Volume 16, Issue 3 (9-2025)
Res Anim Prod 2025, 16(3): 83-94 |
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Fathi M, Mardani P. (2025). The Effects of Chlorogenic Acid on Antioxidant Status, Immunity, Inflammatory Cytokines, some Biochemical Parameters, Mortality, and Performance in Broilers. Res Anim Prod. 16(3), 83-94. doi:10.61882/rap.2025.1488
URL: http://rap.sanru.ac.ir/article-1-1488-en.html
URL: http://rap.sanru.ac.ir/article-1-1488-en.html
1- Department of Animal Science, Payame Noor University, Tehran, Iran
2- Department of Biology, Payame Noor University, Tehran, Iran
2- Department of Biology, Payame Noor University, Tehran, Iran
Abstract: (1116 Views)
Extended Abstract
Background: In today's conditions of industrial poultry breeding, especially broilers, stress is a critical problem that affects poultry performance and brings economically significant mortality. Environmental, nutritional, and internal stressors often cause a decrease in performance and endanger health. Stresses often lead to oxidative stress, and through the induction of lipid peroxidation, reduction of antioxidant capacity, inflammation, etc., it ultimately decreases performance and increases mortality in broiler chickens. Therefore, they have suggested that the administration of antioxidants during stress is probably effective in reducing the problems caused by the stress of stressful factors to maintain the health and production performance of flocks in broiler farms. Chlorogenic acid, as one of the most abundant phenolic acid compounds in nature, is an ester of caffeic acid with quinic acid, which is found naturally in various plant species and has several biological functions, such as antioxidant, antimicrobial, antiviral, anti-inflammatory, anti-diabetic, and anticancer activities. The special phenolic structure endows chlorogenic acid with good free radical-scavenging activities, and can effectively scavenge various free radicals, effectively inhibit cellular lipid peroxidation, and beneficially regulate cell membrane stability. Experimental and clinical evidence has confirmed the antioxidant effects of chlorogenic acid in vivo and in vitro through direct antioxidant activity and/or its regulation on signal transduction pathways involved in cellular antioxidant defense. The antioxidant property of chlorogenic acid makes it possible to use it as a promising and green antioxidant in animal feed. It has been shown that food supplements with chlorogenic acid can improve the growth performance and antioxidant capacity of intestinal mucosa by increasing the activity of antioxidant enzymes, preventing lipid peroxidation, and activating antioxidant signaling pathways. In broilers, chlorogenic acid supplementation has been reported to increase growth performance, reduce inflammatory response, prevent intestinal damage, improve intestinal mucosal barrier function, and improve oxidative damage. However, little is known about the antioxidant activity of chlorogenic acid in broilers. Therefore, the current research was designed to evaluate chlorogenic acid effects on growth performance, antioxidant status and immunity, and some biochemical parameters of broiler chickens.
Methods: A total of 300 one-day-old male broiler chickens (45 ± 1.2 g) from the commercial strain Ross (308) were distributed in four experimental treatments (five replicates and 15 chickens per experimental unit). Experimental treatments includeed 1- a control group (fed with a basic diet), 2- CGA-500 group (fed with a basic diet + 500 mg/kg chlorogenic acid), 3- CGA-1000 group (fed with a basic diet + 1000 mg/kg of chlorogenic acid), and 4- CGA-1500 group (fed with a basic diet + 1500 mg/kg of chlorogenic acid). The chlorogenic acid used in this research was produced by Shanghai China Company with a purity of 98.12%, which was added to flour feed in the form of powder. The light, temperature, ventilation, humidity, and hygiene programs were set the same for all the experimental treatments and according to the recommendations of the Ross 308 strain for different periods. The feed intake and body weight of the birds in each experimental unit were measured at the age of 42 days, and performance indicators (food intake, weight gain, and feed conversion ratio) were calculated from the age of 1 to 42 days. At the age of 42 days, two chickens were randomly selected from each cage, and a 2 ml blood sample was used to prepare serum from the wing vein using special syringes to measure blood biochemical indices. Liver tissue samples were immediately washed in distilled water, and after drying, they were immediately placed in clear freezer plastic bags and frozen at -20 °C.
Serum antioxidant parameters, including glutathione peroxidase and superoxide dismutase enzymes, were also measured using Nonad Salamat brand kits. To determine the concentration of malondialdehyde in serum, the amount of light absorption in the samples was determined by the colorimetric method using the device. Liver levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), and TNF-α (tumor necrosis factor-α) were quantitatively measured by the commercial kits of Crystal De Shanghai, China, and by the sandwich ELISA method. Serum enzymes, including aspartate aminotransferase and alanine aminotransferase, were measured using Padco brand kits. Serum levels of G and M immunoglobulin were measured using Diasorin S.P.A., Italy kits. Serum levels of lipid parameters, triglyceride, cholesterol, LDL, and HDL were measured using a quantitative diagnosis kit (Pars Azmoun Company). The collected data were statistically analyzed using the GLM procedure of SAS software version 1/9 (SAS, 2003) in the form of a completely randomized design. Mortality data were transformed using the √(X+1) square root transformation before analysis. To compare the means, Duncan's multi-range test was used at a significance level of 5%. The method of independent comparisons (Orthogonal) was used to determine the linear and quadratic effects of different CGA levels.
Results: The inclusion of chlorogenic acid caused a significant increase in feed intake and weight gain and a significant decrease in the feed conversion ratio and total mortality (P < 0.05). The best performance belonged to the 1000 mg/kg of chlorogenic acid treatment compared to the other treatments. In addition, chlorogenic acid supplementation at levels of 500 and 1000 mg significantly improved the antioxidant capacity (increased activity of glutathione peroxidase and decreased malondialdehyde activity) in serum and the liver (P < 0.05). The administration of 500 and 1000 mg of chlorogenic acid significantly reduced serum levels of inflammatory cytokines IL-1β, IL-6, and TNF-α (P < 0.05).
Conclusion: Decreased serum levels of aspartate aminotransferase, alanine aminotransferase, triglyceride, cholesterol, and LDL, and increased HDL and immunoglobulin G and M were also observed in CGA-1000 and CGA-1500 treatments (P < 0.05). In general, the results of this research show that using 1000 mg/kg of chlorogenic acid in the diet of chickens may improve both growth performance and antioxidant and inflammatory indices.
Background: In today's conditions of industrial poultry breeding, especially broilers, stress is a critical problem that affects poultry performance and brings economically significant mortality. Environmental, nutritional, and internal stressors often cause a decrease in performance and endanger health. Stresses often lead to oxidative stress, and through the induction of lipid peroxidation, reduction of antioxidant capacity, inflammation, etc., it ultimately decreases performance and increases mortality in broiler chickens. Therefore, they have suggested that the administration of antioxidants during stress is probably effective in reducing the problems caused by the stress of stressful factors to maintain the health and production performance of flocks in broiler farms. Chlorogenic acid, as one of the most abundant phenolic acid compounds in nature, is an ester of caffeic acid with quinic acid, which is found naturally in various plant species and has several biological functions, such as antioxidant, antimicrobial, antiviral, anti-inflammatory, anti-diabetic, and anticancer activities. The special phenolic structure endows chlorogenic acid with good free radical-scavenging activities, and can effectively scavenge various free radicals, effectively inhibit cellular lipid peroxidation, and beneficially regulate cell membrane stability. Experimental and clinical evidence has confirmed the antioxidant effects of chlorogenic acid in vivo and in vitro through direct antioxidant activity and/or its regulation on signal transduction pathways involved in cellular antioxidant defense. The antioxidant property of chlorogenic acid makes it possible to use it as a promising and green antioxidant in animal feed. It has been shown that food supplements with chlorogenic acid can improve the growth performance and antioxidant capacity of intestinal mucosa by increasing the activity of antioxidant enzymes, preventing lipid peroxidation, and activating antioxidant signaling pathways. In broilers, chlorogenic acid supplementation has been reported to increase growth performance, reduce inflammatory response, prevent intestinal damage, improve intestinal mucosal barrier function, and improve oxidative damage. However, little is known about the antioxidant activity of chlorogenic acid in broilers. Therefore, the current research was designed to evaluate chlorogenic acid effects on growth performance, antioxidant status and immunity, and some biochemical parameters of broiler chickens.
Methods: A total of 300 one-day-old male broiler chickens (45 ± 1.2 g) from the commercial strain Ross (308) were distributed in four experimental treatments (five replicates and 15 chickens per experimental unit). Experimental treatments includeed 1- a control group (fed with a basic diet), 2- CGA-500 group (fed with a basic diet + 500 mg/kg chlorogenic acid), 3- CGA-1000 group (fed with a basic diet + 1000 mg/kg of chlorogenic acid), and 4- CGA-1500 group (fed with a basic diet + 1500 mg/kg of chlorogenic acid). The chlorogenic acid used in this research was produced by Shanghai China Company with a purity of 98.12%, which was added to flour feed in the form of powder. The light, temperature, ventilation, humidity, and hygiene programs were set the same for all the experimental treatments and according to the recommendations of the Ross 308 strain for different periods. The feed intake and body weight of the birds in each experimental unit were measured at the age of 42 days, and performance indicators (food intake, weight gain, and feed conversion ratio) were calculated from the age of 1 to 42 days. At the age of 42 days, two chickens were randomly selected from each cage, and a 2 ml blood sample was used to prepare serum from the wing vein using special syringes to measure blood biochemical indices. Liver tissue samples were immediately washed in distilled water, and after drying, they were immediately placed in clear freezer plastic bags and frozen at -20 °C.
Serum antioxidant parameters, including glutathione peroxidase and superoxide dismutase enzymes, were also measured using Nonad Salamat brand kits. To determine the concentration of malondialdehyde in serum, the amount of light absorption in the samples was determined by the colorimetric method using the device. Liver levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), and TNF-α (tumor necrosis factor-α) were quantitatively measured by the commercial kits of Crystal De Shanghai, China, and by the sandwich ELISA method. Serum enzymes, including aspartate aminotransferase and alanine aminotransferase, were measured using Padco brand kits. Serum levels of G and M immunoglobulin were measured using Diasorin S.P.A., Italy kits. Serum levels of lipid parameters, triglyceride, cholesterol, LDL, and HDL were measured using a quantitative diagnosis kit (Pars Azmoun Company). The collected data were statistically analyzed using the GLM procedure of SAS software version 1/9 (SAS, 2003) in the form of a completely randomized design. Mortality data were transformed using the √(X+1) square root transformation before analysis. To compare the means, Duncan's multi-range test was used at a significance level of 5%. The method of independent comparisons (Orthogonal) was used to determine the linear and quadratic effects of different CGA levels.
Results: The inclusion of chlorogenic acid caused a significant increase in feed intake and weight gain and a significant decrease in the feed conversion ratio and total mortality (P < 0.05). The best performance belonged to the 1000 mg/kg of chlorogenic acid treatment compared to the other treatments. In addition, chlorogenic acid supplementation at levels of 500 and 1000 mg significantly improved the antioxidant capacity (increased activity of glutathione peroxidase and decreased malondialdehyde activity) in serum and the liver (P < 0.05). The administration of 500 and 1000 mg of chlorogenic acid significantly reduced serum levels of inflammatory cytokines IL-1β, IL-6, and TNF-α (P < 0.05).
Conclusion: Decreased serum levels of aspartate aminotransferase, alanine aminotransferase, triglyceride, cholesterol, and LDL, and increased HDL and immunoglobulin G and M were also observed in CGA-1000 and CGA-1500 treatments (P < 0.05). In general, the results of this research show that using 1000 mg/kg of chlorogenic acid in the diet of chickens may improve both growth performance and antioxidant and inflammatory indices.
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