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SoilContents of risk metals in soilsLands of localities from which soil and plant samples were taken belong to agricultural lands.Soil reaction is one of the factors that most affects the behaviour of heavy metals in soil. Low pH values pose a risk of reduced nutrient intake and increase the availability of heavy metals for plants29,30.The presence of risk elements in the soil was evaluated based on their contents in bioavailable form (mobile forms), determined in soil extracts NH4NO3, and the total contents of risk elements were determined in soil extract by aqua regia (Table 1).Table 1 The contents of risk elements (Cu, Ni, Pb, Cd, Hg, Mn) in soil (mg/kg).Full size tableAccessible heavy metals for plants are those which are present in the soil solution as soluble components or those which are easily dissolved by root exudates31. The highest Cu contents determined in soil extract by NH4NO3, were in the cadastre of Gemerská Poloma (max. 0.390 mg/kg) (Table 1). However, even the highest determined concentration of Cu in its bioavailable form did not exceeded the determined critical value for this element18. Nickel is a beneficial element for plants. Elevated Ni concentrations in soils have a potential negative effect on plants32. Content of bioavailable forms of nickel is lower than the determined critical value in all analysed samples. Cadmium and lead present a risk to agricultural activity in this area. Cadmium in soil is highly bioavailable and has higher mobility in plants compared to other heavy metals. It is easily transported by roots to shoots. In contrast, lead is one of the least mobile heavy metals. It is naturally concentrated in the upper layers of the soil33. The contents of the available forms of cadmium and lead exceed the critical values for these elements. In case of lead, the determined contents are from 0.257 Henckovce to 0.676 Gemerská Poloma. Takáč et al.34 determined in 20 soil samples from the Central Spiš region 7.2–257.6 mg Cu/kg soil and 1.0–84.8 mg Pb/kg in their potentially mobilizable form and 0.4–1.4 mg Cu/kg soil and 4.3–7.1 mg Pb/kg in their mobile form. In comparison with our results, Vilček et al.35 determined a lower content of Cd (0.04), Pb (0.17), Ni (0.15) and higher Cu content (0.48) mg/kg in forms accessible to plants in 16 soil samples from locality Nižná Slaná in the years 2006–2008. However, high concentrations of metals in soil do not necessarily mean the availability of metals for plants36. As a result, extractable Mn is often a better indicator of Mn availability. Mn2+ is generally considered to be bioavailable22. The highest concentration of Mn was measured in soil samples from the cadastre of Nižná Slaná. On the contrary, the lowest concentrations were detected in samples from Gemerská Poloma cadastre, which is the furthest cadastre from the source. No critical limit is set up for manganese according to Slovak legislation, it is not possible to classify these soils as contaminated/uncontaminated. For comparison, the EDTA-extractable content of Mn ranged from 22.7 to 127 mg/kg dry soil (China)29; the mobile concentrations between 0.32 and 202.0 mg/kg and the available concentrations from 5.4 to 126.3 mg/kg (Egypt)37.Based on results of statistical analysis, significant higher content of Cu, Pb and Cd can be stated in samples from Gemerská Poloma cadastre. These soils are classified as gley fluvisols, soils from the other two localities are cambisols (from medium heavy to light) and acid cambisols (Henckovce), cambisols from medium heavy to light and typically acid cambisols (Nižná Slaná). The soil profile of fluvisols is repeatedly disrupted by floods, which often enriches them with a new layer of sludge sediments2.Another method for determination of metal content in soil is mineralisation using aqua regia, which dissolves most of the soil constituents except those strongly bound in silicate minerals. This content is sometimes referred to as pseudototal (determined in aqua regia). In this way, all elements that are likely to become bioavailable in the long term are determined38.Pseudototal contents of risk metals (Table 1) determined in soil extract using aqua regia were higher than their limit value in case of Cu (Gemerská Poloma cadastre), Cd (all cadastres) and Hg (cadastre of Henckovce and Gemerská Poloma).Due to the fact that the hygienic condition of agricultural soils is assessed according to the exceeding of the limit values of at least one risk substance, the monitored plots can be classified as contaminated (Cu  > 60.0, Cd  > 0.7, Hg  > 0.5 mg/kg soil).Manganese is not classified as risk element in Slovak legislation.Tóth et al.39 classified European soils into four categories: (1) no detectable content of HM, (2) the concentration of the investigated element is above the threshold value (Hg 0.5, Cd 1, Cu 100, Pb 60 and Ni 50 mg/kg), but below the lower guideline value (Hg 2, Cd 10, Cu 150, Pb 200 and Ni 100 mg/kg), (3) concentration of one or more element exceeds the lower guideline value but is below the higher guideline value (Hg 5, Cd 20, Cu 200, Pb 750 and Ni 150 mg/kg), (4) samples having concentrations above the higher guideline value.In comparison with the threshold and guideline values, soils in cadastres of Gemerská Poloma (Cu), Henckovce, Nižná Slaná, Gemerská Poloma (Cd, Hg) represent the ecological risk. Threshold and guideline values for Mn were not defined.The Spiš region and the northern part of the Gemer region belong to the most polluted areas in Slovakia in terms of soil contamination due to mining and metallurgical activities that have been carried out here in the past. Soils near the sludge in Nižná Slaná contain 3.17–53.3 (14.2–301, 0.71–20.6, 3.33–177, 12.9–223 and 675–11,510, respectively) mg Cd (Cu, Hg, Ni, Pb and Mn, respectively)/kg of soil14. In loaded area of Dongchuan, (China), contained Cd (Cu, Hg, Ni and Pb, resp.) 0.20–3.57 (45.38–2026, 0.02–0.23, 24.06–95.9 and 6.83–146.6, resp.) mg/kg40. In contrast, in the agricultural area of Punjab of the India, the soil contamination was caused by an excessive use of agrochemicals and polluted irrigation sources. Increased Cu (Pb and Cd) contents were determined in the soil samples: 9.0–48.5 (5.5–9.67 and 0.516–1.58, resp.) mg/kg41.However, in most cases, a large portion of the total element content is not available for immediate uptake by plants. Available forms represent a small proportion of this total content which is potentially available to plants. Availability is affected by many factors, including pH, redox state, macronutrient levels, available water content and temperature29,33,36,38.Indicators of soil contaminationContamination factors and degree of contaminationThe contamination character may be described in a uniform, adequate and standardised way by means of the contamination factor and the degree of contamination. Hakanson24 reported four Contamination degrees of individual metal (({mathrm{C}}_{mathrm{f}}^{mathrm{i}})) – low (({mathrm{C}}_{mathrm{f}}^{mathrm{i}}) < 1), moderate (1 ≤ ({mathrm{C}}_{mathrm{f}}^{mathrm{i}})   More

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