Bioavailability and bioaccumulation characterization of essential and heavy metals contents in R. acetosa, S. oleracea and U. dioica from copper polluted and referent areas.

BACKGROUND: Bioavailability of metals occurring in soil is the basic source of its accumulation in vegetables and herbs. The impact of soil pollution (due to urban and mining areas) on the food chain presents a challenge for many investigations. Availability of metals in a potentially polluted soil and their possible transfer and bioaccumulation in sorrel (Rumex acetosa), spinach (Spinacia oleracea) and common nettle (Urtica dioica), were examined. METHODS: Microwave digestion was applied for total digestion of the plant tissues, while on the soil samples open wet digestion with a mixture of acids was applied. Three extraction methods were implemented for the bioavailable metals in the soil. Atomic emission spectrometry with inductively coupled plasma was used for determination of the total contents of 21 elements. RESULTS: Significant enrichments in agricultural soil for As, Pb and Zn (in urban area), Cd, Cu and Ni (in a copper mine area), compared with the respective values from European standards were detected. On the basis of three different extraction methods, higher availability was assumed for both lithogenic and anthropogenic elements. Translocation values >1 were obtained for As, Cd, Cu, Ni, Pb and Zn. Higher bioconcentrating value was obtained only for Cd, while the bioaccumulation values vary from 0.17 for Cd to 0.82 for Zn. CONCLUSIONS: The potential availability of hazardous metals in urban and mining soils is examined using DTPA-TEA-CaCl2 (urban) and HCl (Cu-mines areas). Our results suggested that S. oleracea and R. acetosa have a phytostabilization potential for Cd, Cu, Ni and Pb, while U. dioica only for Cu. R. acetosa has a potential for phytoextraction of Cd in urban and copper polluted areas.

Distribution of chemical elements in soils and stream sediments in the area of abandoned Sb-As-Tl Allchar mine, Republic of Macedonia.

The aim of this study was to investigate the distribution of some toxic elements in topsoil and subsoil, focusing on the identification of natural and anthropogenic element sources in the small region of rare As-Sb-Tl mineralization outcrop and abandoned mine Allchar known for the highest natural concentration of Tl in soil worldwide. The samples of soil and sediments after total digestion were analyzed by inductively coupled plasma-mass spectrometry (ICP-MS) and inductively coupled plasma-atomic emission spectrometry (ICP-AES). Factor analysis (FA) was used to identify and characterize element associations. Six associations of elements were determined by the method of multivariate statistics: Rb-Ta-K-Nb-Ga-Sn-Ba-Bi-Li-Be-(La-Eu)-Hf-Zr-Zn-In-Pd-Ag-Pt-Mg; Tl-As-Sb-Hg; Te-S-Ag-Pt-Al-Sc-(Gd-Lu)-Y; Fe-Cu-V-Ge-Co-In; Pd-Zr-Hf-W-Be and Ni-Mn-Co-Cr-Mg. The purpose of the assessment was to determine the nature and extent of potential contamination as well as to broadly assess possible impacts to human health and the environment. The results from the analysis of the collected samples in the vicinity of the mine revealed that As and Tl elements have the highest median values. Higher median values for Sb are obviously as a result of the past mining activities and as a result of area surface phenomena in the past.

Biomonitoring of air pollution with mercury in Croatia by using moss species and CV-AAS.

Moss samples from four dominant species (Hypnum cupressiforme, Pleurozium schreberi, Homalothecium sericeum and Brachythecium rutabulum) were collected during the summer and autumn of 2010 from 121 sampling sites evenly distributed over the territory of Croatia. Samples were totally digested by using microwave digestion system, whilst mercury was analysed by using cold vapour atomic absorption spectrometry (CV-AAS). Descriptive statistics were done from analyses of mercury in all moss samples. The content of mercury ranged from 0.010 to 0.145 mg kg(-1) with a median value of 0.043 mg kg(-1). Hg distribution map shows the sites of the country with higher levels of this element. High contents of Hg were found in moss samples collected from the regions of Podravina and Istria as a result of anthropogenic pollution. Comparison of median values and ranges with those found in moss samples in 2006 shows slight reduction of mercury air pollution. When compared to the results obtained from recent studies conducted in Slovenia, Macedonia and especially in Norway-which serves as a reference considering the fact that it is a pristine area-mercury air pollution in Croatia is insignificant.

Applying of factor analyses for determination of trace elements distribution in water from Vardar and its tributaries, Macedonia/Greece.

A systematic study was carried out to investigate the distribution of fifty-six elements in the water samples from river Vardar (Republic of Macedonia and Greece) and its major tributaries. The samples were collected from 27 sampling sites. Analyses were performed by mass spectrometry with inductively coupled plasma (ICP-MS) and atomic emission spectrometry with inductively coupled plasma (ICP-AES). Cluster and R mode factor analysis (FA) was used to identify and characterise element associations and four associations of elements were determined by the method of multivariate statistics. Three factors represent the associations of elements that occur in the river water naturally while Factor 3 represents an anthropogenic association of the elements (Cd, Ga, In, Pb, Re, Tl, Cu, and Zn) introduced in the river waters from the waste waters from the mining and metallurgical activities in the country.

Multi-elements atmospheric deposition study in Albania.

For the first time, the moss biomonitoring technique and inductively coupled plasma-atomic emission spectrometric (ICP-AES) analytical technique were applied to study multi-element atmospheric deposition in Albania. Moss samples (Hypnum cupressiforme) were collected during the summer of 2011 and September-October 2010 from 62 sites, evenly distributed over the country. Sampling was performed in accordance with the LRTAP Convention-ICP Vegetation protocol and sampling strategy of the European Programme on Biomonitoring of Heavy Metal Atmospheric Deposition. ICP-AES analysis made it possible to determine concentrations of 19 elements including key toxic metals such as Pb, Cd, As, and Cu. Cluster and factor analysis with varimax rotation was applied to distinguish elements mainly of anthropogenic origin from those predominantly originating from natural sources. Geographical distribution maps of the elements over the sampled territory were constructed using GIS technology. The median values of the elements in moss samples of Albania were high for Al, Cr, Ni, Fe, and V and low for Cd, Cu, and Zn compared to other European countries, but generally were of a similar level as some of the neighboring countries such as Bulgaria, Croatia, Kosovo, Macedonia, and Romania. This study was conducted in the framework of ICP Vegetation in order to provide a reliable assessment of air quality throughout Albania and to produce information needed for better identification of contamination sources and improving the potential for assessing environmental and health risks in Albania, associated with toxic metals.

Influence of heavy metal stress on antioxidant status and DNA damage in Urtica dioica.

Heavy metals have the potential to interact and induce several stress responses in the plants; thus, effects of heavy metal stress on DNA damages and total antioxidants level in Urtica dioica leaves and stems were investigated. The samples are sampled from areas with different metal exposition. Metal content was analyzed by Inductively Coupled Plasma-Atomic Emission Spectrometer (ICP-AES), for total antioxidants level assessment the Ferric-Reducing Antioxidant Power (FRAP) assay was used, and genomic DNA isolation from frozen plant samples was performed to obtain DNA fingerprints of investigated plant. It was found that heavy metal contents in stems generally changed synchronously with those in leaves of the plant, and extraneous metals led to imbalance of mineral nutrient elements. DNA damages were investigated by Random Amplified Polymorphic DNA (RAPD) technique, and the results demonstrated that the samples exposed to metals yielded a large number of new fragments (total 12) in comparison with the control sample. This study showed that DNA stability is highly affected by metal pollution which was identified by RAPD markers. Results suggested that heavy metal stress influences antioxidant status and also induces DNA damages in U. dioica which may help to understand the mechanisms of metals genotoxicity.

The study on air pollution with nickel and vanadium in Croatia by using moss biomonitoring and ICP-AES.

Moss samples were collected from 121 sampling sites all over Croatia during the summer and autumn of 2010. They were totally digested by using microwave digestion system and analysed by using atomic emission spectrometry with inductively coupled plasma (ICP-AES). Descriptive statistics and maps of distribution were made. The data obtained in this study were compared with those from the study in 2006 and additionally with the data obtained in the similar studies in neighbouring countries and Norway as pristine area. The median value of nickel is 3.16 mg kg(-1) and the content varies from 1.04 to 14.66 mg kg(-1). The content of vanadium ranges between 0.23 and 37.26 mg kg(-1) with the median value of 2.55 mg kg(-1). High contents of these elements are found in the vicinity of Rijeka, Zagreb and Sisak as a result of their emission from oil refinery, thermal power plant and industrial processes.

Air dispersion of heavy metals in the vicinity of the As-Sb-Tl abounded mine and responsiveness of moss as a biomonitoring media in small-scale investigations.

A systematic study was carried out to investigate air deposition and to explore the natural distribution and enrichment (contamination) with trace elements in the small area (cca. 13 km(2)) of an antimony-arsenic-thallium mineralization outcrop at an abandoned mine "Allchar." The mine is located on the northwestern part of Kozuf Mount, Republic of Macedonia. The locality of Allchar is unique in its mineral composition; besides a very intriguing mineral, lorandite, there are 45 other minerals, some of which are rare. The distribution of 53 elements (with special attention to As, Sb, and Tl) were detected in 69 moss samples from eight various species collected from this area. Moss samples were analyzed following microwave digestion by inductively coupled plasma-mass spectrometry and inductively coupled plasma-atomic emission spectrometry. It was found that the atmospheric deposition for As in the moss samples on or around the Allchar mine is >6.5 times higher and for Tl is 19 times higher compared to values for the samples from the rest of the Allchar area. By the application of multivariate cluster and R-mode factor analyses (FA), five geochemical associations were determined. Cluster and R-mode FA were used to identify and characterize element associations, and five associations of elements were determined by the method of multivariate statistics. F1 (Co, Cr, Fe, Sc, Li, V, Ga, Y, Ni, Mn, Al, La-Lu, Cu, Ge, Be, Bi, and Hf); F2 (As, Tl, Sb, and Mg); F3 (Rb, Cs, and Mo); F4 (Sr, Ba, Hf, Zr, La-Lu, and Bi), and F5 (Cd, Zn, Ag, and Cu).

Influence of a nickel smelter plant on the mineralogical composition of attic dust in the Tikves Valley, Republic of Macedonia.

Mineral phases and their content were determined in attic dust samples collected from 27 houses in the Tikves Valley, Republic of Macedonia. By using quantitative X-ray diffraction, the principal mineral phases were determined to be the serpentinite group (chrysotile, lizardite) and amphibole group of minerals (ribecite, tremolite, actinolite) present in the attic dust samples from this region which are not common constituents of urban dust. Strong correlations existed between these mineral phases in the dust and those in ores processed at a ferronickel smelter plant situated in this region. Spatial distributions of specific mineral phases were made and were consistent with wind directions and predicted deposition (60-70 %) of dust emitted from the metallurgical plant.

Mineral nutrient imbalance, total antioxidants level and DNA damage in common bean (Phaseolus vulgaris L.) exposed to heavy metals.

The present study aimed to analyze the biological effects induced by bioaccumulation of metals in common bean (Phaseolus vulgaris L.). Effects of mineral nutrient imbalance, total antioxidants level and DNA damage induced by accumulation of heavy metals, were investigated in bean seedlings treated with two selected metal concentrations for 7 days. Metal content is analyzed by inductively coupled plasma - atomic emission spectrometer (ICP-AES), for total antioxidants level assessment the Ferric-Reducing Antioxidant Power (FRAP) assay is used and Random Amplified Polymorphic DNA (RAPD) method was applied for investigation of DNA damages. The increasing metal concentration in the treatment medium changed synchronously metal content in samples, and decreased total antioxidant activity in all samples with exception only for samples treated with Ni and Cd. The obtained "DNA fingerprints" demonstrated that the increasing metal concentrations induced changes in RAPD profiles (disappearance and/or appearance of bands in comparison with untreated control samples). The highest number of missing bands was observed in samples treated with zinc (total 4 bands) and nickel (total 4 bands) at both concentrations. These results suggested that mineral nutrient imbalance is involved in changes of antioxidant levels and DNA damages of the seedlings, which may help to understand the mechanism of metal toxicity in plants.

Moss biomonitoring of air pollution with heavy metals in the vicinity of a ferronickel smelter plant.

The purpose of this study was to establish the atmospheric deposition of various elements in the Kavadarci region, Republic of Macedonia (known for its ferronickel mining and metallurgical activities) using moss biomonitoring, and to determine whether the deposition is anthropogenic or from geogenic influences. The sampling network includes 31 moss samples evenly distributed over a territory of about 600 km(2). A total of 46 elements (Ag, Al, As, Au, Ba, Be, Ca, Cd, Ce, Co, Cr, Cs, Cu, Dy, Er, Eu, Fe, Ga, Gd, Ge, Hg, Ho, K, La, Li, Lu, Mg, Mn, Mo, Na, Nb, Ni, P, Pb, Rb, Sb, Sm, Sr, Tb, Th, Ti, U, V, Yb, Zn, Zr) were determined by mass spectrometry with inductively coupled plasma (ICP-MS). Based on a distribution pattern of elements determined in moss, two anthropogenic geochemical associations (Co-Cr-Cu-Fe-Mg-Ni and As-Cd-Cu-Hg-Pb-Zn), were detected. The distribution of these elements shows an increased content (especially Ni, Co and Cr) in the moss samples from the surroundings of the smelter plant compared to the rest of the samples. Thus, the median value of Ni in moss samples from the whole region (40 mg kg(-1)) is much higher than the median for Macedonia (5.82 mg kg(-1)). Moreover, the median content of Ni in the moss samples from the polluted area (around the smelter) is 178 mg kg(-1) with an enrichment ratio in the moss samples of almost 5.5 times higher than the unpolluted areas (32 mg kg(-1)). This fact confirms the influence of the dust from the ferronickel plant to the air pollution in this region.

Assessment of the genotoxicity of heavy metals in Phaseolus vulgaris L. as a model plant system by Random Amplified Polymorphic DNA (RAPD) analysis.

Impact assessments of environmental pollutants are important in eco-genotoxicology. A random amplified polymorphic DNA (RAPD) technique was used to detect genotoxicity-induced DNA damage in Phaseolus vulgaris L. from heavy metals at two different concentrations. The results from six 10-base pair (bp) random RAPD primers with 60-70% GC content used, showed a total of 295 RAPD fragments of 700-4000 bp in molecular size in the seedlings of untreated and treated samples, of which only 163 fragments were polymorphic. Polymorphisms became evident as the disappearance and/or appearance of DNA fragments in treated samples compared to the control. A dendrogram constructed using the Numerical Taxonomy and Multivariate Analysis System (NTSYSps), showed that the control group merged with groups treated with CuSO(4)·5H(2)O (150 mg L(-1)) and MnSO(4)·H(2)O (150 mg L(-1)) in a separate cluster. These groups were linked with all of the other samples treated with metals at concentrations of 150 mg L(-1) and CuSO(4)·5H(2)O and Cd(NO(3))(2) at concentrations of 350 mg L(-1). Finally, the samples treated with metals at concentrations of 350 mg L(-1) together with NiSO(4) at the concentration of 150 mg L(-1), clustered separately. The DNA polymorphism detected by RAPD analysis offered a useful biomarker assay for the detection of toxic chemicals genotoxicity in plant model systems.

Distribution of chemical elements in attic dust as reflection of their geogenic and anthropogenic sources in the vicinity of the copper mine and flotation plant.

The main aim of this article was to assess the atmospheric pollution with heavy metals due to copper mining Bucim near Radovis, the Republic of Macedonia. The open pit and mine waste and flotation tailings are continually exposed to open air, which leads to winds carrying the fine particles into the atmosphere. Samples of attic dust were examined as historical archives of mine emissions, with the aim of elucidating the pathways of pollution. Dust was collected from the attics of 29 houses, built between 1920 and 1970. Nineteen elements (Ag, Al, As, Ba, Ca, Cd, Co, Cr, Cu, Li, Fe, K, Mg, Mn, Na, Ni, Pb, Sr, and Zn) were analyzed by atomic emission spectrometry with inductively coupled plasma. The obtained values of the investigated elements in attic dust samples were statistically processed using nonparametric and parametric analysis. Factor analysis revealed three factors governing the source of individual chemical elements. Two of them grouping Ca, Li, Mg, Mn, and Sr (Factor 1) and Co, Cr, and Ni (Factor 2) can be characterized as geogenic. The third factor grouping As, Cu, and Pb is anthropogenic and mirrors dust fallout from mining operation and from flotation tailings. Maps of areal deposition were prepared for this group of elements, from which correlation of these anthropogenic born elements was confirmed.

Assessment of heavy metal pollution in Republic of Macedonia using a plant assay.

Different plant organs (leaves, flowers, stems, or roots) from four plant species-Urtica dioica L. (Urticaceae), Robinia pseudoacacia L. (Fabaceae), Taraxacum officinale (Asteraceae), and Matricaria recutita (Asteraceae)-were evaluated as possible bioindicators of heavy-metal pollution in Republic of Macedonia. Concentrations of Pb, Cu, Cd, Mn, Ni, and Zn were determined in unwashed plant parts collected from areas with different degrees of metal pollution by ICP-AES. All these elements were found to be at high levels in samples collected from an industrial area. Maximum Pb concentration was 174.52 ± 1.04 mg kg⁻¹ in R. pseudoacacia flowers sampled from the Veles area, where lead and zinc metallurgical activities were present. In all control samples, the Cd concentrations were found to be under the limit of detection (LOD <0.1 mg kg⁻¹) except for R. pseudoacacia flowers and T. officinale roots. The maximum Cd concentration was 7.97 ± 0.15 mg kg⁻¹ in R. pseudoacacia flowers from the Veles area. Nickel concentrations were in the range from 1.90 ± 0.04 to 5.74 ± 0.03 mg kg⁻¹. For U. dioica leaves and R. pseudoacacia flowers sampled near a lead-smelting plant, concentrations of 465.0 ± 0.55 and 403.56 ± 0.34 mg kg⁻¹ Zn were detected, respectively. In all control samples, results for Zn were low, ranging from 10.2 ± 0.05 to 38.70 ± 0.18 mg kg⁻¹. In this study, it was found that the flower of R. pseudoacacia was a better bioindicator of heavy-metal pollution than other plant parts. Summarizing the results, it can be concluded that T. officinale, U. dioica, and R. pseudoacacia were better metal accumulators and M. recutita was a metal avoider.

Content of toxic and essential metals in medicinal herbs growing in polluted and unpolluted areas of Macedonia.

The aim of this study was to determine and compare Ba, Cr, Cd, Fe, Sr, Pb, and Zn content in medicinal herbs Urtica dioica L., Taraxacum officinale, and Matricaria recutita growing in polluted and unpolluted areas of the Republic of Macedonia. The metal content was determined using inductively coupled plasma atomic emission spectroscopy (ICP-AES). In the unpolluted area of Mt. Plackovica the metal content in Taraxacum officinale was in the descending order: Fe>Sr>Zn>Ba>Cr, while Pb and Cd were below the limit of detection. In the polluted area of Veles, the order was as follows: Fe>Zn>Sr>Pb>Ba>Cd>Cr. Our results suggest that quality assurance and monitoring of toxic metals is needed for plants intended for human use and consumption. Medicinal plants should be picked in areas free of any contamination sources.

Biomonitoring of atmospheric pollution with heavy metals in the copper mine vicinity located near Radovis, Republic of Macedonia.

This investigation was undertaken to determine the atmospheric pollution with heavy metals due to copper mining Bucim near Radovis, the Republic of Macedonia. Moss samples (Hyloconium splendens and Pleurozium schrebery) were used for biomonitoring the possible atmospheric pollution with heavy metals in mine vicinity. Sixteen elements (Al, As, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Mn, Na, Ni, Pb, Sr, and Zn) were analysed by application of flame and electrothermal atomic absorption spectrometry (FAAS and ETAAS) and atomic emission spectrometry with inductively coupled plasma (ICP-AES). The obtained values were statistically processed using nonparametric and parametric analysis. The median value for copper obtained from moss samples (10 mg kg(-1)) was much lower compared with the same values for the whole territory of the Republic of Macedonia (22 mg kg(-1)). The range of values (2.1-198 mg kg(-1)) shows much higher content of this element in the samples taken from the study area compared to the appropriate values for the whole territory of Macedonia. The association of elements As, Cd, Cu, Fe, Pb, and Zn was singled out by factor analysis as a characteristic anthropogenic group of elements. Maps of area deposition were made for this group of elements, wherefrom correlation of these anthropogenic born elements was confirmed.