (2006) Effects of spray-dried porcine plasma and plant extracts on intestinal morphology and on leukocyte cell subsets of weaned pigs. J Anim Sci 84:2735–2742CrossRefPubMedGoogle Scholar
  30. O’Loughlin A, McGee M, Doyle S, Earley B (2014) Biomarker responses to weaning stress in beef calves. Res Vet Sci 97:459–464Google Scholar
  31. Pacher P, Beckman JS, Liaudet L (2007) Nitric oxide and peroxynitrite in health and disease. Physiol Rev 87:315–424CrossRefPubMedPubMedCentralGoogle Scholar
  32. Palone F, Vitali R, Cucchiara S, Pierdomenico M, Negroni A, Aloi M, Nuti F, Felice C, Armuzzi A, Stronati L (2014) Role of HMGB1 as a suitable biomarker of subclinical intestinal inflammation and mucosal healing in patients with inflammatory bowel disease. Inflam Bowel Dis 20:1448–1457CrossRefGoogle Scholar
  33. Pie S, Lalles JP, Blazy F, Laffitte J, Seve B, Oswald IP (2004) Weaning is associated with an upregulation of expression of inflammatory cytokines in the intestine of piglets. J Nutr 134:641–647PubMedGoogle Scholar
  34. Pluske JR, Hampson DJ, Williams IH (1997) Factors influencing the structure and function of the small intestine in the weaned pig: a review. Livest Prod Sci 51:215–236CrossRefGoogle Scholar
  35. Pomorska-Mól M, Kwit K, Markowska-Daniel I (2012) Major acute phase proteins in pig serum from birth to slaughter. Bull Vet Inst Pulawy 56:553–557Google Scholar
  36. Salak-Johnson JL, McGlone JJ, Norman RL (1996) In vivo glucocorticoid effects on porcine natural killer cell activity and circulating leukocytes. J Anim Sci 74:584–592CrossRefPubMedGoogle Scholar
  37. Sauerwein H, Schmitz S, Hiss S (2005) The acute phase protein haptoglobin and its relation to oxidative status in piglets undergoing weaning-induced stress. Redox Rep 10:295–302CrossRefPubMedGoogle Scholar
  38. Shinde PL, Dass RS, Garg AK, Chaturvedi VK (2007) Immune response and plasma alpha tocopherol and selenium status of male buffalo (Bubalus bubalis) calves supplemented with vitamin E and selenium. Asian Australas J Anim Sci 20:1539–1545CrossRefGoogle Scholar
  39. Spreeuwenberg MA, Verdonk JM, Gaskins HR, Verstegen MW (2001) Small intestine epithelial barrier function is compromised in pigs with low feed intake at weaning. J Nutr 131:1520–1527PubMedGoogle Scholar
  40. Sunderman SW Jr, Nomoto S (1970) Measurement of human serum ceruloplasmin by its p-phenylenediamine oxidase activity. Clin Chem Acta 16:903–910Google Scholar
  41. Sutherland MA, Niekamp SR, Rodriguez-Zaz SL, Salakjohnson JL (2006) Impacts of chronic stress and social status on various physiologi-cal and performance measures in pigs of different breeds. J Anim Sci 84:588–596CrossRefPubMedGoogle Scholar
  42. Tang D, Shi Y, Kang R, Li T, Xiao W, Wang H, Xiao X (2007) Hydrogen peroxide stimulates macrophages and monocytes to actively release HMGB1. J Leukoc Biol 81:741–747CrossRefPubMedGoogle Scholar
  43. Villena J, Kitazawa H (2014) Modulation of intestinal TLR4-inflammatory signaling pathways by probiotic microorganisms: lessons learned from Lactobacillus jensenii TL2937. Front Immunol 4:512CrossRefPubMedPubMedCentralGoogle Scholar
  44. Vitali R, Stronati L, Negroni A, Di-Nardo G, Pierdomenico M, del Giudice E, Rossi P, Cucchiara S (2011) Fecal HMGB1 is a novel marker of intestinal mucosal inflammation in pediatric inflammatory bowel disease. Am J Gastroenterol 106:2029–2040CrossRefPubMedGoogle Scholar
  45. Wang J, Chen L, Li P, Li X, Zhou H, Wang F, Li D, Yin Y, Wu G (2008) Gene expression is altered in piglet small intestine by weaning and dietary glutamine supplementation. J Nutr 138:1025–1032CrossRefPubMedGoogle Scholar
  46. Westly HJ, Kelley KW (1984) Physiologic concentrations of cortisol suppress cell-mediated immune events in the domestic pig. Proc Soc Exp Biol Med 177:156–164CrossRefPubMedGoogle Scholar
  47. Wu F, Xiong X, Yang H, Yao K, Duan Y, Wang X, Tan B, Li T, Xiao L, Hou Y, Wu G, Yin Y (2017) Expression of proteins in intestinal middle villus epithelial cells of weanling piglets. Front Biosci 22:539–557CrossRefGoogle Scholar
  48. Xiao K, Song ZH, Jiao LF, Ke YL, Hu CH (2014) Developmental changes of TGF-b1 and Smads signaling pathway in intestinal adaption of weaned pigs. PLoS One 9:e104589CrossRefPubMedPubMedCentralGoogle Scholar
  49. Yin J, Wu MM, Xiao H, Ren WK, Duan JL, Yang G, Li TJ, Yin YL (2014) Development of an antioxidant system after early weaning in piglets. J Anim Sci 92:612–619CrossRefPubMedGoogle Scholar
  50. Yucel AA, Gulen S, Dincer S, Yucel AE, Yetkin GI (2012) Comparison of two different applications of the Griess method for nitric oxide measurement. J Exp Integr Med 2:167–171CrossRefGoogle Scholar
  51. Zhu LH, Zhao KL, Chen XL, Xu JX (2012) Impact of weaning and an antioxidant blend on intestinal barrier function and antioxidant status in pigs. J Anim Sci 90:2581–2589CrossRefPubMedGoogle Scholar
  52. Zuo J, Ling B, Long L, Li T, Lahaye L, Yang C, Feng D (2015) Effect of dietary supplementation with protease on growth performance, nutrient digestibility, intestinal morphology, digestive enzymes and gene expression of weaned piglets. Anim Nutr 1:276–282CrossRefGoogle Scholar

Copyright information

© Springer-Verlag London 2016

Authors and Affiliations

  • U. K. De
    • 1
  • S. Nandi
    • 2
  • Reena Mukherjee
    • 1
  • G. K. Gaur
    • 3
  • M. R. Verma
    • 4
  1. 1.Division of MedicineIndian Veterinary Research InstituteIzatnagarIndia
  2. 2.CADRADIndian Veterinary Research InstituteIzatnagarIndia
  3. 3.Livestock Production and Management SectionIndian Veterinary Research InstituteIzatnagarIndia
  4. 4.Division of Livestock Economics, Statistics and Information TechnologyIndian Veterinary Research InstituteIzatnagarIndia

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