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VETERINARY SCIENCE / «COLLMUWM-JMrMaL» #22(109), 2©2U
Veterinary science
Huberuk V. O., Gufrij D. F., Gutyj B. V., Hunchak V. M.
Hariv 1.1., Vasiv R. O., Leskiv Kh. Ya., Guta Z. A.
Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv, Ukraine
DOI: 10.24412/2520-6990-2021-22109-8-14
THE INFLUENCE OF URSOVIT-ADES AND SODIUM SELENITE ON BIOCHEMICAL AND MORPHOLOGICAL PARAMETERS OF BULL'S BLOOD IN ACUTE NITRATE-NITRITE
TOXICOSIS
Abstract
The article presents studies of the Ursovit-ADES and sodium selenite effect on the biochemical and morphological parameters of the bull's blood in acute nitrate-nitrite toxicosis. The experiments were performed on bulls of six months of age, black-spotted breed. The combined use of chromosmone with Ursovit-ADES and sodium selenite generally had a positive effect on the morphological and biochemical indicators of the blood under conditions of acute nitrate-nitrite toxicosis. It affected the improvement of the functional state of the test animals and their rapid recovery. Furthermore, introducing the investigated drugs to animals helped reduce nitrates, nitrites, methemoglobin, ammonia, and leukocytes in the blood. Under these conditions, an increase in erythrocytes, hemoglobin, and urea was found. Therefore, the combined use of chromosmone, Ursovit-ADES, and sodium selenite in bulls to a greater extent has a curative and therapeutic effect, which helps to reduce the manifestations of endogenous intoxication.
Key words: nitrates, nitrites, toxicosis, blood, Ursovit-ADES, sodium selenite, chromosmone.
Introduction
Contamination of the environment with nitrates and nitrites and their negative impact on the body of animals make the problem of studying the mechanism of nitrate-nitrite toxicosis especially relevant, which has theoretical and practical significance [1, 2].
Against the background of published scientific works on the study of nitrate-nitrite toxicosis in farm animals, some reports claim that the toxic effects of nitrates are manifested in two interrelated directions. In the first stage, there is methemoglobin formation and activation of free radicals, which in the second stage initiate the processes of lipid peroxidation [3-5].
The problem of acute nitrate-nitrite toxicosis in young cattle is relevant today, as the violation of the ecological situation in the country has led to the poisoning of animals with nitrates, even in small quantities, which is associated with reduced natural resistance and to various factors. [6, 8]
A significant number of scientific works of both domestic and foreign researchers are devoted to the study of the etiology, pathogenesis, diagnosis, treatment, and prevention of acute nitrate-nitrite toxicosis. Scientific studies have been performed on animals to reveal the multifaceted pathogenesis of nitrate-nitrite toxicosis in farm animals. Various methods of treatment and prevention of animals from subclinical chronic and acute toxicosis have been proposed. Nitrate poisoning of animals has shown high therapeutic efficacy of vitamin preparations: ascorbic acid, pyridoxine hydrochloride, and thiamine chloride [7-11].
Our studies aimed to determine the effect of Urso-vit ADES and sodium selenite on the biochemical and morphological indicators of the blood of bulls in acute nitrate-nitrite toxicosis.
Material and methods of research.
The experiments were performed on bulls of six months of age, a black-spotted breed formed into 3 groups of 5 animals each.
The bulls of the control group were fed with feed once sodium nitrate at a dose of 0.45 NO3~/kg body weight. The bulls of the E1 group were fed sodium nitrate once at a dose of 0.45 NO3~/kg body weight, and three hours later, after 3 hours, iv was injected with chromosmon eat a dose of 0.1 ml/kg body weight iv was administered Ursovit-ADES in the dose of 4 ml per animal. The bulls of the second experimental group were fed sodium nitrate once at a dose of 0.45 NO3~/kg body weight. After three hours, intravenously was injected with chromosmone of 0.1 ml/kg body weight. IV was administered Ursovit-ADES at a dose of 4 ml per animal and a 0.5% sodium selenite solution dose of 1 ml per animal.
Venous blood was collected at the beginning of the trial and 3 hours after feeding bovine sodium nitrate and 1, 2, 3, 6, 8 hours after administering vitamin preparations.
Analysis of research results was performed using Statistica 6.0. Student's t-test assessed the probability of differences; the results were considered probable at P<0.05, the data in the figures are given as mean and standard deviation.
Research results
After examining the adverse effects of nitrates in toxic doses on the bulls, we concluded that for effective treatment and correction of morphological, biochemical parameters of blood and balance between LPO processes and antioxidant system, it is necessary to use an-tioxidants along with standard antidotes. In our case, we used Ursovit-ADES and sodium selenite along with the chromosmone. The effect of these antioxidants on the body of bulls fed sodium nitrate at a dose of 0.45 g NO3~/kg is shown in table 1.
When bulls were fed sodium nitrate at a dose of 0.45 gNO3~/kg body weight, a high level of nitrates was
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found in their blood. At the third hour of research, their level was in the range of 2.76±0.095 - 2.90±0.095 mg/l. After introducing chromosmones into the body of ani-
The concentration of nitrates in the blood of bulls
For the first hour of the trial in the experimental groups of animals injected intramuscularly with Urso-vite ADES and sodium selenite, the level of nitrates decreased by 60%. In group Ei for the second and third hours, the nitrates level felt, respectively, by - 71 and 80%, and group E2 - by 74 and 81% relative to the control group of bulls. At the eighth hour of the investigation, the level of nitrates in the test groups was the lowest.
In the research of nitrite level in the blood of experimental bulls, which were subjected to nitrate loading, before the introduction of antidote and antioxidant drugs, it was high for the third hour of the trial, where it was in the range of 0.084±0.0018 - 0.088±0.0016 mg/l (Table 2).
The nitrite concentration probably reduced throughout the investigation after applying chro-mosmone, Ursovit-ADES, and sodium selenite to the pilot bulls. Thus, for the first hour after the use of anti-
mals commonly with antioxidants in acute nitrate-nitrite toxicosis, the level of nitrates in the blood of experimental bulls decreased relative to the control group (Table 1).
Table 1
er the use of Ursovit-ADES and sodium selenite in
oxidant medicines, the concentration of the studied indicator in the blood of bulls of groups E1 and E2 was 0.076±0.0020 and 0.069±0.0020 mg/l, respectively. After applying Ursovit-ADES, the concentration of nitrites in the second hour decreased by 71%, in the third hour - by 90% concerning animals that were not used vitamin D preparations. After using Ursovit-ADES and sodium selenite, the nitrites concentration reduced in these periods by 72 and 90%, respectively.
At the eighth hour, the nitrites level in the blood of bulls of the experimental group, which was used with the chromosmone, Ursovit-ADES, was 0.008±0.0003 mg/l. In bulls, which were used Ursovit-ADES with sodium selenite, the nitrite concentration was within 0.007±0.0003 mg/l.
Therefore, the use of the above drugs in bulls, which were carried out nitrate load in toxic doses, prevented the development of poisoning by nitrates and nitrites, as indicated by their low level in the blood of test animals.
acute nitrate-nitrite toxicosis, mg/l (M±m, n=5)
Animal blood counts (hours) Groups of animals
Control Experimental (E1) Experimental (E2)
Before feeding sodium nitrate
Control 0.025±0.0015 0.029±0.0013 0.031±0.0013
After feeding sodium nitrate
Third hour 2.76±0.095 2.90±0.095 2.83±0.090
— After the introduction of chromosmone
Ursovit-ADES Ursovit-ADES + 0.5% solution of sodium selenite
First hour 3.39±0.080 1.35±0.010** 1.37±0.010**
Second hour 3.64±0.070 1.05±0.009** 0.94±0.009**
Three hour 4.03±0.080 0.81±0.005** 0.78±0.005**
Six hour 3.24±0.080 0.56±0.001** 0.51±0.001**
Eighth hour 1.95±0.060 0.32±0.001** 0.31±0.001**
Note: the degree of probability compared with the control group - P<0.05 - *, P<0.001 - **
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Note: the degree of probability compared with the control group - P<0.05 - *, P <0.001 - **
A decrease of the nitrates and nitrites level in the a toxic dose, the level of methemoglobin in the blood blood was accompanied by methemoglobin reduction. of these animals probably progressed and in the control As shown in table 3, the methemoglobin level at the be- group was the maximum value of 31.06±0.31%. ginning of the test was within physiological values. After feeding bulls with sodium nitrate compound feed in
Table 3
The methemoglobin level in the blood of bulls after the use of Ursovit-ADES and sodium selenite in acute
nitrate-nitrite toxicosis, % (M±m, n = 5)
Table 2
The nitrites concentration in the blood of bulls after the use of Ursovit-ADES and sodium selenite in acute nitrate-nitrite toxicosis, mg/l ^ (M ± m, n = 5)_
Animal blood counts (hours) Groups of animals
Control Experimental (Ei) Experimental (E2)
Before feeding sodium nitrate
Control 0 0 0
After feeding sodium nitrate
Third hour 0.088±0.0016 0.084±0.0018 0.087±0.0015
— After the introduction of chromosmone
Ursovit-ADES Ursovit-ADES + 0.5% solution of sodium selenite
First hour 0.139±0.0019 0.076±0.0020** 0.069±0.0020**
Second hour 0.187±0.0016 0.054±0.0015** 0.051±0.0015**
Three hour 0.216±0.0032 0.021±0.0010** 0.020±0.0010**
Six hour 0.131±0.0030 0.011±0.0005** 0.009±0.0005**
Eighth hour 0.076±0.0020 0.008±0.0003** 0.007±0.0003**
Animal blood counts (hours) Groups of animals
Control Control Experimental (E1) Experimental (E2)
Before feeding sodium nitrate
4.2±0.08 4.5±0.05 4.2±0.05
After feeding sodium nitrate
Third hour 18.60±0.30 18.67±0.32 18.62±0.35
— After the introduction of chromosmone
Ursovit-ADES Ursovit-ADES + 0.5% solution of sodium selenite
First hour 23.42±0.30 17.24±0.15** 16.85±0.14**
Second hour 27.51±0.28 13.63±0.10** 13.12±0.10**
Three hour 31.06±0.31 9.35±0.11** 9.21±0.11**
Six hour 25.87±0.28 7.93±0.10** 7.74±0.10**
Eighth hour 22.54±0.30 7.19±0.10** 7.11±0.10**
Note: the degree of probability compared with the control group - P<0.05 - *, P<0.001 - **
After intramuscular administration of chromosmone and Ursovit-ADES, the methemoglobin level decreased by 26.4% in the first hour and by 50% in the second hour. After the combined use of Ursovit-ADES and sodium selenite, the methemoglobin level in the first hour was reduced by 28%. The second hour decreased by 52% relative to the control group. At the eighth hour of the investigation, the methemoglobin level in the blood of experimental groups, which were
used antioxidant drugs, was 7.19±0.10 and 7.11±0.10%, respectively.
Thus, chromosmone and Ursovit-ADES and sodium selenite administration helped to reduce the methemoglobin level in their blood.
The effect of Ursovit-ADES and sodium selenite on the level of hemoglobin in the blood of bulls after feeding sodium nitrate is shown in table 4.
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Table 4
The level of hemoglobin in the blood of bulls after the use of Ursovit-ADES and sodium selenite in acute _ nitrate-nitrite toxicosis, g/l (M±m, n = 5)_
Animal blood counts (hours) Groups of animals
Control Experimental (Ei) Experimental (E2)
Before feeding sodium nitrate
Control 100.9±1.40 101.4±1.45 101.1±1.42
After feeding sodium nitrate
Third hour 84.1±1.45 86.2±1.30 85.5±1.41
— After the introduction of chromosmone
Ursovit-ADES Ursovit-ADES + 0.5% solution of sodium selenite
First hour 80.2±1.55 88.7±1.40* 88.9±1.40*
Second hour 78.3±1.40 90.5±1.34** 91.1±1.34**
Three hour 76.2±1.41 94.8±1.35** 94.9±1.35**
Six hour 79.1±1.50 99.2±1.30** 99.7±1.30**
Eighth hour 84.9±1.45 100.9±1.34** 101.4±1.34**
Note: the degree of probability compared with the control group - P<0.05 - *, P<0.001 - **
After nitrate loading, in the blood of bulls of experimental groups, the hemoglobin level probably declined relative to the initial values and fluctuated within 84.1±1.45 - 86.2±1.30 g/l. After introducing chro-
mosmone and Urosvit-ADES, the bulls' blood's hemoglobin levels ranged from 88.7±1.40 to 100.9±1.34 g/l. Finally, after using chromosmone, vitamin preparation, and sodium selenite, the hemoglobin level at the eighth hour reached the limits of physiological values.
Table 5
The urea content in the serum of bulls after using Ursovit-ADES and sodium selenite in acute nitrate-nitrite toxicosis, mmol/l (M±m, n = 5)
Animal blood counts (hours) Groups of animals
Control Experimental (E1) Experimental (E2)
Before feeding sodium nitrate
Control 3.99±0.081 4.00±0.080 4.03±0.072
After feeding sodium nitrate
Third hour 3.40±0.061 3.39±0.072 3.40±0.070
— After the introduction of chromosmone
Ursovit-ADES Ursovit-ADES + 0.5% solution of sodium selenite
First hour 3.23±0.060 3.34±0.044 3.35±0.052
Second hour 3.02±0.054 3.49±0.061** 3.49±0.070**
Three hour 2.81±0.072 3.70±0.054** 3.74±0.054**
Six hour 3.02±0.063 3.89±0.070** 3.92±0.062**
Eighth hour 3.64±0.070 3.98±0.054** 3.99±0.053*
Note: the degree of probability compared with the control group - P<0.05 - *, P<0.001 - **
It is known that acute nitrate-nitrite toxicosis in the blood of urea lowers the urea content, as shown in table 5. After using chromosmone with Ursovit-ADES and sodium selenite, the urea content in the blood of bulls of the experimental groups normalized throughout the experiment.
The combined use of the above experimental drugs in acute nitrate-nitrite toxicosis contributed to a better normalizing effect on the urea content in sick bulls with nitrate-nitrite toxicosis. At the eighth hour of the search, the urea content in the experimental animals reached the initial values and were respectively 3.98±0.054 and 3.99±0.053 mmol/l.
The ammonia level in the blood of bulls before nitrate load at the beginning of the experiment was within physiological values.
After nitrate loading, by feeding sodium nitrate in a toxic dose, the ammonia level increased. For the third hour of the research it was; in animal groups C 1.56±0.045 mg/l, in group Ei - 1.53±0.036 mg/l, in group E2 - 1.55±0.040 mg/l. After administration of Ursovit-ADES and sodium selenite to sick bulls, the level of ammonia in their blood probably reduced throughout the trial, where at the eighth hour, it was 1.08±0.060 mg/l, respectively.
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Thus, with the development of acute nitrate-nitrite toxicosis, the introduction of chromosmone and antioxidants contributed to the normalization of ammonia levels in their serum.
Table 6
The level of ammonia in the serum of bulls after the use of Ursovit-ADES and sodium selenite in acute ni_ trate-nitrite toxicosis mg/l (M±m, n=5)_
Animal blood counts (hours) Groups of animals
Control Experimental (Ei) Experimental (E2)
Before feeding sodium nitrate
Control 1.02±0.042 1.08±0.030 1.06±0.025
After feeding sodium nitrate
Third hour 1.56±0.045 1.53±0.036 1.55±0.040
— After the introduction of chromosmone
Ursovit-ADES Ursovit-ADES + 0.5% solution of sodium selenite
First hour 1.61±0.036 1.47±0.065* 1.44±0.054*
Second hour 1.79±0.035 1.32±0.061** 1.29±0.061**
Three hour 1.92±0.045 1.19±0.055** 1.18±0.055**
Six hour 1.85±0.040 1.14±0.050** 1.11±0.050**
Eighth hour 1.52±0.050 1.10±0.060** 1.08±0.060**
Note: the degree of probability compared with the control group - P<0.05 - *, P<0.001 - **
Table 7 shows the effect of standard antidote and antioxidant drugs on the erythrocytes number in the blood of animals under nitrate load. For the third hour of the search under nitrate loading in the bull's blood, the erythrocytes number fluctuated in the range of 6.10±0.112 - 6.05±0.120 T/l. Throughout the analysis, the erythrocytes number of the control group, which was not injected with antidote and antioxidants, ranged from 7.01±0.10 - 5.15±0.12 T/l.
The erythrocytes number gradually increased in bulls who were injected with chromosmone and Urso-vit-ADES in acute nitrate-nitrite toxicosis. A probable raise in this indicator was detected firstly, increasing by 2% relative to sick animals. At the second hour of the experiment, the erythrocytes number in the blood of animals of the experimental group Ei was 6.20±0.12 T/l. A probable decrease was observed at the sixth and eighth test hours; it declined by 14 and 13%, respectively, relative to the control group.
Table 7
The number of erythrocytes in the blood of bulls after the use of Ursovit-ADES and sodium selenite in
acute nitrate-nitrite toxicosis T / l (M±m, n = 5)
Animal blood counts (hours) Groups of animals
Control Experimental (Ei) Experimental (E2)
Before feeding sodium nitrate
Control 7.01±0.102 7.02±0.110 7.04±0.111
After feeding sodium nitrate
Third hour 6.10±0.112 6.05±0.120 6.07±0.104
— After the introduction of chromosmone
Ursovit-ADES Ursovit-ADES + 0.5% solution of sodium selenite
First hour 5.90±0.102 6.01±0.130 6.08±0.130
Second hour 5.61±0.101 6.20±0.122* 6.29±0.133*
Three hour 5.15±0.124 6.61±0,141** 6.65±0.142**
Six hour 6.02±0.123 6.85±0.152* 6.89±0.160**
Eighth hour 6.16±0.112 6.91±0.133* 6.98±0.144*
Note: the degree of probability compared with the control group - P<0.05 - *, P<0.001 - **
After the combined use of chromosmone, Ursovit- the test, this indicator raised by 3%, in the second - by ADES, and sodium selenite, the erythrocytes number in 12%, in the third - 29% relative to the group of animals the blood of bulls increased. Thus, in the first hour of that were not treated.
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At the sixth and eighth hours of the test, the eryth-rocytes number reached the level of physiological values and was, severally, 6.89±0.160 and 6.98±0.144 T/l.
Table 8
The number of leukocytes in the blood of bulls after the use of Ursovit-ADES and sodium selenite in acute _nitrate-nitrite toxicosis G / l (M±m, n = 5)_
Animal blood counts (hours) Groups of animals
Control Experimental (E1) Experimental (E2)
Before feeding sodium nitrate
Control 7,27±0,180 7,29±0,191 7,25±0,190
After feeding sodium nitrate
Three o'clock 7,90±0,183 7,94±0,182 7,92±0,164
— After the introduction of chromosmone
Ursovit-ADES Ursovit-ADES + 0.5% solution of sodium selenite
First hour 8,31±0,190 7,85±0,162 7,81±0,160*
Second hour 8,38±0,200 7,61±0,153* 7,57±0,161*
Three hour 8,45±0,232 7,52±0,190* 7,47±0,180*
Six hour 8,14±0,230 7,46±0,154* 7,39±0,173*
Eighth hour 7,45±0,214 7,34±0,170 7,31±0,190
Note: the degree of probability compared with the control group - P<0.05 - *, P<0.001 - **
Leukocytes number grew in the blood of bulls under conditions of acute nitrate-nitrite toxicosis, and for the third hour of the investigation, it was in the range of 7.90±0.183 - 7.94±0.182 G/l. This is due to inflammatory processes in the digestive tract.
The application of chromosmone with Ursovit-ADES and sodium selenite helped reduce the leukocytes quantity in the blood throughout the search.
In the first hour after drugs treatment, the leukocytes number in the blood of groups E1 and E2 decreased by 5.5 and 6% relative to the values of the control group, and for the second hour by 9 and 9.7%, respectively. Subsequently, the number of leukocytes continued to decrease again, and at the eighth hour, group E1 7.34 ± 0.170 G/l, and group E2 - 7.31±0.190 G/l i.e., reached the initial values of the research.
Conclusions
The use of chromosmone together with Ursovit-ADES and sodium selenite generally had a positive effect on the morphological and biochemical parameters of the blood under acute nitrate-nitrite toxicosis conditions. It affected the improvement of the functional state of the experimental animals and their rapid recovery. The introduction of the examined drugs to bulls helped reduce the nitrates, nitrites, methemoglobin, ammonia, and the number of leukocytes concentration in the blood. Under these conditions, an increase in erythrocytes, hemoglobin, and urea was found in the blood.
References
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UDC 577.1
B. Gutyj1, I. Khariv1, V. Khalak2 M. Khariv1, R. Vasiv1, Kh. Leskiv1, T. Martyshuk1, Z. Guta1
Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv,
Pekarska Str., 50, Lviv, 79010, Ukraine 2State Institution Institute of grain crops of NAAS, V. Vernadsky Str., 14, Dnipro, 49027, Ukraine DOI: 10.24412/2520-6990-2021-22109-14-18 THE EFFECT OF LIPOSOMAL DRUG "BUTAINTERVIT" ON THE ACTIVITY OF THE T-SYSTEM OF CELLULAR IMMUNITY OF RATS UNDER OXIDATIVE STRESS
Abstract
The results of investigations on the effect of the liposomal drug "Butaintervit" on rats' T-cell immune system activity under carbon tetrachloride poisoning are presented. We detected that administrating to rats of the experimental groups of 50% carbon tetrachloride at a dose of 0.25 ml per 100 g of body weight causes oxidative stress. It negatively affects cellular immunity and functional activity performance immunocompetent blood cells. The immunosuppressive impact of oxidative stress led to a decrease in the rats' blood of the first and second experimental groups, the T-lymphocytes number and their immunoregulatory subpopulations, mainly on the 2nd and 5 th days of the trial. At the equal background, the obtained data indicate a positive effect of butaphosphane, interferon, injectable milk thistle and vitamins A, D3, E in the liposomal drug "Butaintervit" on the relative number of T-lymphocytes in the rats' blood and on the redistribution of avidity in strengthening receptor membranes cells. It was found that the normalization on the 2nd day of studies of the level in the blood of rats of the second experimental group of total T-lymphocytes occurred due to medium-avid forms and active T-lymphocytes by changing the number of low-avid forms.
Key words: T-lymphocytes, blood, rats, oxidative stress, tetrachloroethane, butaphosphane, interferon, milk thistle, vitamins.
It is known that oxidative stress causes significant disorders of physiological functions and metabolic processes in animals and adversely affects the immune system [2, 7, 15-18]. Accordingly, the action of various chemical factors that contribute to oxidative pressure is relevant to study the immune system activity because adrenal cortex hormones play the key role in the development of stress syndrome - glucocorticoids, the level of which in the blood under these conditions increases significantly [1, 3, 10]. Under the glucocorticoids influence, peroxide processes are intensified, that leads to a significant supply in the formation of lipid peroxidation products (LPO), which have a destructive effect on the cellular immune system of animals [6, 8, 11, 26]. In this regard, the search for effective means would develop animals' adaptability and have a regulatory effect on various parts of the body's immunobiological reactivity to free radicals.
To date, scientists have found that among the many drugs that have a positive influence on the immune system due to stress on the body play an essential role, butaphosphane, interferon, milk thistle, as well as fat-soluble vitamins A, D3, E [12-14, 18, 2025]. In this regard, this work aimed to elucidate the impact of a complex immunotropic drug-containing butaphosphane, interferon, milk thistle, and vitamins A, D3, E in the form of a liposomal emulsion, on the activity of the cellular immune system of rats under conditions effects of carbon tetrachloride oxidative stress.
Materials and methods of research
The experimental part of the work was performed on young white male Wistar rats, weighing 180-200 g, kept in the vivarium of the State Research Control Institute of Veterinary Drugs and Feed Additives.
