Green solution to riparian pollution: Populus alba L. potential for phytoremediation and bioindication of PTEs along the Sava river.

This study addresses the potential of Populus alba L. for bioindication and phytoremediation of the contaminated lower part of the Sava River. The main objectives are to assess soil contamination with potentially toxic elements (PTEs: As, B, Cd, Cr, Cu, Li, Ni, Pb, Sr, and Zn), evaluate their availability, and assess the phytoremediation and bioindication potential of Populus alba. Quantification of the PTE contents was performed using inductively coupled plasma optical emission spectroscopy (ICP-OES), while bioindication and phytoremediation potential were evaluated using accumulation indices. The study revealed phytotoxic contents of Cr, Cu, Ni and Zn in the riparian soils of the lower Sava River. The percentage of available Cd was high, but due to its low total content, its phytotoxic potential is limited. According the metal accumulation index, Populus alba exhibits significant potential to accumulate the PTEs studied (with accumulated toxic contents of B, Cr, Li, Sr, and Zn). The ability of Populus alba to accumulate and bioindicate Cd, Cr, and Ni is promising, as is its ability to potentially remediate B, Cd, and Zn. Copper deficiency in leaves resulted in a reduction in photosynthetic performance, but without visible morphological symptoms. The reduced photosynthetic capacity serves as an adaptive strategy for this species in response to toxic levels of PTEs. Since Populus alba is widely distributed in European riparian forests, it is a good candidate to address soil contamination through phytoremediation and bioindication techniques.

Ethnobotanical and ethnomedicinal research into medicinal plants in the Mt Stara Planina region (south-eastern Serbia, Western Balkans).

BACKGROUND: Ethnobotanical research in Southeast Europe-one of the most important European hotspots for biocultural diversity-is significant for the acquisition of Traditional Ecological Knowledge related to plants as well as for encouraging the development of local environments. The current ethnobotanical research was conducted in the region of Mt Stara Planina (south-eastern Serbia), which is characterised by rich phytodiversity with a large number of endemic and relict plant species. The aim of the study was to document the diversity of uses of medicinal plants and of traditional knowledge on their therapeutic uses. METHODS: Ethnobotanical data was collected through both open and semi-structured interviews with locals. Fifty-one inhabitants were interviewed (26 men and 25 women), aged 30-91, and data was analysed by means of use reports, citation frequency, use values (UV), and the informant consensus factor (ICF). RESULTS: The study identified 136 vascular medicinal plant taxa and one lichen species belonging to 53 families and 116 genera. Lamiaceae (19), Rosaceae (18), and Asteraceae (17) had the highest species diversity. The plant parts most commonly used to make a variety of herbal preparations were the aerial parts (54 citations), leaves (35 citations), fruits (20 citations), flowers (18 citations), and roots (16 citations), while the most common forms of preparation were teas (60.78%), consumption of fresh tubers, leaves, roots, and fructus (6.86%), compresses (5.88%), juices (5.39%), decoctions (3.92%), 'travarica' brandy (3.92%), and syrups (2.45%). Of the recorded species, 102 were administered orally, 17 topically, and 18 both orally and topically. The plants with a maximum use value (UV = 1) were Allium sativum, Allium ursinum, Gentiana asclepiadea, Gentiana cruciata, Gentiana lutea, Hypericum perforatum, Thymus serpyllum and Urtica dioica. The highest ICF value (ICF = 0.95) was recorded in the categories of Skin and Blood, Blood Forming Organs, and Immune Mechanism. CONCLUSIONS: This study shows that medicinal plants in the research area are an extremely important natural resource for the local population as they are an important component of their health culture and provide a better standard of living.

Ecophysiological response of Populus alba L. to multiple stress factors during the revitalisation of coal fly ash lagoons at different stages of weathering.

The enormous quantities of fly ash (FA) produced by thermal power plants is a global problem and safe, sustainable approaches to reduce the amount and its toxic effects are still being sought. Vegetation cover comprising long-living species can help reduce FA dump-related environmental health issues. However, the synergistic effect of multiple abiotic factors, like drought, low organic matter content, a deficit of essential nutrients, alkaline pH, and phytotoxicity due to high potentially toxic element (PTE) and soluble salt content, limits the number of species that can grow under such stressful conditions. Thus, we hypothesised that Populus alba L., which spontaneously colonised two FA disposal lagoons at the 'Nikola Tesla A' thermal power plant (Obrenovac, Serbia) 3 years (L3) and 11 years (L11) ago, has high restoration potential thanks to its stress tolerance. We analysed the basic physical and chemical properties of FA at different weathering stages, while the ecophysiological response of P. alba to multiple stresses was determined through biological indicators [the bioconcentration factor (BCF) and translocation factor (TF) for PTEs (As, B, Cr, Cu, Mn, Ni, Se, and Zn)] and by measuring the following parameters: photosynthetic efficiency and chlorophyll concentration, non-enzymatic antioxidant defence (carotenoids, anthocyanins, and phenols), oxidative stress (malondialdehyde (MDA) concentrations), and total antioxidant capacity (IC50) to neutralise DPPH free radical activity. Unlike at L3, toxic As, B, and Zn concentrations in leaves induced oxidative stress in P. alba at L11, shown by the higher MDA levels, lower vitality, and reduced synthesis of chlorophyll, carotenoids, and total antioxidant activity, suggesting its stress tolerance decreases with long-term exposure to adverse abiotic factors. Although P. alba is a fast-growing species with good metal accumulation ability and high stress tolerance, it has poor stabilisation potential for substrates with high As and B concentrations, making it highly unsuitable for revitalising such habitats.

The Phytoremediation Potential and Physiological Adaptive Response of Tamarix tetrandra Pall. Ex M. Bieb. during the Restoration of Chronosequence Fly Ash Deposits.

The challenging process of identifying and selecting plant species suited to the phytoremediation of fly ash (FA) dumps involves studying their functional properties and physiological response to a deficit of essential elements and toxicity from heavy metal(loid)-induced oxidative stress. We hypothesised that Tamarix tetrandra has high potential to be used for the phytoremediation of FA deposit sites thanks to its secretion strategy and antioxidative system. In this study, this hypothesis was examined by determining the bioconcentration and translocation factors for As, B, Cr, Cu, Mn, Ni, Se and Zn at the FA disposal lagoons at the 'Nikola Tesla A' thermal power plant in Obrenovac, Serbia, three (lagoon L1) and eleven (lagoon L2) years after the phytoremediation process had begun, and by measuring parameters of photosynthetic efficiency and chlorophyll concentration, non-enzymatic antioxidant defence (carotenoids, anthocyanins and phenolics), oxidative stress (concentration of malondialdehyde-MDA) and total antioxidant capacity to neutralise DPPH free radical activity. Tamarisk not only showed the ability to phytostabilise As, Cr and Ni and to accumulate low-availability Mn, Zn and Cu, but also the potential to maintain the structural and functional integrity of cell membranes and stable vitality at L1 under multiple stress conditions due to the high synthesis of phenols and tolerance to increased salinity. However, toxic concentrations of B and Se in leaves induced oxidative stress in tamarisk at L2 (reflected in higher MDA content and lower vitality) and also decreased the synthesis of chlorophyll, carotenoids, anthocyanins and total antioxidant activity. In addition, the prooxidative behaviour of phenols in the presence of spin-stabilising metals from FA could also have resulted in their weaker antioxidant protection at L2. These findings indicate that the choice of tamarisk was justified, but only at the beginning of the phytoremediation process because its presence contributed to an improvement in the harsh conditions at FA deposit sites and the creation of more favourable conditions for new plant species. This knowledge can be of great importance when planning sustainable ash deposit site management worldwide.

Fractionation of Potentially Toxic Elements (PTEs) in Urban Soils from Salzburg, Thessaloniki and Belgrade: An Insight into Source Identification and Human Health Risk Assessment.

Concentrations of potentially toxic elements (PTEs) (Al, As, Cd, Cr, Cu, Ni, Pb, and Zn) were measured in topsoil samples collected from parks in the cities of Salzburg (Austria), Thessaloniki (Greece), and Belgrade (Serbia) in order to assess the distribution of PTEs in the urban environment, discriminate natural (lithogenic) and anthropogenic contributions, identify possible sources of pollution, and compare levels of pollution between the cities. An assessment of the health risks caused by exposure to PTEs through different pathways was also conducted. The study revealed that, with the exception of Pb in Salzburg, levels of PTEs in the soils in polluted urban parks were higher than in unpolluted ones, but still lower than those recorded in other European soils. Results of sequential analyses showed that Al, Cr, and Ni were found in residual phases, proving their predominantly lithogenic origin and their low mobility. In contrast, the influence of anthropogenic factors on Cu, Pb, and Zn was evident. Site-dependent variations showed that the highest concentrations of As, Cu, Pb, and Zn of anthropogenic origin were recorded in Salzburg, while the highest levels of Al, Cr, and Ni of lithogenic origin were recorded in Belgrade and Thessaloniki, which reflects the specificity of the geological substrates. Results obtained for the health risk assessment showed that no human health risk was found for either children or adults.

An Assessment of the Phytoremediation Potential of Planted and Spontaneously Colonized Woody Plant Species on Chronosequence Fly Ash Disposal Sites in Serbia-Case Study.

In this study, the potential of planted (Tamarix tetrandra Pall. ex M.Bieb. and Robinia pseudoacacia L.) and spontaneously colonized (Amorpha fruticosa L. and Populus alba L.) woody species for the phytoremediation of potentially toxic trace elements (TEs) such as As, B, Cr, Cu, Mn, Ni, Se, and Zn, from the chronosequence fly ash (FA) deposit lagoons (L1 and L2) at the 'Nikola Tesla A' Thermal Power Plant (TENT-A) in Serbia were analyzed. The differences in the pseodototal and bioavailable (DTPA-extractable) concentrations and mobility (AR index) of TEs in FA at the examined lagoons are a result of the time-conditioned influence of weathering (3 and 11 years respectively) and vegetation development on changing the basic physical and chemical properties of FA (texture, pH, EC, CEC, C, N, and bioavailable P and K) and its toxicity. This resulted in differences in the concentration of TEs in the roots and leaves of the examined plants at L1 and L2. All examined species accumulated Cr the most in the root (BAF > 1 and TF < 1), which suggests that they are good stabilizers of this element. Biological indices for As (BAF > 1 and TF < 1) identified T. tetrandra and A. fruticose as good stabilizers of As. P. alba stood out as accumulating the highest levels of B, Ni, and Zn, T. tetrandra the highest levels of Cu, Mn, and Se, and R. pseudoacacia the highest levels of As and B in leaves (BAF > 1; TF > 1), which makes them good extractors of these elements from the FA at TENT-A. However, due to toxic concentrations of As, B, Se, and Zn in their leaves, they are not recommended for the phytoremediation of the investigated lagoons through the process of phytostabilization. Under conditions of elevated total Cu and Ni concentration in FA, the content of these elements in the leaves of A. fruticosa at both lagoons were within the normal range. This, in addition to a good supply of essential Zn, the stabilization of As and Cr in the roots, an increase in BAF, and a decrease in TF for B with a decrease in its mobility in ash over time, singles this invasive species out as the best candidate for the phytostabilization of TEs in FA at the TENT-A ash deposit site.

Phytoremediation Potential, Photosynthetic and Antioxidant Response to Arsenic-Induced Stress of Dactylis glomerata L. Sown on Fly Ash Deposits.

Arsenic (As) from coal fly ash can be released into soil/groundwater, presenting a global threat to the environment and human health. To overcome this environmental problem, phytoremediation represents an urgent need, providing 'green' cleanup of contaminated lands. The present study focused on As concentrations in fly ash and plants, evaluation of phytoremediation potential of Dactylis glomerata sown on fly ash deposits together with its photosynthetic activity, and oxidative and antioxidative response to As stress. Field research was carried out on fly ash deposits at the thermal power plant "Nikola Tesla", Obrenovac (TENT-A, Serbia) and the control site. Fly ash is characterized by alkaline pH reactions, small amounts of organic matter, a large amount of available phosphate, and total and available As concentrations. Results in this study indicate that phosphate application can ameliorate As toxicity, uptake and root-shoot transport. Furthermore, D. glomerata can be considered as good As phytostabilizator, because it retains more As in roots than in leaves. Excess As in leaves decreases photosynthetic efficiency (Fv/Fm) and concentrations of chlorophylls, carotenoids, and anthocyanins, whereas high content of malondialdehyde (MDA) can be a signal for biosynthesis phenolics and ascorbic acid, providing cellular redox homeostasis and recovery of photosystem II (PSII) photochemistry. In the roots, low oxidative stress under high concentrations of As is related to intense antioxidant biosynthesis. Taken together, the results in this study indicate a high adaptive potential of D. glomerata to As stress. These findings may suggest that physiological and metabolic tools can be used as a way forward in the 'real field' scenario, phytomanagement of fly ash and ecosystem services providing sustainable phytoremediation of As-contaminated sites around the globe.

The potential of elm trees (Ulmus glabra Huds.) for the phytostabilisation of potentially toxic elements in the riparian zone of the Sava River.

The use of trees to immobilise potentially toxic elements (PTEs) is a low-cost and effective method of soil remediation. Thus, the objective of this study was to assess the content of total and bioavailable As, Cd, Cr, Cu, Ni, Pb and Zn in soil samples, as well as their levels in the roots and leaves of elm (Ulmus glabra Huds.) in order to evaluate its potential for the phytostabilisation of PTEs in the riparian zone of the Sava River. Analysis of soils showed that the availability of PTEs ranged from low to medium, while the pollution load index (PLI) and potential ecological risk index (RI) showed that the examined soil fell into the category of uncontaminated to moderately contaminated, as well as into the category of low risk of PTEs contamination. However, the levels of Cr, Cu and Ni in soils were above the critical range for plants. The content of As and Cr measured in roots and leaves was in the toxic range for plants, while the content of Cd and Ni was elevated but not in the toxic range. Bioaccumulation (BCF) and translocation (TF) factors indicated that U. glabra is suitable for the phytostabilisation of As, Cu, Cr, Ni and Pb. Additionally, this species displayed the ability to transport most of the acquired Cu and Zn to the leaves. Correlation analysis showed that PTE content in U. glabra roots was significantly positively correlated to their respective levels in soil (total and DTPA-extractable), except for Cu, indicating that PTE levels in soil strongly influence those in plants. This research into a successful phytoremediating species provides new possibilities when selecting PTE-tolerant native trees in riparian zones of large regional rivers such as the Sava.

Fractionation, Mobility, and Contamination Assessment of Potentially Toxic Metals in Urban Soils in Four Industrial Serbian Cities.

The main soil properties, concentrations of selected elements (As, Cd, Cr, Cu, Fe, Mn, Ni, Pb, and Zn), and the chemical speciation of each element were determined in urban soil samples taken from urban parks in four Serbian cities (Belgrade, Pancevo, Obrenovac, and Smederevo) exposed to different sources of pollution. Pollution indices (PI, PIN) and factors (MF, ICF, GCF) also were evaluated. The study revealed As and Cd concentrations below the detection limit, whereas the content of Cr, Cu, Fe, Mn, Ni, Pb, and Zn at some sites exceeded the limits established by local regulations, as well as the background values, which may represent an environmental threat. Sequential extraction results show that Fe, Cr, Cu, and Ni were predominantly in the residual fraction at most sites; however, Ni from Pancevo and Smederevo also was bound to the reducible fraction. The presence of Pb at all sites and Zn in Smederevo and Belgrade was mainly associated with the reducible and residual fractions. The highest Mn content was found in the reducible fraction, followed by the acid soluble/exchangeable and residual fractions. Based on the obtained indices and factors, the overall soil status at the selected sampling sites was found to range from the warning limit to slightly polluted, whereby Smederevo had the highest risk, and Pancevo and the control site the lowest risk of contamination by toxic metals.

Traditional wound-healing plants used in the Balkan region (Southeast Europe).

ETHNOPHARMACOLOGICAL RELEVANCE: The geographical and ecological specificity of the Balkan Peninsula has resulted in the development of a distinct diversity of medicinal plants. In the traditional culture of the Balkan peoples, plants have medicinal, economic and anthropological/cultural importance, which is reflected in the sound knowledge of their diversity and use. This study analyses the traditional use of medicinal plants in the treatment of wounds and the pharmacological characteristics of the most frequently used species. MATERIALS AND METHODS: A detailed analysis of the literature related to ethnobhe uses of medicinal plants in the Balkan region was carried out. Twenty-five studies were analysed and those plants used for the treatment of wounds were singled out. RESULT: An ethnobotanical analysis showed that 128 plant species (105 wild, 22 cultivated and 1 wild/cultivated) are used in the treatment of wounds. Their application is external, in the form of infusions, decoctions, tinctures, syrups, oils, ointments, and balms, or direct to the skin. Among those plants recorded, the most commonly used are Plantago major, Hypericum perforatum, Plantago lanceolata, Achillea millefolium, Calendula officinalis, Sambucus nigra, Tussilago farfara and Prunus domestica. The study showed that the traditional use of plants in wound healing is confirmed by in vitro and/or in vivo studies for P. major and P. lanceolata (3 laboratory studies for P. major and 2 for P. lanceolata), H. perforatum (5 laboratory studies and 3 clinical trials), A. millefolium (3 laboratory studies and one clinical trial), C. officinalis (6 laboratory studies and 1 clinical trial), S. nigra (3 laboratory studies) and T. farfara (one laboratory study). CONCLUSION: The beneficial effects of using medicinal plants from the Balkan region to heal wounds according to traditional practices have been proven in many scientific studies. However, information on the quantitative benefits to human health of using herbal medicines to heal wounds is still scarce or fragmented, hindering a proper evaluation. Therefore, further studies should be aimed at isolating and identifying specific active substances from plant extracts, which could also reveal compounds with more valuable therapeutic properties. Furthermore, additional reliable clinical trials are needed to confirm those experiences encountered when using traditional medicines. A combination of traditional and modern knowledge could result in new wound-healing drugs with a significant reduction in unwanted side effects.

An ethnobotanical survey of traditionally used plants on Suva planina mountain (south-eastern Serbia).

ETHNOPHARMACOLOGICAL RELEVANCE: This study documents the ethnobotanical and ethnomedicinal importance of plants in the Suva planina mountain region (south-eastern Serbia). It is reflected in their high diversity and their wide range of uses in the treatment of the local population. The aim of this study was a comparative analysis of data collected in the Suva planina region with relevant data from the Western Balkans, which included identifying the 'most popular' plants, as well as those species which are used specifically for treatment solely in the research area. MATERIALS AND METHODS: Ethnobotanical research was carried out between 2012 and 2014 and data was collected through both open and semi-structured interviews with locals. A total of 66 people were interviewed (37 women and 29 men), aged between 49 and 90 (with a mean age of 71). RESULTS: This study identified 128 plants and 2 fungi which are used in ethnomedicine, 5 plant species used in ethnoveterinary medicine, and 16 plants used for 'other' purposes. Lamiaceae (20), Asteraceae (17), Rosaceae (16), Brassicaceae (5), Alliaceae (4) and Apiaceae (4) have the greatest diversity of species. Results showed that Achillea mellefolium, Allium cepa, Allium sativum, Arctostaphyllos uva-ursi, Gentiana lutea, Hypericum perforatum, Juglans regia, Matricaria chamomilla, Mentha piperita, Plantago lanceolata, Plantago major, Salvia officinalis, Sempervivum tectorum, Tilia cordata and Thymus sepyllum are the 'most popular' medicinal plants (UV=1). Those plants with the most phytotherapeutic uses are Gentiana cruciata (14), H. perforatum (11) and A. sativum (10), while the most common conditions treated with medicinal plants are respiratory (79), urogenital (53), gastrointestinal (51), skin (43) and those relating to the circulatory system (35). A comparative analysis of the data collected in the research area and that from other parts of the Western Balkans showed that there are great similarities within Serbia between Suva planina and the Zlatibor region (37.2%) and Kopaonik Mt. (32.3%), while further afield it is most similar to Bosnia and Herzegovina (40.9%) and Bulgaria (40.6%). Moreover, it was established that 14 plant species and 2 fungi are used only in the Suva planina region, which points to the specificity of the diversity and the sound knowledge of medicinal plants in this region. CONCLUSIONS: Our results confirm that medicinal plants are an invaluable resource of the research area and need to be protected as they contribute to an improvement in living standards and the survival of people threatened by unfavourable demographic trends. However, due to over-exploitation, some plants have become exceptionally rare and are under threat, leading to the need for their rational use and protection so as to ensure they are still around for future generations.

Phytotherapy in medieval Serbian medicine according to the pharmacological manuscripts of the Chilandar Medical Codex (15-16th centuries).

The Chilandar Medical Codex is the most significant and best preserved medieval Serbian manuscript and collects together documents on European medical science from the 12th to 15th centuries. It represents the best-known and most complete example of a large collection of medical manuscripts from the Salerno-Montpellier school, written in the vernacular - something which does not exist among the majority of European nations. This paper presents the section of the Codex that deals with phytotherapy, which is contained within the pharmacological manuscripts. An analysis of their contents shows that out of a total of 167 recorded substances, 135 are of plant origin (81%), 13 animal origin (7.7%) and 19 inorganic (11.3%). The recorded plant species are categorised into 63 families, of which the most frequent are: Apiaceae (8.1%), Lamiaceae (8.1%), Asteraceae (5.9%), Rosaceae (5.9%) and Fabaceae (4.4%). All possible plant parts were used in treatments: the whole plant (6%), underground parts (13.7% - root, rhizome, bulb) and aerial parts (80.3% - stem, leaf, flower, buds, fruit, seeds). Of the plants quoted, the following are mentioned most frequently: Vitis sp. (120), Rosa canina (55), Olea europaea (45), Pistacia lentiscus (25), Saccharum officinarum (23), Artemisia absinthium (16) and Foeniculum vulgare (15). The contents of the pharmacological manuscripts of the Chilandar Medical Codex point to the sound contemporary knowledge of the diversity of plant species, their origins, habitat types, the levels of their healing powers, and when and how to gather them and prepare them, as well as the recommended dose for the treatment of specific illnesses. As these manuscripts contain not only common, lay terms for the plants, but also scientific, botanical ones, we can consider them the precursor to Serbian botany. Based on its contents and the way in which they are presented, it can be viewed not only as the first Serbian pharmacopeia, but first Slavic pharmacopeia, too, because similar manuscripts written in the vernacular did not exist during that period among the other Slavic nations, or even most European nations.

The potential of Festuca rubra and Calamagrostis epigejos for the revegetation of fly ash deposits.

Two grass species, Festuca rubra (sown) and Calamagrostis epigejos (naturally recolonised), were studied at two fly ash deposit lagoons, weathered 5 (L1) and 13 years (L2). Both species were assessed in terms of their invasive ability, photosynthetic efficiency, trace elements accumulation, and damage symptoms, while the characteristics of the habitat were assessed in terms of trace element content, and the pH and EC of the ash. In the ash weathered for 5 and 13 years, B and Mn concentrations fell within the normal range for soils, while there was an As, Mo and Cu excess, and Se and Zn deficit. There was no difference in the photosynthetic efficiency (ns) of the C. epigejos populations at the different-aged ash lagoons, but differences were found between the populations of F. rubra (P<0.001). F. rubra displayed damage symptoms, in the form of leaf tip chlorosis and necrosis, and wilting seedlings--a result of B accumulation in toxic concentrations and a Cu and Mn deficit. After 13 years the sown F. rubra remained over 9.5% of the ash lagoon's area (L2), while over the same period the naturally recolonised C. epigejos had overgrown 87.5% of the area. The greater colonisation and survival potential of C. epigejos is a result of multiple tolerance to the conditions at ash deposits and of the competitive ability of this species to dominate the colonised habitat. Therefore, characteristics of naturally colonised species can be used for modelling future actions of biological restoration of fly ash deposits aimed at binding the ash with minimal investment (the short-term aim), and for providing conditions for revegetation, and shortening the successive phases in the revegetation of ash deposits after the closure of thermal plants (the long-term aim).