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VETERINARY SCIENCE / «€®[LL®qUQUM~J®UrMaL>>#32IM,2©2©
Veterinary science
UDC 577.1:612.015
Martyshuk T. V., Gutyj B.V., Leskiv Ch. Ya., Slobodiuk N.M., Rudenko O.P.
Stepan Gzhytskyi National Univesity of Veterinary Medicine and Biotechnologies,
Lviv, Pekarska str., 50, Lviv, 79010, Ukraine
Khalak V.I.
State Institution Institute of grain crops of NAAS, V. Vernadsky Str., 14,
Dnipro, 49027, Ukraine DOI: 10.24412/2520-2480-2020-3284-8-11 THE EFFECT OF FEED ADDITIVE "BUTASELMEVIT-PLUS" ON RATS' BLOOD'S MORPHOLOGICAL PARAMETERS UNDER CARBON TETRACHLORIDE INTOXICATION
CONDITIONS
Abstract.
The research aimed to study the effect of feed additive "Butaselmevit-plus" on rats' blood's morphological parameters under conditions of carbon tetrachloride intoxication. The study was performed on white sexually mature young male Wistar rats weighing 180-200 g, which were kept on the standard diet of the institute vivarium of the State Research Control Institute of Veterinary Drugs and Feed Additives. The animals were divided into three groups of 20 animals each: Group 1 (C) intact animals; 2nd group (E1) - rats affected by carbon tetrachloride; Group 3 (E2) - rats affected by carbon tetrachloride and used feed additive "Butaselmevit-plus." Experimental intoxication in animals was performed by two (after 48 h) intravesical administration of carbon tetrachloride at a dose of 0.1 ml per 100 g of rat body weight in the form of a 50% oil solution. Group E2 in trial toxicosis for 30 days was supplied a feed additive "Butaselmevit-plus" at a dose of 0.1 g per 100 g of body weight with food. It was found that under conditions of intoxication with carbon tetrachloride in rats, the bone marrow's hematopoietic function is impaired, which is manifested by a decrease in the number of erythrocytes by 26.5%, hemoglobin content by 8.7%. During applying the feed additive "Butaselmevit-plus" in mice, under forms of oxidative stress during the research, the blood normalizes the activity of hematological parameters, namely on the 25th and 30th day within physiological values were indicators of erythrocytes, hemoglobin, leukocytes, and red blood cell indices compared to controls, indicating the restoration of hematopoietic function of the bone marrow.
Keywords: oxidative stress, rats, butaselmevit-plus, blood, milk thistle.
Chemical compounds are environmental factors that are continually acting on the body of animals. There is a certain balance between the external chemical environment and the organism's chemical composition, the violation of which leads to a shift in homeostasis. In recent decades, a toxic situation is emerging globally due to the intensive introduction of chemicals into the industry, agriculture, human and veterinary medicine, and the growing number of acute and chronic poisonings [1, 2, 3].
In recent years, the role of the processes of activation of free radical oxidation of lipids of plasma and intracellular membranes of hepatocytes against the background of depletion of protective antiradical systems has been established in liver pathogenesis diseases [4, 5]. It is known from the literature that excessive accumulation of lipid peroxidation products causes edema of mitochondria, separation of respiratory processes and oxidative phosphorylation, damage to sulfhydryl groups of thiol enzymes [5, 6, 7, 8]. Factors that cause oxidative stress are a violation of the redox balance in the direction of oxidation and the formation of secondary free radicals. Pathological disorders of homeostasis that lead to oxidative stress, in particular, include: change in homeostasis as a result of pathological factors; change in homeostasis as a result
of the violation of genetic information; defect of the regulatory system or target organ [9, 10]. Under the action of a pathological factor, there is a change in the intensity of lipid peroxidation, accumulation in the blood of the concentration of free radical oxidation products and reactive oxygen species, reduction of blood buffer capacity to maintain optimal parameters of the intensity of free radical reactions [11, 12].
The development of methods for integrating the oxidation state in animals under various risk agents is an essential factor in creating safe and highly effective drugs for parenteral and oral use. To increase the organism's adaptability and immunobiological reactivity, enhancing protein-synthesizing and enzyme function in animals in recent years, new complex drugs have been successfully used [13, 14, 15]. Some authors have established the stimulating effect of milk thistle, vitamins, selenium, and butaphosphane on antioxidant and hepatoprotective action in animals [16, 17]. However, the complex application of these drugs on liver function and hematological parameters is currently insufficiently covered in the scientific literature.
Therefore, our work aimed to study the morphological parameters of rats' blood under
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oxidative stress and the action of feed additive "Butaselmevit-plus."
Materials and methods. The study was performed on white sexually mature young male Wistar rats weighing 180-200 g, which were kept on the standard diet of the institute vivarium of the State Research Control Institute of Veterinary Drugs and Feed Additives. Throughout the experiment, the rats were kept on a balanced diet containing all the necessary components; the animal's drinking water was obtained without restrictions from glass drinking bowls with a volume of 0.2 liters.
The animals were divided into three groups of 20 animals each: Group 1 (C) intact animals; 2nd group (Ei) - rats affected by carbon tetrachloride; Group 3 (E2) - rats affected by carbon tetrachloride and used feed additive "Butaselmevit-plus." Experimental intoxication in animals was performed according to the method described by O.Yu. Zhuravlyov and V.M Hunchak, by two with interval injections (after 48 h) of carbon tetrachloride in a dose of 0.1 ml per 100 g of rat body weight in the form of 50% oil solution. Group E2 in for 30 days, was fed a supplied additive "Butaselmevit-plus" at a dose of 0.1 g per 100 g of body weight with food. Blood for biochemical tests was taken under etheric anesthesia from the jugular vein on the fifth, tenth and twentieth, twenty-fifth, and thirtieth days of the experiment.
According to the generally accepted method, the number of erythrocytes and leukocytes was counted on the counting chamber's Goryaev grid. The hemoglobin level in the blood was determined by the cyanhemoglobin method using FEK-M by the technique of G.V. Derviza and A.G. Sparrow. The hematocrit was determined by centrifugation of blood in micropipettes at 3000 v/min. According to the values of erythrocytes, hemoglobin level, and hematocrit, according to the appropriate formulas, by mathematical calculations, calculated the following amounts of red blood indices: the average volume of one erythrocyte (MCV); the average mass of hemoglobin in the erythrocyte (MSH); the average concentration of hemoglobin in erythrocytes (MCNS) [18].
The analysis of research results was performed using the software package Statistica 6.0. The Student's t-test assessed the probability of differences. The results were considered plausible at P < 0.05.
Results and discussion. In trial tetrachloro-methane poisoning, a decrease in the erythrocytes number of the E1 group was found starting from the 5th day of the test; it reduced by 12.2%. On the 10th and 20th days, the erythrocytes number in the E1 group blood was the lowest, relative to the C group; it declined by 22.9 and 26.5%, respectively (Table 1).
Table 1
The number of erythrocytes in the blood of rats under conditions of carbon tetrachloride poisoning and _the action of feed additive "Butaselmevit-plus," T/l (M±m; n=20)_
Test Day Groups of animals
Control Experimental 1 Experimental 2
5th day 6,46±0,149 5,67±0,124** 5,91±0,119*
10th day 4,98±0,099*** 5,98±0,112*
20th day 4,75±0,123*** 5,92±0,085*
25th day 5,04±0,111*** 6,15±0,123
30th day 5,11±0,097*** 6,39±0,130
Degree of probability:* - p < 0,05;** - p < 0,025; *** - p < 0,001
When supplying the feed additive "Butaselmevit-plus," the erythrocytes number in the blood of the E2 group of rats was higher than the E1 group. Still, it reached control values only on the 30th day of the experiment. The lowest erythrocytes number in the blood of the E2 group was on the 5th day of the investigation, where, respectively, it was 5.91 ± 0.119 T / l. On the 20th day of the research, the studied indicator in the E2 group's blood was higher than the E1, but the C group was lower by 8.4%.
In sick rats of the E1 group, hemoglobin level was reduced compared to indicators of intact animals. Thus, on the 10th day of the experiment, the level of hemoglobin in their blood decreased by 5.6%, while on the 20th day - by 8.7% relative to rats' C group. On the 25th and 30th days of the test, the hemoglobin level increased slightly compared with the previous day but remained low compared with the C group (Table 2).
Table 2
The hemoglobin level in the blood of rats under conditions of carbon tetrachloride poisoning and the
Test Day Groups of animals
Control Experimental 1 Experimental 2
5th day 148,8±1,57 150,2±1,85 149,8±1,42
10th day 140,4±2,05* 148,2±1,76
20th day 135,8±0,97*** 153,4±1,24
25th day 140,7±1,11** 150,7±1,59
30th day 141,6±2,17* 150,4±1,24
Degree of probability:* - p < 0,05;** - p < 0,025; *** - p < 0,001.
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0.68 l / l. Nourishing of supplies additive "Butaselmevit-plus" to rats of E2 group in the specified period promoted an increase in hematocrit size to 29,9 ± 1,06 l / l (Tab. 3).
The increase in the leukocytes number in the rats' blood of the experimental groups after introducing carbon tetrachloride is the result of the development of inflammatory processes in the body.
Table 3
The value of hematocrit in the blood of rats under conditions of carbon tetrachloride poisoning and the
Decreases in red blood cells in the experimental group of rats are also associated with the fact that they are particularly sensitive to oxidative stress, which develops due to the introduction of carbon tetrachloride.
On the 10th day of the research in the blood of rats of the E1 group found an extension in hematocrit to 30.7 ± 1.45 l / l, while in the C group, this figure was 28.1 ±
Test Day Groups of animals
Control Experimental 1 Experimental 2
5th day 28,1±0,68 29,4±1,24 29,0±1,10
10th day 30,7±1,45 29,9±1,06
20th day 31,6±1,34 29,2±0,98
25th day 31,3±1,30 28,9±0,91
30th day 30,9±1,46 28,6±1,01
Degree of probability:* - p < 0,05;** - p < 0,025; *** - p < 0,001
In the blood of rats of the E1 group, the leukocytes in these periods was higher by 9.6 and 7.6%. On the number on the 10th day increased by 24.9%, while on 30th day of the research, the studied indicator in the E2 the 20th day - by 34.0% compared with intact mice. In group's blood reached physiological values (Table 4). the blood of rats of the E2 group, the leukocytes number
Table 4
The leukocytes number in the blood of rats under conditions of carbon tetrachloride poisoning and the _action of feed additive "Butaselmevit-plus," G / l (M±m; n=20)_
Test Day Groups of animals
Control Experimental 1 Experimental 2
5th day 9,34±0,760 10,74±0,731 10,13±0,820
10th day 11,67±1,221 10,24±1,106
20th day 12,52±1,357* 10,05±1,310
25th day 12,10±1,621* 9,87±1,754
30th day 11,67±0,987 9,45±1,052
Degree of probability:* - p < 0,05;** - p < 0,025; *** - p < 0,001
Restoration of the hematopoietic function of rat bone marrow in tetrachloromethane intoxication is because the feed additive "Butaselmevit-plus" contains milk thistle fruits have a high level of vitamins A and K, and trace elements, namely: Copper, iron, and Cobalt directly involved in hematopoiesis.
Conclusion
Under conditions of tetrachloromethane intoxication in rats, the bone marrow's hematopoietic function is impaired, manifested by a decrease in the number of erythrocytes 26.5%, the hemoglobin content by 8.7%. When using the feed additive "Butaselmevit-plus" in rats, under circumstances of oxidative stress during the study, the blood normalizes the activity of hematological parameters, namely on the 25th and 30th day within the physiological values of erythrocytes, hemoglobin, and leukocytes compared with control, indicating the restoration of hematopoietic function of the bone marrow.
Research prospects. In the future, there are plans to study the effect of feed additive "Butaselmevit-plus" on the morphological parameters of piglets at weaning from the sow.
References
1. Li, D., Cai, H., Hou, M., Fu, D., Luo, Q., Yuan, X., Lv, M., Zhang, X., Cong, X., & Lv, Z.
(2012) Effects of indoleamine 2,3-dioxygenases in carbon tetrachloride-induced hepatitis model of rats. Cell Biochem Funct, 30(4), 309-314. doi: 10.1002/cbf.2803.
2. Gutiy, B. V. (2012). Vplyv hlorydu kadmiyu na intensivnist protsesiv perekisnogo okysnennya lipidiv ta stan sistemy antyoksydantnogo zahystu organizmu shchuriv. Visnyk Sumskogo natsionalnogo agrarnogo universitetu, 7(31), 31-34 (in Ukrainian).
3. Gutyj, B. V. (2016). Osobennosti funkcion-irovanija sistemy antioksidantnoj zashhity organizma krys pri kadmievom toksikoze. Nauchno-prakticheskij zhurnal. Uchenye Zapiski, 52(2), 24-28 (in Russian).
4. Abragamovich, O. O., Grabovska, O. I., & Terletska, O. I. (2000). Protsesy lipidnoyi peroksidatsiyi pry hronichnyh urazhennyah pechinky. Medichna himiya, 2(1), 5-8 (in Ukrainian).
5. Hariv, M. I., & Gutyj, B. V. (2016). Influence of the liposomal preparation Butaintervite on protein synthesis function in the livers of rats under the influence of carbon tetrachloride poisoning. Visnyk of Dnipropetrovsk University. Biology, medicine, 7(2), 123-126. doi: 10.15421/021622.
6. Gutyj, B. V., Murs'ka, S. D., Gufrij, D. F., Hariv, I. I., Levkivs'ka, N. D., Nazaruk, N. V., Gajdjuk, M. B., Pryjma, O. B., Bilyk, O. Ja., & Guta, Z. A. (2016). Influence of cadmium loading on the state of
«C@yL@qyiym-J®yrMaL»#32i84),2©2© / VETERINARY SCIENCE
11
the antioxidant system in the organism of bulls. Visnyk of Dnipropetrovsk University. Biology, ecology, 24(1), 96-102. doi: 10.15421/011611.
7. Gutyj, B., Lavryshyn, Y., Binkevych, V., Binkevych, O., Paladischuk, O., Strons'kyj, J., & Hariv, I. (2016). Influence of «Metisevit» on the activity of enzyme and nonenzyme link of antioxidant protection under the bull's body cadmium loading. Scientific Messenger LNUVMBT named after S.Z. Gzhytskyj, 18, 2(66), 52-58. doi: 10.15421/nvlvet6612.
8. Khariv, M., Gutyj, B., Butsyak, V., & Khariv, I. (2016). Hematological indices of rat organisms under conditions of oxidative stress and liposomal preparation action. Biological Bulletin of Bogdan Chmelnitskiy Melitopol State Pedagogical University, 6(1), 276-289. doi: 10.15421/201615.
9. Martyshuk, T. V., Gutyj, B. V., & Vishchur, O. I. (2016). Level of lipid peroxidation products in the blood of rats under the influence of oxidative stress and under the action of liposomal preparation of «Butaselmevit», Biological Bulletin of Bogdan Chmelnitskiy Melitopol State Pedagogical University, 6(2), 22-27. doi: 10.15421/201631.
10. Huberuk, V., Gutyj, B., Gufriy, D., Binkevych, V., Hariv, I., Binkevych, O., & Salata, R. (2017). Impact of antioxidants on enzym activities of glutatione system of bulls bodies antioxidant defense under acute nitrate and nitrite toxicity. Scientific Messenger LNUVMBT named after S.Z. Gzhytskyj, 19(77), 220-224.
11. Lavryshyn, Y. Y., Varkholyak, I. S., Mar-tyschuk, T. V., Guta, Z. A., Ivankiv, L. B., Paladischuk, O. R., Murska, S. D., Gutyj, B. V., & Gufriy, D. F. (2016). The biological significance of the antioxidant
defense system of animals body. Scientific Messenger LNUVMBT named after S.Z. Gzhytskyj, 18, 2(66), 100-111. doi: 10.15421/nvlvet6622.
12. Khariv, M., Gutyj, B., Ohorodnyk, N., Vishchur, O., Khariv, I., Solovodzinska, I., Mudrak, D., Grymak, C., & Bodnar, P. (2017). Activity of the Tand B-system of the cell immunity of animals under conditions of oxidation stress and effects of the liposomal drug. Ukrainian Journal of Ecology, 7(4), 536-541. doi: 10.15421/2017_157.
13. Lee, J. Y., Lee, J. H., & Kim, H. J. (2004). The preventive inhibition of chondroitin sulfate against the CCI4-induced oxidative stress of subcellular level. Arch. Pharm. Res., 27(3), 340-345.
14. Skry'pny'k, I. M. (2007). Gepatoprotetorni zasoby' v suchasnij gepatologiyi. Consilium Medicumllkraina, 1(5), 11-15 (in Ukrainian).
15. Saba, A. B., Oyagbemi, A. A., & Azeez, O. I. (2010). Amelioration of carbon tetrachloride-induced hepatotoxicity and haemotoxicity by aqueous leaf extract of Cnidoscolus aconitifolius in rats. Nig. J. Physiol. Sci., 25(2), 139-147.
16. Antonyak, G. L., Babich, N. O., & Sologub, L. I. (2000). Utvorennya aktivnih form kisnyu ta sistema antioksidantnogo zahistu v organizmi tvarin. Biologiya tvaryn, 2(2), 34-43 (in Ukrainian).
17. Belenichev, I. F. Levitskiy, E. L., & Kovalenko, S. I. (2002) Antioksidantna sistema zahistu organizmu (oglyad). Sovremennye problemy toksikologii, 3, 29-31 (in Russian).
18. Vlizlo, V. V., Fedoruk, R. S., & Ratych, I. B. (2012). Laboratory methods of investigation in biology, stock-breeding and veterinary. A reference book. Ed. by VV Vlizlo. Lviv: Spolom (in Ukrainian).
UDC 577.1:612.015
Lavryshyn Y., Gutyj B., Leskiv Ch., Kalyn B., Holub O., Romanovych M.
Stepan Gzhytskyi National Univesity of Veterinary Medicine and Biotechnologies,
Lviv, Pekarska street, 50, Lviv, 79010, Ukraine DOI: 10.24412/2520-2480-2020-3284-11-14
THE EFFECT OF CADMIUM ON THE LEVEL OF VITAMINS A AND E IN THE BLOOD OF
YOUNG CATTLE
Abstract.
The study aimed to examine the effect of cadmium load on the level of vitamins A and E in young cattle's blood. The research was conducted based on a farm in the village of Ivanivtsi, Zhydachiv district, Lviv region, on ten six-month-old bulls, Ukrainian black-and-white dairy breed, which was formed into two groups of five animals each: control (C) and experimental (E). The bulls of the control group were on a regular diet. The (E) group animals were fed cadmium chloride at a dose of 0.04 mg/kg body weight. The test lastedfor 30 days. For research, the rules obligatory for performing zootechnical experiments on selecting and keeping analogous animals in groups, harvesting technology, use, and accounting of consumed feed were followed. The animals' diet was balanced in terms of nutrients and minerals, which provided their need for essential nutrients. The investigations were carried out under the principles of humanity set out in the European Community Directive. The experiment results indicate that Cadmium significantly affects liver cells' metabolism, thus stimulating the processes of lipid peroxidation and inhibiting the activity of the non-enzymatic part of the antioxidant system, the level of vitamins A and E in the blood of young cattle. The lowest level of vitamins A and E was in the blood of the (E) group of bulls on the 20th day of the experiment, respectively, fluctuated between 0.62 ± 0.035 and 2.9 ± 0.10 fimol/l. Only on the 5 th day of the investigation there was a slight increase in the content of vitamins, which may be due to the
