Hostname: page-component-cd9895bd7-gbm5v Total loading time: 0 Render date: 2024-12-27T15:27:42.728Z Has data issue: false hasContentIssue false
  • English
  • Français

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

Published online by Cambridge University Press:  27 March 2013

E. Negrato ,
G. Di Martino ,
M. Vascellari ,
G. Radaelli ,
K. Capello ,
F. Pascoli ,
D. Bertotto  and
L. Bonfanti
E. Negrato
Affiliation:
Dipartimento di Biomedicina Comparata e Alimentazione, University of Padova, Viale dell'Università 16, 35020 Legnaro, Italy
G. Di Martino*
Affiliation:
Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Università 10, 35020 Legnaro, Padova, Italy
M. Vascellari
Affiliation:
Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Università 10, 35020 Legnaro, Padova, Italy
G. Radaelli
Affiliation:
Dipartimento di Biomedicina Comparata e Alimentazione, University of Padova, Viale dell'Università 16, 35020 Legnaro, Italy
K. Capello
Affiliation:
Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Università 10, 35020 Legnaro, Padova, Italy
F. Pascoli
Affiliation:
Dipartimento di Biomedicina Comparata e Alimentazione, University of Padova, Viale dell'Università 16, 35020 Legnaro, Italy
D. Bertotto
Affiliation:
Dipartimento di Biomedicina Comparata e Alimentazione, University of Padova, Viale dell'Università 16, 35020 Legnaro, Italy
L. Bonfanti
Affiliation:
Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Università 10, 35020 Legnaro, Padova, Italy
*
†E-mail: gdimartino@izsvenezie.it
Get access

Abstract

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.

Keywords

HSP70Western blotliverIHCheavy pig
Type
Behaviour, welfare and health
Information
animal , Volume 7 , Issue 8 , August 2013 , pp. 1362 - 1366
Copyright
Copyright © The Animal Consortium 2013 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

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, 459–466.Google Scholar
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, 2900–2912.Google Scholar
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, 248–254.CrossRefGoogle ScholarPubMed
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, 159–163.CrossRefGoogle ScholarPubMed
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.Google Scholar
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, 1558–1570.Google Scholar
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, 118–127.Google Scholar
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, 27–35.Google Scholar
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, 604–616.Google Scholar
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, 571–578.CrossRefGoogle ScholarPubMed
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, 75–76.Google Scholar
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.Google Scholar
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, 1–14.Google Scholar
Feder, ME, Hofmann, GE 1999. Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. Annual Review of Physiology 61, 243–282.Google Scholar
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, 205–213.Google Scholar
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, 1620–1625.Google Scholar
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, 146–148.Google Scholar
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. 3–24. Birkhäuser, Basel, CH.Google Scholar
Kaneko, JJ 2008. Clinical biochemistry of domestic animals, 6th edition. Academic Press, San Diego, CA, USA.Google Scholar
Kiang, JG, Tsokos, GC 1998. Heat shock protein 70 kDa: molecular biology, biochemistry, and physiology. Pharmacology & Therapeutics 80, 183–201.CrossRefGoogle ScholarPubMed
Laemmli, UK 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.Google Scholar
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, 14–22.Google Scholar
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, 10–17.Google Scholar
Morimoto, RI, Tissieres, A, Georgopoulos, C 1990. Stress proteins in biology and medicine. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.Google Scholar
Murata, H, Shimada, N, Yoshioka, M 2004. Current research on acute phase proteins in veterinary diagnosis: an overview. The Veterinary Journal 168, 28–40.Google Scholar
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, 1899–1909.CrossRefGoogle ScholarPubMed
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, 1910–1921.Google Scholar
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, 519–524.Google Scholar
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, 340–348.CrossRefGoogle ScholarPubMed
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, 656–667.Google Scholar
Velazquez, JM, Lindquist, S 1984. hsp70: nuclear concentration during environmental stress and cytoplasmic storage during recovery. Cell 36, 655–662.Google Scholar
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. 121–138. Birkhäuser, Basel, CH.CrossRefGoogle Scholar
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, 4501–4513.Google Scholar
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. 49–78. Welfare Quality® Consortium, The Netherlands.Google Scholar
Supplementary material: Image

Negrato Supplementary Material

Figure S1

Download Negrato Supplementary Material(Image)
Image 5.5 MB
Supplementary material: Image

Negrato Supplementary Material

Figure S2

Download Negrato Supplementary Material(Image)
Image 3.2 MB
Supplementary material: Image

Negrato Supplementary Material

Figure S3

Download Negrato Supplementary Material(Image)
Image 2.8 MB