Acute phase protein mRNA expressions and enhancement of antioxidant defense system in Black-meated Silkie Fowls supplemented with clove (Eugenia caryophyllus) extracts under the influence of chronic heat stress

Abstract

Background: The current study investigates the anti-stress effects of clove (Eugenia caryophyllus) extracts (0, 200, 400, and 600 mg/kg) on serum antioxidant biomarkers, immune response, immunological organ growth index, and expression levels of acute phase proteins (APPs); ovotransferrin (OVT), ceruloplasmin (CP), ceruloplasmin (AGP), C-reactive protein (CRP), and serum amyloid-A (SAA) mRNA in the immunological organs of 63-d-old male black-meated Silkie fowls subjected to 21 d chronic heat stress at 35 ± 2 °C.

Results: The results demonstrated that clove extract supplementation in the diet of Silkie fowls subjected to elevated temperature (ET) improve growth performance, immune responses, and suppressed the activities of glutathion peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT), and thioredoxin reductase (TXNRD); reduced serum malonaldehyde (MDA) and glutathione (GSH) concentrations when compared with fowls raised under thermoneutral condition (TC). Upon chronic heat stress and supplementation of clove extracts, the Silkie fowls showed a linear increase in GSH-Px, SOD, CAT, and TXNRD activities (P = 0.01) compared with fowls fed diets without clove extract. ET decreased (P < 0.05) the growth index of the liver, spleen, bursa of Fabricius and thymus. However, the growth index of the liver, spleen, bursa of Fabricius and thymus increased significantly (P < 0.05) which corresponded to an increase in clove supplemented levels. The expression of OVT, CP, AGP, CRP, and SAA mRNA in the liver, spleen, bursa of Fabricius and thymus were elevated (P < 0.01) by ET compared with those maintained at TC. Nevertheless, clove mitigates heat stress-induced overexpression of OVT, CP, AGP, CRP and SAA mRNA in the immune organs of fowls fed 400 mg clove/kg compared to other groups.

Conclusions: The results showed that clove extracts supplementation decreased oxidative stress in the heat-stressed black-meated fowls by alleviating negative effects of heat stress via improvement in growth performance, antioxidant defense mechanisms, immunity, and regulate the expression of acute phase genes in the liver and immunological organs.

Keywords: Acute phase protein; Antioxidant; Black-meated fowls; Clove; Immunity.

Fig. 1

Effects of ET and clove levels on APP; OVT, CP, AGP, CRP and SAA mRNA expression. Values are Means ± SEM, n = 2. The OVT, CP, AGP, CRP, and SAA mRNA expression of ET (clove, 0 mg/kg) was different from TC, *P < 0.05, **P < 0.01, ***P < 0.001 (ET vs. TC, without clove). a Linear contrasts of 0, 200, 400, & 600 mg clove of OVT mRNA expression of the liver, bursa of fabricius, thymus, and spleen are expressed as P = 0.001, P = 0.002, and P = 0.008, respectively, and the quadratic contrast of clove as P = 0.05, P = 0.172, and P = 0.0046, respectively. b Linear contrasts of 0, 200, 400, and 600 mg clove of CP mRNA expression of the same organs are expressed as P < 0.002, P = 0.004, and P < 0.001, and the quadratic contrasts of clove as P = 0.001, P = 0.471, and P < 0.003. c Linear contrasts of 0, 200, 400, and 600 mg clove of AGP mRNA expression of the same organs are expressed as P < 0.002, P = 0.012, and P < 0.001, and the quadratic contrast of clove as P = 0.001, P = 0.049, and P = 0.001, respectively. d Linear contrasts of 0, 200, 400, and 600 mg clove of CRP mRNA expression of the same organs are expressed as P < 0.001, P = 0.031, and P < 0.003, and the quadratic contrast of clove as P = 0.003, P = 0.033, and P = 0.001, respectively. e Linear contrasts of 0, 200, 400, & 600 mg clove in SAA mRNA expression of the same organs are expressed as P < 0.004, P = 0.054, and P < 0.001, and the quadratic contrast of clove as P = 0.001, P = 0.01, and P = 0.029, respectively