Abstract

Heat stress negatively affects poultry production and animal health. In response, animals invoke a heat stress response by inducing heat shock proteins (HSPs). Scientists are actively seeking natural products that can enhance the heat shock response. The present study aimed at assessing the effects of a purified rosemary extract comprising antioxidant compounds on the heat shock response and HSP expression profile in broiler chickens. The response of broilers to HS in the presence of purified rosemary extract was assessed using an in vivo myocardial cell model. Pathological lesions of heart tissue were examined microscopically. The levels and activities of enzymes associated with heart damage and oxidative damage were detected. Immunohistochemical staining was performed for HSPs in myocardial cells. The results showed that lactate dehydrogenase (LDH), creatine kinase (CK), and myocardial CK (CKMB) levels were reduced by the purified rosemary extract before and during heat stress. Heat stress alone increased CK and CKMB levels. The levels of oxidative damage-associated enzymes were compared between the rosemary + heat stress and heat stress-alone groups. The results indicated that in terms of these enzymes, the purified rosemary extract induced a more antioxidative state. Pathological examinations showed that heat stress caused myocardial fiber fracture, karyopyknosis, and degeneration. The addition of purified rosemary extract ameliorated these lesions to some degree, preserving more of the basic structure. Heat stress decreased the cellular levels of crystallin alpha B (CRYAB) and HSP70. The addition of the purified rosemary extract significantly increased the levels of CRYAB and HSP70 during heat stress (p < 0.0001). Immunohistochemistry showed that after rosemary treatment, CRYAB and HSP70 showed more intense staining compared with the no heat stress control group. In the rosemary + heat group, after 10 hours of heat stress, the staining intensity of these two proteins remained higher than in the heat stress group. Thus, purified rosemary extract could induce high levels of HSP70 and CRYAB in chicken hearts before and during heat stress. Purified rosemary extract could be used to alleviate heat stress in broiler chickens.

Highlights

  • Heat stress (HS) affects animal production worldwide and has a significant impact on animal well-being

  • We confirmed that after a short period of heat stress (1 h), the enzymes related to heart damage, such as lactate dehydrogenase (LDH), creatine kinase (CK), and CKMB, could be detected in the blood serum and supernatant of rat myocardial cells

  • Our previous in vivo and in vitro research in rats confirmed that the cause of heat-induced sudden death was heart cell damage [5, 33]

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Summary

Introduction

Heat stress (HS) affects animal production worldwide and has a significant impact on animal well-being. Animals experiencing HS tend to reduce their heat production by limiting feed intake, with subsequent negative effects on growth performance [1]. Stressful conditions, such as environmental, pathological, and nutritional disturbances, can generate a state of tension that evokes behavioral and physiological chain reactions that generally impair poultry performance. Many radicals and metabolic substances are potentially toxic and are defined as Oxidative Medicine and Cellular Longevity “reactive oxygen/nitrogen/chlorine species”[4, 9]. These substances are highly reactive and can modify several biologically cellular macromolecules, such as proteins, lipids, and nucleic acids (DNA and RNA) [10, 11]. We focused on identifying further natural substances that can ease HS of broilers in vivo

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