УДК 599:574.4+577.15

Ecology. Environmental monitoring.

O.M. Vasilyuk

Oles’ Honchar Dnipropetrovsk National University

Effect of Nickel on Alanine Aminotransferase activity in Glechoma hederacea L. leaves subject to excretory function of Mammalia

Introduction

The ecological methods of dealing with anthropogenic pollution is the great importance in the present. Thus, the environmental stress: salinity, fluctuating temperatures, water deficit (Dzyubak and Vasilyuk, 2009; Vasilyuk and Dzyubak, 2009; Martín et al., 2013; Matějka and Leugner, 2013; Salehi et al., 2013), cold shock (Hébrard et al., 2013) and anthropogenic stress has stimulated to study the impact of these factors on all components of the biota (Malá et al., 2013). Thus, the osmotic and ionic phases of salt stress have been determined (Kulik and Vasilyuk, 2009; Shavrukov, 2013), plant growth regulators (Vasilyuk and Kordin, 2005; Vasilyuk and Grycenko, 2008; Heiskanen et al., 2013), mechanisms of protection from heavy metals (HM) in plants (Varennes A. et al., 1996; Gardea-Torresdey J.L. et al., 2005; Sun et al., 2013), resistance against the organic compounds (Gallie, 2013), the specificity and nonspecific biochemical reactions to these factors have been established (Naji and Devaraj, 2011). The mechanisms of plant resistance to HM (Perfetto B. et al., 2006) involves both mechanical mechanisms (Vasilyuk and Kulik, 2008, 2009, 2011, 2011) protection and biochemical metabolisms (Vestena et al., 2011; Hameed A. et al., 2011; Hasan et al., 2011; Ruscitti et al., 2011; Millaleo et al., 2013; Becerril et al., 2013), plant growth regulators (PGR), phytohormones, vitamins (Clemans, 2001; Bernini et al., 2010; Pakhomov and Vasilyuk, 2012, 2013). One of the environmental measures to increase plant resistance to HM stress may be the use of Bacillus subtilis (Bіlanich, 2008), Jatropha Curcas L. (Ghavri and Singh, 2010), ground vegetation (Heiskanen et al., 2013; Buriánek et al., 2013) as an important factor in the biodiversity. We have studied a variety sensitive and simple methods of environmental impact assessment, leading to creation of measures designed to preserve and restore natural biodiversity in areas where diversity has been depleted by man.

We researched the effect of nickel on plant organisms, nickel being one of the most widespread pollutants in Dnipropetrovs’k region and Nickel belongs to the category of HM (Alexeev, 1987). A reduction of toxicity of Ni ions has been facilitated by such environmental factors as the excretory function of Mammalia (Bulakhov et al., 1997; Bulakhov and Pakhomov, 1998, 2003). The excretory function by Mammalia (Bulakhov and Pakhomov, 2006) affects the physical and chemical properties of soils. Therefore, mammals as biotic factors, play a role in ecological restoration of technologically contaminated and biodiversity - depleted areas. The excretory function of Mammalia directly affected the chemical and biological processes, provided adaptive mechanisms, that was of great practical importance. Using the enzymes of nitrogen metabolism (Pakhomov and Vasilyuk, 2013; Vasilyuk and Pakhomov, 2012), as a highly sensitive indicator for environmental change, we determined the impact of HM (anthropogenic factors) and excretory function of Mammalia (environmental factor) on certain plant specimens.

Materials and methods

The experiment was carried out at the International Biosphere Prysamarsky Station (Andriivka village, Novomoskovsk district, Dnipropetrovsk region) in natural floodplain oak forest in a sward area with lime-ash oak and greater stitchwort (Stellaria holostea L.). We studied the excretory function of Mammalia (as our example mammals such as Sus scrofa L. and Capreolus capreolus L.) under the condition of artificial Ni soil pollution. As a control we selected an area unaffected by mammals and without Ni pollution. Ni was introduced onto the soil surface in the form of a salt solution Ni(NO₃)₂*6H₂O with concentrations of 0,2; 1,0; 2,0 g/m², which was equivalent to the presence of Ni at 1, 5, 10 times the dose of maximum acceptable level concentration (MAC). To prevent contamination of the surrounding soil layers by nickel we used isolated soil blocks for our experiment - on the perimeter of each plot vertically placed plates of inert impermeable material were placed into the ground to a depth of 20 cm. A month later we determined the total enzyme activity (nM pyruvic acid/ml*s) of Alanin aminotransferase (AlT, 2.6.1.2.) by the of method of Polevoy and Maximov (2008), as indicators of the ecological condition of the environment, and the concentration of water-soluble fraction of protein (C, mg/ml) by method of Bradford (1976) in the leaves of Glechoma hederacea L., which dominated in this research area. It was found that ALT and AST (Alspartate Aminotransferase (Ast, 2.6.1.1.)), are the part of the system, which utilizes the primary product of photosynthesis in the C₄ aspartate plants group – aspartate which is formed in the leaf mesophyll of plants. The reliable difference between the samples in this paper was considered to be at p<0,05 (Dospekhov, 1985). The experiment was performed under the following schemes:

- control (the area without pollution of Ni and excretory function of Mammalia), control Ni at a dose 1 МAС, combined effect of excretory function of Sus scrofa L. and Ni at a dose 1 МAС, combined effect of excretory function of Capreolus capreolus L. and Ni at a dose 1 МAС, control Ni at a dose 5 МAС, combined effect of Sus scrofa L. and Ni at a dose 5 МAС, combined effect of Capreolus capreolus L. and Ni at a dose 5 МAС, control Ni at a dose 10 МAС, combined effect of Sus scrofa L. and Ni at a dose 10 МAС, combined effect of Capreolus capreolus L. and Ni at a dose 10 МAС.

Results and discussion

It was found that addition of salts of nickel in the range of concentrations (1 МAС, 5 МAС, 10 МAС) produced a reliable decrease in ALT activity in Glechoma hederacea L. leaves subject by 65%-88% for all variants of the experiment (P≤0,05) relative to control (the area without pollution of Ni and excretory function of Mammalia), which disproves the null hypothesis H₀d=0 (under conditions of criterion of significant differences (P≤0,05). This fact proved the toxic influence of the metal on the growth and development of plants and soils in these concentrations. The concentration of water-soluble protein fraction in the Glechoma hederacea L. leaves under the range of Ni concentrations was inhibited (P≤0,05) in all variants of the experiment by 31%-64% relative to control (the area without pollution of Ni and excretory function of Mammalia), which proved the toxic effect of nickel ions (Table. 1).

Table 1

The effect of nickel on Alanine Aminotransferase total activity and concentration of water-soluble protein fraction in the Glechoma hederacea L. leaves subject (n=12)

The variant of the experiment	X± m	Value of experiment relative to control, %
Alanine Aminotransferase total activity (nM pyruvic acid/ml*s)
Control	1,61±0,445	100,00
Ni 1 МAС	0,43±0,177*	26,47
Ni 5 МAС	0,19±0,102*	11,76
Ni 10 МAС	0,57±0,177*	35,29
Water-soluble protein fraction (C, mg/ml)
Control	1,82±0,285	100,00
Ni 1 МAС	1,26±0,061*	69,40
Ni 5 МAС	0,67±0,033*	36,79
Ni 10 МAС	1,20±0,127*	65,60

Note: X - average; ± m - confidence interval; * – criterion of significant differences when P <0.05;

Н₀d = 0 is rejected under conditions P≥1;

Adding the nickel salts at a dose of 1 МAС under the influence of excretory function of Sus scrofa L. (combined effect of anthropogenic and natural factors) contributed to a reliable increase in ALT activity in Glechoma hederacea L. leaves subject by 4,6 times and contributed to a reliable increase in ALT activity by 2,9 times under the influence of excretory function of Capreolus capreolus L relative to control (сontrol Ni 1 МAС), whereas with an increasing concentration factor (Ni 5) activity of ALT was significantly reduced by 32 % (P≤0,05) under the influence of excretory function of Sus scrofa L. and increased by 48% (P≤0,05) under the influence of excretory function of Capreolus capreolus L. compared with controls (control Ni 5 МAС) accordingly. Adding the nickel salts at a dose of 10 МAС under the influence of excretory function of Sus scrofa L. (combined effect of anthropogenic and natural factors) contributed to a reliable reduction ALT activity by 88% and by 8% under the influence of excretory function of Sus scrofa L and Capreolus capreolus L. accordingly. Thus, the excretory function of mammals promoted the intensification of nitrate metabolism in Glechoma hederacea L. leaves subject under the medium and low concentrations of Ni, but under the high concentrations of the metal, the excretory function of Mammalia did not diminish the toxic effect of Ni, which proved the harmful effects of Ni on the process of self-healing in biological systems (table 2).

Table 2

The effect of combined excretory function of Mammalia and Ni on Alanine Aminotransferase total activity in Glechoma hederacea L. leaves subject (n=12)

The variant of the experiment	X± m	Value of experiment relative to control, %
Alanine Aminotransferase total activity (nM pyruvic acid/ml*s)
Control: Ni 1МРС	0,45±0,270
Sus scrofa L. + Ni 1МРС	1,92±0,353*	468,21
Capreolus capreolus L. + Ni 1МРС	1,21±0,177*	294,80
Control: Ni 5МРС	0,21±0,177
Sus scrofa L. + Ni 5МРС	0,17±0,102*	67,78
Capreolus capreolus L. + Ni 5 МРС	0,28±0,177*	148,89
Control: Ni МРС	0,64±0,177
Sus scrofa L. + Ni 10 МРС	0,09±0,102*	11,85
Capreolus capreolus L. + Ni 10МРС	0,62±0,204	92,59

Note:

see Table 1

It was observed that the excretory function of mammals (Sus scrofa L.) reliably increased the concentration of water-soluble protein fractions in Glechoma hederacea L. leaves subject by 12% relative to control and by 48% (Capreolus capreolus L.) under the nickel salts at a dose of 1 МAС (the combined action of the excretory function of Mammalia and nickel at a dose of 1 MAC) compared to control (Ni 1 МAС). We observed an reliable increase (in the activity of protein metabolism by 32% and 8% (P≤0,05) under Ni 5 MAC and with the excretory function of mammals (Sus scrofa L. and Capreolus capreolus L. accordingly) compared to control (Ni 5 МAС). The excretory function of Mammalia (Sus scrofa L. and Capreolus capreolus L.) under the maximum concentration of nickel (Ni 10 МAС) did not provide a reduction in metal toxicity, there was a reliable (P≤0,05) reduction in the concentration of water-soluble proteins fraction by 10%-21% compared to the control (control Ni 10 МAС ). The excretory function of Mammalia under conditions of nickel salts at the maximum dose did not provide a reduction of metal toxicity due to the inability of biological systems to adapt and restore functional activity of metabolic processes at this concentration. The representatives of Mammalia reduced anthropogenic pressure, produced a reduction of toxicity of metal components in biosystems and the environment under the low and medium doses of nickel (Table 3).

Table 3

The effect of combined excretory function of Mammalia and Ni on the concentration of water-soluble protein fraction in Glechoma hederacea L. leaves (n=12)

The variant of the experiment	X± m	Value of experiment relative to control, %
Water-soluble protein fraction (C, mg/ml)
Control: Ni 1МРС	1,26±0,061	100,00
Sus scrofa L. + Ni 1МРС	2,83±0,257*	223,76
Capreolus capreolus L. + Ni 1МРС	1,87±0,229*	147,96
Control: Ni 5МРС	0,67±0,033	100,00
Sus scrofa L. + Ni 5МРС	0,89±0,052*	132,25
Capreolus capreolus L. + Ni 5 МРС	0,74±0,069*	108,11
Control: Ni МРС	1,20±0,127	100,00
Sus scrofa L. + Ni 10 МРС	0,94±0,117*	78,23
Capreolus capreolus L. + Ni 10МРС	1,08±0,026*	90,02

Note:

see Table 1

Conclusions

Thus, we discovered the fact of the reduction in total activity of ALT by 65%-81% (P≤0,05) and concentrations of water-soluble protein by 31%–64% (P≤0,05) relative to control (the area without pollution of Ni and without excretory function of Mammalia) in leaves of Glechoma hederacea L., which dominated in the area under study, which was subject to considerably toxic concentrations of Ni, at doses of 1 MAC, 5 MAC, 10 MAC. The combined effect of the excretory function of Mammalia (as our example mammals such as Sus scrofa L. and Capreolus capreolus L.) and nickel at doses of 1 MAC, 5 MAC contributed to the increased activity of the enzyme and nickel at doses of 10 MAC contributed to the reduced activity of the enzyme (compared with the control in the corresponding concentration factor). The concentration of water soluble protean fraction under the combined effect of the excretory function of Mammalia and Ni at maximum concentration was reduced by 10%-21% (compared with the corresponding control), because it was difficult for the system to operate the mechanisms of recovery and normalization function, while at low-and medium metal concentration the processes of protein metabolism increased (P≤0,05) by 123%-47% and 8%-32% (under the combined effect of the excretory function of Mammalia Sus scrofa L. and Capreolus capreolus L. separately) accordingly.

So, it was observed, that the excretory function of Mammalia reduced the toxic effects of nickel at small and medium concentrations of the metal. Thus, the use of individual elements of zoocoenosis in the system of nature conservation and improvement of the transformed ecosystems in the Steppe Dnieper region had positive results.

References

1. Alexeev, J.V., 1987. Heavy metals in soil and plants [Tjazhelye metally v pochve i rastenijah]. Leningrad Nauka (in Russian).

2. Becerril, F.R., Juárez-Vázquez, L.V., Hernández-Cervantes, S.C., Acevedo-Sandoval, O.A., Vela-Correa, G., Cruz-Chávez, E., Moreno-Espíndola, I.P., Esquivel-Herrera, A., de León-González, F., 2013. Impacts of manganese mining activity on the environment Interactions among soil, plants and arbuscular mycorrhiza. Environ. Contam. Toxicol. 64(2), 219–227.

3. Bernini, E.S., Carmo, T.M.S., Cuzzuol, do G.R.F., 2010. Spatial and temporal variation of the nutrients in the sediment and leaves of two Brazilian mangrove species and their role in the retention of environmental heavy metals. Braz. J. Plant Physiol. 22(3), 177–187.

4. Bіlanich, M.M., 2008. The current state of research of influence of heavy metals on plant life [Suchasnij stan doslіdzhennja vplivu vazhkih metalіv na roslinnij svіt]. Visn. Carpathian. Univ. Biol.12, 161–176 (in Ukrainian).

5. Bradford, M., 1976. A rapid and sensitive method for the quantification of microgram quantities of protein utilising the principle of protein-dye binding. Analyt. Biochemistry J. 72 248–254.

6. Bulakhov, V.L., Pakhomov, A.Y., Reva, A.A., 1997. Usage of vertebrate animals on technogen ecosystems recultivation and purification. Secotox 97. Abstracts Ecotoxicology and Environment Safety Central Eastern European Conferens, Jurmala, Latvia, 198.

7. Bulakhov, V.L., Pakhomov, A.Y., 1998. Soil burrowers-phytophagans participation in the soil – β radioactivity decrease. Toxicol. Lett Suppl. 60– 61.

8. Bulakhov, V.L., Pakhomov, A.Y., 2003. Teriogenic formation of soil microflora in steppe woods of Ukraine. I FEMS Congress of European Microbiologists. Abstract book, Lubbjax, Slovenia: Cunkazjev Dom, 368.

9. Bulakhov, V.L., Pakhomov, О.Е., 2006. Biological diversity of Ukraine. Dnipropetrovsk region.Mammals (Mammalia) [Biologichne riznomanittja Ukrai'ny. Dnipropetrovs'ka oblast'. Ssavci (Mammalia)]. Dnipropetrovs'k [Dnipropetrovsk University Press.] (in Ukrainian).

10. Clemans S., 2001. Molecular mechanisms of plant metal tolerance and homeostasis. Planta. 212(4), 475–486.

11. Buriánek, V., Novotný, R., Hellebrandová, K., Šrámek, V., 2013. Ground vegetation as an important factor in the biodiversity of forest ecosystems and its evaluation in regard to nitrogen deposition. J. For. Sci., 59 (2013): 230-237.

12. Dospekhov B. A., 1985. Methods of field experience [Metodika polevogo opyta]. Moscaw. [Agroprom Press.] (in Russian).

13. Dzyubak, O.I., Vasilyuk, O.M., 2009. Effect of chloride salinity on morphometric and biochemical indices in the dynamics of plant growth and development [Vplyv hlorydnogo zasolennja na morfometrychni ta biohimichni pokaznyky roslyn u dynamici rostu ta rozvytku]. Materialy I Mizhnarodnoi' Naukovoi' Konferencii' Studentiv, Aspirantiv ta Molodyh Uchenyh Fundamental'ni ta Prykladni Doslidzhennja v Biologii' [Materials of the International Scientific Conference of Students and Young Scientists Fundamental and Applied Research in Biology], Donetsk, Veber, 2, 231–232 (in Ukrainian).

14. Gallie, D.R., 2013. The role of l-ascorbic acid recycling in responding to Environmental stress and in promoting plant growth. Exp. Botany. 64(2), 433–443.

15. Gardea-Torresdey, J.L., Rosa, G. de la, Peralta-Videa, J.R., Montes, M., Cruz-Jimenez, G., Cano-Aguilera, I., 2005. Differential uptake and transport of trivalent and behexavalent chromium by tumbleweed (Salsola kali). Environ. Contam. Toxicol. 48(2), 225–232.

16. Ghavri, S.V., Singh, R.P., 2010. Phytotranslocation of Fe by biodiesel plant Jatropha curcas L. grown on iron rich wasteland soil. Braz. J. Plant Physiol. 22(4), 235–243.

17. Hameed, A., Mahmooduzzafar, T.N.Q, Siddiqi, T.O., Iqbal, M., 2011. Differential activation of the enzymatic antioxidant system of Abelmoschus esculentus L. under CdCl₂ and HgCl₂ exposure. Braz. J. Plant Physiol. 23(1), 46–54.

18. Hasan, S.A., Hayat, S., Wani, A.S, Ahmad, A., 2011. Establishment of sensitive and resistant variety of tomato on the basis of photosynthesis and antioxidative enzymes in the presence of cobalt applied as shotgun approach. Braz. J. Plant Physiol. 23(3), 175–185.

19. Hébrard, C., Trap-Gentil, M.V., Lafon-Placette, C., Delaunay, A., Joseph, C., Lefèbvre, M., Barnes, S., Maury, S., 2013. Identification of differentially methylated regions during vernalization revealed a role for RNA methyltransferases in bolting. Exp. Botany. 64(2), 651–663.

20. Heiskanen, J., Tammeorg, P., Dumroese, R.K., 2013. Growth of Norway spruce seedlings after transplanting into silty soil amended with biochar: a bioassay in a growth chamber. J. For. Sci., 59, (3): 117-124.

21. Kulik, A.F., Vasilyuk, O.M., 2009. Catalase activity in soils of forest biocenosis in Samara Region [Aktyvnist' katalazy v g'runtah lisovyh biogeocenoziv Prysamar’ja]. Visn. Dnipropetr. Univ. Biol. Ecol. [Dnipropetrovsk University Press.] 17(2), 63–68 (in Ukrainian).

22. Lefcort, Н., Wehner, E.A., Cocco, P.L., Lefcort, H., 2013. Look inside get aaccess pre-exposure to heavy metal pollution and the odor of predation decrease the ability of snails to avoid stressors. Environ. Contam. Toxicol. 64(2), Р.273–280.

23. Malá, J., Cvikrová, M., Hrubcová, M., Máchová, P., 2013. Influence of vegetation on phenolic acid contents in soil. J. For. Sci., 59, (7): 279-287.

24. Martín, J.A.R., Carbonell, G., Nanos, N., Gutiérrez, S.C., 2013. Identification of soil mercury in the Spanish Islands. Environ. Contam. Toxicol. 64(1), 171–179.

25. Matějka, K., Leugner, J., 2013. Growth of Norway spruce (Picea abies [L.] Karsten) from artificial and natural regeneration in the Krkonoše Mts. and air temperature variability. For. Sci., 59, (4): 137-149.

26. Millaleo, R., Reyes-Díaz, M., Alberdi, M., Ivanov, A.G., Krol, M., Hüner, N.P.A., 2013. Excess manganese differentially inhibits photo system I versus II in Arabidopsis thaliana L. Exp. Botany. 64(91), 343–354.

27. Naji, K.M., Devaraj, V.R., 2011. Antioxidant and other biochemical defense responses of Macrotyloma uniflorum (Lam.) Verdc. (Horse gram) induced by high temperature and salt stress. Braz. J. Plant Physiol. 23(3), 187–195.

28. Pakhomov, A.Y., Bulakhov, V.L., 1993. Migration of some micro- and microelements in environment under influence of digging activity of mammals. Central and Eastern European Regional Meeting. Environmental toxicology: Pathways of Anthropogenic Pollutants in the Environment and their Toxic Effect, Porabka-Kozubnik, Poland, 82.

29. Pakhomov, O.Е., Vasilyuk, O.М., 2011. Activity of trans - amination enzymes as the indicator of biological revegetation of soils Mammalia in transformed ecosystems. The Abstracts NATO Advanced Research Workshop (ARW): Environmental Security for South-East Europe and Ukraine, NATO Science Series Book, Dnipropetrovs’k, 74–75.

30. Pakhomov, O.Е., Vasilyuk, O.M., 2012. The influence of anthropogenic factors on the activity of transferases in the background environment generating function of mammals [Vplyv antropogennyh faktoriv na aktyvnist' transferaz na foni seredovyshhetvirnoi' funkcii' ssavciv]. Visn. Dnipropetr. Univ. Biol. Ecol. [Dnipropetrovsk University Press.] 20(2), 64–70 (in Ukrainian).

31. Pakhomov, O.E., Vasilyuk, O.M., 2013. Activity of transamination enzymes as the indicator of Environmental forming function of Mammalia representatives in transformed anthropogenic ecosystem. Nauk. Visn. Cherniv. Univ. Biol. (Biol. systems). 21(5), 54–61.

32. Perfetto, B., Lamberti, M., Giuliano, M.T., 2006. Analysis of the signal transduction pathway of nickel-induced matrix metalloproteinase-2 expression in the human keratinocytes in vitro: preliminary findings. Cutaneous Pathology. 34(6), 441–447.

33. Polevoy, V., Maximov, G. (eds.), 2008. Methods of analysis biochemically of plants, 2008. [Metody biohimicheskogo analiza rastenij]. Leningrad University Press (in Russian).

34. Ruscitti, M., Arango, M., Ronco, M., Beltrano, J., 2011. Inoculation with mycorrhizal fungi modifies proline metabolism and increases chromium tolerance in pepper plants (Capsicum annuum L.), Braz. J. Plant Physiol. 23(1), 15–25.

35. Salehi, A., Ghorbanzadeh, N., Kahneh, E. J., 2013. Earthworm biomass and abundance, soil chemical and physical properties under different poplar plantations in the north of Iran. .For. Sci., 59, (6): 288-294.

36. Shavrukov, Y. (2013). Salt stress or salt shock: which genes are we studying. J. Exp. Bot., 64(1), 119–127.

37. Sun, J., Cui, J., Luo, C., Gao, L., Chen, Y., Shen, Z., 2013. Contribution of cell walls, nonprotein thiols, and organic acids to cadmium resistance in two cabbage varieties. Environ. Contam. Toxicol., 64(2), 243–252.

38. Varennes, A., Torres, M. O., Coutinho, J. F., Rocha, M.M.G.S., Neto, M.M.P. M., 1996. Effects of heavy metals on the growth and mineral composition of a nickel hyper accumulator. Plant Nutr. 19(5), 669–676.

39. Vasilyuk, O.M., Dzyubak, О.І., 2009. Physiological and biochemical parameters of plants as markers of a condition of environment (Materials of the International Scientific Conference of Students and Young Scientists Fundamental and Applied Research in Biology), Donetsk, Veber, 2, 348–349.

40. Vasilyuk, O.M., Grycenko, P.V., 2008. Effect of growth regulators on the activity of transamination enzymes in the leaves and roots of Salix alba L. [Vplyv reguljatoriv rostu na aktyvnist' fermentiv pereaminuvannja v lysti ta korenjah Salix alba L.]. Visn. Dnipropetr. Univ. Biol. Ecol. [Dnipropetrovsk University Press.] 16(1), 34–40 (in Ukrainian).

41. Vasilyuk, O.M., Kordin, O.I., 2005. Morphometric indices and yields of maize hybrids as markers of plant adaptation. [Morfometrychni pokaznyky ta vrozhajnist' gibrydiv kukurudzy jak markery adaptacii' roslyn] Visn. Dnipropetr. Univ. Biol. Ecol. [Dnipropetrovsk University Press.] 3-1, 13–18 (in Ukrainian).

42. Vasilyuk, O.M., Kulik, A.F., 2008. Aspartate Aminotransferase activity in Zea Maize L. shoots under the cesium ions influence. Visnyk zaporіz'kogo nacіonal'nogo unіversitetu, (Bulletin of the Zaporizhzhya National University, 1, 238.

43. Vasilyuk, O.M., Kulik, A.F., 2009. Invertase and urease activity in soils reclaimed areas of the Western Donbass. [Aktyvnist' invertazy ta ureazy v g'runtah rekul'tyvovanyh terytorij Zahidnogo Donbasu]. Visn. Dnipropetr. Univ. Biol. Ecol. [Dnipropetrovsk University Press.] 17(3), 10–15 (in Ukrainian).

44. Vasilyuk, О.M., Kulik A.F., 2011. Transamination enzyme activity in leaves Sambucus nigro L. in high salinity [Aktyvnist' fermentiv pereaminuvannja v lystkah Sambucus nigro L. v umovah pidvyshhenoi' mineralizacii']. Visn. Dnipropetr. Univ. Biol. Ecol. [Dnipropetrovsk University Press.] 19(1), 10–15 (in Ukrainian).

45. Vasilyuk, O. М., Kulik, A. F., 2011. The trans-amimation enzyme activity in the leaves of Sambucus Nigra L. under high minaralization of small rivers of Steppe Dnieper The Abstracts NATO Advanced Research Workshop (ARW): Environmental Security for South-East Europe and Ukraine, NATO Science Series Book, Dnipropetrovs’k, 84–85.

46. Vasilyuk, O.M., Pakhomov, O.E., 2012. Effect of nickel ions on the functional activity of enzymes in the leaves of Glechoma hederatia L. in digging activity of Mammalia. [Vplyv ioniv nikelju na funkcional'nu aktyvnist' transaminaz v lystkah Glechoma hederatia L. v umovah ryjnoi' aktyvnosti Mammalia]. Materialy VIІI Mezhdunarodnoj Nauchno-Prakticheskoj Konferencii Dostizhenija Vysshej Shkoly-2012. (Proceedings of the 8th International Scientific and Practical Conference Investigation of High Society Schools-2012). Bjal GRAD-BG, Sofija, Bolgarija, 1, 25–32 (in Ukrainian).

47. Vestena, S., Cambraia, J., Ribeiro, C., Oliveira, J.A, Oliva, M.A., 2011. Cadmium induced oxidative stress and antioxidative enzyme response in water hyacinth and salvinia. Braz. J. Plant Physiol. 23(2), 131–139.