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veterinary sciences
UDC 619: 612.397:612.176:636.5
Stoyanovskyy V.G., Shevchuk M.O., Kolomiiets I.A.
Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv
Lviv, Ukraine DOI: 10.24411/2520-6990-2020-12172 LIPID PEROXIDE OXIDATION PROCESSES AND THE STATE OF THE ANTIOXIDANT PROTECTION SYSTEM OF CHICKEN BROILERS DURING COMBINED STRESS
Abstract.
The article presents the results of a study characterizing the intensity of lipid peroxidation processes and the state of the antioxidant defense system of broiler chickens at different stages of stress syndrome. It was found that the intensity of lipid peroxidation processes in erythrocytes of broiler chickens of the control group 3 days after stress is characterized by a low content of intermediate andfinal products against the background of high activity of the enzymatic link of the antioxidant system. At different stages of development of the stage of resistance in the erythrocytes of broiler chickens of the control group, the content of lipid hydroperoxides and TBA-active products increases by 1.8-3.6 and 2.4 times compared with the stage of anxiety with a decrease in superoxide dismutase by 27.8% and glutathione peroxidase 2.5-5.4 times. It was found that the use of "Reasil Humic Vet" + "Laktin" in the diet of broiler chickens at different stages of development of the resistance stage inhibits certain stages of lipid peroxidation in erythrocytes by reducing the concentration of lipid hydroperoxides and TBA-active products by 32.8 and 42 5% (P <0.05) and increases the activity of superoxide dismutase - by 46.6% (P <0.05), catalase - by 36.9-52.9% (P <0.05), glutathione peroxidase - by 33.4-80.7% (P <0.05) compared with the control. With the use in the diet of chickens "Reasil Humic Health " in the late stages of the resistance stage revealed a decrease in the content of TBA-active products by 1.6 times (P <0.05) against the background of increasing activity of superoxide dismutase and glutathione peroxidase by 35.8 and 22.7 % (P <0,05) compared with the control.
Key words: lipid peroxidation, antioxidant defense system, broiler chickens, stress, supplements.
Topicality. It is known that lipid peroxidation is a normal physiological process that occurs in all tissues of living organisms, but at a low level with a stable concentration of radicals, while the body's antioxidant defense system controls and inhibits all stages of free radical formation, which helps maintain homeostasis. [1, 2]. At the cellular level, the influence of stress factors initiates excessive activation of the intensity of lipoperoxidation with the formation of a pathologically large number of aggressive and dangerous reactive oxygen species [4, 5, 10]. Therefore, the endogenous antioxidant system, which subtly regulates the neutralization of prooxidants and free radicals, as well as the detoxification and excretion of toxic products of lipid peroxidation, is important in the overall protection system [6, 8, 11]. It is important to study the content of intermediate and final products of oxidation of polyunsaturated fatty acids and the main parameters of the enzymatic link of the antioxidant defense of broiler chickens at different stages of technological stress, systema-tization of results, generalization and improvement of the evaluation system. 3 9].
Aim of the research: to find out the intensity of lipid peroxidation processes and the state of the antiox-idant defense system of broiler chickens against the background of combined stress when included in the diet "Reasil Humic Vet" + "Laktin" and "Reasil Humic Health".
Materials and methods of the research. All manipulations with chickens were carried out in compliance with the requirements of the European Convention
for the Protection of Vertebrate Animals Used for Experimental and Scientific Purposes (Strasbourg, 1986), the First National Congress on Bioethics (Kyiv, 2001) - General Ethical Principles of Animal Experiments and in compliance with the principles of humanity set out in the European Community Directive. The experiment was conducted on the basis of the vivarium of the State Research Control Institute of Veterinary Drugs and Feed Additives in young broiler chickens cross "Kobb-500", of which at 5 days of age formed three groups - control (C) and two experimental (R1, R2 ) 25 individuals in each group, selected on the principle of analogues. Prior to the experiment, all poultry were vaccinated against Marek's disease, Newcastle disease and infectious bronchitis according to the timing of the respective vaccinations. Poultry keeping met the generally accepted technological requirements of floor keeping with free access to drinkers and feeders. From 11 days of life and until the end of the experiment (45 days of life) in addition to the standard diet of chickens R1 group fed feed "Reasil Humic Vet" in liquid form at the rate of 100 ml / 100 l of water and probiotic feed additive "Laktin" at the rate of 1-2 l / 100 l of water; chickens of group R2 were fed the feed preparation "Reasil Humic Health" in dry form at the rate of 1-2 kg / 1 ton of feed according to the instructions. At day 13 of life, all clinically healthy poultry were exposed to combined stress - revaccination (intranasal Newcastle disease) + cold stress (for 60 minutes by air conditioning and 5 ° C in the vivarium). Material for research was selected for 3, 13, 20 and 26 days after the action of the stress
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factor: these periods of implementation of stress reactions are proposed on the basis of research and development of the Department of Normal and Pathological Physiology named after S.V. Stoyanovskyi [7]. Slaughter was performed before morning feeding after light ether anesthesia by decapitation of 5 chickens from each group. The state of lipid peroxidation in the erythrocytes of broiler chickens was assessed by the level of intermediate products - lipid hydroperoxides and final products - TBA-active products. In erythrocyte hemol-ysates, the state of the enzymatic part of the antioxidant defense system was determined by the activity of superoxide dismutase, catalase, glutathione peroxidase [12].
Results of the research and discussion. As a result of the conducted researches it was established that in the blood of group C chickens 3 days after the action of stress, which corresponded to the stage of anxiety, the content of intermediate and final products of lipid peroxidation was 0.17 ± 0.03 OE / ml and 1.85 ± 0 , 44 nmol / mL. Since the elimination of primary reactive forms of Oxygen primarily involves enzymes of the an-tioxidant defense system, analyzing their activity in the erythrocytes of chickens of group C, it should be noted that the value of superoxide dismutase was 7.94 ± 0.65 IU / mg protein, catalase - 4, 13 ± 0.52 mmol / min x mg of protein, glutathione peroxidase - 21.64 ± 1.40 nmol / min x mg of protein, which indicated a sufficient level of protection of cellular structures from endogenous reactive oxygen species. In the chickens of the experimental groups it is necessary to note some increase in the content of lipid hydroperoxides and TBA-active products without any significant intergroup differences compared to the control. In the R1 group of birds there was a slight decrease in superoxide dismutase activity and a probable decrease in glutathione peroxidase activity by 29.3% (p <0.05) against the background of a slight increase in catalase activity. In chickens of group R2, a decrease in superoxide dismutase activity by 30.4% (P <0.05) was registered against the background of an increase in catalase and glutathione peroxidase activity by 17.0% compared with the control, which indicated a different mechanism of action of the additives used. In the initial stages of the development of the resistance stage, which occurred on the 13 th day after stress, the content of the studied products of lipid peroxidation - lipid hydroperoxides and TBA-active products in the erythrocytes of broiler chickens C increased 1.8 and 2.4 times compared with stage of anxiety, which indicated an increase in the activity of free radical oxidation, but in physiological conditions could be considered as an adaptive response of the body to stress factors [13]. With regard to the enzymatic link of the antioxidant defense system in chickens of group C, it is necessary to note a decrease of superoxide dismutase by 27.8% and glutathione peroxidase by 2.5 times without changes in catalase activity, which can be considered a negative consequence of cold stress and revaccination. protection protect cells and the body as a whole from the toxic effects of oxygen radicals and li-pid peroxides, as well as neutralize toxic products that have a membrane-destructive effect.
In chickens of group R1, 13 days after exposure to stress, lower rates of lipid peroxidation were recorded than in group C: the difference in the concentration of lipid hydroperoxides and TBA-active products was 12.9 and 42.1%, respectively (P <0.05). Against the background of a decrease in erythrocytes intermediate and final products of lipid peroxidation, an increase in superoxide dismutase activity by 48.8% (P <0.05) was observed, while the value of catalase and glutathione peroxidase activity approached the level of control. Investigating the dynamics of the content of lipid peroxidation products in the erythrocytes of chickens of group R2 under the influence of complex stress, a slight increase in the content of lipid hydroperoxides was recorded, but the content of TBA-active products decreased by 2.0 times (P <0.05). An increase in the value of superoxide dismutase activity by 40.0% (P <0.05), catalase - by 9.9% against the background of a decrease in glutathione peroxidase activity compared with the control.
On the 20th day after exposure to cold stress and revaccination (development of the resistance stage) in chickens of group C recorded a significant increase in the number of intermediate and final products of lipid peroxidation compared to the anxiety stage: the content of lipid hydroperoxides increased 2.9 times and the content of TBA-active products 2.4 times. The obtained data testified to the sensitivity of lipid peroxidation processes in broilers to complex stress, and lipid peroxida-tion indicators, first of all, the content of TBA-active products, can be used as a characteristic of the state of the bird under stress. With regard to the system of antioxidant protection in chickens of group C, it should be noted an increase in superoxide dismutase by 12.2%, a further decrease in glutathione peroxidase by 5.4 times without changes in catalase activity. The increase in the content of lipid peroxidation products against the background of reduced glutathione peroxidase in the C group of birds could be due to the body's response to cold stress and vaccination, which can be used to indicate the functional state of broilers and the strength of the stress stimulus. During the development of the adaptation syndrome, the processes of peroxidation of li-pids and the activity of the enzymatic part of the anti-oxidant defense system of broilers of group R1, which used "Reasil Humic Vet" + "Laktin", were characterized by positive dynamics of changes. 1.8 times (P <0,05), increase in superoxide dismutase activity - by 46,6% (P <0,05), catalase - by 52,9% (p <0,05), glutathione peroxidase - by 80, 7% (P <0.05) compared to control. From these data it follows that the use of supplements in the diet of broiler chickens inhibits certain stages of lipid peroxidation, primarily intermediate and final, and has a stimulating effect on the activity of the antioxidant system in their body. When used in the diet of chickens R2 group "Reasil Humic Health" 20 days after exposure to stress, the content of lipid hydroperoxides was reduced by 1.4 times and the content of TBA-active products by 1.6 times (p <0.05) compared with the control. In the study of the activity of enzymes of the antioxidant defense system - superoxide dis-mutase, catalase, glutathione peroxidase, a tendency to increase by 5-9% in broilers R2 group relative to the
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control without significant intergroup differences. In general, the obtained results indicated a stimulating effect of the additive on the inhibition of lipid peroxidation processes against the background of lower antiox-idant action.
At the day 26 of the resistance stage in the erythrocytes of group C chickens, the content of LPO products changed: the content of lipid hydroperoxides continued to increase, which increased 3.6 times relative to the stage of anxiety, and the content of TBA-active products decreased compared to previous stages of resistance. anxiety remained 1.6 times higher. Since the processes of lipid peroxidation occur primarily in biomembranes, which leads to a violation of their functional properties, and the marker of intensification of these processes - TBA-active products, the results showed a decrease in the intensity of lipid peroxidation, which is an important step in studying - and antioxidant processes of the body. There was an increase in superoxide dismutase activity by 18.8%, a decrease in catalase activity by 6.1% and an increase in glutathione peroxidase compared to previous stages of resistance, but relative to the stage of anxiety, its activity remained lower by 23.8%. The obtained results indicated the stabilization of the enzyme unit of the antioxidant protection system of group C broilers. In chickens of group R1 26 days after exposure to stress revealed a decrease in the content of lipid hydroperoxides and TBA-active products by 32.8 and 42.5% (P <0.05) against the background of increased activity of catalase and glutathione peroxidase by 36.9 and 33, 4% (P <0.05) compared to control. These data indicate the inhibitory effect of the additives used on the content of intermediate and final products of lipid peroxidation, the level of which is largely regulated by increasing the activity of the enzymatic link of the antioxidant defense system. For use in the diet of chickens R2 group "Reasil Humic Health" 26 days after exposure to stress decreased lipid hydroperoxides by 49.2% (P <0.05) and TBA-active products by 35.5% against the background of increased superoxide dismutase activity on 35.8% (P <0.05), catalase - by 20.1%, glutathione peroxidase by 22.7% (P <0.05) compared with the control, which indicated a decrease in the activation of lipid peroxidation and increased structural functional changes in erythrocytes and their membranes.
Conclusions.
1. The intensity of lipid peroxidation processes in erythrocytes of broiler chickens 3 days after exposure to stress is characterized by low content of intermediate and final products against the background of high activity of the enzymatic link of the antioxidant system. At different stages of development of the stage of resistance in the blood erythrocytes of broiler chickens, the content of lipid hydroperoxides and TBA-active products increases by 1.8-3.6 and 2.4 times compared with the stage of anxiety with a decrease in superoxide dismutase by 27.8% and glutathione peroxidase in 2.55.4 times.
2. It was found that the use of "Reasil Humic Vet" + "Laktin" in the diet of broiler chickens at different stages of development of the resistance stage inhibits certain stages of lipid peroxidation in erythrocytes by
reducing the concentration of lipid hydroperoxides and TBA-active products by 32.8 and 42.5% (p <0.05) and increases the activity of superoxide dismutase - by 46.6% (p <0.05), catalase - by 36.9-52.9% (p <0.05), glutathione peroxidase - by 33.4-80.7% (p <0.05) compared with the control. When used in the diet of chickens "Reasil Humic Health" in the late stages of the resistance stage revealed a decrease in the content of TBA-active products by 1.6 times (p <0.05) against the background of increasing activity of superoxide dis-mutase and glutathione peroxidase by 35.8 and 22.7 % (p <0.05) compared with the control.
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