THE EFFECT OF MILK THISTLE, METHIPHEN AND SILIMEVIT ON THE VITAMINS A AND E LEVEL IN THE BLOOD OF LAYING HENS IN EXPERIMENTAL CHRONIC CADMIUM TOXICOSIS Текст научной статьи по специальности «Животноводство и молочное дело»

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Ostapyuk A. Y., Gutyj B. V., Hunchak V. M., Leskiv Kh. Ya., Khariv 1.1., Vasiv R. O., Kamratska O. I., Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv, Ukraine

DOI: 10.24412/2520-2480-2020-3082-17-20 THE EFFECT OF MILK THISTLE, METHIPHEN AND SILIMEVIT ON THE VITAMINS A AND E LEVEL IN THE BLOOD OF LAYING HENS IN EXPERIMENTAL CHRONIC CADMIUM

TOXICOSIS

Abstract

The research aimed to investigate the effect of milk thistle, methiphene and silimevit on the vitamins A and E level in the blood of laying hens in experimental chronic cadmium toxicosis. 32 chickens, 78 weeks old, were selectedfor the analysis. Four groups were formed: control and three experimental. Chickens of the control group (C) and three experimental groups were administrated with water cadmium sulfate at a dose of 4 mg/kg body weight. Poultry of the experimental group (Ei) were supplied the fruits of milk thistle at a dose of 2.0 g / kg offeed once a day for 30 days. Birds of the experimental group (E2) were fed metifen at a dose of 0.28 g / kg offeed once a day for 30 days. Chickens of experimental group E3 were added silimevit at a dose of 0.36 g / kg offeed once a day for 30 days. It was found that with the development of chronic cadmium toxicosis in poultry, the vitamins A and E content in their blood decreases throughout the test. The administration to chickens under cadmium load of milk thistle, methiphen and silimevit increases the vitamins A and E level in their blood. When silymevit was used on laying hens, the level of vitamins in their blood was the highest compared with the use of methefen and milk thistle.

Key words: toxicology, cadmium, bird, blood, silymevit, methiphen, milk thistle.

Introduction

Cadmium is one of the most toxic heavy metals [2, 7]. It belongs to the second class of danger - highly dangerous substances [1, 11, 23]. Like many other heavy metals, it has a pronounced tendency to accumulate in the body: its half-life is 10-35 years [12, 19]. Cadmium accumulates in the kidneys (30-60% of the total) and liver (20-25%). This heavy metal is also found in the pancreas, spleen, tubular bones, other organs, and tissues [8, 14].

Cadmium adversely affects the vital systems of humans and animals, causing pathological changes in tissues and organs (kidneys, lungs, bone, reproductive and endocrine systems), inhibiting the process of eryth-ropoiesis [13, 15, 18]. Disorders caused by long-term intake of this heavy metal in the body are determined by the level of Cd2+ accumulation in cells and the manifestation of cumulative toxicity [4, 6, 17]. The vulnerability of cells to the action of Cadmium largely depends on the level of expression in them of genes of metal-binding proteins - metallothioneins [5, 9, 20].

Cadmium and its compounds are immunotoxi-cants that cause disturbances in the functioning of the body's immune system, reduce resistance to infections, contribute to the formation of allergic, autoimmune and oncological pathologies [16, 21]. A feature of the biological action of cadmium is its ability to adversely affect animal health with prolonged exposure to low levels of pollution due to the high rate of biological accumulation (up to 40 years) [3, 10].

The research aimed to study the effect of milk thistle, methiphen, and silimevit on the level of vitamins A and E in the blood of laying hens in experimental chronic cadmium toxicosis.

Material and methods of research

32 chickens, 78 weeks old, were selected for the analysis. Four groups were formed: control and three experimental. Chickens of the control group (C) and three experimental groups were administrated with water cadmium sulfate at a dose of 4 mg/kg body weight. Poultry of the (E1) group were supplied the fruits of milk thistle at a dose of 2.0 g / kg of feed once a day for 30 days. Birds of the (E2) group were fed metifen at a

dose of 0.28 g / kg of feed once a day for 30 days. Chickens of E3 group were added silimevit at a dose of 0.36 g / kg of feed once a day for 30 days.

The conditions of keeping chickens and the parameters of the microclimate in the room for all groups were similar. The amount of feed and water consumed was taken into the record during the experiment.

All experimental interventions and slaughter of animals 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, 1985) and the First National Congress on Bioethics (Kyiv, 2001).

Blood from laying hens was taken from the axillary vein in the periods: before the administration of drugs and cadmium sulfate, on the first, seventh, fourteenth, twenty-first, and thirtieth days of the test.

Vitamins A and E were determined in blood plasma by high-performance liquid chromatography [22].

The experiments used the following drugs:

Cadmium sulfate - Cadmium sulfate, an inorganic compound with the chemical formula CdSO4. It is well soluble in water, so it is well absorbed in the digestive tract, blood and is known for its toxic effects on living organisms.

Milk thistle (MT) Silybum marianum is a family of compound flowers, grows wild in wastelands, along roads, in abandoned fields, and is cultivated in medicinal gardens. For the treatment of milk thistle fruits are used. They contain protein 17-18%, fats 10-11%, fla-volignans 2-3%, essential oil 0.08%, vitamins A, E, K, nutrients, quartcetin.

Metifen (M) is a white crystalline powder, sweet in taste, with the smell of sulfur. Poorly soluble in cold water, better - in hot water (1:20). Thermostable. The drug contains phenarone and methionine.

Silimevit (C) - feed additive, which includes the fruits of milk thistle, selenium, methiphen, vitamins A, E, and C.

The analysis of research results was performed using the software package Statistica 6.0. The probability

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31%, on day 21 of the test - by 39% relative to the initial quantities. Starting from the first day of the research, a gradual increase in the vitamin A content of the E1, E2 and E3 groups of hens relative to the control group was installed. On the 14th day of the experiment, the vitamin A content of the E1 group was 0.41 ± 0.015 ^g / ml, in the E2 group, respectively, was 0.40 ± 0.020 ^g / ml, in the D3 group, respectively, was 0.57 ± 0.016 ^g / ml, whereas in the control group of birds this figure was 0.34 ± 0.015 ^g / ml. On the 21st day of the research, the vitamin A content increased by 43, 27 and 97% relative to laying hens of the control group.

Table 1

The vitamin A level in the blood of laying hens after the administration of milk thistle, methiphen and sili-

mevit in chronic cadmium toxicosis; (M±m, n = 8)

of differences was assessed by Student's t-test. The results of the mean values were considered statistically significant at * - P < 0,05, ** - P < 0,01, ***- P < 0,001 (ANOVA).

Results and discussion

It was found that with the development of chronic cadmium toxicosis in laying hens, the vitamin A content in blood decreases throughout the research. As can be seen from Table 1, the vitamin A content after administration of cadmium sulfate began to decrease on the first day by 6%, on day 7 - by 12%, on day 14 - by

Blood test time (days) Vitamin A (^g / ml)

Groups of chickens

Control (cadmium) Experimental 1 (cadmium + MT) Experimental 2 (cadmium + M) Experimental 3 (cadmium + S)

At the beginning of the experiment 0.49±0,011 0.48±0,010 0.45±0,012 0.47±0,017

the first day 0.46±0.012 0.47±0.016 0.46±0.022 0.51±0.019 *

7th day 0.43±0.011 0.44±0.022 0.42±0.017 0.53±0.010 ***

14th day 0.34±0.015 0.41±0.015 * 0.40±0.020 * 0.57±0.016 ***

21th day 0.30±0.025 0.43±0.019 ** 0.38±0.011 * 0.59±0.014 ***

30th day 0.33±0.020 0.45±0.014 ** 0.41±0.017 ** 0.55±0.012 ***

The use of silimevit contributed to an increase in the vitamin A content in the poultry under conditions of cadmium loading throughout the investigation. From the first day of the experiment, a probable rise in the vitamin A content in the blood of the experimental group of chickens relative to the control group. On the 14th day of the test, the vitamin A content in the experimental group of hens E3 grew by 67% relative to the control group of birds. On day 21 of the test, the vitamin A level in the blood of this group was the highest in comparison with the control and other experimental

Table 2

The vitamin E level in the blood of laying hens after the administration of milk thistle, methiphen and sili-

mevit in chronic cadmium toxicosis; (M±m, n = 8)

groups of chickens which were fed milk thistle and me-thiphen.

Vitamin E, which protects cell membranes from attack by free radicals and reactive oxygen species, is important in the antioxidant defense system of chickens. The content of the above vitamin in the blood of hens with chronic cadmium toxicosis is shown in table 2. It was found that in the blood of the control group of chickens, the vitamin E level decreased from 1.11 ± 0.039 to 0.70 ± 0.011 ^g / ml.

Blood test time (days) Vitamin E (^g / ml)

Groups of chickens

Control (cadmium) Experimental 1 (cadmium + MT) Experimental 2 (cadmium + M) Experimental 3 (cadmium + S)

At the beginning of the experiment 1,11±0,039 1,15±0,028 1,17±0,024 1,14±0,020

the first day 1,06±0,034 1,11±0,022 1,13±0,034 * 1,16±0,026 *

7th day 0,97±0,043 1,07±0,026 * 1,05±0,040 * 1,19±0,031 **

14th day 0,85±0,045 1,02±0,041 ** 0,96±0,025 * 1,21±0,039 ***

21th day 0,70±0,011 0,95±0,031 *** 0,90±0,033 *** 1,20±0,027 ***

30th day 0,78±0,040 1,03±0,019 *** 0,97±0,028 *** 1,17±0,040 ***

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In the blood of laying hens under cadmium toxins, was found the raise of the vitamin E level on the first day by 4.7%, on the 7th day of the experiment - by 10.3%, on the 14th day of the experiment - by 20%, on the 21st day of the experiment - by 35, 7% and on the 30th day of the experiment - 32% according to the supplementary feeding with milk thistle.

An increase in vitamin E in the blood of chickens compared with the control group was found when admin administreted with feed metifen, however, the level of this vitamin remained slightly lower in the experimental groups. Thus, on days 14 and 21 of the test, the vitamin E level in the blood of the E2 group ranged from 0.96 ± 0.025 - 0.90 ± 0.033 ^g / ml, while in the E3 group the level of the vitamin was significantly higher.

Feeding silimevit to laying hens under a cadmium load contributed to an increase in vitamin E in their blood throughout the test, where vitamin levels ranged from 1.16 ± 0.026 to 1.21 ± 0.039 ^g/ml. The highest vitamin E level was on the 14th and 21st day of the experiment, compared with the control group, it increased by 42 and 71%, respectively.

Conclusions

The use of laying hens under cadmium load, milk thistle, methiphen and silimevit increases the level of vitamins A and E in their blood. When sylimevit was used in chickens, the level of vitamins in their blood was the highest compared with the use of methefen and milk thistle.

Acknowledgments

This scientific work was financially supported by the Ministry of Education and Science of Ukraine (0120U101999).

References

1. Al-Azemi, M., Omu, F.E., Kehinde, E.O., Anim, J.T., Oriowo, M.A., & Omu, A.E. (2010). Lithium protects against toxic effects of cadmium in the rat testes. J. Assist. Reprod. Genet, 27 (8), 469-476. doi: 10.1007/s10815-010-9426-3.

2. Ali, M. M., Murthy, R. C., & Chandra, S. V. (1986). Developmental and longterm neurobehavioral toxicity of low-level in utero Cd exposure in rats. Neurobehavioral Toxicology and Teratology, 8 (5), 463468. https://www.ncbi.nlm.nih.gov/pubmed/3785508.

3. Antoniak, H. L., Biletska, L. P., & Babych, N. A. (2010). Kadmii v orhanizmi liudyny i tvaryn. Nad-khodzhennia do klityn i yikh akumuliatsiia. Biolohichni Studii. Studia Biologica, 4 (2), 127-140 (in Ukrainian).

4. El-Shahat, A. E., Gabr, A., Meki, A. R., & Me-hana, E. S. (2009). Altered testicular morphology and oxidative stress induced by cadmium in experimental rats and protective effect of simultaneous green tea extract. Int. J. Morphol., 27 (3), 757-764. doi: 10.4067/S0717-95022009000300020.

5. Fregoneze, J. B., Marinho, C. A., Soares, T., Castro, L., Sarmento, C., Cunha, M., Gonzalez, V., Oliveira, P., Nascimento, T., Luz, C.P., Santana, Jr. P., De-Oliveira, I.R., & e-Castro-e-Silva, E. (1997). Lead (Pb2+) and cadmium (Cd2+) inhibit the dipsogenic ac-

19_

tion of central beta-adrenergic stimulation by isopro-terenol. Brazilian Journal of Medical and Biological Research, 30 (3), 419-423. doi: 10.1590/S0100-879X1997000300018.

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 the anti-oxidant system in the organism of bulls. Visnyk of Dnipropetrovsk University. Biology, ecology, 24(1), 96-102. doi: 10.15421/011611.

7. Hutyi, B. V. (2013). Vmist vitaminiv A i E u krovi bychkiv za umov kadmiievoi intoksykatsii. Visnyk Sumskoho natsionalnoho ahrarnoho univer-sytetu. Seriia: Veterynarna medytsyna, 2, 31-33. Rezhym dostupu: http://nbuv.gov.ua/UJRN/Vsna_vet_2013_2_10 (in Ukrainian).

8. Hutyi, B. V. (2013). Vplyv E-selenu na vmist vitaminiv A i E u krovi bychkiv za umov kadmiievoi intoksykatsii. Naukovyi visnyk Lvivskoho natsionalnoho universytetu veterynarnoi medytsyny ta bio-tekhnolohii im. Gzhytskoho, 15, 3 (3), 311-314. Rezhym dostupu:

http://nbuv.gov.ua/UJRN/nvlnu_2013_15_3(3)_55

(in Ukrainian).

9. Hutyi, B. V. (2013). Vplyv khlorydu kadmiiu u riznykh dozakh na aktyvnist aminotransferaz syrovatky krovi buhaitsiv. Naukovyi visnyk Lvivskoho natsionalnoho universytetu veterynarnoi medytsyny ta biotekhnolohii im. Gzhytskoho, 15, 1 (1), 49-52. Rezhym dostupu:

http ://nbuv. gov.ua/UJRN/nvlnu_2013_15_1(1)_11

(in Ukrainian).

10. Lavryshyn, Y., & Gutyj, B. (2019). Protein synthesize function of bulls liver at experimental chronic cadmium toxicity. Scientific Messenger of LNU of Veterinary Medicine and Biotechnologies. Series: Veterinary Sciences, 21 (94), 92-96. doi: 10.32718/nvl-vet9417.

11. Lavryshyn, Y., Gutyj, B., Palyadichuk, O., & Vishchur, V. (2018). Morphological blood indices of bulls in experimental chronic cadmium toxicosis. Scientific Messenger of LNU of Veterinary Medicine and Biotechnologies. Series: Veterinary Sciences, 20 (88), 108-114. doi: 10.32718/nvlvet8820.

12. Lavryshyn, Y., Gutyj, B., Paziuk, I., Lev-kivska, N., Romanovych, M., Drach, M., & Lisnyak, O. (2019). The effect of cadmium loading on the activity of the enzyme link of the glutathione system of bull organism. Scientific Messenger of LNU of Veterinary Medicine and Biotechnologies. Series: Veterinary Sciences, 21 (95), 107-111. doi: 10.32718/nvlvet9520.

13. Liu, J., Qian, S.Y., Guo, Q., Jiang, J., Waalkes, M.P., Mason, R.P., & Kadiiska, M.B. (2008). Cadmium generates reactive oxygen- and carbon-centered radi-calspecies in rats: Insights from in vivo spin-trap-pingstudies. Free Radic Biol Med., 45 (4), 475-481. doi: 10.1016/j.freeradbiomed.2008.04.041.

14. Lu, J., Jin, T., Nordberg, G., & Nordberg, M. (2005). Metallothionein gene expression in peripheral lymphocytes and renal dysfunction in a population environmentally exposed to cadmium. Toxicol. Appl.

20

VETERINARY SCIENCES / <<€®LL®qUQUM~J®U©MaL>>#30®2),2©2©

Pharmacol., 206 (2), 150-156. doi: 10.1016/j.taap.2004.12.015.

15. Melnychuk, D. O., Melnykova, N. M., & Derkach, Ye. A. (2004). Vikovi osoblyvosti kumuliatsii kadmiiu v orhanakh otruienykh shchuriv i zminy pokaznykiv kyslotnoluzhnoho stanu krovi za riznykh umov antyoksydantnoho zakhystu orhanizmu. Ukrain-skyi biokhimichnyi zhurnal, 95-99 (in Ukrainian).

16. Nazaruk, N., Gutyj, B. V., & Hufriy, D. (2015). Influence of metifen and vitamix se on the activity of aminotransferases of bulls blood serum at cadmium nitrate loading. Scientific Messenger of LNU of Veterinary Medicine and Biotechnologies, 17 (1), 121126. Retrieved from https://nvlvet.com.ua/in-dex.php/journal/article/view/231.

17. Ostapyuk, A. Y., & Gutyj, B. V. (2018). Influence of cadmium loading on morphological parameters of blood of the laying hens. Scientific Messenger of Lviv National University of Veterinary Medicine and Biotechnologies, 20 (88), 48-52. doi: 10.32718/nvl-vet8808.

18. Peng, L., Wang, X., Huo, X., Xu, X., Lin, K., Zhang, J., Huang, Y., & Wu, K. (2015). Blood cadmium burden and the risk of nasopharyngeal carcinoma: a case-control study in Chinese Chaoshan population. Environmental Science and Pollution Research, 22 (16), 12323-12331. doi: 10.1007/s11356-015-4533-4.

19. Rodriguez, E. M., Bigi, R., Medesani, D. A., Stella, V. S., Greco, L. S. L., Moreno, P. A. R., Monser-rat, J. M., Pellerano, G. N., & Ansaldo, M. (2001). Acute and chronic effects of cadmium on blood homeostasis of an estuarine crab, Chasmagnathus granulata, and the modifying effect of salinity. Brazilian Journal of Medical and Biological Research, 34 (4), 509-518. https://www.ncbi.nlm.nih.gov/pubmed/11285463.

20. Salvatori, F., Talassi, C. B., Salzgeber, S. A., Sipinosa, H. S., & Bernardi, M. M. (2004). Embryo-toxic and long-term effects of cadmium exposure during embryogenesis in rats. Neurotoxicology and Teratology, 26 (5), 673-680. doi: 10.1016/j.ntt.2004.05.001.

21. Uetani, M., Kobayashi, E., Suwazono, Y., Okubo, Y., Honda, R., Kido, T., & Nogawa, K. (2005). Selenium, Cadmium, Zinc, Copper, and Iron Concentrations in Heart and Aorta of Patients Exposed to Environmental Cadmium. Bulletin of Environmental Contamination and Toxicology, 75 (2), 246-250. doi: 10.1007/s00128-005-0744-6.

22. Vlizlo, V. V. (2012). Laboratorni metody doslidzhen u biolohiyi, tvarynnytstvi ta veterynarniy medytsyni. Spolom, Lviv (in Ukrainian).

23. Vorozhenko, V. V., Skalskyi, Yu. M. (2011). Hihiienichna otsinka ryzykiv vplyvu neradiatsiinykh antropohennykh chynnykiv na stan zdorovia naselennia Ukrainy. Odeskyi medychnyi zhurnal, 5 (127), 4-8 (in Ukrainian).

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 (E1), (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