УДК 599:574.4+577.15

Ecology. Environmental monitoring.

O.M. Vasilyuk

Oles’ Honchar Dnipropetrovs’k National University

Effect of lead on Alanine aminotransferase activity in Glechoma hederacea leaves subject to digging function of Mammalia

Introduction

Mn, with median toxic concentrations of (μM): 0.30 Pb, 0.47 Hg, 2.0 Cu, 5.0 Cd, 9.0 As, 17 Co, 19 Ni, 25 Zn, and 46 Mn. (Kopittke, at el., 2010). The regulatory roles of ubiquitin during the plant’s response to abiotic stress are summarized and highlighted in the review (Lyzenga and Stone, 2012) was discussed. The mechanisms for the alleviation of Cu toxicity in plants by coexistent cations (e.g. Al³⁺, Mn²⁺, Ca²⁺, Mg²⁺, H⁺, Na⁺, and K⁺) was investigated, because suggests risk assessment of copper in natural waters and soil (Wang, at el., 2012). The effect of tolerances to ferrous (Fe²⁺), sulphide (S^2–), and zinc (Zn²⁺) ions were investigated in seedlings of eight species of mangrove, including three pioneer species, three rhizophoraceous and two landward semi-mangrove species (Cheng, at el., 2012). The autorths (Degenhardt and Gimmler, 2000) reported that lignin was more sensitive to heavy metals. The amount of lignin in roots of maize increased significantly, when exposed to a metal slag (containing more than 1000 mg kg⁻¹ Pb, Zn, and Cu), but suberization in both endo- and hypodermal cell walls was not affected. Was performed to investigate (Takamatsu, at el., 2010) the weathering and dissolution rates of Pb on open grassland and under canopies of Japanese cedar (Cryptomeria japonica) and bamboo-leafed oak (Quercus myrsinaefolia) and accumulation Cd²⁺, Zn²⁺ and Pb²⁺inEuglena gracilis (Mendoza-Cózatl, at el., 2006). The content of Cd, Pb, and Cu in Odonate larvae and their responses to heavy metals (equimolar concentrations of cadmium, lead, or copper) was studied. Larvae were tolerant of high concentrations of cadmium and lead (0.893 and 2.232 mM, respectively), more sensitive to copper exposure, as low as 2.360 mM (Tollett, at el., 2009). Twenty-seven urban topsoil samples (kitchen garden and lawn) were collected and analysed for Cd, Pb and Zn. Such, the quantities of Cd, Pb and Zn extracted from soils indicated, on average, 68, 62 and 47% bioaccessibility, respectively, in the gastric phase and 31, 32 and 23% bioaccessibility, respectively, in the gastro-intestinal phase, what is important for health (Roussel, at el., 2010). We explained the effect of Pb on the plant organism. Lead is one of the common pollutants of Dnipropetrovs’k region. Reduction of toxicity Pb ions also contributes to Environmental factors as digging activity of Mammalia. Digging activity of Mammalia affected to physical and chemical properties of soils (Bulakhov and Pakhomov, 2006). The Environmental forming role of Mammalia is a biotic factor. It was in creation of ecological buffer to anthropogenic load to biota, in the leveling of anthropogenic press through the processes of self-purification of soils to restore biodiversity under anthropogenic impact on the biota. The digging activity of Mammalia indirectly affected to chemical and biological processes, provided adaptive mechanisms to support state homeostasis under the effect of the factor below the acceptable level, which was a great practical value. To determine the degree of influence of heavy metals (HM) to the Environment, we need very sensitive indicators, which were enzymes of nitrogen metabolism (Pakhomov and Vasilyuk, 2012, 2013).

Materials and methods

The experiment was carried out at the International Biosphere Prysamarsky fixed practice bases (Andriїvka village, Novomoskovs’k district, Dnipropetrovs’k region) in natural floodplain oak forest in a sward area with lime-ash oak and greater stitchwort (Stellaria holostea L.). We have detected an environment forming role played by one species of mammal, Talpa europaea L. (European mole), under the condition of artificial Pb soil pollution. As a control we selected an area unaffected by mammals and without Pb pollution. Pb was introduced onto the soil surface in the form of a salt solution Pb (NO₃)₂with concentrations of 1.6, 8, 16 g/m², which was equivalent to the presence of Pb at 1, 5, 10 times the dose of maximum acceptable level concentration (MAC). To prevent contamination of the surrounding soil layers by lead 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 indicator 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 Glechoma hederacea L. leaves subject of 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 Pb and digging function of T. europaea L), control Pb at a dose 1 МAС, combined effect of digging function of T. europaea L. and Pb at a dose 1 МAС, control Pb at a dose 5 МAС, combined effect of T. europaea L. and Pb at a dose 5 МAС, control Pb at a dose 10 МAС, combined effect of T. europaea L. and Pb at a dose 10 МAС.

Results and discussion

It was found that addition of salts of Pb in the range of concentrations (1, 5, 10 МAС) produced a reliable increase in ALT activity by 39% (P<0, 05) for variants of the experiment with Pb in the 10 MAC relative to control (the area without Pb pollution and without digging activity of T. europaea L.) in Glechoma hederacea L. leaves subject. It 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 Pb concentrations was inhibited in variants of the experiment (Pb 5 МAС and Pb 10 МAС) by 19 - 36%, which proved the toxic effect of Pb ions (table 1) and reliable increase by 58% (P<0, 05) to control (the area without Pb pollution and without digging activity of T. europaea L.) in variant of the experiment, where Pb at a dose 1 МAС, which proved the nontoxic effect of Pb ions in the concentration (Tab. 1).

Table 1

The effect of lead 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 experience to control, %
АLТ
Control	1,61 ± 0,270	100,00
Pb 1 МAС	1,40 ± 0,270	92,19
Pb 5 МAС	1,56 ± 0,177	103,13
Pb 10 МAС	2,11 ± 0,368*	139,06
Water-soluble proteins fraction
Control	1,82 ± 0,285	100,00
Pb 1 МAС	2,89 ± 0,341*	158,36
Pb 5 МAС	1,48 ± 0,056*	81,19
Pb 10 МAС	1,15 ± 0,361*	63,06

Note: X - average; ± m - confidence interval; Н₀d = 0 is rejected under conditions P <0.05;

It was found that digging activity of T. europaea L. produced a total reduction of ALT activity by 76% in leaves of Glechoma hederacea L., the concentration of water-soluble protein fraction decreased by 8% relative to control (the area without pollution of Pb and without digging activity of T. europaea L.) during one month. This can be explained by mechanical damage to the soil and plant and insufficient temporal self-healing for the biochemical processes of plants and soil (Tab. 2, 3).

Under the combined action of the digging activity of T. europaea L. and lead at a dose of 1 MAC we observed an reliable increase (compared to control Pb 1 МAС) in the activity of protein metabolism by 34% and at 4, 7 times at combined action of Pb 5 MAC and digging activity (P<0, 05). The digging activity of small rodents under the maximum concentration of lead (Pb 10 МAС) provided an increasing (P<0, 05) in the concentration of water-soluble proteins fraction by 71% compared to the control (control Pb 10 МAС). The representatives of Mammalia reduced anthropogenic pressure, produced a reduction of toxicity of metal components in bio systems and the environment under the doses of lead (Tab. 2).

Table 2

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

The variant of the experiment	X± m	Value of experience to control, %
Control	1,61 ± 0,445	100,00
Digging activity	0,43 ± 0,177*	26,47
Control: Pb 1 МAС	1,04 ± 0,270	100,00
Digging activity + Pb 1 МAС	1,40 ± 0,317*	134,09
Control: Pb 5 МAС	0,33 ± 0,102	100,00
Digging activity + Pb 5 МAС	1,56 ± 0,177*	471,43
Control: Pb 10 МAС	1,23 ± 0,541	100,00
Digging activity + Pb 10 МAС	2,11 ± 0,368*	171,15

Note:

see Table 1

It was observed that the digging activity of T. europaea L reliably decreased the concentration of water-soluble protein fractions in Glechoma hederacea L. leaves subject by 8% relative to control (the area without pollution of lead and digging activity of Mammalia). Under the combined action of the digging activity of T. europaea L. and Pb at a dose of 1 MAC we observed a reliable increase (compared to control Ni 1 МAС) in the activity of protein metabolism by 141% and at 10% at combined action of Ni 5 MAC and digging activity (P<0, 05). The digging activity of small rodents under the maximum concentration of lead (Pb 10 МAС) did not provide a reduction in metal toxicity. There was an unreliable (P<0, 05) compared to the control (control Pb 10 МAС) increasing in the concentration of water-soluble proteins fraction by 2% (Tab.3).

Table 3

The effect of combined digging function of Mammalia and Pb on the concentration of water-soluble protein fraction in Glechoma hederaceaL. leaves subject (n=12)

The variant of the experiment	X± m	Value of experience to control, %
Control	1,82 ± 0,285	100,00
Digging activity	1,69 ± 0,209	92,69
Control: Pb 1 МAС	1,19 ± 0,114	100,00
Digging activity + Pb 1 МAС	2,89 ± 0,340*	241,69
Control: Pb 5 МAС	1,34 ± 0,033	100,00
Digging activity + Pb 5 МAС	1,48 ± 0,056*	110,68
Control: Pb 10 МAС	1,18 ± 0,113	100,00
Digging activity +Pb 10 МAС	1,21 ± 0,186	102,89

Note:

see Table 1

Conclusions

The digging function of this species decreased the level of toxicity from lead at low (1 MAC), medium (5 MAC) and high (10 MAC) concentrations of the metal (on the example of the enzyme activity of ALT and on the example of the concentration of water-soluble protein fractions), but proved ineffective on the example of the concentration of water-soluble protein fractions in Glechoma hederacea L. leaves subject, when the levels of lead were high (10 MAC). The mammals as biotic factors play a role in ecological restoration of technologically contaminated and biodiversity-depleted areas. A creative environmental role in the creation of ecological buffers against anthropogenic impact on biota, in the process of self-purification of soil and the restoration of biodiversity in conditions of human impacts on biota is played by Mammals. Thus, the use of this element of zoocoenosis in the system of conservation measures for natural and transformed ecosystems in the Steppe Dnieper region can produce positive results.

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