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Slobodian S. O., Gutyj B. V., Leskiv Kh. Ya., Khariv I.I.
Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies, Pekarska, Str., 50, Lviv, 79010, Ukraine
Paziuk I. S.
Vyshnia College of Lviv National Agricultural University, Rudki, Ukraine
DOI: 10.24412/2520-2480-2020-3082-20-24 THE SODIUM SELENITE AND FEED ADDITIVE "METISEVIT PLUS" EFFECT ON THE MORPHOLOGICAL PARAMETERS OF THE BLOOD OF RATS IN CADMIUM AND LEAD
INTOXICATION
Abstract
Lead and cadmium are among the influential environmental pollutants today. The results of many experimental studies indicate that in mammals Cadmium and Lead have a toxic effect on many organs and systems. The research aimed to study the impact of sodium selenite and feed additive "Metisevit Plus" on the morphological parameters of the blood of rats under cadmium and lead intoxication. The analyses were performed on male Wistar rats weighing 200-220 g, from which 3 groups of animals were formed: control (C) and two experimental (Ei), (E2). Mice of the (C) and (E1), (E2) groups were injected with 16.6% lead acetate aqueous solution at a dose of 100 mg/kg and 0.029% cadmium chloride aqueous solution at a dose of 2.0 mg/kg. Rats of the (E1) group were additionally administered sodium selenite at a dose of 0.2 mg/kg body weight. Animals of the (E2) group were additionally given a feed additive "Metisevit plus" at a dose of 0.5 mg/kg body weight. Sodium selenite and feed additive "Metisevit plus" contributes to the normalization of morphological parameters of the blood in spite of the development of chronic lead-cadmium toxicosis. However, it should be noted that the better normalizing effect of the blood is feed additive "Metisevit Plus" on the morphological parameters of the blood than the sodium selenite in animals.
Key words: toxicology, rats, cadmium, lead, sodium selenite, Metisevit plus.
Introduction
Heavy metals in the biosphere (water, soil, plants) have a double meaning: as the trace elements, they are necessary for the normal course of physiological processes, but at the same time are toxic in high concentra-
tions, which adversely affects health, animal productivity, and agricultural quality [3, 7, 14]. Contamination of agricultural lands with heavy metals usually occurs due to the atmospheric emissions from enterprises, waste from livestock farms, and the use of mineral fertilizers and pesticides [6]. Toxic chemical elements that enter
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the human and animal body (with food, feed) are excreted from it slowly. In the body, they accumulate individual organs and tissues [5, 15].
Some of the influential environmental pollutants today are Lead and Cadmium [4, 11, 12, 18]. The results of many studies indicate that in mammals Cadmium and Lead have a toxic effect on several organs and systems [2, 8, 16]. Prolonged contact with these metals and their entry into the body, even in small concentrations, leads to suppression of immunity, reduced resistance to infections, the development of allergic, autoimmune, and oncological pathology [9].
Lead and cadmium are thiol poisons due to their ability to bind to CH groups of proteins [1]. The mechanism of action of these poisons is to inhibit the activity of enzyme systems by blocking carboxyl, amine and SN groups of protein molecules [10, 17].
It is known that one of the manifestations of the toxic effects of heavy metals is the activation of the LPO processes of cell membranes [11]. Lipid peroxidation (LPO) is one of the most important oxidative processes in animals. Currently, LPO is considered one of the main causes of cell damage and death due to the action of reactive oxygen species (ROS) and free radicals [19]. It is proved that any sufficiently powerful environmental impact on the body of animals can initiate the processes of LPO [20].
Therefore, our work aimed to study the effect of sodium selenite and feed additive "Metisevit Plus" on the morphological parameters of the blood of rats under cadmium and lead intoxication.
Materials and methods.
The analyses were performed on male Wistar rats weighing 200-220 g, from which 3 groups of animals were formed: control (C) and two experimental (E1), (E2). Mice of the (C) and (E1), (E2) groups were injected
Table 1
The sodium selenite and feed additive "Metisevit plus" effect of on the erythrocytes number in the blood of rats with cadmium and lead intoxication, T / l (M ± m, n = 6)
with 16.6% lead acetate aqueous solution at a dose of 100 mg/kg and 0.029% cadmium chloride aqueous solution at a dose of 2.0 mg/kg. Rats of the (E1) group were additionally administered sodium selenite at a dose of 0.2 mg/kg body weight. Animals of the (E2) group were additionally given a feed additive "Metisevit plus" at a dose of 0.5 mg/kg body weight.
Throughout the test, the rats were kept on a diet that was balanced in all the necessary components and received drinking water without restrictions from drinking glass bowls with a volume of 0.2 liters.
All animal manipulations were performed by the European Convention for the Protection of Vertebrate Animals Used for Experimental and Scientific Purposes (Strasbourg, 1986).
Blood for hematological tests was taken under ether anesthesia from the jugular vein on days 1, 7, 14, 21, and 28 of the experiment.
The following parameters were determined in the blood: the erythrocytes number - photoelectrocolomet-rically according to the method of E. S Gavrilets et al. (1966); the leukocytes number - using the Gorev grid in the counting chamber; hemoglobin concentration -by the method of L. M Pimenova et al. (1975) [21].
The analysis of research results was performed using the software package Statistica 6.0. The probability of differences was assessed by Student's t-test. The results were considered plausible at P < 0.05.
Results and discussion
The erythrocytes number decreased throughout the experiment in case of intoxication with heavy metals in rats. The lowest erythrocytes number was in the blood of mice of C group on 21th day compared with the initial values reduced by 41.3%, respectively (Table 1).
Groups of animals Output data day of the experiment
7 14 21 28
Control (Cd2+ + Pb2+) 7.48±0.08 6.51±0.11 5.23±0.09 4.39±0.16 4.41±0.22
Experimental 1 (Cd2+ + Pb2+ + sodium selenite) 7.39±0.05 6.84±0.15 6.56±0.18*** 6.28±0.09*** 6.70±0.21***
Experimental 2 (Cd2+ + Pb2+ + Metisevit plus) 7.43±0.10 7.15±0.20* 6.99±0.17*** 6.73±0.20*** 7.40±0.15***
The erythrocytes number in rats of E:group on the 7th experiment day, relative to the initial values decreased to 6.84 ± 0.15 T/l, however, compared with sick mice, this figure was higher by 5.1%. Subsequently, this indicator on the 14th and 21st day of the experiment was higher by 25.4 and 43.1% relative to the C group. However, it should be noted that the erythrocytes number in the blood of the E1 group of animals treated with sodium selenite did not reach the initial values taken before the start of heavy metal loading.
The introduction of rats of the E2 group of feed additive "Metisevit plus" contributed to the normalization of the erythrocytes number on 21th and 28th days of the
experiment, where, respectively, this figure was higher by 53.3 and 67.8% relative to the C group.
It was found that the hemoglobin level decreased to 90.3 ± 2.63 g/l. in rats of C group. It was found that the hemoglobin level increased by 7th and 14th days of the test by 1.1 and 6.8% during sodium selenite introduction and feed additive "Metisevit plus" for intoxicated mice relative to the blood parameters of rats of C group. On the 21st day of the research, the application of feed additive to rats of the E2 group contributed to a more possible growth in hemoglobin than the treatment of sodium selenite (Table 2).
Thus, the hemoglobin level in the blood of the E2 group in this period grew by 33.2%, while in the E2
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group - by 12.3%. The highest hemoglobin level in the the E2 group, which under heavy metal intoxication a blood of rats was on the 28th day of the experiment in feed additive "Metisevit Plus" was used.
Table 2
The sodium selenite and feed additive "Metisevit plus" effect of on the on the hemoglobin level in the _blood of rats with cadmium and lead intoxication, g/l (M ± m, n = 6)_
Groups of animals Output data day of the experiment
7 14 21 28
Control 1 (Cd2+ + Pb2+) 118,9±2,57 103,4±1,58 95,7±4,03 90,3±2,63 95,4±1,84
Experimental 1 (Cd2+ + Pb2+ + sodium selenite) 117,5±0,99 104,5±2,10 102,7±3,36 101,4±1,84** 110,4±4,05**
Experimental 2 (Cd2+ + Pb2+ + Metisevit plus) 115,1±1,99 110,4±3,57 115,2±2,43** 120,3±3,12 *** 126,1±2,49 ***
Simultaneously with the decrease in the hemoglobin level and the erythrocyte number, an increase in the average content of hemoglobin in one erythrocyte of blood to 20.57 ± 1.20 pg in intoxicated rats was observed (Table 3).
In rats of experimental groups, a reduce in the average hemoglobin content in one erythrocyte on the 21st day of the experiment by 21.5 and 13.1% relative to the control group of rats was installed.
As a result of the development of lead-cadmium intoxication in animals, leukopenia is observed, which, in our opinion, is associated with systemic damage to the hematopoietic organs of animals by cadmium and lead, which suppress leukopoiesis. It was found that the leukocytes number in rats of C group decreased by 36.1% compared to baseline, even before the start of heavy metal loading (Table 4).
Table 3
The sodium selenite and feed additive "Metisevit plus" effect of on the on the average content of hemoglo-
Groups of animals Output data day of the experiment
7 14 21 28
Control 1 (Cd2+ + Pb2+) 15,89±1,33 15,88±1,12 18,30±1,35 20,57±1,20 21,63±1,29
Experimental 1 (Cd2+ + Pb2+ + sodium selenite) 15,90±1,06 15,28±1,25 15,66±0,99 16,15±1,32* 16,48±1,21*
Experimental 2 (Cd2+ + Pb2+ + Metisevit plus) 15,49±1,22 15,44±1,30 16,48±1,08 17,88±1,30 17,04±1,09
Table 4
The sodium selenite and feed additive "Metisevit plus" effect of on the the leukocytes number in the blood _of rats intoxicated with Cadmium and Lead, G / l (M ± m, n=6)_
Groups of animals Output data day of the experiment
7 14 21 28
Control 1 (Cd2+ + Pb2+) 8,12±0,75 7,63±0,82 6,42±0,55 5,19±0,85 5,30±1,00
Experimental 1 (Cd2+ + Pb2+ + sodium selenite) 7,85±0,56 7,71±0,37 7,32±0,46 7,02±0,56 7,10±0,51
Experimental 2 (Cd2+ + Pb2+ + Metisevit plus) 7,92±0,81 7,74±0,71 7,54±0,52 7,69±0,80* 7,80±0,56*
In the rats of the experimental groups, which, together with heavy metals, were fed antioxidant remedies: sodium selenite and metisevit plus, the leukocytes number in their blood probably grew from the 14th day of the search. It was found that on the 21st test day the leukocytes number in the blood of the E1 group increased by 35.3% in the E2 group this indicator progressed by 48.2% relative to the C group of rats.
On the 28th research day, the leukocytes number in the blood of the Ei group increased but did not reach the initial values. Only the feeding of Metisevit Plus
feed additives improved the leukocytes number to physiological levels.
Conclusions
Sodium selenite and Metisevit Plus feed additive contribute to the normalization of blood morphological parameters in the development of chronic lead-cadmium toxicosis in the blood of rats. However, the better normalizing effect was of the feed additive "Metisevit plus" on the morphological parameters of the blood of rats than the use of sodium selenite in animals.
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Acknowledgments
This scientific work was financially supported by the Ministry of Education and Science of Ukraine (0120U101999).
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Dovhiy Y. Y., Kot T.F., Rudik O. V., Dovhiy M. Y., Rudik V. V.
Polissia National University, Zhytomyr, Ukraine Staryi Blvd. 7, Zhytomyr, Zhytomyr Oblast, Ukraine, 10008, Ukraine DOI: 10.24412/2520-2480-2020-3082-24-30 CHANGES IN CLINICAL CHARACTERISTICS AND BIOCHEMICAL INDICES OF FUR ANIMALS AND POULTRY BLOOD UNDER EIMERIA INVASION
Abstract
The research has revealed the invasion agents like: Eimeria perforans, E. magna, E. stiedae in rabbits and nutrias, in a small amount- E. coecicola, E. pisiformis and E. intestinalis in rabbits, E. maxima, E. tenella, E. necatrix and E. acervulina in chickens and quails. Rapidly infected fur animals (280 rabbits and 180 nutrias), 3468 chickens and 1800 quails were examined while conducting a research. Under the localization of a pathalogic process, the disease appeared in poultry in a mixed pattern; in rabbits and nutrias in an intestinal pattern (11.9 % and 11 %), in a hepatic pattern (8.1 % and 11 %) and mostly in a mixed pattern (80 % and 78 % respectively).
The research showed that sick rabbits and nutrias were weak, mucous membranes of eyes and nose were pale (50.5 and 49 %), yellowish (38 and 42 %). Occasional watery feces (80 and 91 %), flatus were observed, the abdomen was increased and infirm (87.1 and 85 %). Under complications the rabbits had intermittent fever (5%), acute bouts of cramps (7.14 %). Death was observed in rabbits under a severe disease in two weeks time (10%) and in nutrias in 3-5 weeks (7%). Digestive unsets were a characteristic feature under an asymptomatic disease course.
From among the blood biochemical indices of nutrias and rabbits, a decrease in the count of hemoglobuline, in the amount of albumen, crude protein as compared to a control group was registered.
A depressed state, thirst, (27th day), 1-2 °C temperature rise, drowsiness, awkward gait (35th day), watery feces, feces with blood admixtures were registered in chickens, and the quails manifested unwillingness to eat, gathered in warm places, they also had white-and-green watery feces, labored respiration and accelerated cardiac rate.
Some decrease in the amount of red blood cells, pseudoeosinophils as well as some increase in the amount of white blood cells and eosonophils were registered in the blood of sick chickens and quails. Some decrease in the count of hemoglobulin, in the amount of creatinine and urine was registered in blood serum of the infected chickens and quails.
Thus, some changes in blood indices of chickens and quails which suffered from protozoal invasion testified to a toxic and trophic effects of eimerias and their metabolites on the organisms.
Key words: rabbits, nutrias, chickens, quails, invasion, agent, eimerias, metabolites, blood.
Introduction. Eimeriosis is a widely spread animal disease caused by monocellular Eimeria parasites. Eimeriosis of rabbits, nutrias and chickens results in great economic losses in animal breeding. (Yatusevych, 2005, 2006; Dalloul, & Lillehoj, 2005; Manzhos, 2006; Yevstafyeva and others, 2010; Stybel and others, 2012, 2014; Abbas, Colwell, & Gilleard, 2012; Avdosieva, & Mysko, 2015; Arafat, & Abbas, 2018).
The disease causes digestive unset, organism intoxication, lowers down the animal immunity against infectious and invasive diseases (Bogach and others ,2008; Peredera, 2009; Dovhiy and others., 2018, 2019).
National and foreign researchers (Nakai, Tsuchiya, & Takahashi, 1992; Berezovskiy and others, 2002; Teixeira, Teixeira Filho, & Lopes, 2004; Pavlenko and others, 2008; Michels, Bertolini, Esteves, Moreira, & Franca, 2011; Sokol, Gesek, Ras-Norynska, & Michalczyk, 2014; Umar, Lawal, Okubanjo, & Wakawa, 2014; El-Morsy, Abou El-Azm, & Awad,
2016; Arafat, & Abbas, 2018) report about the extension of invasive diseases (helminthic infection and eimeriosis) in poultry, rabbits and nutrias.
Some difficulties in fighting the disease are caused by high resistance of eimerias to the environmental factors as well as to desinfectants ( Zaikina and others., 2008). These agents cause 90-100% death of rabbits, nutrias and poultry, as well as hypovitaminose, natural resistance weakening, growth inhibition and delaying in progression (Gawel ab., 2005; Glechyk and others., 2005; Golubtsova and others., 2012, 2016. It should be mentioned that eimeriosis treatment and prevention in chickens and quails are closely connected with chemotherapy and immunocorrection (Nagorna 2017). Thus, one of the most perspective ways of fighting the parasites resistance is using effective, ecologically safe and available medicines (Zaikina, 2013)
Materials and methods. Scientific and production studies were conducted on both private and spe-
