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Few studies are focused on correlations between the concentrations of PAHs in mosses and other bioindicator plant species. This study was carried out to investigate the potential of the joint use of devitalized H. cupressiforme transplants and R. pseudoacacia leaves as cost effective biomonitors for the assessment of PAHs in the air. The test was performed in a land historically devoted to agriculture, where recurrent waste burnings randomly occur, especially in the season we chose for the investigation. The presence of 20 PAHs was assessed following EPA 3550 C 2007 and EPA 8270 D 2014 protocols. R. pseudoacacia was able to accumulate both LMW and HMW PAHs, while moss prevalently collected the latter. It is suggested that R. pseudoacacia combined chronic pyrogenic and petrogenic PAH inputs, while moss transplants reflected PAH depositions from recent pyrogenic events. Our approach revealed long and short-term pollution footprints, with R. pseudoacacia recording the chronic input of PAH compounds loaded along its vegetative growth, and moss bags reflecting acute pollution inputs occurred during the exposure duration.

In bioaccumulation studies, the interpretation of pollutant contents in the target biomonitor has to be performed by assessing a deviation from an unaltered reference condition. A common strategy consists in the comparison with background element content (BEC) values, often built up by uncritically merging methodologically heterogeneous data. In this respect, the acid digestion of samples was identified as a major step affecting BEC data. Here, the analytical outcomes of two acid mixtures were compared on a set of matched paired samples of the lichen Pseudevernia furfuracea, a widely used biomonitor for which BEC values based on partial digestion were previously provided. The standard reference material BCR 482 (P. furfuracea) was used to validate analytical procedures consisting of either a HF total mineralization or an aqua regia partial one, both associated to ICP-MS multi-element analysis. In particular, the performance of the procedures was evaluated by comparing analytical results of field samples with the accuracy obtained on BCR aliquots (measured-to-expected percentage ratio). The total digestion showed a better performance for Al, As, Ba, Ca, Cd, Cu, Fe, Mn, Ni, Se, Sn, and Zn, whereas the opposite was found for Cr, Co, P, and S. Moreover, new BEC values were provided for P. furfuracea using a consolidated statistical approach, after a total sample digestion with hydrofluoric acid. The multivariate investigation of the background variability of 43 elements in 57 remote Italian sites led to the identification of geographically homogeneous areas for which BEC values are provided for use as reference in biomonitoring applications.

In this study, we used amplified fragment length polymorphism analysis to investigate the genetic structure of the terrestrial moss Pseudoscleropodium purum (Hedw.) M. Fleish. naturally exposed to different levels of atmospheric deposition of heavy metals. We also determined the heavy metal concentrations in samples of this moss to evaluate whether there was a relationship between atmospheric pollution and population genetic diversity. A low level of genetic diversity and a limited gene flow among populations were observed which is in accordance to the prevalence of asexual reproduction in this species. In addition, no significant correlation was found between metal content and gene diversity in P. purum, probably because of the common history of the sampled populations and/or to the lack of a drastic reduction of the size of the population; nonetheless, a clear genetic structure was evident in relation to the existing pollution gradient. Thus, based on the results of the principal coordinate analysis and Bayesian analysis of the genotypes, the mixed structure of the second most polluted population would suggest an ongoing differentiation of metal-tolerant genotypes in the most polluted sites of the sampling area.

In this paper we investigated the possibility to use moss bags to detect pollution inputs - metals, metalloids and polycyclic aromatic hydrocarbons (PAHs) - in sites chosen for their different land use (agricultural, urban/residential scenarios) and proximity to roads (sub-scenarios), in a fragmented conurbation of Campania (southern Italy). We focused on thirty-nine elements including rare earths. For most of them, moss uptake was higher in agricultural than in urban scenarios and in front road sites. Twenty PAHs were analyzed in a subset of agricultural sites; 4- and 5-ringed PAHs were the most abundant, particularly chrysene, fluoranthene and pyrene. Overall results indicated that investigated pollutants have a similar spatial distribution pattern over the entire study area, with road traffic and agricultural practices as the major diffuse pollution sources. Moss bags proved a very sensitive tool, able to discriminate between different land use scenarios and proximity to roads in a mixed rural-urban landscape.

Inventory of emission sources and biomonitoring with moss transplants are two different methods to evaluate air pollution. In this study, for the first time, both these approaches were simultaneously applied in five municipalities in Campania (southern Italy), deserving attention for health-oriented interventions as part of a National Interest Priority Site. The pollutants covered by the inventory were CO, NOx, particulate matter (PM10), volatile organic compounds (VOCs), and some heavy metals (As, Cd, Cr, Cu, Hg, Ni, Pb, Se, and Zn). The biomonitoring survey was based on the use of the devitalized moss Hypnum cupressiforme transplanted into bags, following a harmonized protocol. The exposure covered 40 agricultural and urban/residential sites, with half of them located in proximity to roads. The pollutants monitored were Al, As, Cd, Cr, Cu, Fe, Hg, Ni, Pb, Se, and Zn, as well as total polycyclic aromatic hydrocarbons (PAHs) only in five sites. Using the emission inventory approach, high emission loads were detected for all the major air pollutants and the following heavy metals: Cr, Cu, Ni, Pb and Zn, over the entire study area. Arsenic, Pb, and Zn were the elements most accumulated by moss. Total PAH postexposure contents were higher than the preexposure values (~20–50% of initial value). Moss uptakes did not differ substantially among municipalities or within exposure sites. In the five municipalities, a similar spatial pattern was evidenced for Pb by emission inventory and moss accumulation. Both approaches indicated the same most polluted municipality, suggesting their combined use as a valuable resource to reveal contaminants that are not routinely monitored.

Investigating the nature of PM10 is crucial to differentiate sources and their relative contributions. In this study we compared the levels, and the chemical and mineralogical properties of PM10 particles sampled in different seasons at monitoring stations representative of urban background, urban traffic and suburban traffic areas of Naples city. The aims were to relate the PM10 load and characteristics to the location of the monitoring stations, to investigate the different sources contributing to PM10 and to highlight PM10 seasonal variability. Bulk analyses of chemical species in the PM10 fraction included total carbon and nitrogen, δ13C and other 20 elements. Both natural and anthropogenic sources were found to contribute to the exceedances of the EU PM10 limit values. The natural contribution was mainly related to marine aerosols and soil dust, as highlighted by X-ray diffractometry and SEM-EDS microscopy. The percentage of total carbon suggested a higher contribution of biogenic components to PM10 in spring. However, this result was not supported by the δ13C values which were seasonally homogeneous and not sufficient to extract single emission sources. No significant differences, in terms of PM10 load and chemistry, were observed between monitoring stations with different locations, suggesting a homogeneous distribution of PM10 on the studied area in all seasons. The anthropogenic contribution to PM10 seemed to dominate in all sites and seasons with vehicular traffic acting as a main source mostly by generation of non-exhaust emissions Our findings reinforce the need to focus more on the analysis of PM10 in terms of quality than of load, to reconsider the criteria for the classification and the spatial distribution of the monitoring stations within urban and suburban areas, with a special attention to the background location, and to emphasize all the policies promoting sustainable mobility and reduction of both exhaust and not-exhaust traffic-related emissions.

In biomonitoring, the knowledge of background element content (BEC) values is an essential pre-requisite for the correct assessment of pollution levels. Here, we estimated the BEC values of a highly performing biomonitor, the epiphytic lichen Pseudevernia furfuracea, by means of a careful review of literature data, integrated by an extensive field survey. Methodologically homogeneous element content datasets, reflecting different exposure conditions across European and extra-European countries, were compiled and comparatively analysed. Element content in samples collected in remote areas was compared to that of potentially enriched samples, testing differences between medians for 25 elements. This analysis confirmed that the former samples were substantially unaffected by anthropogenic contributions, and their metrics were therefore proposed as a first overview at supra-national background level. We also showed that bioaccumulation studies suffer a huge methodological variability. Limited to original field data, we investigated the background variability of 43 elements in 62 remote Italian sites, characterized in GIS environment for anthropization, land use, climate and lithology at different scale resolution. The relationships between selected environmental descriptors and BEC were tested using Principal Component Regression (PCR) modelling. Elemental composition resulted significantly dependent on land use, climate and lithology. In the case of lithogenic elements, regression models correctly reproduced the lichen content throughout the country at randomly selected sites. Further descriptors should be identified only for As, Co, and V. Through a multivariate approach we also identified three geographically homogeneous macro-regions for which specific BECs were provided for use as reference in biomonitoring applications.

The effects of cadmium and lead were investigated in Cynara cardunculus L. Plant uptake by root and shoot, changes in cell ultrastructure and photosynthetic efficiency, photosynthetic key protein levels, as well as regulation of stress-induced Hsp70 were examined. Cynara cardunculus accumulated Cd and Pb in their tissue, with a different trend for the two metals. The prompt translocation of Cd to the shoot may justify the ultrastructural injuries, especially observed in chloroplasts. However, Cd- treated plants did not show any decline in photochemistry; it is likely that Cd in shoot tissue triggers defense mechanisms, increasing the level of proteins involved in photosynthesis (i.e., Rubisco and D1 increased 7 and 4.5 fold respectively) as a compensatory response to neutralize chloroplast damage. The accumulation of Pb mainly in root, can explain the increase in Hsp70 level (23 folds) in this tissue. Pb reached the shoots, even at low amounts, causing an overall significant change in some photochemical parameters (QY and NPQ decreases and increases of 25%, respectively). The results suggest a higher sensitivity of C. cardunculus to Pb than Cd, although maximal photochemical efficiency suggests that this species seems to tolerate Pb and Cd and hence, it is a suitable candidate for phytoremediation.

Although a large body of literature exists on the use of transplanted mosses for biomonitoring of air pollution, no article has addressed so far the use and the accumulation performance of a cloned moss for this purpose. In this work, a direct comparison of metal accumulation between bags filled with a Sphagnum palustre L. clone or with native Pseudoscleropodium purum Hedw., one of the most used moss species in biomonitoring surveys, was investigated. The test was performed in sites with different atmospheric contamination levels selected in urban, industrial, agricultural and background areas of Italy and Spain. Among the eighteen elements investigated, S. palustre was significantly enriched in 10 elements (Al, Ba, Cr, Cu, Fe, Hg, Pb, Sr, V and Zn), while P. purum was enriched only in 6 elements (Al, Ba, Cu, Hg, Pb and Sr), and had a consistently lower uptake capacity than S. palustre. The clone proved to be more sensitive in terms of metal uptake and showed a better performance as a bioaccumulator, providing a higher accumulation signal and allowing a finer distinction among the different land uses and levels of pollution. The excellent uptake performance of the S. palustre clone compared to the native P. purum and its low and stable baseline elemental content, evidenced in this work, are key features for the improvement of the moss bag approach and its large scale application.

The epiphytic lichen Pseudevernia furfuracea is widely used as biomonitor of airborne trace elements and other contaminants and consists of two taxonomic varieties (var. furfuracea and var. ceratea). Here, we assessed the occurrence of inter-varietal differences in the elemental composition of paired samples of var. furfuracea and var. ceratea collected in 20 remote sites of Italian mountains. The concentration of 40 elements was measured by inductively coupled plasma mass spectroscopy, after digestion with HNO3 and aqua regia. The magnitude of inter-varietal differences compared to the effect of large-scale site-dependent environmental factors (i.e., lithological substrate, host tree species, and altitude) on overall element content was explored by multivariate analysis techniques and tested by generalized linear mixed modeling (GLMM). Further GLMMs were separately fitted for each element testing taxonomic-related variability against uncertainty associated to the analytical procedure. Inter-varietal differences were statistically significant only for Hg and P, with higher content in var. ceratea at most sites, and for Mg and Zn, showing the opposite pattern. Since the elemental composition of P. furfuracea in remote sites was mostly affected by local lithology and climatic conditions, our results confirm that lichen material for active biomonitoring should be collected in a single ecologically homogeneous remote area. We also indicate sites in the Eastern Alps where P. furfuracea showed the minimum content of most elements, which are suggested as locations to collect lichen material for transplants. Besides the context-dependency at large spatial scale, variations of elemental composition apparently related to taxonomy, could possibly be due to unequal incidence of morphological traits of the collected material. Further research is needed to clarify this issue, and how it affects bioaccumulation phenomena.

In the present work, the genotoxic effect of cadmium and lead supplied in a laboratory trial, was investigated for the first time in the moss Sphagnum palustre, by ISSR molecular markers. A total of 169 reproducible bands were obtained with 12 primers, ten of which gave polymorphisms (i.e., appearance/disappearance of bands), indicating a clear genotoxic effect induced by the metals. Both metals induced a decrease of the genome template stability in a dose dependent manner. At concentration >10−5 Cd also induced a general toxic effect in S. palustre, leading to chlorophyll degradation and moss death. Moreover, we followed the fate of supplied heavy metals into the moss tissue by SEM-EDX to see if they entered the cells. SEM-EDX observations on moss cultures treated with equimolar concentrations of the two metals showed that most Pb precipitated in form of particles on moss surface, while Cd did not aggregate in particles and was not found on moss surface. In light of these findings, we concluded that probably Pb induced a genotoxic effect at lower intracellular concentrations than Cd.

Source at https://doi.org/10.1002/ece3.2579. The PREDICTS project—Projecting Responses of Ecological Diversity In Changing Terrestrial Systems (www.predicts.org.uk)—has collated from published studies a large, reasonably representative database of comparable samples of biodiversity from multiple sites that differ in the nature or intensity of human impacts relating to land use. We have used this evidence base to develop global and regional statistical models of how local biodiversity responds to these measures. We describe and make freely available this 2016 release of the database, containing more than 3.2 million records sampled at over 26,000 locations and representing over 47,000 species. We outline how the database can help in answering a range of questions in ecology and conservation biology. To our knowledge, this is the largest and most geographically and taxonomically representative database of spatial comparisons of biodiversity that has been collated to date; it will be useful to researchers and international efforts wishing to model and understand the global status of biodiversity.

Gli studi di biomonitoraggio attivo e passivo di elementi in traccia basati sul lichene epifita Pseudevernia furfuracea sono molto numerosi ma caratterizzati da una importante eterogeneità metodologica: si è ritenuto perciò opportuno procedere alla sua caratterizzazione mediante la revisione critica di 62 lavori di rilevanza nazionale e internazionale. I dati di contenuto elementare derivanti dal processo di revisione, integrati da quelli ottenuti dalla campagna di campionamento condotta in Italia nel 2014/15 dagli autori, sono stati raccolti in 3 dataset distinti, rispettivamente per (i) talli nativi raccolti in aree di background (BG), (ii) talli nativi utilizzati in studi di biomonitoraggio passivo (P), e (iii) talli trapiantati (T). Il confronto inter-dataset effettuato su 25 elementi tramite test non parametrici ha evidenziato valori significativamente inferiori per il dataset BG, proposto come riferimento preliminare per studi di biomonitoraggio a livello europeo. Al fine di fornire dei valori di riferimento nazionali relativi a macroaree omogenee e per analizzare le relazioni tra variabili ambientali e contenuto elementare baseline, i siti di raccolta sono stati caratterizzati in ambiente GIS in termini di antropizzazione, uso del suolo, clima e litologia. Una cluster analysis della matrice “siti di raccolta  elementi” (62  43) ha consentito l’identificazione di 3 gruppi di siti omogenei per i quali sono stati forniti i valori di background nazionale. La predittività delle variabili ambientali sul contenuto elementare baseline è stata testata tramite Principal Component Regression: una PCA della matrice “siti di raccolta  variabili ambientali” (62  13) ha permesso di utilizzare combinazioni lineari delle variabili di partenza in modelli di regressione multipla applicati al contenuto dei 43 elementi analizzati. I modelli hanno prodotto risultati statisticamente significativi per molti elementi litogenici ma non per elementi di origine antropica.

Particulate matter has to be constantly monitored because it is an important atmospheric transport form of potentially harmful contaminants. The cost-effective method of the moss-bags can be employed to evaluate both loads and chemical composition of PM. PM entrapped by the moss Pseudoscleropodium purum exposed in bags in 9 European sites was characterized for number, size and chemical composition by SEM/EDX. Moreover, moss elemental uptake of 53 elements including rare earth elements was estimated by ICP-MS analysis. All above was aimed to find possible relations between PM profile and moss uptake and to find out eventual element markers of the different land use (i.e. agricultural, urban, industrial) of the selected sites. After exposure, about 12,000 particles, mostly within the inhalable fraction, were counted on P. purum leaves; their number generally increased from the agricultural sites to the urban and industrial ones. ICP analysis indicated that twenty-three elements were significantly accumulated by mosses with different element profile according to the various land uses. The PM from agricultural sites were mainly made of natural/crustal elements or derived from rural activities. Industrial-related PM covered a wider range of sources, from those linked to specific industrial activities, to those related to manufacturing processes or use of heavy-duty vehicles. This study indicates a close association between PM amount and moss element-uptake, which increases in parallel with PM amount. Precious metals and REEs may constitute novel markers of air pollution in urban and agricultural sites, respectively.

00002 ăWOS:000388157900039; International audience; This work aimed to define the molecular and chemical signature of a S. palustre clone developed in the framework of the EU-FP7 Mossclone project to improve the standardization and reliability of the moss bag technique. The molecular characterization was performed by a set of DNA molecular markers (RAPD, ISJ, PCR-RFLP, sequencing and microsatellites) to tag the clone produced within the project. Molecular characterization also provided new DNA markers that can be applied in systematic analyses of Sphagnum, and gave new insights to implement well established techniques. The elemental composition of the clone was measured by ICP-MS analysis of 54 major and trace elements, with and without commonly applied pre-exposure treatments (oven devitalization and EDTA washing). Concentrations of almost all analyzed elements were significantly lower (from 10 to 100 times) in the clone than in conspecific field moss, apart from some elements (K, Mo, P and Na) deriving from the culture medium or EDTA treatment. Oven devitalization and EDTA washing did not significantly affect the clone composition. A comparison between the elemental composition of the clone with that of naturally growing Sphagnum species proved the particularly low elemental content of the clone. Therefore, in view of a rigorously standardized moss-bag protocol for the monitoring of persistent atmospheric pollutants, the use of the S. palustre clone, a biomaterial with very low and constant element composition, and homogenous morphological characteristics is strongly recommended. (C) 2016 Elsevier Ltd. All rights reserved.

To develop an internationally standardized protocol for the moss bag technique application, the research team participating in the FP7 European project “MOSSclone” focused on the optimization of the moss bags exposure in terms of bag characteristics (shape of the bags, mesh size, weight/surface ratio), duration and height of exposure by comparing traditional moss bags to a new concept bag, “Mossphere”. In particular, the effects of each variable on the metal uptake from the air were evaluated by a systematic experimental design carried out in urban, industrial, agricultural and background areas of three European countries with oceanic, Mediterranean and continental climate. The results evidenced that the shape, the mesh size of the bags and the exposure height (in the tested ranges), did not significantly influence the uptake capacity of the transplanted moss. The aspects more affecting the element uptake were represented by the density of the moss inside the bags and the relative ratio between its weight and the surface area of the bag. We found that, the lower the density, the higher the uptake recorded. Moreover, three weeks of exposure were not enough to have a consistent uptake signal in all the environments tested, thus we suggest an exposure period not shorter than 6 weeks, which is appropriate in most situations. The above results were confirmed in all the countries and scenarios tested. The adoption of a shared exposure protocol by the research community is strongly recommended since it is a key aspect to make biomonitoring surveys directly comparable, also in view of its recognition as a monitoring method by the EU legislation.

Twenty nine South European specimens of Sphagnum centrale, S. palustre, S. papillosum and S. magellanicum were studied with 15 microsatellite markers. In contrast with eastern North American populations, our analysis showed a genetic overlapping between S. centrale and S. palustre in mixed populations. Moreover, Mediterranean species showed a genetic richness (total number of alleles) higher than that calculated in conspecific American samples. As Mediterranean Sphagnum bogs are remnant populations, microsatellites could well work for selecting source populations in order to recover Mediterranean peatlands.

When terrestrial mosses are used to monitor atmospheric contamination, the samples are usually analysed without first being washed, so that the contaminants deposited on the moss can be quantified. However, when the aim is to evaluate phyto-toxic effects and the risks associated with increased concentrations of contaminants, it is the bioconcentrated fraction that is of interest. In such cases, the samples are washed to remove deposited material adhering to the surface of the moss. In the present study, the efficacy of the washing step was evaluated, by electron microscopy and microanalysis, in samples exposed to different levels of contamination. The results obtained show that washing is not effective at removing particles deposited on the moss surface, so that an alternative cleaning method is required. Until such a method becomes available, use of the Sequential Elution Technique is recommended for precise determination of the bioconcentrated contaminants.

In the present study, scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM–EDS) was used in conjunction with the sequential elution technique (SET) to evaluate the efficacy of extraction of Zn from the moss Pseudoscleropodium purum (Hedw.) M. Fleisch. Moss samples were collected from two areas, a contaminated area (with high concentrations of Zn) in the surroundings of a steel works, and an uncontaminated area. Some samples from the uncontaminated area were used as controls, and others were incubated in a solution of ZnCl2 for subsequent analysis of uptake and location of Zn. The concentrations of Zn in the different cell compartments of the moss were determined by use of the SET, and different points on the surface of the moss and in deposited particles were analysed by SEM–EDS. The results showed that the extracellular fraction obtained by SET includes the Zn bound to cation exchange sites in the moss and Zn bound to particles on the moss surface, which leads to overestimation of extracellular Zn. After extracellular extraction, Zn was not detected on the surface of the moss or in the particles. To avoid problems associated with the presence of particles on the moss surface, a washing procedure should be used to remove such particles before application of the SET. Another possibility is to use the intracellular concentration, as this is not affected by the metal load in the particles and better reflects those contaminants that affect the moss metabolism.

In the present article we characterized the emissions at the exhaust of a Common Rail (CR) diesel engine, representative of light-duty class, equipped with a catalyzed diesel particulate filter (CDPF) in controlled environment. The downstream exhausts were directly analyzed (for PM, CO, CO2, O2, HCs, NOx) by infrared and electrochemical sensors, and SEM-EDS microscope; heavy metals were chemically analyzed using mosses and lichens in bags, and glass-fibre filters all exposed at the engine exhausts. The highest particle emission value was in the 7-54 nm size range; the peak concentration rose until one order of magnitude for the highest load and speed. Particle composition was mainly carbonaceous, associated to noticeable amounts of Fe and silica fibres. Moreover, the content of Cu, Fe, Na, Ni and Zn in both moss and lichen, and of Al and Cr in moss, was significantly increased. Glass-fibre filters were significantly enriched in Al, B, Ba, Cu, Fe, Na, and Zn. The role of diesel engines as source of carbonaceous nanoparticles has been confirmed, while further investigations in controlled environment are needed to test the catalytic muffler as a possible source of silica fibres considered very hazardous for human health. © 2010 The Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences.

This paper focuses on chemical composition of the epiphytic moss Leptodon smithii, gathered on Quercus ilex bark, assessed in seven sites located in urban and extra-urban/remote areas of southern Italy, a poorly surveyed geographic area. The concentrations of Cr, Cu, Pb and Zn in moss tissue are generally more abundant in moss gathered in the urban sites; among extra-urban/remote sites Valle delle Ferriere showed the highest metal concentrations, mostly related to an industrial activity occurred in the past. L. smithii chemistry seems influenced by airborne dust locally enhanced by erosion phenomena, longrange transport of pollutants and marine aerosols. Element content in moss is compared with genetic variability of L. smithii estimated in the same sites. Pearson’s correlation coefficient between gene diversity and total element load (r = 0.851; p = 0.03) suggests that anthropogenic pressure, determining habitat disturbance and fragmentation, leads both to genetic impoverishment consequent to population shrink, and to a higher accumulation in moss tissues, as a consequence of increased airborne major/trace elements. Thus, the coupled evaluation of chemical composition in mosses and gene diversity may prove a useful tool to highlight environmental disturbance in a gradient of land use.

Mosses and lichens are widely used to detect changes in the environmental concentrations of potential contaminants. Several studies have shown the usefulness of transplanted mosses and/or lichens to monitor air depositions of inorganic and organic pollutants at different scales. Here, we present the results of a biomonitoring study carried out in two cities, London (United Kingdom) and Naples (Italy), using four biomonitors (two lichens and two mosses). The lichens, Pseudevernia furfuracea (L.) Zopf var. furfuracea and Parmelia sulcata Taylor, and mosses, Sphagnum capillifolium (Ehrh.) Hedw. and Hypnum cupressiforme Hedw., were exposed in bags in urban streets of Naples and London and in semi-rural and rural areas. Samples were analysed for Cd, Cr, Cu, Fe, Ni, Pb, V and Zn by atomic absorption spectrometry after acid digestion and for 16 EPA polycyclic aromatic hydrocarbons (PAHs) by gas chromatography coupled to mass spectrometry after matrix solid-phase dispersion. For heavy metals, the comparison between the selected mosses indicated that, in all exposure sites, S. capillifolium had a better accumulation performance than H. cupressiforme, whereas for the lichens, it was P. furfuracea which accumulated higher concentrations of metals. Also for total PAHs, S. capillifolium showed a good accumulation capability compared to the other biomonitors investigated, especially compared to H. cupressiforme. It was observed an increasing heavy metal and PAH uptake by biomonitors from rural to urban sites, in both cities.

As builders and major components of peatlands, Sphagnopsida (peat mosses) are very important organisms for ecosystems and world’s climate. Nowadays many Sphagnum species as well as their habitats are largely protected, while their scientific and economic relevance remains considerable. Advanced methods of in vitro cultivation provide the potential to work in a sustainable way with peat mosses and address aspects of basic research as well as biotechnological and economical topics like biomonitoring or the production of renewable substrates for horticulture (Sphagnum farming). Here, we describe the establishment of axenic in vitro cultures of the five peat moss species Sphagnum fimbriatum Wils. and Hook., Sphagnum magellanicum Brid., Sphagnum palustre L., Sphagnum rubellum Wils. and Sphagnum subnitens Russ. and Warnst. with specific focus on large-scale cultivation of S. palustre in bioreactors. Axenic, clonal cultures were established to produce high quantities of biomass under standardized laboratory conditions. For advanced production of S.palustre we tested different cultivation techniques, growth media and inocula, and analyzed the effects of tissue disruption. While cultivation on solid medium is suitable for long term storage, submerse cultivation in liquid medium yielded highest amounts of biomass. By addition of sucrose and ammonium nitrate we were able to increase the biomass by around 10- to 30-fold within 4 weeks. The morphology of in vitro-cultivated gametophores showed similar phenotypic characteristics compared to material from the field. Thus the tested culture techniques are suitable to produce S. palustre material for basic and applied research. Electronic supplementary material The online version of this article (doi:10.1007/s11240-014-0658-2) contains supplementary material, which is available to authorized users.

This paper provides an analysis of genetic variability in local populations of the clonal moss Pleurochaete squarrosa, a Mediterranean moss typical of post-fire recovery, and characterised by asexual reproduction. ISSR (Internal Simple Sequence Repeats) primers and trnLUAA (intron of plastid gene for Leu tRNA) length polymorphism were employed to evaluate genetic structure in five southern Italy populations of this moss. Both molecular tools highlight high values of genetic diversity with geographic structure of sampled populations and a low gene flow among the investigated sites. Gene diversity is significant at every hierarchical level of sampling, and generally increases from lower to higher hierarchical levels. Among other factors, the ubiquitous genetic variability detected in P. squarrosa can be related to continuous, occasionally massive, shortrange recruitment of propagules, and to the high degree of intermingling, both favoured by the modality of vegetative reproduction and growth occurring in the species.

The results of trace element content analysed in Sphagnum capillifolium and Pseudevernia furfuracea exposed in bags in 1999 are reconsidered to evaluate the reliability of moss and lichen transplants to detect urban trace element atmospheric pollution, using Naples as a case example. After 4 months' exposure, trace element concentrations were at least twice as high as the pre-exposure values and in general higher in Sphagnum than in Pseudevernia. Moss samples were enriched in the following order: As = Cu > Mo > Pb > V > Co > Cr > Zn; lichen samples in the order: Mo > Cu > As = Co = Ni > V > Pb. Based on the calculation of a cumulative load factor, all sites located along the coast had higher trace element loads compared to sites in the hilly inland area. Complementary SEM, TEM and EDS observations showed, despite significant damage to tissue and cell integrity, the recurrent presence of particulate matter in moss and lichen, indicating the considerable presence of dust in the urban atmosphere which, according to chemical composition, may be due both to anthropogenic and natural sources such as volcanic rock and soil and sea salts.

This paper presents the results of a bioaccumulation study of trace elements in the Naples urban area based on the use of the moss Sphagnum capillifolium (Ehrh.) Hedw. and the lichen Pseudevernia furfuracea (L.) Zopf exposed in bags in 23 sites. Moss and lichen bags were exposed for 4 months starting from the beginning of July 1999. Bags gathering was carried out after 10 weeks of exposure, at the end of the dry season, and after 17 weeks, during the wet season. The elements Al, As, Ca, Cd, Cr, Co, Cu, Fe, K, Mg, Mn, Mo, Ni, Pb, Ti, V and Zn were analysed by inductively coupled plasma-mass spectrometry ICP-MS in both biomonitors. For the majority of the elements the total amounts found in S. capillifolium were higher than in P. furfuracea whether considering the whole period of exposure or the weekly uptake. It was observed that there was a much greater difference in metal accumulation by P. furfuracea between the dry and wet seasons compared with S. capillifolium. In the wet period, the lichen seems to accumulate a larger quantity of metals. With the exception of Mn, trace element concentrations did not appear to be significantly affected by the washing away of rainfall. K loss during exposure suggested cell membrane damage in both organisms. For P. furfuracea the K leakage was limited to the dry period of exposure. A clear distinction between "lithophilic" and "anthropogenic" elements was achieved by cluster analysis. Significant correlations were found among Fe-Cu-Cr-Ni, Pb-Cd-Co, V-Cr-Ni, Zn-Ni-Pb, suggesting a common source for each group of elements.

We determined genetic variation and metal and polycyclic aromatic hydrocarbon concentrations in Leptodon smithii moss collected in holm oak stands at cities, outskirts and remote areas of Campania and Tuscany (Italy) to investigate if anthropogenic pressure (pollutant emissions and land use change) affects moss genetic richness. In both regions, metal and polycyclic aromatic hydrocarbon concentrations reflected the trend urban > outskirts > remote areas, excepting Tuscany remote site. In both regions, themoss gene diversity increased fromurban to remote areas. The findings suggest the extent and the fragmentation of urban green areas, as drivers of moss genetic richness.

In recent years, the number of studies concerning population genetics and phylogenetics in mosses using molecular markers has remarkably grown. This review summarizes the main features of the most used molecular techniques based on PCR and their application in bryology, with particular attention to mosses. This work also provides a bibliographic guide to 108 molecular studies focused on mosses to assist in choosing the most suitable markers according to the main aims of a specific research. Population ecology / phylogeny / phylogeography / molecular markers / taxonomy / mosses

Accumulation of lead and zinc was studied in the moss Funaria hygrometrica Hedw. collected from mine tailings. Heavy metal accumulation in gametophytes and sporophytes was quantified by graphite furnace atomic absorption spectrometry (GFAAS) and inductively coupled plasma-atomic emission spectrometry (ICP-AES). Pb and Zn accumulation in the placental zone was analysed by x-ray scanning electron microscopy (SEM) and transmission electron microscopy (TEM) microanalysis. Spectrometry showed that while moss gametophytes accumulated considerable concentrations of heavy metals, sporophytes accumulated only small concentrations of metals. X-ray SEM and TEM showed that the two metals accumulated in placental transfer cells on both the gametophytic and sporophytic sides. To investigate the uptake pattern for both metals under controlled conditions, F. hygrometrica plants collected from a non-polluted site were treated in the laboratory with separate solutions of Pb and Zn at two concentrations (10−2and 10−4 M ) for 24 or 168 h. Metal accumulation was analysed separately in gametophytes and sporophytes using GFAAS and ICP–AES. Each generation had a different accumulation quotient for both metals, and gametophytes accumulated significantly more metal than sporophytes. Concentrations of Zn in sporophytes were always higher than concentrations of Pb. The findings are discussed in relation to the role performed by the gametophyte and the placenta in the accumulation and sequestration of Pb and Zn.

This paper presents data on the induction by alpha-D-oligogalacturonides (OG) of antibiotic activity in vitro by the fern Nephrolepis sp. The extracts from the fern grown aseptically, partly in a medium containing a mixture of OG and partly in a medium lacking OG, as control, were tested against several bacterial strains. The results show that the OG mixture promotes the production of antibiotic compounds. Comparing the present results with those on the antimicrobial properties of the same fern grown in a greenhouse, we discuss the hypothesis that the production of antibiotic substances can be elicited by different factors, such as products of synthesis or degradation of the biotic component of the soil or by OG (in axenic culture) that can mimic the effect of natural elicitors. (C) 1997 Elsevier Science B.V.

Mosses, lichens and cellulose filters were exposed for 17 weeks at four urban monitoring stations in Naples (S Italy) to assess the accumulation of airborne Al, As, Ba, Ca, Cd, Co, Cr, Cu, Fe, Hg, K, Mg, Mn, Mo, Ni, Pb, Ti, V, and Zn. In each site, the element accumulation was significantly higher in the moss Hypnum cupressiforme than in the lichen Pseudevernia furfuracea. Acid washed mosses accumulated the highest amount of trace elements, but the differences in element concentrations among the moss samples exposed after water washing and different devitalisation treatments (acid washing, oven drying and water boiling) and between the lichen samples exposed with and without the nylon bag were not statistically significant. The cellulose filters showed the lowest accumulation capability. The reciprocal ordination of sites and exposed materials showed an increasing contamination gradient (especially for Pb, Cu and Zn) from the background site to the trafficked city streets; this pattern was undetectable from PM10 data recorded by the automatic monitoring devices operating in the four exposure sites. The element profile in exposed materials did not change substantially throughout the urban area and particles of polluted urban soils seem the main source of airborne metals in Naples. Through a comprehensive evaluation of the results from this and previous studies, a protocol is suggested for the moss-bag monitoring of trace element deposition in urban environments.

For the first time until now, the results from a prediction model (Atmospheric Dispersion Modelling System (ADMS)-Road) of pollutant dispersion in a street canyon were compared to the results obtained from biomonitors. In particular, the instrumental monitoring of particulate matter (PM10) and the biomonitoring of 14 polycyclic aromatic hydrocarbons (PAHs) and 11 metals by Quercus ilex leaves and Hypnum cupressiforme moss bags, acting as longand short-term accumulators, respectively, were carried out. For both PAHs and metals, similar bioaccumulation trends were observed, with higher concentrations in biomonitors exposed at the leeward canyon side, affected by primary air vortex. The major pollutant accumulation at the leeward side was also predicted by the ADMS-Road model, on the basis of the prevailing wind direction that determines different exposure of the street canyon sides to pollutants emitted by vehicular traffic. A clear vertical (3, 6 and 9 m) distribution gradient of pollutants was not observed, so that both the model and biomonitoring results suggested that local air turbulences in the street canyon could contribute to uniform pollutant distributionat different heights.

Although the moss bag technique has been used for active biomonitoring for the past 40 years, there is still no standardized protocol that enables application of the technique as a tool to monitor air quality. The aim of this review paper is to evaluate the degree of standardization of each of the variables that must be considered in applying the technique (i.e. the variables associated with preparation of the moss and moss bags, exposure of the bags, and post-exposure treatment). For this purpose, 112 scientific papers that report the methods used in applying the moss bag technique were consulted. Finally, on the basis of the conclusions reached, we propose a protocol that will enable each of these variables to be investigated separately, with the final aim of developing a standardized methodology

In urban environments, high levels of pollutants, as polycyclic aromatic hydrocarbons (PAHs) and trace elements, coming mainly from anthropogenic sources as vehicular exhausts, industrial emissions and waste incineration, impose serious environmental and biological problems. These pollutants cause concern, especially for their potential toxicity and the adverse health effects. In this work the results of a passive biomonitoring study, concerning the accumulation of trace elements and PAHs in Quercus ilex leaves and in the epiphytic moss Leptodon smithii, are presented. Both biomonitors were sampled in two Italian Regions (Campania and Toscana) along a gradient of environmental disturbance in i) urban, ii) periurban and iii) extraurban areas. Fresh leaves and mosses were extracted by sonication in a mix of methane dichloride and acetone (1:1=v:v) and analysed for 16 EPA PAHs by GC-MS. Oven-dried (75 °C) plant samples were mineralized with acids (HF:HNO3=1:2=v:v) and analysed for elements (Al, B, Ba, Cd, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, Pb, V and Zn) by AAS and ICP-OES. In both Regions, trace element and PAH concentrations were generally higher in leaves and mosses sampled in urban areas than in periurban and extraurban ones, confirming the suitability of the selected native species to monitor air pollutants in the Mediterranean areas. The high concentrations of some elements in leaves (Mn, Na and Zn) and in mosses (B, Ba, Cd, Mn, Ni and V) from the extraurban site of Toscana, likely due to the influence of marine aerosols and of an industrial settlement located about 40 Km from the sampled area, put in evidence the importance of selecting suitable “reference” sites in order to find out situations at risk and properly evaluate the degree of pollution in urban areas.

Peat-forming floating mires could provide an exceptional resource for palaeoenvironmental and environmental monitoring studies, as much of their own history, as well as the history of their surrounds, is recorded in their peat deposits. In his Naturalis historia (AD 77–79), Pliny the Elder described floating islands on Lake Vadimonis (now Posta Fibreno Lake, Italy). Actually, a small floating island (ca. 35 m of diameter and 3 m of submerged thickness) still occurs on this calcareous lake fed by karstic springs at the base of the Apennine Mountains. Here the southernmost Italian populations of Sphagnum palustre occur on the small surface of this floating mire known as “La Rota”, i.e., a cup-formed core of Sphagnum peat and rhizomes of Helophytes, erratically floating on the water-body of a submerged doline, annexed to the easternmost edge of the lake, characterised by the extension of a large reed bed. Geological evidence point out the existence in the area of a large lacustrine basin since Late Pleistocene. The progressive filling of the lake caused by changing in climatic conditions and neotectonic events, brought about the formation of peat deposits in the area, following different depositional cycles in a swampy environment. Then, a round-shaped portion of fen, originated around lake margins in waterlogged areas, was somehow isolated from the bank and started to float. Coupling data about concentrations and fluxes of several major and trace elements of different origin (i.e., dust particles, volcanic emissions, cosmogenic dusts and marine aerosols), with climate records (plant micro- and macrofossils, pollens, isotopic ratios), biomolecular records (e.g., lipids), detailed age-depth modelling (i.e., 210Pb, 137Cs, 14C), and humification indexes, the present work is hoped to identify and better understand the reliability of this particular “archive”, and thus possible relationships between biogeochemical processes occurring in this floating bog and environmental changes.

Biomonitoring of air pollutants by moss and lichen transplants has been widely used over the past years, especially to monitor trace elements, since it gives useful information on their deposition, particularly in those urban and industrial areas lacking of indigenous moss and lichen species. Moreover, this technique provides information on contaminants associated to airborne particles which in European urban areas are not routinely checked, hindering the identification of pollutant sources crucial to undertake effective policies to knock down air pollutants. Nevertheless, the current lack of standardization at international level, and the scarce knowledge on the factors influencing element uptake make this technique still open to criticism. In this experiment moss and lichen were exposed for 17 weeks at four urban air quality monitoring stations of Naples city. Trace element uptake was evaluated in moss material before and after different devitalisation treatments (acid washing, oven drying and the novel water boiling) and in lichen exposed with and without the nylon bag. A major uptake was overall recorded in moss materials. Uptake differences among moss materials were not significant (P = 0.378), as well as the difference between lichen exposed with and without nylon bag (P=0.532). The ordination of sites/materials in relation to element uptake indicates a contamination gradient of the investigated sites, but element profiles did not differentiate substantially in the different areas of the city suggesting that atmospheric deposition in Naples urban area is qualitatively homogeneous only differing quantitatively in diverse monitoring sites. The overall high and significant correlations observed among and between geogenic and antropogenic elements suggest that polluted urban soil could represent a massive source for most elements. The implementation of devitalized moss bag could represent a ready-to-use robust tool for routine monitoring of air pollutants.

Taxonomic arrangement of Hypnum species has been controversial since the beginning of the nineteenth century. The molecular pattern and phylogenetic relationships within the Hypnum cupressiforme complex, in relation to other pleurocarps species, were investigated by study of nuclear ribosomal ITS1/ITS2 and chloroplast trnLUAA intron sequences. Seven Hypnum species from different continents were sequenced and analysed, with a relatively large set of pleurocarpous mosses. Sequences indicate a low level of genetic differentiation within the complex. Both maximum parsimony and maximum likelihood analyses indicate that Hypnum s.l. is distributed in several clades of pleurocarps; in addition, H. cupressiforme sensu strictu, and the complex, are not monophyletic. Our results clearly circumscribe only two species, H. jutlandicum and H. imponens; phylogenetic analyses, in particular, highlight the isolation of the latter from the complex. Genetic lineages with a clear geographic structure were found in Eurasia; these probably originated during the last glaciations. The presence of H. cupressiforme in some related clades flanking the other species examined supports the occurrence of an ongoing speciation within the complex.

The conservation status of endangered Italian populations of the peatmoss Sphagnum palustre L. was evaluated through the study of their intraspecific genetic variation and structure. One-hundred-thirty-eight shoots from four populations were analyzed by 35 inter-simple sequence repeat (ISSR) loci. Molecular diversity indices highlight a general low level of variation, which is higher in centre-northern populations than in the southernmost one. AMOVA indicates that most variation is partitioned among populations, supporting a low level of gene flow and a severe isolation among them (FST= 0.7071–0.9198; Nm = 0.106). Mantel test gave a significant correlation (0.8634, P = 0.0083) between genetic and geographical distances. A strong association among loci suggests that recombination due to sexual reproduction contributes poorly to genetic variation observed, supported by the lack of sporophytes in the study areas. Both experimental results and theoretical predictions suggest a concrete risk of local extinction for the studied populations, further increased by fragile habitats hosting the species in Italy. The southernmost population is the most threatened, due to combined effects of lower genetic variation and geographic isolation.

In this study cytological ultrastructure, total content of C, N and S, and cellular location of major and trace elements (K, Ca, Mg, Cu, Pb and Zn) were investigated in the moss Hypnum cupressiforme and in the lichen Pseudevernia furfuracea exposed in bags for a spring–summer 12-weeks period in the urban area of Naples city. In the moss, severe ultrastructural damages, such as membrane interruptions and dehydration, developed after exposure supporting the occurrence of a dead biomonitor. In the lichen, the post-exposure stress marks, such as the development of lysosome-like vesicles and concentric bodies, or the production of melanin, were overall compatible with life. With exposure, N, S, major and trace element contents all increased in both biomonitors, while C remained substantially unchanged. Copper and Pb were mainly retained in extracellular and particulate forms. Intracellular concentration of Zn consistently increased in both biomonitors, irrespective of their vitality. In transplants, cellular location of elements can better reflect the form in which they occur in the environment.

A biomonitoring of airborne trace elements was performed in 2006 in Naples urban area through the exposure of devitalised Hypnum cupressiforme for 10 weeks at 4 m height. In one street, the moss was exposed at different heights to assess vertical gradients of element concentrations. Results were compared with those of a 1999 biosurvey. Correlations among Al, Fe and Ti suggested a soil particles contribution to element uptake. Cu, Mo and Fe were related with traffic flows. Long-range transport contributed to Cd, Cu and Mo accumulation in moss at higher heights. As in 1999, the airborne element load was higher in coastal sites, more affected by marine aerosols and traffic. In all sites, contents of Cd, Fe, Pb, Ni and V in moss were remarkably lower than in 1999, indicating a positive effect of actions set up in recent years to reduce the traffic and to improve the city air quality.

To define a harmonized methodology for the use of moss and lichen bags as active monitoring devices of airborne trace elements in urban areas, we evaluated the element accumulation in bags exposed in Naples in different spring weather conditions for 6- and 12-weeks. Three different pre-exposure treatments were applied to moss and lichen materials: water-washing, acid-washing and oven-drying. During the different exposure periods in the Naples urban environment the moss accumulated always higher amounts of elements (except Hg) than lichens and the element accumulation increased during wetter weather and higher PM(10) conditions. The oven pre-treatment did not substantially modify the morphology and element composition of moss and the exposure in bags of this material for 6-weeks was sufficient to detect the pattern of airborne trace elements.

The mineralogy and geochemistry (major and trace elements) of particulate matter collected from 14 April to 29 May 2003 by automatic device (PM10) and entrapped by moss and lichen exposed in bags in a monitoring site of Naples urban area were studied with the aim to obtain useful information for risk assessment and control measures feasibility. PM10 concentrations were generally above the threshold values fixed by the 1999/30/EC directive. Constant and low intensity winds enhanced re-suspension of fine particles and dispersion of gaseous pollutants. PM10 samples contained trace elements in relatively lower amounts compared to literature reports from other Mediterranean monitoring sites. Significant correlations between Al, Ca, Cu, Fe, K, Mg and Mn indicated that soil dust largely contributed to the accumulation of fine particles on filters and exposed mosses and lichens. Highly significant correlations were also found between Ni and V, indicative of oil combustion processes, Fe, Cu and Cr, indicative of vehicle emissions and mechanical components abrasion, and Na and Mg, indicative of marine aerosols. Lead and Hg did not correlate significantly with any other element. Comparison of element EFs, calculated with respect to the composition of Naples surface soils, indicated higher contribution of soil dust to PM and moss chemical composition compared with lichens, which in turn, according with their better preserved vitality, intercepted/absorbed more efficiently anthropogenic particles and elements of metabolic interest. Crystalline and amorphous detrital components (quartz, calcite, feldspars, volcanic glass, mica, kaolinite and smectite) and sea-bearing salts phases (halite, gypsum, Mg–K sulphates, Mg–Ca carbonates) were the main minerals in PM10, along with silica fibers and tuff particles.

To enhance the reliability of the moss and lichen transplant technique for active biomonitoring of trace metals in urban environments, we evaluated the natural variability in the chemical composition of the (epilithic and epiphytic) moss Hypnum cupressiforme and the epiphytic lichen Pseudevernia furfuracea from two reference areas in NE Italy. Green shoots of epilithic mosses and lobes of epiphytic lichens from larch branches showed rather homogenous composition and were selected for the exposure in nylon bags. As different physico-chemical pre-treatments are usually applied to selected cryptogamic material before its exposure, we also evaluated the effects of oven-drying at 120 °C for 24 h, washing in 1 N HNO 3 solution, and in 0.5% NH 4 oxalate solution at 85 °C for 15 h on the chemical composition and morphology of water-washed moss shoots and lichen lobes. Pre-treatments remarkably changed the chemical composition of selected materials but not their surface morphology.

Several lines of evidence suggest that recent long-distance dispersal may have been important in the evolution of intercontinental distribution ranges of bryophytes. However, the absolute rate of intercontinental migration and its relative role in the development of certain distribution ranges is still poorly understood. To this end, the genetic structure of intercontinental populations of six peatmoss species showing an amphi-Atlantic distribution was investigated using microsatellite markers. Methods relying on the coalescent were applied (IM and MIGRATE) to understand the evolution of this distribution pattern in peatmosses. Intercontinental populations of the six peatmoss species were weakly albeit significantly differentiated (average F(ST) = 0.104). This suggests that the North Atlantic Ocean is acting as a barrier to gene flow even in bryophytes adapted to long-range dispersal. The im analysis suggested a relatively recent split of intercontinental populations dating back to the last two glacial periods (9000-289,000 years ago). In contrast to previous hypotheses, analyses indicated that both ongoing migration and ancestral polymorphism are important in explaining the intercontinental genetic similarity of peatmoss populations, but their relative contribution varies with species. Migration rates were significantly asymmetric towards America suggesting differential extinction of genotypes on the two continents or invasion of the American continent by European lineages. These results indicate that low genetic divergence of amphi-Atlantic populations is a general pattern across numerous flowering plants and bryophytes. However, in bryophytes, ongoing intercontinental gene flow and retained shared ancestral polymorphism must both be considered to explain the genetic similarity of intercontinental populations.

Bryophytes have sometimes ambiguous morphological diagnostic features, so molecular markers can prove a useful tool in systematics, even more in a well known morphologically variable moss such as Hypnum cupressiforme. This paper focuses on genetic diversity in the H. cupressiforme complex inferred by ISSR markers and ITS and trnL sequences. Shoots from nine Italian populations of H. cupressiforme and from five other species included in the complex were analysed. Sequence divergence among the analysed species is very low, with the exception of Hypnum jutlandicum. The neighbour joining tree based on ISSR data shows that all the H. cupressiforme samples do not merge into a distinct cluster, but spread on the whole dendrogram. Molecular variance is partitioned more within- than between-species. Therefore, the genetic variation detected by ISSR at intra- and inter-specific level appears to be, to a large extent, the result of the individual variation, suggesting that the polymorphism detected appears p...

Lobes of the lichen Pseudevernia furfuracea (L.) Zopf and shoots of the moss Hypnum cupressiforme Hedw. were subjected to different treatments (water washing, oven drying, HNO3 washing, NH4-oxalate extraction) to assess the influence of vitality on accumulation efficiency, during a 6-week exposure in bags in two Italian cities, Trieste and Naples. No trend emerged between treatments, in terms of accumulation ability, for major and trace elements. Only water-washed lichens showed an increased C and N content after exposure in both cities. Element concentrations generally reached higher values in mosses than in lichens, especially for Al, Fe, and Zn (both cities), and for Cu, Mg and Na (Naples). Surface development strongly influenced accumulation capacity of the biomonitors. Quartzose and cation exchange filters revealed, on a weight basis, a poor performance. In urban environments, surface interception of atmospheric particulate seems to play a major role in accumulation, irrespective of organism vitality.

Bryophytes seem particularly suitable to investigate genetic diversity in relation to habitat disturbance due to their large employment as bioindicators and to the recent application of molecular markers to moss population studies. Genetic variation and structure were analysed in seven urban, extraurban and remote populations of Leptodon smithii, an epiphytic moss of Quercus ilex, a phanerogamic species of Mediterranean climax vegetation. A total of 210 individual shoots were DNA extracted and amplified with internal simple sequence repeat (ISSR) primers, and 54 haplotypes were identified. An uneven distribution of haplotype number and frequencies was observed among sites, with a higher number of haplotypes and more homogeneous haplotype frequencies in the extraurban/remote populations. Molecular diversity indices were overall higher in the extraurban sites than in the urban ones. Multilocus linkage disequilibrium values were in line with the occurrence of sexual/asexual reproduction in the seven populations. The isolation-by-distance model was not supported by Mantel test among sites; however, within-population fixation index (F(ST)) highlighted a clear relation between genetic and physic distances among trees, suggesting a limited dispersal range for L. smithii's spores. The genetic structure was mainly affected by population size, wood structure and extent, and genetic drift consequent to habitat fragmentation and human-induced disturbance.