Atmospheric pollution by potentially toxic elements: measurement and risk assessment using lichen transplants.

The lichen Usnea articulata collected from an unpolluted area was exposed for 6 months at 26 sites for the sample chosenusing a stratified random design, and the content of potentially toxic elements (PTEs) including As, Cd, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, Sn, V, and Zn, was assessed using ICP-MS. The health risk for both adults and children was then calculated using the PTEs concentrations. The results showed that despite the hostile urban conditions, transplanted lichens depicted clear deposition patterns of airborne PTEs, mostly associated with industrial sites, where As and other elements showed remarkably high values. The cumulative hazard index was below the risk threshold, both for adults and children. For the entire population (particularly children) residing in areas surrounding industrial sites, As and Cr appeared to be potentially carcinogenic elements.

Resilience of Xanthoria parietina under Mars-like conditions: photosynthesis and oxidative stress response.

MAIN CONCLUSION: Xanthoria parietina survivability in Mars-like conditions was supported by water-lysis efficiency recovery and antioxidant content balancing with ROS production after 30 days of exposure. Xanthoria parietina (L.) Th. Fr. is a widespread lichen showing tolerance against air pollutants and UV-radiation. It has been tested under space-like and Mars-like conditions resulting in high recovery performances. Hereby, we aim to assess the mechanisms at the basis of the thalli resilience against multiple space stress factors. Living thalli of X. parietina were exposed to simulated Martian atmospheric conditions (Dark Mars) and UV radiation (Full Mars). Then, we monitored as vitality indicator the photosynthetic efficiency, assessed by in vivo chlorophyll emission fluorescence measurements (FM; FV/F0). The physiological defense was evaluated by analyzing the thalli antioxidant capacity. The drop of FM and FV/F0 immediately after the exposure indicated a reduction of photosynthesis. After 24 h from exposure, photosynthetic efficiency began to recover suggesting the occurrence of protective mechanisms. Antioxidant concentrations were higher during the exposure, only decreasing after 30 days. The recovery of photosynthetic efficiency in both treatments suggested a strong resilience by the photosynthetic apparatus against combined space stress factors, likely due to the boosted antioxidants at the beginning and their depletion at the end of the exposure. The overall results indicated that the production of antioxidants, along with the occurrence of photoprotection mechanisms, guarantee X. parietina survivability in Mars-like environment.

Air quality in post-mining towns: tracking potentially toxic elements using tree leaves.

In this study, leaves of the evergreen holm oak Quercus ilex were used to assess airborne contamination of potentially toxic elements (PTEs) at five towns located on the slopes of the Mt. Amiata (central Italy), an area with a long history of mining and, more recently, an important district for the industrial exploitation of geothermal energy. PTE composition and covariance of washed and unwashed Q. ilex leaves of three different ages (6, 12 and 24 month-old) were used to identify atmospheric inputs of PTEs at residential areas, evaluate long-term adsorption and retention of PTEs by the leaves, thus providing an indication of potential human exposure. Moreover, the determination of foliar concentrations of major elements (C, N, S and P) allowed an assessment of the nutritional status of the investigated urban tree stands which excluded the existence of stress condition caused by air pollution or other disturbances. Results indicated that overall Pb, Cu, and Cd concentration were low in the investigated urban sites, if compared with similar studies conducted in larger Italian cities, denoting a low contribution of vehicular traffic to the atmospheric pathway. The five urban settlements were characterized by a specific profile of elements (Al, Ba, Hg and Sb) enriched in unwashed leaves, resulting from the distinct geochemical characteristics of the area and from diffuse (i.e., urban activity) and point sources of PTEs emission (i.e., brownfields, geothermal power plants). The latter sources primarily govern the distribution of Hg, whose contamination was found to be very localized close to a major abandoned mining area. Our data provided quantitative evidence of the spectrum of PTEs potentially impacting resident population and may prove useful in support of follow-up instrumental monitoring campaigns of air quality, as well as for human health and ecological risk assessments.

Selecting the species to be used in lichen transplant surveys of air pollution in Tunisia.

This study was undertaken with the aim of selecting one or more lichen species that are the most suitable for transplant-based surveys of air pollution in Tunisia, in areas where the local native lichen vegetation is scanty or missing at all. To this purpose, four epiphytic (tree inhabiting) lichen species (Evernia prunastri, Flavoparmelia caperata, Parmotrema perlatum, Ramalina farinacea) were collected from the Babouch forests, a remote and unpolluted area of NW Tunisia, and analyzed for their content of potentially toxic elements (PTEs), namely Al, As, Cd, Cr, Cu, Fe, Ni, Pb, Sb, and Zn, by ICP-MS. Moreover, also the physiological status of the lichen samples was evaluated by measuring their chlorophyll content, photosynthetic efficiency, and spectral reflectance. The results indicated a remarkable contribution of airborne soil and dust particles to the total PTE content, especially for the foliose species F. caperata and P. perlatum. The fruticose lichens E. prunastri and R. farinacea had a lower and similar content of PTEs, and hence were regarded as more suitable to be used in transplant studies, since are able to detect even minimal accumulation amounts. All lichen species were healthy, as emerged from the analysis of physiological parameters.

Effects of wood distillate (pyroligneous acid) on sensitive bioindicators (lichen and moss).

Wood distillate (pyroligneous acid) can be successfully applied in agriculture to increase crop quality and productivity with a lower risk for the environment respect to synthetic chemical herbicides, pesticides or fertilizers. However, the effects of wood distillate on the environment and biota are still under investigation, depending on biological attributes of potentially influenced organisms. The potential toxicological effects of wood distillate on sensitive non-target organisms, lichens and mosses, are studied for the first time. The physiological parameters (chlorophyll a fluorescence emission FV/FM and PI(ABS), chlorophyll content, spectral reflectance, antioxidant power, and dehydrogenase activity) and eventual bioaccumulation of selected elements (As, Ba, Cd, Cr, Cu, Fe, Ni, Pb, Zn) were investigated in the lichen Xanthoria parietina and the moss Hypnum cupressiforme after short-term treatments over a range of wood distillate solutions (1:300, 1:500, 1:700) to detect potential early stress responses. Overall, the lichen did not show changes after the treatments, while in the moss wood distillate caused only modest alterations in FV/FM and PI(ABS) and progressive increasing of antioxidant activity according to the dose supplied. The bioaccumulation of toxic elements was low and did not show any pattern of uptake with increasing concentrations of wood distillate.

Functional and structural biomarkers to monitor heavy metal pollution of one of the most contaminated freshwater sites in Southern Europe.

This study evaluated the biological effects of highly polluted freshwater environment (Regi Lagni channels, S Italy) on the aquatic moss Leptodictyum riparium, exposed in bags at three sites representative of different environmental conditions and characterized by different heavy metal burdens. Bioaccumulation, ultrastructural alterations, Reactive Oxygen Species (ROS) production, antioxidant enzymes activity and DNA damage were assessed. To better evaluate the biological response of the moss species to heavy metals, the same biological parameters were assessed also in L. riparium samples cultured in vitro using metal mixtures at the same concentrations as measured at the 3 field exposure sites. Heavy metals were accumulated into the moss tissues causing severe ultra-structural damages at higher concentration case studies, and the ROS production as well as the activity of the enzyme followed a concentration-dependent increase. However, the DNA damage trend suggested a threshold effect that changed between field and in vitro experiment. The enrichment factor suggests that the concentration in the most polluted site is close to the upper limit of L. riparium to accumulate metals. Overall, combining measures of the morpho-functional traits at different level contribute to improving the knowledge about the tolerance of L. riparium to heavy metal stress, suggesting that this moss could be suitable for biomonitoring activity in field conditions.

Toxicity of Diclofenac in the Fern Azolla filiculoides and the Lichen Xanthoria parietina.

This study investigated the occurrence of toxicity, expressed as damage to the photosynthetic apparatus, in the aquatic fern Azolla filiculoides and the lichen Xanthoria parietina following treatments with diclofenac at different concentrations (0.1, 1, 10 and 100 mg/L) and different exposure times (24, 48, 72 and 240 h). Measurements of photosynthetic efficiency, chlorophyll content and chlorophyll degradation indicated dose- and time-dependent toxicity, since significant differences with control samples as well as among treatments, emerged mainly for the highest concentration (100 mg/L) and the longest time (240 h). In addition, also the mycobiont of the lichen X. parietina showed similar toxic effects, expressed as ergosterol content. The absence of relevant alterations at the lowest concentration (0.1 mg/L) suggested a very limited susceptibility of these species to environmentally relevant levels of this pharmaceutical.

Impact of mechanical mowing and chemical treatment on phytosociological, pedochemical and biological parameters in roadside soils and vegetation.

Many chemical and non-chemical strategies have been applied to control weeds in agricultural and industrial areas. Knowledge regarding the effects of these methods on roadside vegetation is still poor. A 2-year field experiment was performed along a road located near Livorno (Tuscany, central Italy). Eight plots/strips were identified, of which four were subjected to periodical mechanical mowing and the remaining four were treated with a chemical herbicide based on glyphosate (the producer's recommended rates were used for the selective control of broad-leaved weeds). Our results clearly showed that roadside soil and vegetation are a significant reservoir of anthropogenic activities which have a strong negative effect on several phytosociological, pedochemical and biological parameters. Compared with conventional mechanical mowing, chemical treatment induced (i) a significant increase in organic matter in the upper plot layers (+18%), and (ii) a marked reduction in weed height throughout the entire period of the experiment. Irrespectively of the kind of treatment, no significance differences were detected in terms of (i) biological quality of soil (the abundance and diversity of arthropod communities did not change), and (ii) plant elemental content (bulk concentrations of analysed trace elements had a good fit within ranges of occurrence in the "reference plant"). The glyphosate partially controlled broad-leaved weeds and this moderate efficacy is dependent upon the season/time of application. In conclusion, the rational and sustainable use of chemical herbicides may be a useful tool for the management of roadside vegetation.

Spatial Variation in the Accumulation of Elements in Thalli of the Lichen Pseudevernia furfuracea (L.) Zopf Transplanted Around a Biomass Power Plant in Italy.

Thalli of the lichen Pseudevernia furfuracea were transplanted for 3 months at 32 sites located in and around an industrial area of S Italy whose main anthropogenic sources of atmospheric trace elements are a biomass power plant and vehicular emissions. Meteorological stations were deployed at four sites for finer detection of local wind patterns. The station near the biomass power plant showed a significant S-SE wind component not detectable by measurements made at the regional scale or by the other local meteorological stations. Sb, Sn, and Mo showed a very high degree of covariance and a statistically significant correlation with traffic rate. No element concentrations in the exposed thalli were correlated with distance from the biomass power plant, although Ti and Co concentrations showed a significant correlation with the "Potential Number of Times the Winds coming from the biomass power plant Reach each exposure Site" (PNTWRS). This value is calculated dividing the time (minutes) during the experimental trimester that the wind blows from the power plant into each of the four geographical sides by the time (minutes) the winds passing through the power plant take to reach the exposure sites in each of the four geographical sides.) during the period of thalli transplantation. Moreover, there were significant differences among clusters of sites with different levels of enrichment of Ti, Co, Al, V, and Cu and a "local control" group. These results, together with the high covariance of the Al-Ti and V-Co pairs, indicate an association between the biomass power plant and spatial variation of Ti, Co, Al, and V levels in the transplanted lichens. The nature of the fuels used in the biomass power plant explains the spatial variation of As, Cr, Cu, and Zn concentrations.

Uptake and toxicity of glyphosate in the lichen Xanthoria parietina (L.) Th. Fr.

This study investigated if treatment of the lichen Xanthoria parietina (L.) Th. Fr. with glyphosate caused uptake of this herbicide as well as physiological alterations. Samples were treated with Glifene SL®, a common commercial glyphosate-based herbicide, at the lowest recommended doses (3.6g/L) as well as with doses slightly higher than the highest suggested (36 g/L). The results clearly showed glyphosate uptake in X. parietina proportionally to the dose provided. Adverse physiological effects were evident on the photosynthetic apparatus (photosynthetic efficiency, chlorophyll a content, chlorophyll degradation) as well as on the fungal respiration rates and cell membrane integrity (ergosterol content, dehydrogenase activity) already after 24h from treatment, also at the low application dose. It is concluded that lichens are suitable organisms for monitoring unwanted biological effects from the application of glyphosate-based herbicides, as well as for detecting the accumulation of this compound in the biota, thus screening for its environmental fate.

Effects of acute NH3 air pollution on N-sensitive and N-tolerant lichen species.

Lichens are sensitive to the presence of ammonia (NH3) in the environment. However, in order to use them as reliable indicators in biomonitoring studies, it is necessary to establish unequivocally the occurrence of certain symptoms following the exposure to NH3 in the environment. In this paper, we simulated an episode of acute air pollution due to the release of NH3. The biological effects of acute air pollution by atmospheric NH3 have been investigated using N-sensitive (Flavoparmelia caperata) and N-tolerant (Xanthoria parietina) species. Lichen samples were exposed to ecologically relevant NH3 concentrations for 8 weeks, simulating three areas of impact: a control area (2 µg/m(3)), an area of intermediate impact (2-35 µg/m(3)) and an area of high impact (10-315 µg/m(3)), with a peak of pollution reached between the fourth and fifth week. Ammonia affected both the photobiont and the mycobiont in F. caperata, while in X. parietina only the photosynthetic performance of the photobiont was altered after exposure to the highest concentration. In the photobiont of F. caperata we recorded chlorophyll degradation as indicated by OD435/415 ratio, decrease of the photosynthetic performance (as reflected by the maximum quantum yield of primary photochemistry FV/FM and the performance index PIABS); in the mycobiont, ergosterol reduction, membrane lipid peroxidation (as reflected by the increase of thiobarbituric acid reactive substances), alteration (decrease) of the secondary metabolite usnic acid. No effects were detected on caperatic acid and dehydrogenase activity. In X. parietina, the only signal determined by NH3 was the alteration of FV/FM and the performance index PIABS. The results suggest that physiological parameters in N-sensitive lichens well reflect the effects of NH3 exposure and can be applied as early indicators in monitoring studies.

Antiproliferative, antibacterial and antifungal activity of the lichen Xanthoria parietina and its secondary metabolite parietin.

Lichens are valuable natural resources used for centuries throughout the world as medicine, food, fodder, perfume, spices and dyes, as well as for other miscellaneous purposes. This study investigates the antiproliferative, antibacterial and antifungal activity of the acetone extract of the lichen Xanthoria parietina (Linnaeus) Theodor Fries and its major secondary metabolite, parietin. The extract and parietin were tested for antimicrobial activity against nine American Type Culture Collection standard and clinically isolated bacterial strains, and three fungal strains. Both showed strong antibacterial activity against all bacterial strains and matched clinical isolates, particularly against Staphylococcus aureus from standard and clinical sources. Among the fungi tested, Rhizoctonia solani was the most sensitive. The antiproliferative effects of the extract and parietin were also investigated in human breast cancer cells. The extract inhibited proliferation and induced apoptosis, both effects being accompanied by modulation of expression of cell cycle regulating genes such as p16, p27, cyclin D1 and cyclin A. It also mediated apoptosis by activating extrinsic and intrinsic cell death pathways, modulating Tumor Necrosis Factor-related apoptosis-inducing ligand (TRAIL) and B-cell lymphoma 2 (Bcl-2), and inducing Bcl-2-associated agonist of cell death (BAD) phosphorylation. Our results indicate that Xanthoria parietina is a major potential source of antimicrobial and anticancer substances.

Spatial variation of eco-physiological parameters in the lichen Pseudevernia furfuracea transplanted in an area surrounding a cement plant (S Italy).

Thalli of the lichen Pseudevernia furfuracea were transplanted for 3 months (November 2010-January 2011) at 61 monitoring sites around a cement plant near Castrovillari (Calabria, southern Italy). NH3, NO x and SO2 concentrations were monitored monthly in a subarea of 10 sites (SA10) where the cement plant was located. At the end of the exposure period, the integrity of cell membranes; membrane lipid peroxidation (thiobarbituric acid reactive substances, TBARS level); vitality (cell respiration); chlorophyll a; chlorophyll b; carotenoids; phaeophytization quotient; photosynthetic efficiency and thalli concentrations of Al, Ca, Mg, V and Fe were measured. NO x concentrations correlated with the site distance from the cement plant while NH3 concentrations correlated with lichen vitality within SA10. For the monitoring area as a whole, only Fe and Mg concentrations correlated with membrane lipid peroxidation, while TBARS levels showed a significant increase and chlorophyll a, chlorophyll b and carotenoids a significant decrease with respect to the lichen origin area. Multivariate analysis (detrended correspondence analysis, cluster analysis and multi-response permutation procedure) of the eco-physiological parameters × monitoring sites data set resulted in four clusters termed C1, C2, C3 and C4. The eco-physiological parameters were compared among the four clusters and lichen origin area by one-way ANOVA. An index of environmental favourableness (IEF) to lichens was calculated to evaluate the spatial recovery of impaired values of TBARS, chlorophyll a, chlorophyll b, xanthophylls + carotenoids and phaeophytization quotient. The results indicate that there is no clear spatial trend in mycobiont impairment even though the IEF values suggest a higher number of sites with low levels of membrane lipid peroxidation in the 2--3-km distance band from the cement plant (the outermost) than in the two other distance bands (0-1 and 1-2 km). The photobiont seems to be damaged mainly in the inner distance band of the study area as suggested by the gradual but significant recovery trend of pigment levels and phaeophytization quotient from the inner distance band to the outer one (as shown by the IEF values). Conversion of chlorophyll to phaeophytin probably is not the only process affecting pigment levels.

Estimating atmospheric mercury concentrations with lichens.

The uptake kinetics of elemental gaseous Hg (Hg(0)) in three species of epiphytic lichens (Pseudevernia furfuracea, Evernia prunastri, and Xanthoria parietina) were investigated under four different Hg concentrations (10, 15, 30, and 45 µg/m(3)) and three different temperatures (10, 20, and 30 °C) with the aim of evaluating the lichen efficiency for Hg(0) accumulation and their potential use in the estimate of atmospheric concentrations of this metal in the field. The results showed that under our experimental conditions the lichens accumulated Hg according to exposure time and that the metal is not released back to the atmosphere after Hg(0) was removed from the air (clearance). Pseudevernia furfuracea showed the highest Hg accumulation capacity and Evernia prunastri showed the lowest, but in these species the metal uptake kinetics was affected by temperature. Xanthoria parietina showed an intermediate metal accumulation capacity and a Hg accumulation rate independent of temperature (in the range 10-30 °C). The use of first-order kinetics equations for Hg uptake in X. parietina and available field data on Hg bioaccumulation in this species allowed reliable estimates of atmospheric Hg concentrations in the environment.

Influence of sample cleaning prior to the analysis on the elemental content of the lichen Xanthoria parietina (L.) Th.Fr.

In this paper we investigated whether a different accuracy in sample cleaning prior to the analysis may affect the elemental composition of unwashed samples of the lichen Xanthoria parietina. To this purpose, samples of this lichen were collected from a remote area of Sardinia (Italy) and randomly divided into two aliquots, one cleaned very carefully (ca. 3-4 h to obtain ca. 200 mg of material for analysis) and the other cleaned with a lower efficiency (ca. 1-2 h). Samples were analysed by ICP-MS for the content of Al, As, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Mg, Mn, Ni, Pb, V and Zn. The results indicated that an accurate cleaning is effective for reducing the content of Al and Fe, i.e. soil-related elements, but that it has no effect on the concentrations of other elements less represented in the soil.

Wood Distillate Promotes the Tolerance of Lettuce in Extreme Salt Stress Conditions.

Soil salinization is an adverse phenomenon in agriculture that severely affects crop growth and yield. The use of natural products, such as wood distillate (WD, derived from the pyrolysis of woody biomass), could be a sustainable approach to enhance the tolerance of plants cultivated in the saline soils. Hence, this study aimed to evaluate the potential of WD, a foliar sprayed at 0.2% (v/v), in lettuce plants subjected to grow under both moderate and high soil sodium chloride (NaCl) concentrations (ranging from 0 to 300 mM). The changes in the physiological and biochemical responses of these plants to the varying salt stress conditions allowed the identification of a maximum tolerance threshold (100 mM NaCl), specific to lettuce. Beyond this threshold, levels related to plant defense antioxidant power (antiradical activity) were lowered, while those indicative of oxidative stress (malondialdehyde content and electrolyte leakage) were raised, causing significant losses in leaf fresh biomass. On the other hand, WD significantly improved plant growth, enabling plants to survive high salt conditions >200 mM NaCl. Collectively, these observations highlight that treatments with WD could be of paramount importance in coping with current environmental challenges to have better yields under soil conditions of high salt concentrations.

Suitability of XRF for Routine Analysis of Multi-Elemental Composition: A Multi-Standard Verification.

This study investigated the suitability of X-ray fluorescence (XRF) analysis for routine multi-elemental composition analysis, checking its analytical capabilities by measuring a wide array of certified reference materials of soil and plant origin. A portable XRF analyzer was used to evaluate 32 soil and 12 plant standard materials, using both the Soil and Geochem mode, with sequential beams, allowing the detection of a wide range of elements. Recovery rates were calculated by comparing XRF measurements with certified values, and their correlations were verified through the Spearman coefficient. The results demonstrated the reliability of XRF measurements for soil samples, with a large number of elements showing a good or very good recovery and strong correlations with certified values. For plant samples, XRF largely overestimated the certified values, but the strong statistically significant correlations for almost all tested elements allowed us to correct this systematic bias, using the reported median value for dividing the value obtained via XRF. The Geochem mode emerged as more reliable for a larger number of elements. It was concluded that XRF may be a suitable alternative to ICP-MS in routine multi-elemental composition analysis.

Hormetic Effect of Wood Distillate on Hydroponically Grown Lettuce.

The addition of biostimulants to nutrient solutions of hydroponically grown crops to speed up plant growth and improve plant yield and quality has been attracting more and more attention. This study investigated the effects of wood distillate (WD) addition to hydroponically grown lettuce (Lactuca sativa L.) plants. Two concentrations of WD, 0.2% and 0.5%, were added to the nutrient solution, and biometric (i.e., leaf fresh weight, root fresh weight, root length and root surface area), photosynthetic (i.e., chlorophyll a, chlorophyll b, and carotenoid content) and biochemical (i.e., electrolyte leakage, total polyphenols, total flavonoids, and total antioxidant power content) parameters were evaluated. The effects of WD were hormetic, as the 0.2% concentration stimulated biometric and biochemical parameters, while the 0.5% concentration inhibited plant growth. Based on these results, it can be suggested that the addition of 0.2% WD to the nutrient solution has a stimulating effect on the growth of lettuce plants, and could be a successful strategy to boost the yield of crops grown hydroponically.

Integration of ecological indicators to assess a multitemporal impact of cement industries.

The present study evaluated an integrated biomonitoring approach based on three different bioindicators: tree rings, lichens, and beetles in a complex environment (urban-industrial-forest). In Central Italy, four sampling sites were selected to assess the anthropogenic impact of cement plants taking into account (1) long-term exposure (1988-2020) through the analysis of trace elements in tree rings of Quercus pubescens; (2) medium-term exposure (2020-2021) through the analysis of trace elements in thalli (outermost portions) of the lichen Xanthoria parietina; (3) short-term exposure in spring 2021 through the bioaccumulation and evaluation of sample vitality in transplants of the lichen Evernia prunastri and a periodic survey of entomological biodiversity carried out during spring and summer 2021. Trace elements of industrial origin were found in tree rings, with different levels of accumulation between 1988 and 2020 and a maximum in 2012. Native thalli of the lichen X. parietina showed an overall low bioaccumulation of trace elements except for Cr, probably reflecting the influence of national lockdown measures. The transplants of E. prunastri showed a weak stress response in the industrial and urban sites, but not in the forest, and identified Tl and V as the main elements contributing to atmospheric contamination, with peaks at the industrial sites. Concerning the beetles, a significantly lower number of species was found at the Semonte industrial site.