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. 2025 Apr 7;15(7):1066.
doi: 10.3390/ani15071066.

Physiological and Histological Responses of Awassi Lambs to High Dietary Organic Copper Supplementation

Osama A Saeed  1 , Mohanad A Shareef  2 , Hassan M Alnori  1 , Teik K Leo  3 , Mohammed A Al-Bayar  1 , Idham A Abed  4 , Omar K Attallah  1
Affiliations

Physiological and Histological Responses of Awassi Lambs to High Dietary Organic Copper Supplementation

Osama A Saeed et al. Animals (Basel). .

Abstract

This study investigated the effects of high dietary organic copper supplementation on growth performance, mineral metabolism and organ histology in male Awassi lambs. Fifteen lambs with similar body weight (16 ± 1.53 kg) were assigned to three groups: a control group (T1) receiving a basal diet, and two treatment groups (T2 and T3) supplemented with 0.5 g and 1 g of organic copper per kg DM, respectively. The total dietary copper concentrations were 6.18, 81.68 and 156.75 ppm in T1, T2, and T3 groups, respectively. The basal diet, composed of soybeans, wheat, and barley, provided 2.4 Mcal/kg metabolizable energy and 14% crude protein. High dietary copper supplementation had no significant impact on growth performance and body biometric measurements. Testicular length was increased in T3 lambs. Mineral analysis showed increased hepatic copper concentrations in T2 and T3 lambs. Kidney copper levels remained within normal ranges, while renal zinc concentrations declined in T2 and T3 lambs. Gene expression analysis demonstrated upregulation of ATP7A and ATP7B genes in T3 lambs, indicating a cellular adaptation to elevated copper levels, while IGF1 expression remained unchanged. Histological assessments, however, revealed hepatic and renal changes in T3 lambs. Overall, high dietary organic copper supplementation, particularly at 1 g Cu/kg DM, increases copper retention and may support reproductive health in Awassi lambs. However, excessive intake poses risks of liver and kidney damage, highlighting the need for careful dietary management.

Keywords: Awassi sheep; copper supplementation; gene expression; histology; physiology.

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Conflict of interest statement

T.K.L., an employee of Imperium Grp Sdn Bhd, contributed to the software and manuscript writing for this study. The company was not involved in the conduct of the study or the publication of the manuscript. All authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Histological analysis of liver tissues from Awassi lambs supplemented with organic copper. (A) Lambs supplemented with 0 g of organic copper (T1). (B) Lambs supplemented with 0.5 g of organic copper (T2). (C) Lambs supplemented with 1 g of organic copper (T3). The liver tissues were stained with hematoxylin and eosin and observed under 40× magnification using a light microscope.
Figure 2
Figure 2
Histological analysis of kidney tissues from Awassi lambs supplemented with organic copper. (A) Lambs supplemented with 0 g of organic copper (T1). (B) Lambs supplemented with 0.5 g of organic copper (T2). (C) Lambs supplemented with 1 g of organic copper (T3). The kidney tissues were stained with hematoxylin and eosin and observed under 40× magnification using a light microscope.
Figure 3
Figure 3
Histological analysis of testes tissues from Awassi lambs supplemented with organic copper. (A) Lambs supplemented with 0 g of organic copper (T1). (B) Lambs supplemented with 0.5 g of organic copper (T2). (C) Lambs supplemented with 1 g of organic copper (T3). The testes tissues were stained with hematoxylin and eosin and observed under 40× magnification using a light microscope.
Figure 4
Figure 4
Expression of IGF1, ATP7A, and ATP7B genes in the blood serum of Awassi lambs. The mRNA expression levels of IGF1, ATP7A, and ATP7B were quantified using qRT-PCR, with GAPDH as the reference gene for normalization. Treatment groups include T1 (control: 0 g Cu/kg DM), T2 (0.5 g Cu/kg DM), and T3 (1 g Cu/kg DM). Bars with different letters (a, b) indicate significant differences (p < 0.05) among the groups.
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