Tree-rings analysis to reconstruct atmospheric mercury contamination at a historical mining site.

Mercury (Hg) is a global environmental concern due to its toxicity (especially high in methylated form) and the long-range distribution of its gaseous elemental form (GEM). Hg-contaminated areas, such as abandoned mining sites, pose intrinsic difficulties for their management and heavy monitoring costs. In these environments, plant-based solutions may play a key role in the ecosystem quality assessment and support remediation strategies, combining reliability and cost-effectiveness. In this study, we adopted a biomonitoring approach by using tree rings of four different species collected in the proximity of the mining-metallurgical area of Abbadia San Salvatore, central Italy, a major former Hg mining district whose reclamation is currently in progress. Our dendrochemical analysis was aimed at identifying the historical changes of local atmospheric Hg contamination and at singling out, for the first time in the study area, other potentially toxic elements (PTEs) associated with the past mining activity. Collected cores dated back to early as 1940 and provided the temporal patterns of atmospheric Hg emission vs the produced liquid quantities, so reconstructing the historical impact of the mining site on nearby terrestrial ecosystems and resident human population. Current GEM contamination was found about twenty times lower than that of the fully operational mine periods. From a first survey on other PTEs, thallium (Tl) and lead (Pb) appeared to be potentially associated with the mining activity, thus suggesting new working assumptions for further dendrochemical analyses and for the inclusion of Pb in human biomonitoring surveys of the Mt. Amiata area, actually not present in the control list. The results prompt a more thorough assessment by tracking for a longer time span a critical site that is an ideal open-field lab to study the ecophysiology of different tree species in relation to environmental behavior of PTEs for better-assessing wildlife and human exposures.

First evidence of legacy chlorinated POPs bioaccumulation in Antarctic sponges from the Ross sea and the South Shetland Islands.

Antarctica is no longer pristine due to the confirmed presence of anthropogenic contaminants like Persistent Organic Pollutants (POPs). Benthic organisms are poorly represented in contamination studies in Antarctica although they are known to bioaccumulate contaminants. Sponges (Phylum Porifera) are dominant members in Antarctic benthos, both in terms of abundance and biomass, and are an important feeding source for other organisms, playing key functional roles in benthic communities. To the best of our knowledge, legacy chlorinated POPs such as polychlorinated biphenyls (PCBs), hexachlorobenzene (HCB), and dichlorodiphenyltrichloroethane (DDT) and their metabolites have never been investigated in this Phylum in Antarctica. The aim of this work was to evaluate the bioaccumulation of PCBs, HCB, o,p'- and p,p'-DDT and their DDE and DDD isomers in 35 sponge samples, belonging to 17 different species, collected along the coast of Terra Nova Bay (Adèlie Cove and Tethys Bay, Ross Sea), and at Whalers Bay (Deception Island, South Shetland Islands) in Antarctica. Lipid content showed a significant correlation with the three pollutant classes. The overall observed pattern in the three study sites was ΣPCBs>ΣDDTs>HCB and it was found in almost every species. The ΣPCBs, ΣDDTs, and HCB ranged from 54.2 to 133.7 ng/g lipid weight (lw), from 17.5 to 38.6 ng/g lw and from 4.8 to 8.5 ng/g lw, respectively. Sponges showed contamination levels comparable to other Antarctic benthic organisms from previous studies. The comparison among sponges of the same species from different sites showed diverse patterns for PCBs only in one out of four cases. The concentration of POPs did not vary significantly among the three sites. The predominance of lower chlorinated organochlorines in the samples suggested that long-range atmospheric transportation (LRAT) could be the major driver of contamination as molecules with a high long range transport potential (e.g. low chlorinated PCBs, HCB) prevails on heavier ones.

Aryl hydrocarbon reporter gene bioassay for screening polyhalogenated dibenzo-p-dioxins/furans and dioxin-like polychlorinated biphenyls in hydrochar and sewage sludge.

The suitability of the AhR reporter gene bioassays to screen the presence of polychlorinated dibenzo-p-dioxins/furans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (dl-PCBs) in sewage sludge (SL) and related hydrochar (HC) was here investigated. Samples of SL obtained from six WWTPs were processed by hydrothermal carbonization to obtain the resultant HCs and both tested with DR-CALUX® bioassay. Levels of PCDD/Fs and dl-PCBs were also determined analytically in the same samples by GC-MS/MS. Bioanalytical Toxicity Equivalent values (BEQ) resulted in one order of magnitude higher in HC compared to SL samples and those obtained from the dl-PCBs fraction higher than those from PCDD/Fs. BEQ and TEQWHO values, the latter obtained by GC-MS/MS analysis on the same matrices, were highly correlated showing also a similar trend in the six WWTPs (RS= 0.8252, p < 0.001; Pearson's R RP =0.8029, p < 0.01). The suitability of AhR bioassays and in particular of the DR-CALUX® to screen the presence and biological activity of legacy organohalogen compounds in both SL and HC matrices was demonstrated for the first time which support their usage for the assessment of potential risks associated with their further environmental applications.

Legacy and novel flame retardants from indoor dust in Antarctica: Sources and human exposure.

The air humidity in Antarctica is very low and this peculiar weather parameter make the use of flame retardants in research facilities highly needed for safety reasons, as fires are a major risk. Legacy and novel flame retardants (nFRs) including polybrominated diphenyl ethers (PBDEs), hexabromocyclododecanes (HBCDs), 1,2-bis(2,4,6-tribromophenoxy) ethane (BTBPE), Dechlorane Plus (DP), and other nFRs were measured in indoor dust samples collected at research Stations in Antarctica: Gabriel de Castilla, Spain (GCS), Julio Escudero, Chile (JES), and onboard the RRS James Clark Ross, United Kingdom (RRS JCR). The GC-HRMS and LC-MS-MS analyses of dust samples revealed ∑7PBDEs of 41.5 ± 43.8 ng/g in rooms at GCS, 18.7 ± 11.6 ng/g at JES, and 27.2 ± 37.9 ng/g onboard the RRS JCR. PBDE pattern was different between the sites and most abundant congeners were BDE-183 (40%) at GCS, BDE-99 (50%) at JES, and BDE-153 (37%) onboard the RRS JCR. The ∑(4)HBCDs were 257 ± 407 ng/g, 14.9 ± 14.5 ng/g, and 761 ± 1043 ng/g in indoor dust collected in rooms at GCS, JES, and RRS JCR, respectively. The ∑9nFRs were 224 ± 178 ng/g at GCS, 14.1 ± 13.8 ng/g at JES, and 194 ± 392 ng/g on the RRS JCR. Syn- and anti-DP were detected in most of the samples and both isomers showed the highest concentrations at GCS: 163 ± 93.6 and 48.5 ± 61.1 ng/g, respectively. The laboratory and living room showed the highest concentration of HBCDs, DPs, BTBPE. The wide variations in FR levels in dust from the three research facilities and between differently used rooms reflect the different origin of furnishing, building materials and equipment. The potential health risk associated to a daily exposure via dust ingestion was assessed for selected FRs: BDEs 47, 99, and 153, α-, ß-, and γ-HBCD, BTBPE, syn- and anti-DP. Although the estimated exposures are below the available reference doses, caution is needed given the expected increasing use of novel chemicals without a comprehensive toxicological profile.

PBDEs and PCBs in terrestrial ecosystems of the Victoria Land, Antarctica.

The Victoria Land (East Antarctica) is characterized by the presence of lakes and ponds where rare water is found during the Summer months. These freshwater ecosystems are an important resource for migrating seabirds that use them and leave there their droppings and feathers, contributing to enrich the water with organic matter. Persistent organic pollutants (POPs) are long-range transported to polar regions and their detection in the Antarctic ecosystems date back to the 1960s. Most studies have been related to POP concentrations in marine environment, and terrestrial ecosystems have been poorly investigated. This study reports the concentration of PBDEs (23 congeners) and PCBs (21 among non-, mono-, di-, and ortho congeners) in lake sediment, soil and vegetation mat (community of algae, cyanobacteria, bryophites) collected close to six lakes in the Victoria Land (74°31'S-74°97'S, 165°07'E-162°51'E): Edmonson Point 14 and 15A, Carezza, Enigma, Tarn Flat 20, Inexpressible Island 10B. The ∑PBDEs averaged 0.09-0.28 ng/g and BDEs 28, 47 and 154 were higher in mat and soils, while BDEs 183 and 47 in sediment samples. PCBs ranged <0.003-0.807 ng/g and congeners nos. 114, 138, and 187 were the most abundant. In addition, TEQs were derived for the non-and mono-ortho PCBs detected and values were very low in each matrix (0.010 pg/g in the soil, 0.012 pg/g in sediment and mat). The long-range atmospheric transport can be confirmed as the most important POP source in Antarctica, although the scientific stations and seabird colonies may be potential local sources and contribute to contaminant release.

Dioxin-like compounds bioavailability and genotoxicity assessment in the Gulf of Follonica, Tuscany (Northern Tyrrhenian Sea).

The Gulf of Follonica (Italy) is impacted by the chemical pollution from ancient mining activity and present industrial processes. This study was aimed to determine the bioavailability of dioxin-like compounds (DLCs) in coastal marine environment and to assess the genotoxic potential of waste waters entering the sea from an industrial canal. Moderately high levels of DCLs compounds (∑ PCDDs + PCDFs 2.18­29.00 pg/g dry wt) were detected in Mytilus galloprovincialis transplanted near the waste waters canal and their corresponding Toxic Equivalents (TEQs) calculated. In situ exposed mussels did not show any genotoxic effect (by Comet and Micronucleus assay). Otherwise, laboratory exposure to canal waters exhibited a reduced genomic template stability (by RAPD-PCR assay) but not DNA or chromosomal damage. Our data reveal the need to focus on the levels and distribution of DLCs in edible species from the study area considering their potential transfer to humans through the consumption of sea food.

Biomonitoring of chemicals in biota of two wetland protected areas exposed to different levels of environmental impact: results of the "PREVIENI" project.

The PREVIENI project (funded by the Ministry of Environment) investigated the exposure to endocrine disrupters in samples of human population and environmental biota in Italy. The environmental biomonitoring considered two Italian WWF Oasis, with the aim to compare the presence and effects of endocrine disruptors in organisms from two protected natural areas, respectively, upstream and downstream a chemical emission site. Chemical analysis of pollutants' tissue levels was made on tissues from earthworm, barbell, trout, and coot, selected as bioindicator organisms. The contaminants considered were as follows: the perfluorinated compounds perfuoroctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), polychlorinated biphenyls (PCBs 58 congeners), polybrominated diphenyl ethers (PBDEs, 13 congeners), polycyclic aromatic hydrocarbons (PAHs, 16 compounds), toxic trace elements, the phthalate di-2-ethylexyl phthalate (DEHP) and its primary metabolite, bisphenol A, synthetic musk compounds (musk xylene, musk ketone, tonalide, and galaxolide), and p-nonylphenol. The analyses showed low concentrations of most pollutants in all species from both areas, compared to available literature; noticeable exceptions were the increases of DEHP's primary metabolite, PBDE, PAHs, Hg, and Pb in barbells, and of PCB and Cd in earthworms from the downstream area. The results showed the presence of endocrine disruptors, including those considered as "non-persistent," in bioindicators from protected areas, albeit at low levels. The results provide a contribution to the evaluation of reference values in biota from Mediterranean Europe and support the relevance of monitoring exposure to pollutants, in particular for freshwater environment, also in protected areas.

Behavioural responses of the supralittoral amphipod Talitrus saltator (Montagu) to trace metals contamination.

We investigated behavioural responses (locomotor activity and substrate selection) of the amphipod Talitrus saltator after laboratory exposure to different concentrations of Hg, Cu and Cd. Locomotor activity, measured with a microwave radar device, was assessed in animals exposed to contaminated sand and in sandhoppers previously kept for 48 h in contaminated seawater and tested in clean sand. The contents of Hg, Cu and Cd in T. saltator tissues were measured at the end of pre-exposure to contaminated seawater and after 7 days of exposure to contaminated sand. Substrate selection tests were carried out in a binary choice arena. Tests in contaminated sand showed that sandhopper locomotor activity varied in a dose-dependent manner in the presence of Cu and Hg (at lower Hg concentration they were more active during daytime) but did not show significant changes in Cd-exposed animals except for disappearance of the typical circadian activity pattern. Pre-exposure to trace metals in seawater induced a significant decrease of movements for all metals, although the effects varied according to the toxicity of the metal. Trace metals analyses showed that tissue concentrations of Hg, Cu and Cd were related to their concentrations in the test medium. Substrate selection tests showed avoidance of contaminated sand only when sandhoppers were tested in the presence of both concentrations of Hg and at the higher concentration of Cu. In conclusion, locomotor activity and avoidance behaviour of T. saltator could be used as behavioural biomarkers of trace metals exposure.

Trace metal contamination of Tuscan and eastern Corsican coastal supralittoral zones: The sandhopper Talitrus saltator (Montagu) as a biomonitor.

The aim of this study was to determine trace metal concentrations (Hg, Cd, Cu, Zn, Al, Fe, Mn, Cr and Pb) in samples of Talitrus saltator collected at 14 localities along the Tuscan and eastern Corsican coasts. This talitrid amphipod is a well-known biomonitor of trace metal contaminations of the supralittoral zone of European sandy beaches. The results show the ability of T. saltator to accumulate Cu, Cd, Zn, Hg, Al, Fe and Mn, but not Cr or Pb. Moreover, the sites can be distinguished on the basis of the accumulated metals. Finally, comparison with literature data regarding the same species and the same sampling sites shows a general decrease in Cd, Zn and Hg contamination. In conclusion, this study confirms that T. saltator can be used as an indicator of the spatial distribution of trace metals contamination.

Collection of trace evidence of explosive residues from the skin in a death due to a disguised letter bomb. The synergy between confocal laser scanning microscope and inductively coupled plasma atomic emission spectrometer analyses.

In most deaths caused by explosive, the victim's body becomes a depot for fragments of explosive materials, so contributing to the collection of trace evidence which may provide clues about the specific type of device used with explosion. Improvised explosive devices are used which contain "homemade" explosives rather than high explosives because of the relative ease with which such components can be procured. Many methods such as chromatography-mass spectrometry, scanning electron microscopy, stereomicroscopy, capillary electrophoresis are available for use in the identification of explosive residues on objects and bomb fragments. Identification and reconstruction of the distribution of explosive residues on the decedent's body may give additional hints in assessing the position of the victim in relation to the device. Traditionally these residues are retrieved by swabbing the body and clothing during the early phase, at autopsy. Gas chromatography-mass spectrometry and other analytical methods may be used to analyze the material swabbed from the victim body. The histological examination of explosive residues on skin samples collected during the autopsy may reveal significant details. The information about type, quantity and particularly about anatomical distribution of explosive residues obtained utilizing confocal laser scanning microscope (CLSM) together with inductively coupled plasma atomic emission spectrometer (ICP-AES), may provide very significant evidence in the clarification and reconstruction of the explosive-related events.

DNA damage, severe organ lesions and high muscle levels of As and Hg in two benthic fish species from a chemical warfare agent dumping site in the Mediterranean Sea.

The aim of the present study was to evaluate the environmental threat to benthic species from chemical weapons dumped in the southern Adriatic Sea. An ecotoxicological approach using chemical analysis and biological responses was applied, in two sentinel species: the Blackbelly rosefish Helicolenus dactylopterus and European conger Conger conger. Specimen were collected in a stretch of sea, where had been dumped war materials and from a reference site free of ordnance. Residues of yperite, Hg and As were measured in fish fillets. Skin, liver, kidney and spleen were examined for histopathological and macroscopical lesions. Liver detoxifying capacities (EROD and UDPGT) and genotoxicity (comet assay) were also investigated. As and Hg levels were three-four times higher than those from the reference site in both species (p<0.001). Both species captured in dumping site showed clear signs of chronic illness according to the health assessment index (HAI). Deep ulcers and nodules were observed on skin and external organs. Histological lesions such as periportal and bile duct fibrosis, pericholangitis, steatosis, granuloma and elevated splenic MMCs were detected in liver and spleen. Significantly higher EROD activities were also found in both species from dumping site (p<0.01). Comet assay revealed genotoxicty in gills of C. conger from dumping site, indicating uptake of chemical warfare agents through fish gills. European conger was found to be a more sensitive bioindicator of this type of contamination than the Blackbelly rosefish.

The amphipod Talitrus saltator as a bioindicator of human trampling on sandy beaches.

The present paper assesses the use of the supralittoral amphipod Talitrus saltator as a bioindicator of the effects of human trampling on the supralittoral sandy band. Samplings in delimited areas were carried out at sites subjected to different human impact. The results showed a strong negative correlation between the number of swimmers and the sandhopper population density, while there was no clear relationship between sandhopper abundance and the other factors considered: granulometry, compactness and organic carbon content of the sand, and trace metal contents in the sand and sandhoppers. A field test of trampling conducted in a confined space showed its direct negative effect on sandhopper survival. However, trace metal analysis confirmed the ability of T. saltator to bioaccumulate some elements (Hg, Zn, Cu, Cd). Our study demonstrates that T. saltator is a good bioindicator of human impact in the supralittoral zone of sandy shores.