Assessment of the effectiveness of a novel BioFilm-Membrane BioReactor oil-polluted wastewater treatment technology by applying biomarkers in the mussel Mytilus galloprovincialis.

Petrochemical industries and oil refineries are sources of hazardous chemicals into the aquatic environments, and often a leading cause of reduced oxygen availability, thus resulting in adverse effects in biota. This study is an expansion of our previous work on the assessment of the BioFilm-Membrane Bioreactor (BF-MBR) to mitigate the impact of oil-polluted wastewater on marine environments. Specifically, this study evaluated the reduction of selected chemical constituents (hydrocarbons and trace metals) and toxicity related to hypoxia and DNA damage to mussels Mytilus galloprovincialis, before and after treatment of oil-polluted wastewater with the BF-MBR. The application of a multidisciplinary approach provided evidence of the efficiency of BF-MBR to significantly reducing the pollutants load from oily contaminated seawaters. As result, the health status of mussels was preserved by a hypoxic condition due to oily pollutants, as evidenced by the modulation in the gene expression of HIF-1α and PHD and changes in the level of hypotaurine and taurine. Moreover, ameliorative effects in the energy metabolism were also found in mussel gills showing increased levels of glycogen, glucose and ATP, as well as a mitigated genotoxicity was revealed by the Micronucleus and Comet assays. Overall, findings from this study support the use of the BF-MBR as a promising treatment biotechnology to avoid or limiting the compromise of marine environments from oil pollution.

Drawing the baseline of trace element levels in the vulnerable Mediterranean osprey Pandion haliaetus: variations by breeding location, habitats, and egg components.

Due to its peculiarity to accumulate environmental contaminants, the osprey Pandion haliaetus is a sentinel species for the biomonitoring of contaminants in aquatic ecosystems. Despite this, no information on trace element concentration exists for the Mediterranean area, where relict and vulnerable osprey populations exist. We evaluated the geographical patterns of heavy metals and selenium in osprey eggs from three different populations of the Mediterranean basin (Balearic Islands, Corsica, and Tuscany), to identify any possible contaminant sources. Pattern of metal concentration followed the order: Fe > Zn > Cu > Se > Hg > Pb > Cd. Differences in contaminant concentrations between habitats and among egg components were found. Egg content and inner membrane showed higher mercury levels (1.06 ± 0.89 and 0.67 ± 0.62 mg/kg dw, respectively) than those recorded in the eggshell. Mercury concentration was ca. two times higher in marine than in wetland samples, and even higher (3.6 times) when referred to the eggshell. Cu, Fe, Zn, and Se had higher concentration in the inner membrane. We stress how the choice of the biological material can have significant implications for the correct evaluation of contamination. Our study represents a first regional scale survey for the vulnerable Mediterranean osprey populations and provides baseline data for their long-term biomonitoring.

Trace elements levels in muscle and liver of a rarely investigated large pelagic fish: The Mediterranean spearfish Tetrapturus belone (Rafinesque, 1810).

We determined levels of mercury, cadmium, lead and selenium in muscle and liver of 29 specimens of a large pelagic fish rarely investigated, the Mediterranean spearfish Tetrapturus belone (Raf., 1810). The following element concentration ranking (mean ± S.D.; espressed in mg/kg dry weight) was recorded in muscle: Hg (3.401 ± 1.908) > Se (1.727 ± 0.232) > Pb (0.532 ± 0.322) > Cd (0.019 ± 0.015), and Se (6.577 ± 1.789) > Cd (5.815 ± 3.038) > Hg (2.698 ± 2.214) > Pb (0.661 ± 1.334) in liver. Levels of Hg, Se and Cd were compared to those reported for other Istiophoridae from oceanic areas and for other large predators of Mediterranean Sea, like swordfish and tuna. Organotropism of trace elements and their relation to size was discussed. Ecophysiological considerations regarding the Se-Hg relationship as well as Se-Cd indicate a possible detoxification mechanism. The implications for human consumption are briefly discussed.

Assessing the Effect of Contaminated and Restored Marine Sediments in Different Experimental Mesocosms Using an Integrated Approach and Mytilus galloprovincialis as a Model.

The research presented here was conducted to ascertain the effectiveness of recovery technologies in remediating a compromised marine environment. The multidisciplinary approach aims to integrate traditional chemical-physical analysis and to assess the biological parameters of Mytilus galloprovincialis within different experimental mesocosms (W, G, and B). In particular, this system was designed to reproduce sediment resuspension in a marine environment, which is thought to be one cause of contaminant release. The study combined morphological and ultrastructural observations with DNA damage assessment and mRNA expression of those genes involved in cellular stress responses. The tissues of mussels maintained in the polluted mesocosm showed a higher accumulation of Pb and Hg than in those maintained in restored mesocosm. This observation correlates well with mRNA expression of MT10 and data on DNA damage. The outcome of the biological evaluation consolidates the chemical characterization and supports the concept that the remediation method should be evaluated at an early stage, both to analytically determine the reduction of toxic components and to assess its ultimate impact on the biological system.

Mercury, lead and copper in feathers and excreta of small passerine species in relation to foraging guilds and age of feathers.

In the present study excreta and feathers of five species of adult passerine birds from Montepulciano wetland (Siena, Italy) were assayed for trace elements between January and August 2006. Lead concentrations varied from 16.31 to 26.50 mg/kg and were found strictly related to the age of feathers. Copper levels were found to be high mainly in insectivorous birds (9.68 mg/kg) and were probably influenced by local use of copper-based agricultural fungicides. Mercury accumulation in feathers varied between species from 0.08 to 0.73 mg/kg. The role offeeding habits on trace elements levels in feathers and excreta is discussed.

Heavy metals in flamingos (Phoenicopterus ruber) from Italian wetlands: the problem of ingestion of lead shot.

Waterbirds are exposed to many contaminants, including lead from ingestion of shot and fishing sinkers. Lead poisoning had never been reported in flamingos wintering in Italian wetlands. Our investigation stems from a case of four flamingos found dead in Tuscany in 2002 with numerous lead shot in their gizzards. We therefore considered other specimens found dead in different Italian wetlands. Many lead shot found in gizzards and lead tissue concentrations confirmed the hypothesis of lead poisoning in two of the seven specimens analysed: concentrations in liver, kidney, and bone were 361.3, 265.09, and 43.31mug/g d.w., respectively. Lead organotropism was typical of acute poisoning. Cadmium and mercury were also determined, and found to be in line with what little data are available on this species in the literature. Although Italy recently endorsed the African-Eurasian Waterbird Agreement (AEWA) prohibiting use of lead shot for hunting in wetlands, our results reveal a first case of lead shot poisoning in flamingos wintering in Italian wetlands. This evidence sounds a further warning of the problem of spent lead shot in countries where hunting in wetlands is not strictly regulated.

Cadmium, lead, and mercury levels in feathers of small passerine birds: noninvasive sampling strategy.

Bird feathers have been widely used as a nondestructive biological material for monitoring heavy metals. Sources of metals taken up by feathers include diet (metals are incorporated during feather formation), preening, and direct contact with metals in water, air, dust, and plants. In the literature, data regarding the origin of trace elements in feathers are not univocal. Only in the vast literature concerning mercury (as methyl mercury) has endogenous origin been determined. In the present study, we investigate cadmium, lead, and mercury levels in feathers of prey of Falco eleonorae in relation to the ecological characteristics (molt, habitat, and contamination by soil) of the different species. Cluster analysis identified two main groups of species. Differences and correlations within and between groups identified by cluster analysis were then checked by nonparametric statistical analysis. The results showed that mercury levels had a pattern significantly different from those of cadmium and lead, which in turn showed a significant positive correlation, suggesting different origins. Nests of F. eleonorae proved to be a good source for feathers of small trans-Saharan passerines collected by a noninvasive method. They provided abundant feathers of the various species in a relatively small area--in this case, the falcon colony on the Isle of San Pietro, Sardinia, Italy.

Application of a suite of biomarkers in Posidonia oceanica (L.) delile to assess the ecotoxicological impact on the coastal environment.

The main aim of this study was to develop and validate a suite of sensitive responses (biomarkers) for monitoring conservation status and ecotoxicological impact in Posidonia oceanica meadows. Analytical methods were developed for NADPH cytochrome c reductase, ethoxycoumarin-o-deethylase (ECOD), guaiacol peroxidase (GPOX) and superoxide dismutase (SOD) assays. A preliminary proteomic approach using 2-D electrophoresis was also proposed as a biomarker. These techniques were initially tested on samples of posidonia exposed experimentally to various contaminants. Once validated, this approach was applied to posidonia in a field study. Samples of the seagrass were collected at four sites with potentially different environmental impact along the northern Tyrrhenian coast. The results showed that reductase activity was significantly induced in the various sampling areas with respect to the reference site. GPOX and SOD showed a similar trend; the highest activities were found in samples collected off a chlor-alkali plant and near a river estuary. Analysis of trace elements, polycyclic aromatic hydrocarbons (PAHs) and organochlorine compounds (OCs) in posidonia leaves showed differences between sites. A significant correlation was found between Hg concentrations and GPOX activity and between Cr, Al and As concentrations and reductase activity. The results validated these biomarkers in posidonia for the assessment of ecotoxicological impact on the coastal ecosystem.

The effects of heavy metal contamination on the soil arthropod community of a shooting range.

Soils in clay pigeon shooting ranges can be seriously contaminated by heavy metals. The pellets contained in ammunition are composed of Pb, Sb, Ni, Zn, Mn and Cu. The total concentrations of these metals in soils, and the effects of their increasing levels on the arthropod community were investigated at seven sampling sites in a clay pigeon shooting range and compared with two controls. Research revealed that the spatial distribution of Pb and Sb contamination in the shot-fall area was strongly correlated with the flight path of the pellets. Ordination obtained through Redundance Analysis showed that Collembola, Protura and Diplura were positively correlated with major detected contaminants (Pb, Sb), while Symphyla showed a negative correlation with these pollutants. Determination of the soluble lead fraction in soil, and of its bioaccumulation in the saprophagous Armadillidium sordidum (Isopoda) and the predator Ocypus olens (Coleoptera), showed that a significant portion of metallic Pb from spent pellets is bioavailable in the soil and can be bioaccumulated by edaphic organisms, entering the soil trophic network, but without biomagnification.

Posidonia oceanica as a biomonitor of trace elements in the Gulf of Naples: temporal trends by lepidochronology.

Levels of Hg, Cd, Pb, Cu, Zn, Fe, and Mn were assessed by using Posidonia oceanica as a bioindicator in the Gulf of Naples (southern Italy). Lepidochronology, which enables retroactive dating of scales and rhizomes of this seagrass, was combined with atomic spectrometry to assess temporal trends of trace elements in dated scales and rhizomes over a 10-year period. Lepidochronology occasionally has been used to monitor Hg in dated scales, but never has been used to determine concentrations of other trace elements in dated rhizomes. Data were compared between various structural parts of seagrass and between sampling sites. Concentrations of some elements found in dated scales or rhizomes showed a similar trend in most of the sites; increasing for Hg and Cu and decreasing for Pb, Fe, and Zn. Contaminant levels indicated by seagrass meadows varied from metal to metal. Concentrations of Hg, Cd, and Pb fell within the ranges measured in other areas considered to have low levels of heavy metal pollution, whereas Cu, Mn, and Zn had higher concentrations only in some years. However, on the basis of other studies, none of the sampling sites are considered to have high contamination levels.

Assumption and elimination of trace elements in Adélie penguins from Antarctica: a preliminary study.

The concentration and distribution of some trace elements were determined in stomach contents, excreta and feathers of Adélie penguins (Pygoscelis adeliae) breeding at Edmonson Point (Terra Nova Bay-Antarctica). In these materials obtained by non-destructive methods, concentrations of mercury, cadmium and lead measured were used to establish natural levels of these metals in the penguins' diet. An intake-excretion hypothesis was formulated.

Assessment of toxicological status of a SW Mediterranean segment population of striped dolphin (Stenella coeruleoalba) using skin biopsy.

Various studies have revealed high concentrations of contaminants such as organochlorines (OCs) and heavy metals in Mediterranean cetaceans. A geographical trend of contamination (PCBs and DDTs) has been found for striped dolphin (Stenella coeruleoalba). In this study we used a non-lethal approach (skin biopsy) to investigate bioaccumulation of OCs, including polychlorobiphenyls (PCBs), DDTs, polychlorodibenzo-p-dioxins (PCDDs), polychlorodibenzofurans (PCDFs), trace elements (Hg, Cd, Pb) and CYP1A activity (BPMO) in nine striped dolphins sampled in the Aeolian area (Sicily - Italy) in summer 2002. The arithmetic mean value of BPMO activity in this group was 43.46 AUF/g tissue/h. This value is approximately 3 times and 5 times lower, respectively, than the value found in the Ionian and in the Ligurian groups. Skin biopsies of striped dolphins emerged as a suitable material for assessing the toxicological status of the various Mediterranean groups.

Organotropism of persistent organic pollutants and heavy metals in the Greenland shark Somniosus microcephalus in NE Greenland.

The Greenland shark Somniosus microcephalus is an opportunistic feeder, a top predator, and a very long-lived species. The brain, liver, red and white muscle, gonads, fat, skin, pancreas, and spleen of Greenland sharks from NE Greenland fjords were analysed for PCBs, PCDDs/DFs, PBDEs; DDT isomers; HCH isomers; dieldrin; endrin; HCB; Cd, Hg, Pb, and Se. PCBs (2.01-103 ng/g wet wt) and PBDEs (7.9-3050 pg/g wet wt) were detected in most of the samples. PCDDs/DFs showed high values when detected. DDTs, HCB and HCHs were only detected in some tissues. The ΣTEQ was 5.76 pg/g in muscle. Cadmium mainly accumulated in the pancreas and liver (19.6 and 10.7 mg/kg dry wt, respectively); mercury in red muscle (4.10-6.91 mg/kg dry wt); selenium in the pancreas (3.57 mg/kg dry wt) and spleen (1.95 mg/kg dry wt); lead in the skin (0.358 mg/kgd ry wt). The selenium-mercury ratio in the liver was also evaluated.

Effects of sublethal, environmentally relevant concentrations of hexavalent chromium in the gills of Mytilus galloprovincialis.

Hexavalent chromium Cr(VI) is an important contaminant released from both domestic and industrial effluents, and represents the predominant chemical form of the metal in aquatic ecosystems. In the marine bivalve Mytilus galloprovincialis exposure to non-toxic, environmentally relevant concentrations of Cr(VI) was shown to modulate functional parameters and gene expression in both the digestive gland and hemocytes. In this work, the effects of exposure to Cr(VI) (0.1-1-10 µg L(-1) animal(-1) for 96 h) in mussel gills were investigated. Gill morphology and immunolocalization of GSH-transferase (GST), of components involved in cholinergic (AChE and ChAT), adrenergic (TH) and serotoninergic (5-HT(3) receptor) systems, regulating gill motility, were evaluated. Total glutathione content, activities of GSH-related enzymes (glutathione reductase - GSR, GST), of catalase, and of key glycolytic enzymes (phosphofructokinase - PFK and pyruvate kinase - PK) were determined. Moreover, mRNA expression of selected Mytilus genes (GST-π, metallothionein isoforms MT10 and MT20, HSP70 and 5-HT receptor) was assessed by RT-q-PCR. Cr(VI) exposure induced progressive changes in gill morphology and in immunoreactivity to components involved in neurotransmission that were particularly evident at the highest concentration tested, and associated with large metal accumulation. Cr(VI) increased the activities of GST and GSR, and total glutathione content to a different extent at different metal concentrations, this suggesting Cr(VI) detoxication/reduction at the site of metal entry. Cr(VI) exposure also increased the activity of glycolytic enzymes, indicating modulation of carbohydrate metabolism. Significant changes in transcription of different genes were observed. In particular, the mRNA level for the 5-HTR was increased, whereas both decreases and increases were observed for GST-π, MT10, MT20 and HSP70 mRNAs, showing sex- and concentration-related differences. The results demonstrate that Cr(VI) significantly affected functional and molecular parameters in mussel gills, and indicate that this tissue represents the major target of exposure to environmentally relevant concentrations of the metal.

Sometimes sperm whales (Physeter macrocephalus) cannot find their way back to the high seas: a multidisciplinary study on a mass stranding.

BACKGROUND: Mass strandings of sperm whales (Physeter macrocephalus) remain peculiar and rather unexplained events, which rarely occur in the Mediterranean Sea. Solar cycles and related changes in the geomagnetic field, variations in water temperature and weather conditions, coast geographical features and human activities have been proposed as possible causes. In December 2009, a pod of seven male sperm whales stranded along the Adriatic coast of Southern Italy. This is the sixth instance from 1555 in this basin. METHODOLOGY/PRINCIPAL FINDINGS: Complete necropsies were performed on three whales whose bodies were in good condition, carrying out on sampled tissues histopathology, virology, bacteriology, parasitology, and screening of veins looking for gas emboli. Furthermore, samples for age determination, genetic studies, gastric content evaluation, stable isotopes and toxicology were taken from all the seven specimens. The animals were part of the same group and determined by genetic and photo-identification to be part of the Mediterranean population. Causes of death did not include biological agents, or the "gas and fat embolic syndrome", associated with direct sonar exposure. Environmental pollutant tissue concentrations were relatively high, in particular organochlorinated xenobiotics. Gastric content and morphologic tissue examinations showed a prolonged starvation, which likely caused, at its turn, the mobilization of lipophilic contaminants from the adipose tissue. Chemical compounds subsequently entered the blood circulation and may have impaired immune and nervous functions. CONCLUSIONS/SIGNIFICANCE: A multi-factorial cause underlying this sperm whales' mass stranding is proposed herein based upon the results of postmortem investigations as well as of the detailed analyses of the geographical and historical background. The seven sperm whales took the same "wrong way" into the Adriatic Sea, a potentially dangerous trap for Mediterranean sperm whales. Seismic surveys should be also regarded as potential co-factors, even if no evidence of direct impact has been detected.

Review: porphyrins as biomarkers for hazard assessment of bird populations: destructive and non-destructive use.

In this review the biochemical, metabolic and toxicological significance of porphyrins in birds is examined, and their use as biomarkers of exposure to xenobiotics and heavy metals is explored. Laboratory studies pinpointing the main classes of compounds that alter porphyrin profiles are described, as well as those which defined the resulting porphyrin profiles and target organs. Field studies in which the biomarker was validated on natural populations of several species of birds are then reviewed. We finally illustrate their potential as a nondestructive biomarker suitable for situations in which destructive sampling of natural populations must be avoided, suggesting the implementation of this approach.