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VETERINARY SCIENCES / «g®LL®q(U]UM~J®U®MaL» #18(141), 2022
Veterinary sciences
Stybel V.V.1, Gytyj B. V.1, Frejuk D. V.1, Khalak V.I. 2, Kuljaba O. V.1, Martyshuk T. V.1, Adamiv S.S. 3, Pavliv O. V. 3, Leskiv Kh. Ya.1
DOI: 10.24412/2520-6990-2022-18141-7-10
'Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies of Lviv, Ukraine 2State Institution Institute of Grain Crops NAAS of Ukraine, Dnipro, Ukraine 3Separated subdivision of National University of Life and Environmental Sciences of Ukraine "Berezhany
Agrotechnical Institute ", Berezhany, Ukraine
ANTIOXIDANT STATUS OF COWS BODY IN EXPERIMENTAL FASCIOLOSIS AND THE ACTION
OF CORRECTIVE FACTORS
Abstract
The article presents the results of studies on the effect of Clozaverm A and Butaselmevit on the antioxidant status of cows in experimental fasciolosis. In the blood of cows, an increase in lipid peroxidation processes is shown. It is indicated by elevated levels of lipid hydroperoxides and TBA-active products. Fasciola inhibits the activity of enzymes of the antioxidant defense system in the liver of the hosts. A decrease in the catalase activity indicates it by 23.4% and superoxide dismutase by 24.6%. The normalization of redox processes reduces the level of products of lipid peroxidation, and, consequently, the intoxication of the body occurs under the influence of Clozaverm A and Butaselmevit. Clozaverm A and Butaselmevit treatment in cows with experimental fasciolosis improved the antioxidant status of their body. It is accompanied by an increase in blood superoxide dismutase activity by 31.6%, catalase by 30.1%, inhibition of the processes of peroxide oxidation by 5%, and TBA-active products by 18.1%. It is due to the activation of metabolic processes in the body, which involve enzymes, including antioxidant enzymes that catalyze the oxidation and phosphorylation functions, and bone marrow.
Keywords: fasciolosis, Clozaverm A, Butaselmevit, cattle, antioxidants.
Introduction
Fasciolosis is one of the dangerous helminthiases that causes significant economic damage to farms. Under these conditions, the milk productivity of cows and weight gain of young animals decreases. There is a negative impact on the reproduction of the herd. A significant problem is the development of resistance of fasciolosis to anthelmintics [1]. This confirms the relevance of studying various treatment regimens and, in particular, complex therapy of animals in this invasion.
In recent years, scientists have drawn attention to studying free radical oxidation of membrane lipids as a regulator of physiological processes [2]. One of the universal types of damage and the cause of cell death of various organs is the excessive intensification of lipid peroxidation (LPO) processes and damage to biological membranes. Excessive activation of LPO disrupts conformation and increases the permeability of biological membranes, releases enzymes from mitochondria and lysosomes, inactivation of aerobic oxidation enzymes, separation of oxidative phosphorylation and DNA mutations [13, 14, 19].
According to the literature, under the influence of various negative factors on the body, the reactions of free radical oxidation in cells are sharply enhanced [3, 8]. During free radical peroxidation, at almost all stages, many products are formed, resulting from the interaction of free radicals with biological macromole-cules [5, 15, 21]. It is important to note that the increased formation of primary free radicals is a side effect of increasing the intensity of biochemical reactions in response to the action of extreme factors [6, 13, 16, 18].
Based on previous studies, we found that in experimental fasciolosis in cows, parasites secrete metabolic products that promote the formation of free radicals. It enhances the initiation of lipid peroxidation. This is indicated by the growth of lipid peroxidation products: diene conjugates, lipid hydroperoxides, and TBA-active products [4, 10-12].
The study aimed to investigate the effect of Clozaverm A and Butaselmevit on the antioxidant status of cows in experimental fasciolosis.
Material and methods of research
Fifteen 4-5-year-old cows of the black and speckled breed were selected for the research. Three groups were formed, with five animals in each. Animals of the control group (C) were experimentally infected with adolescents. Cows of the first group (E1) in experimental fasciolosis were administered Clozaverm A at a dose of 0.5 ml per 10 kg of body weight intramuscularly. In animals of the second group (E2), in experimental fasciolosis, Clozaverm A was administered intramuscularly at a dose of 0.5 ml of the drug per 10 kg of body weight and Butaselmevit [7] at a dose of 10 ml of the drug per animal.
During the research, the rules of compulsory experiments were followed - selecting and keeping analogous animals in groups. The cows' diet was balanced in terms of nutrients and minerals.
The content of lipid hydroperoxides and TBA-active products in blood plasma was determined according to the methods [22]. In the serum studied: catalase activity (CA 1.11.1.6) - by the method of MA Korolyuk (1988); superoxide dismutase activity was determined (SDA 1.15.1.1.) - by the method of E.E.
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Dubinina [22]. Blood for analysis was taken from the jugular vein before infection and on the experiment's 7th, 14th, 21st, and 28th days.
Experimental research was conducted per the Law of Ukraine "On Protection of Animals from Cruelty" of March 28, 2006, and the European Convention for the Protection of Vertebrate Animals Used for Experimental and Other Scientific Purposes of November 13, 1987.
The analysis of research results was performed using the software package Statistica 6.0. Student's t-test assessed the probability of differences. The results of the mean values were considered statistically significant at * - P<0.05, ** - P<0.001 (ANOVA).
Results and discussion
In experimental fasciolosis in cows' blood was an increase in LPO processes, as indicated by elevated lipid hydroperoxides and TBA-active products.
Table 1
The Clozaverm A and Butaselmevit effect on the antioxidant status of cows in experimental fasciolosis (M ±
m; n = 5)
After the Clozaverm A treatment in the E1 group, a decrease in the intensity of LPO processes was found. On the 14th day, the lipid hydroperoxides' level was 11.6%, and the level of TBA-active products was 8.9% relative to the control group. Subsequently, the level of LPO products in the E1 group continued to decline, and on the 28th day, it was 2.02±0.06 units/ml, TBA-active products 6.03±0.13 nmol/ml, while in the C group, these indicators were 2.60±0.03 units E/ml and 7.11±0.11 nmol/ml.
The use of Butaselmevit and Clozaverm A for treating cows with experimental fasciolosis significantly reduced the intensity of LPO processes in their body. In addition, a probable decrease in LPO products was detected from the 14th day of the trial, where these indicators were reduced by 17.2% (LHP) and 12.1% (TBA-active products).
Groups Term of research (days)
of Before infection 7 14 21 28
animals
Lipid hydroperoxides, units E/ml
C 1.82±0.02 2.18±0.03 2.33±0.03 2.54±0.04 2.60±0.03
Ei 1.80±0.02 2.01±0.05* 2.06±0.06* 2.11±0.02*** 2.02±0,06**
E 2 1.81±0.03 1.85±0.05** 1.93±0.03*** 1.89±0.05*** 1.78±0.05***
TBA-active products, nmol/ml
D 5,82±0.11 6.29±0.10 6.70±0.10 6.95±0.10 7.11±0.11
E 1 5.80±0.10 6.02±0.13 6.10±0.15* 6.17±0.14** 6.03±0.13***
E 2 5.83±0.11 5.92±0.14* 5.89±0.16** 5.85±0.14*** 5.82±0.15***
Catalase, mcat/l
C 47.15±1.12 40.06±1.12 38.72±1.11 36.11±1.12 38.02±1.11
E 1 47.17±1.10 43.31±1.13* 44.78±1.12** 45.32±1.13*** 45.90±1.15**
E 2 47.20±1.11 45.24±1.12* 46.52±1.10** 46.99±1.15*** 47.51±1.16***
Superoxide dismutase, c.u/mg protein
C 1.30±0.01 1.19±0.02 1.11±0.03 0.98±0.03 1.01±0.01
E 1 1.32±0.02 1.22±0.03 1.13±0.02 1.14±0.02** 1.25±0.03**
E 2 1.30±0.02 1.26±0.01* 1.27±0.03* 1.29±0.04*** 1.32±0.02***
The obtained data indicate that under the influence of Butaselmevit and Clozaverm A in the body of cows with experimental fasciolosis, there is a normalization of redox processes and reduces the level of LPO products and, consequently, the intoxication of the body.
On the 21st day in the C group, the catalase activity was lower by - 23.4% than in clinically healthy animals (before infection). After treating cows with the drug "Clozaverm A", we note the normalization of enzyme activity in serum from 14 days of the test. On the 21st day, the catalase activity of the E1 group increased by 25.5%, on the 28th day by 20.7% relative to the C group.
The use of Butaselmevit in the E2 group of animals contributed to a faster increase in catalase activity from the 7th day. However, on 21 and 28 days, the activity of the studied enzyme fluctuated within physiological values.
We also found that at the beginning and end of the experiment, the activity of superoxide dismutase ranged from 1.30±0.01 - 0.98±0.03 c.u /mg protein.
Clozaverm A and Butaselmevit effects in cows contributed to the activation of superoxide dismutase activity in their serum. On the 21st day, the enzyme activity in the E1 group increased by 16.3%, and in the E2 group - by 31.6% relative to the C group. The highest activity of SOD was in the E2 group on the 28th day of the research, it was 1.32±0.02 c.u /mg protein, which is 30.7% higher than the C group.
Conclusions
Fasciola inhibits the activity of enzymes of the antioxidant defense system in the liver of hosts. A decrease in catalase activity indicated by 23.4%, superoxide dismutase by 24.6%, an increase in lipid hydroperoxides by 42.9%, and TBA products by 22.2%.
Using Clozaverm A and Butaselmevit in cows with experimental fasciolosis improved the antioxidant status of their body. The growth of SOD accompanied it by 31.6% and, catalase by 30.1% and, suppression of lipid peroxidation by 31.5%, TBA-active products by 18.1%.
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VETERINARY SCIENCES / «g®LL®(MUM~J®U®MaL» #18(141), 2022
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Said W.S.1, Gytyj B. V. 1, Kushnir I.M. 2, Hunchak V.M.1, Hunchak A. V. 3, Khalak V.l. 4, Kushnir V.l. 2, Martyshuk T. V.1, Leskiv Kh. Ya.1, Gyta Z. A. 1
'Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies of Lviv, Ukraine 2State Research Control Institute of veterinary medicinal products and feed additives, Lviv, Ukraine
3Institute of Animal Biology NAAS, Lviv, Ukraine 4State Institution Institute of Grain Crops NAAS of Ukraine, Dnipro, Ukraine
DOI: 10.24412/2520-6990-2022-18141-10-16 MORPHOLOGICAL PARAMETERS OF DOGS' BLOOD, WITH EXPERIMENTAL INFESTATION
WITH TOXOCARIASIS AND "FENBENZYL"
Abstract
The study aimed to determine the effect of "Fenbenzyl" on the morphological parameters of dogs' blood during experimental infestation with toxocariasis. The tests were performed on 18 dogs aged 2-4 months. It was found that with the use of fenbenzyl in infected dogs, the erythrocytes number and hemoglobin content on 25 and 30 days of the trial ranged within physiological values. The drug "Fenbenzyl" decreases the number of leukocytes within 10 days of the experiment. In the second group, the leukocyte number was more likely to decline compared with the control group. Such changes are due to the antiparasitic effect of the "Fenbenzyl" and the cessation of Toxocara toxins on the bone marrow. Eosinophils, neutrophils, and monocytes were significantly reduced in all groups when "Fenbenzyl" was administered to animals compared to untreated infected dogs. The lymphocyte number of the experimental dogs probably increased from the 15th day of the trial. On 25 and 30 test days, the lymphocyte number grew to physiological values in dogs treated with "Fenbenzyl". In our opinion, this is due to the reduction of the immunosuppressive effect of Toxocara and hepatoprotective, immunostimulatory and antiox-idant effects of milk thistle.
Keywords: toxocariasis, dogs, "Fenbendazole", "Fenbenzyl", leukogram, milk thistle.
Introduction
Among the invasive diseases of dogs, the most common in our country and abroad are helminths of the digestive tract, among which the top place is toxocariasis - nematode infestation of the order Ascaridata [1-3, 6, 24]. Infection of puppies mainly occurs in utero by transplacental transmission of larvae from mother to fetus in the second half of pregnancy or the first days of life with milk [7, 26]. The direct route of infection is by swallowing Toxocara eggs directly from the soil or contaminated soil. In the stomach or small intestine, larvae emerge from the eggs, similar to the larvae of A. suum in pigs [10, 11]. Through the intestinal mucosa, second-stage larvae enter the intestine and portal vein system venous vessels and enter the liver. Then into the inferior vena cava, right half of the heart, and pulmonary artery to the pulmonary capillaries, where the second molt occurs. From pulmonary capillary vessels, larvae of the third stage actively reach bronchioles, bronchial tubes, and trachea. Then, with mucus, they get to an oral cavity and are swallowed with saliva. In the small intestine, there are two more molts. In 4-5 weeks after infection, the parasites reach sexual maturity. This migration path is mainly registered in puppies under five weeks [2, 8, 25].
In adult dogs (older than one year), the larvae do not fully develop. It stops in the larvae of the second stage. Some larvae, reaching the lungs, and penetrating
the pulmonary vein, migrate through the heart into the great circle of blood circulation. The arterial system enters various organs and tissues (lungs, liver, kidneys, muscles), where they incise, maintaining viability for many years. In the case of reduced resistance or changes in hormone levels (pregnancy, lactation), the larvae leave their habitats [9, 22].
For treating carnivorous animals with toxocariasis are widely used drugs based on fenbendazole [15, 16, 20, 21]. They do not have sensitizing, carcinogenic, embryotoxic, mutagenic, teratogenic, and allergenic effects, do not irritate the skin and mucous membranes of animals and do not affect the course of pregnancy [18, 21].
Scientists have established the stimulating effect of milk thistle on animals' antioxidant and immune status under various negative factors [5, 17]. However, the complex application of milk thistle and fenbendazole on morphological and biochemical parameters of the blood of infected dogs is currently insufficiently covered in the scientific literature.
The study aimed to determine the effect of "Fenbenzyl" on the morphological parameters of dogs' blood during experimental infestation with toxocariasis.
