Skip to main content Accessibility help

Expression of heat shock protein 70 in the liver of extensively and intensively kept heavy pigs

  • E. Negrato (a1), G. Di Martino (a2), M. Vascellari (a2), G. Radaelli (a1), K. Capello (a2), F. Pascoli (a1), D. Bertotto (a1) and L. Bonfanti (a2)...


The objective of this work was to investigate the expression of heat shock protein 70 (HSP70) by Western blot (WB) in swine liver. Subsequently, the study aimed to apply this method to two experimental groups of heavy pigs raised in different confinement systems: intensive/indoor (Group A) and extensive/outdoor (Group B). Thirty-six crossbred commercial heavy pigs were divided as follows: Group A (eight castrated males and eight females) was equally distributed into two single-sex indoor pens (1.02 m2/pig); Group B (11 castrated males and nine females) was kept in one single (partially grassy and partially wooded) open area of about 6000 m2. Group A was slaughtered at 41 weeks of age (170 ± 9 kg) and Group B at 48 weeks of age (172 ± 13 kg). At the abattoir the livers of all the animals were collected and analyzed by WB assay in order to quantify the levels of HSP70. Moreover, a further liver sample was taken from the same animals in order to investigate the cellular localization of HSP70 by immunohistochemistry (IHC). The interaction between sex and group resulted statistically significant (P = 0.001). When stratified by sex, Group A showed significantly higher HSP70 values compared with Group B for both male and female subjects (P < 0.001). Stratifying by group, males showed significantly higher HSP70 values than females in Group A (P < 0.001), whereas no statistical differences were observed between sexes for Group B (P = 0.653). The IHC results evidenced cytoplasmic immunoreactivity in a granular pattern in both groups. The different expression pattern observed by WB could prove to be a useful tool in the assessment of pig health and welfare.


Corresponding author


Hide All
Bao, E, Sultan, KR, Nowak, B, Hartung, J 2008. Expression and distribution of heat shock proteins in the heart of transported pigs. Cell Stress and Chaperones 13, 459466.
Bausero, MA, Gastpar, R, Multhoff, G, Asea, A 2005. Alternative mechanism by which IFN-gamma enhances tumor recognition: active release of heat shock protein 72. Journal of Immunology 175, 29002912.
Bradford, MM 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Analytical Biochemistry 72, 248254.
Brambilla, G, Civitareale, C, Ballerini, A, Fiori, M, Amadori, M, Archetti, LI, Regini, M, Betti, M 2002. Response to oxidative stress as a welfare parameter in swine. Redox Report 7, 159163.
Brocchieri, L, Conway de Macario, E, Macario, AJ 2008. Hsp70 genes in the human genome: conservation and differentiation patterns predict a wide array of overlapping and specialized functions. BMC Evolutionary Biology 8, 19.
Ciocca, DR, Osterreich, S, Chamness, GC, McGuire, WL, Fuqua, SAW 1993. Biological and clinical implications of heat shock protein 27000 (HSP27): a review. Journal of the National Cancer Institute 85, 15581570.
Couret, D, Otten, W, Puppe, B, Prunier, A, Merlot, E 2009. Behavioural, endocrine and immune responses to repeated social stress in pregnant gilts. Animal 3, 118127.
Cvoro, A, Korać, A, Matić, G 2004. Intracellular localization of constitutive and inducible heat shock protein 70 in rat liver after in vivo heat stress. Molecular and Cellular Biochemistry 265, 2735.
D'Eath, RB, Turner, SP, Kurt, E, Evans, G, Thölking, L, Looft, H, Wimmers, K, Murani, E, Klont, R, Foury, A, Ison, SH, Lawrence, AB, Mormède, P 2010. Pigs’ aggressive temperament affects pre-slaughter mixing aggression, stress and meat quality. Animal 4, 604616.
de Jong, C, Prelle, IT, van de Burgwal, JA, Lambooij, E, Korte, SM, Blokhuis, HJ 2000. Effects of environmental enrichment on behavioral responses to novelty, learning, and memory, and the circadian rhythm in cortisol in growing pigs. Physiology & Behavior 68, 571578.
De Rui, S, Buffon, L, De Lucchi, D, Vio, P, Favretti, M, Cereser, A 2010. Project of sanitary safe-guarding of “Piccole Produzioni Locali” (PPL). Italian Journal of Food Safety 7, 7576.
Di Martino, G, Scollo, A, Capello, K, Stefani, A, Schiavon, E, Rampin, F, Marangon, S, Gottardo, F, Bonfanti, L 2011. Effect of straw provision on the welfare status of Italian heavy pigs. Proceedings of 15th International Congress of the International Society for Animal Hygiene, 3–7 July 2011, Wien, Austria, volume 1, pp. 423–426.
European Food Safety Authority (EFSA) 2007. Scientific opinion of the Panel on Animal Health and Welfare on a request from the Commission on Animal Health and Welfare in fattening pigs in relation to housing and husbandry. The EFSA Journal 564, 114.
Feder, ME, Hofmann, GE 1999. Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. Annual Review of Physiology 61, 243282.
Filipović, D, Gavrilović, L, Dronjak, S, Demajo, M, Radojcić, MB 2008. Liver glucocorticoid receptor and heat shock protein 70 levels in rats exposed to different stress models. Physiological Research 57, 205213.
Foury, A, Lebret, B, Chevillon, P, Vautier, A, Terlouw, C, Mormède, P 2011. Alternative rearing systems in pigs: consequences on stress indicators at slaughter and meat quality. Animal 5, 16201625.
Fukudo, S, Abe, K, Hongo, M, Utsumi, A, Itoyama, Y 1997. Brain-gut induction of heat shock protein (HSP) 70 mRNA by psychophysiological stress in rats. Brain Research 757, 146148.
Hartman, D, Gething, MJ 1996. Normal protein folding machinery. In Stress-inducible cellular responses (ed. U Feige, RI Morimoto, I Yahara and BS Polla), pp. 324. Birkhäuser, Basel, CH.
Kaneko, JJ 2008. Clinical biochemistry of domestic animals, 6th edition. Academic Press, San Diego, CA, USA.
Kiang, JG, Tsokos, GC 1998. Heat shock protein 70 kDa: molecular biology, biochemistry, and physiology. Pharmacology & Therapeutics 80, 183201.
Laemmli, UK 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680685.
Lebret, B, Prunier, A, Bonhomme, N, Foury, A, Mormède, P, Dourmad, JY 2011. Physiological traits and meat quality of pigs as affected by genotype and housing system. Meat Science 88, 1422.
Marruchella, G, Di Leonardo, M, Di Guardo, G, Romanucci, M, Marà, M, Tiscar, PG, Mosca, F, Della Salda, L 2004. Heat shock proteins (HSPs) 27, 72 and 73 in normal and pre-ulcerative mucosa of the gastric pars oesophagea in swine. Journal of Comparative Pathology 131, 1017.
Morimoto, RI, Tissieres, A, Georgopoulos, C 1990. Stress proteins in biology and medicine. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.
Murata, H, Shimada, N, Yoshioka, M 2004. Current research on acute phase proteins in veterinary diagnosis: an overview. The Veterinary Journal 168, 2840.
O'Connor, EA, Parker, MO, McLeman, MA, Demmers, TG, Lowe, JC, Cui, L, Davey, EL, Owen, RC, Wathes, CM, Abeyesinghe, SM 2010. The impact of chronic environmental stressors on growing pigs, Sus scrofa (Part 1): stress physiology, production and play behaviour. Animal 4, 18991909.
Parker, MO, O'Connor, EA, McLeman, MA, Demmers, TG, Lowe, JC, Owen, RC, Davey, EL, Wathes, CM, Abeyesinghe, SM 2010. The impact of chronic environmental stressors on growing pigs, Sus scrofa (Part 2): social behaviour. Animal 4, 19101921.
Sepponen, K, Pösö, AR 2006. The inducible form of heat shock protein 70 in the serum, colon and small intestine of the pig: comparison to conventional stress markers. The Veterinary Journal 171, 519524.
Shichijo, K, Ihara, M, Matsuu, M, Ito, M, Okomura, Y, Sekine, I 2003. Overexpression of heat shock protein 70 in stomach of stress-induced gastric ulcer resistant rats. Digestive Diseases and Sciences 48, 340348.
Temple, D, Courboulay, V, Manteca, X, Velarde, A, Dalmau, A 2012. The welfare of growing pigs in five different production systems: assessment of feeding and housing. Animal 6, 656667.
Velazquez, JM, Lindquist, S 1984. hsp70: nuclear concentration during environmental stress and cytoplasmic storage during recovery. Cell 36, 655662.
Voellmy, R 1996. Sensing stress and responding to stress. In Stress-inducible cellular responses (ed. U Feige, RI Morimoto, I Yahara and BS Polla) pp. 121138. Birkhäuser, Basel, CH.
Welch, WJ, Feramisco, JR 1984. Nuclear and nucleolar localization of the 72,000-dalton heat shock protein in heat-shocked mammalian cells. The Journal of Biological Chemistry 259, 45014513.
Welfare Quality ® 2009. Welfare Quality® applied to growing and finishing pigs. In Welfare Quality® assessment protocol for pigs (ed. A Dalmau, A Velarde, K Scott, S Edwards, I Veissier, L Keeling and A Butterworth), pp. 4978. Welfare Quality® Consortium, The Netherlands.


Type Description Title
Supplementary materials

Negrato Supplementary Material
Figure S1

 Unknown (5.5 MB)
5.5 MB
Supplementary materials

Negrato Supplementary Material
Figure S2

 Unknown (3.2 MB)
3.2 MB
Supplementary materials

Negrato Supplementary Material
Figure S3

 Unknown (2.8 MB)
2.8 MB


Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed