"Communicative competence assessment of the person with aphasia caregiver: Standardization of the ACCA-CHECKLIST".

Aphasia constitutes a very complex clinical entity that requires a "competent" caregiver to interact with the person with aphasia (PWA). The literature lacks a valid and reliable set of standardized tools which can offer objective and quantifiable data of a caregiver's communicative competence. The aim of the study was to develop, standardize and validate an evaluation tool suited to measur the caregivers' competence in communicating with family members affected by aphasic disorders. Forty-two patients with aphasia and their respective caregivers were enrolled in the study. Caregivers' communicative competence was assessed through a new evaluation tool called ACCA-cl. Aphasia severity and functional communication abilities of the PWA were also investigated. Our data showed encouraging results regarding the reliability and the validity of the ACCA-cl scale in detecting the caregiver's communicative competence, especially as far as verbal content was considered. This scale can also be used to assess the improvement achieved by the caregiver after a communication training. The analysis provides encouraging findings for verbal content scales of the ACCA-cl and its possible use in clinical settings as a quantitative tool for detecting changes induced by the educational method of the caregiver. The tool is currently available in Italian.

Bone Mineral Density and Trabecular Bone Score Changes throughout Menopause in Women with HIV.

OBJECTIVE: The objectives of this study were to describe the trajectories of bone mineral density (BMD) and trabecular bone score (TBS) changes throughout pre-menopause (reproductive phase and menopausal transition) and post-menopause (early and late menopause) in women with HIV (WWH) undergoing different antiretroviral therapies (ARTs) and explore the risk factors associated with those changes. METHODS: This was an observational longitudinal retrospective study in WWH with a minimum of two DEXA evaluations comprising BMD and TBS measurements, both in the pre-menopausal and post-menopausal periods. Menopause was determined according to the STRAW+10 criteria, comprising four periods: the reproductive period, menopausal transition, and early- and late-menopausal periods. Mixed-effects models were fitted to estimate the trajectories of the two outcomes (BMD and TBS) over time. Annualized lumbar BMD and TBS absolute and percentage changes were calculated in each STRAW+10 time window. A backward elimination procedure was applied to obtain the final model, including the predictors that affected the trajectories of BMD or TBS over time. RESULTS: A total of 202 WWH, all Caucasian, were included. In detail, 1954 BMD and 195 TBS data were analyzed. The median number of DEXA evaluations per woman was 10 (IQR: 7, 12). The median observation periods per patient were 12.0 years (IQR = 8.9-14.4) for BMD and 6.0 years (IQR: 4.3, 7.9) for TBS. The prevalence of osteopenia (63% vs. 76%; p < 0.001) and osteoporosis (16% vs. 36%; p < 0.001) increased significantly between the pre-menopausal and post-menopausal periods. Both BMD (1.03 (±0.14) vs. 0.92 (±0.12) g/cm2; p < 0.001) and TBS (1.41 (IQR: 1.35, 1.45) vs. 1.32 (IQR: 1.28, 1.39); p < 0.001) decreased significantly between the two periods. The trend in BMD decreased across the four STRAW+10 periods, with a slight attenuation only in the late-menopausal period when compared with the other intervals. The TBS slope did not significantly change throughout menopause. The delta mean values of TBS in WWH were lower between the menopausal transition and reproductive period compared with the difference between menopause and menopausal transition. CONCLUSIONS: Both BMD and TBS significantly decreased over time. The slope of the change in BMD and TBS significantly decreased in the menopausal transition, suggesting that this period should be considered by clinicians as a key time during which to assess bone health and modifiable risk factors in WWH.

Spatial scale influences the distribution of viral diversity in the eukaryotic virome of the mosquito Culex pipiens.

Our knowledge of the diversity of eukaryotic viruses has recently undergone a massive expansion. This diversity could influence host physiology through yet unknown phenomena of potential interest to the fields of health and food production. However, the assembly processes of this diversity remain elusive in the eukaryotic viromes of terrestrial animals. This situation hinders hypothesis-driven tests of virome influence on host physiology. Here, we compare taxonomic diversity between different spatial scales in the eukaryotic virome of the mosquito Culex pipiens. This mosquito is a vector of human pathogens worldwide. The experimental design involved sampling in five countries in Africa and Europe around the Mediterranean Sea and large mosquito numbers to ensure a thorough exploration of virus diversity. A group of viruses was found in all countries. This core group represented a relatively large and diverse fraction of the virome. However, certain core viruses were not shared by all host individuals in a given country, and their infection rates fluctuated between countries and years. Moreover, the distribution of coinfections in individual mosquitoes suggested random co-occurrence of those core viruses. Our results also suggested differences in viromes depending on geography, with viromes tending to cluster depending on the continent. Thus, our results unveil that the overlap in taxonomic diversity can decrease with spatial scale in the eukaryotic virome of C. pipiens. Furthermore, our results show that integrating contrasted spatial scales allows us to identify assembly patterns in the mosquito virome. Such patterns can guide future studies of virome influence on mosquito physiology.

Trace elements in the muscle tissue of Hemiculter leucisculus and Abramis brama orientalis from the Anzali International wetland, south-west of Caspian Sea: An exposure risk assessment.

High levels of trace elements in aquatic environments can affect the quality of seafood. We analyzed the concentrations of As, Cd, Co, Hg, Mn, Ni and Pb, in the edible muscles of Hemiculter leucisculus and A. brama orientalis caught in four different areas of southwest of the Caspian Sea. We estimated the potential risks for human health deriving by the oral consumption of these two species, and the Target Hazard Quotient (THQ) according to the US-EPA approach. THQ by adults and children was always below 1 for all stations. The greater contribution was given by Co, followed by Cd, Hg, Pb, Ni, As and Mn. Total-THQ was 0.538 and 0.246 for children and adults, respectively. Trace elements detected by our study were not elevated, highlighting a positive picture of the studied area and a lower risk of developing chronic systemic effects deriving from the consumption of local fish products.

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.

Bioaccumulation and health risk assessment of exposure to potentially toxic elements by consuming agricultural products irrigated with wastewater effluents.

Potentially toxic elements (PTEs) have many adverse health effects due to bioaccumulation capability and their long persistence in the environment. As a renewable water source, the effluents of municipal wastewater treatment systems have been used to irrigate agricultural products widely. However, the evidence on the bioaccumulation of PTEs in crops irrigated with these effluents is still scarce, with no available study in low and middle-income countries. Therefore, this study aimed to assess the PTEs concentration in the soil and crops irrigated with effluents of Sabzevar wastewater treatment plant and the related health risk by that. The clustered method was used to determine the soil and craps samples. Seventy cumulative samples were randomly prepared in summer and autumn 2016 and 2017 from crops, soil and effluent. Inductively coupled plasma mass spectrometry (ICP-MS) was used to measure PTEs. The health risk of exposure to PTEs was assessed using Monte Carlo simulation technique. Kruskal Wallis test and Posthoc Tukey HSD test were used to assess the mean difference of PTEs between soil, effluent and crops as well as between crops together. The bioaccumulation factor (BAF) magnitude order in different crop samples was Cd > Sr > Cu > Pb > Zn > Co > As > Cr > Ni, respectively. The Cd accumulation in Sugar beet plant was significantly higher than in other samples. The highest hazard quotient (HQ) based on single PTEs was observed for As (mineral) (mean: 5.62 × 10-1 and percentile 95th: 2.13) in Okra. Regarding total HQ (THQ), the highest and lowest mean (percentile 95th) values were 1.50 (3.22) and 2.40 × 10-1 (4.01 × 10-1) for Okra and Watermelon, respectively. The mean concentrations of Co, Cr, Ni and Zn were significantly higher in crops compared to soil and influent samples. Posthoc tests indicated that the concentration of PTEs between investigated crop samples were not statistically significant different (p > 0.05). Overall, our study suggested that irrigation with the effluent of stabilization pond wastewater treatment system exerts a potential health risk due to bioaccumulation of PTEs in crops.

The Genome Segments of Bluetongue Virus Differ in Copy Number in a Host-Specific Manner.

Genome segmentation is mainly thought to facilitate reassortment. Here, we show that segmentation can also allow differences in segment abundance in populations of bluetongue virus (BTV). BTV has a genome consisting in 10 segments, and its cycle primarily involves periodic alternation between ruminants and Culicoides biting midges. We have developed a reverse transcription-quantitative PCR (RT-qPCR) approach to quantify each segment in wild BTV populations sampled in both ruminants and midges during an epizootic. Segment frequencies deviated from equimolarity in all hosts. Interestingly, segment frequencies were reproducible and distinct between ruminants and biting midges. Beyond a putative regulatory role in virus expression, this phenomenon could lead to different evolution rates between segments.IMPORTANCE The variation in viral gene frequencies remains a largely unexplored aspect of within-host genetics. This phenomenon is often considered to be specific to multipartite viruses. Multipartite viruses have segmented genomes, but in contrast to segmented viruses, their segments are each encapsidated alone in a virion. A main hypothesis explaining the evolution of multipartism is that, compared to segmented viruses, it facilitates the regulation of segment abundancy, and the genes the segments carry, within a host. These differences in gene frequencies could allow for expression regulation. Here, we show that wild populations of a segmented virus, bluetongue virus (BTV), also present unequal segment frequencies. BTV cycles between ruminants and Culicoides biting midges. As expected from a role in expression regulation, segment frequencies tended to show specific values that differed between ruminants and midges. Our results expand previous knowledge on gene frequency variation and call for studies on its role and conservation beyond multipartite viruses.

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.

Evaluation of automotive shredder residues (ASR) landfill behavior through lysimetric and traditional leaching tests.

With regards to European waste catalog, automotive shredder residues (ASR) can be classified both as a hazardous or non-hazardous waste according to its hazardous properties (H1-H14). It is thus important to carry out an adequate chemical-physical characterization to identify the presence and concentration of those substances able to give, to this extremely heterogeneous material, the hazardousness character of. The issue of waste characterization, to identify the proper site for appropriate waste disposal, is based, according to the relevant laws, to the use of leaching tests. The analysis of the potential effects of landfilled waste in laboratory, however, run into several difficulties in reproducing phenomena depending both on the characteristics of small, heterogeneous quantity of waste and on the local boundary conditions. These difficulties are much more significant as the waste is heterogeneous at the small scale of the laboratory. This is one of the main problems often leading to scattered results even when starting from the same waste parcel. Present research aimed to overcome the above-mentioned difficulties deriving from waste heterogeneity and was based on a lysimetric simulation. Experimentation with lysimeter has shown it effectiveness in the comparison between leachate from the lysimeter and an ASR landfill leachate, from which similar distribution of metal mass ratios, close values for both BOD5 and COD, as well as the absence in both the fluids of organochlorinated compounds, emerge.

Vector Competence of Italian Populations of Culicoides for Some Bluetongue Virus Strains Responsible for Recent Northern African and European Outbreaks.

The distribution of Bluetongue virus (BTV) in Europe can be represented by two distinct and interconnected epidemiological systems (episystems), each characterized by different ecological characteristics and vector species. This study investigated the vector competence of Italian populations of Culicoides imicola and Culicoides obsoletus/scoticus to some representative BTV strains after artificial oral infection. The BTV strains were selected according to their ability to spread to one or both episystems and included BTV-4 ITA, responsible of the recent Italian and French BTV-4 outbreaks; the BTV-2 strain which caused the first BTV incursion in Italy, Corsica, and Balearic Islands; BTV-4 MOR, responsible for the epidemic in Morocco; and BTV-8, the strain which spread through Europe between 2006 and 2008. Blood-soaked cotton pledgets and Hemotek membrane feeder using Parafilm® membrane were used to artificially feed midges. For each population/strain, recovery rates (positive/tested heads) were evaluated using serogroup- and serotype-specific RT-PCR. The trial demonstrated that, except for the Abruzzo population of C. obsoletus/C. scoticus, which was refractory to BTV-4 MOR infection, all the investigated Culicoides populations are susceptible to the selected BTV strains and that, if prompt vaccination programs and restriction measures had not been implemented, BTV-2 and BTV-4 MOR could have spread all over Europe.

Mesocosm System to Evaluate BF-MBR Efficacy in Mitigating Oily Wastewater Discharges: an Integrated Study on Mytilus galloprovincialis.

This work presents the results of recovery efficacy of the system "BioFilm Membrane BioReactor" (BF-MBR), in the treatment of oily contaminated seawaters. To this aim, we proposed a multidisciplinary approach that integrates traditional chemical-physical measures together with the assessment on biological sentinel Mytilus galloprovincialis, maintained in a medium-scale artificial system named mesocosm. The setup included: (1) a mesocosm consisting of uncontaminated seawater; (2) a mesocosm composed of an untreated oily wastewater discharge; and (3) a mesocosm receiving the same oily wastewater previously treated by a BF-MBR pilot scale plant. The multidisciplinary approach that included traditional chemical measures on mesocosms together with the evaluation of morphological organization, mRNA expression of those genes involved in cellular stress response, immunohistochemistry and metabolomic analysis on mussel tissues, was able to provide a robust and holistic evidence of how the proposed treatment is able to reduce the overall impact of oily wastewater discharges on the marine ecosystem.

Defining ecological regions in Italy based on a multivariate clustering approach: A first step towards a targeted vector borne disease surveillance.

Ecoregionalization is the process by which a territory is classified in similar areas according to specific environmental and climatic factors. The climate and the environment strongly influence the presence and distribution of vectors responsible for significant human and animal diseases worldwide. In this paper, we developed a map of the eco-climatic regions of Italy adopting a data-driven spatial clustering approach using recent and detailed spatial data on climatic and environmental factors. We selected seven variables, relevant for a broad set of human and animal vector-borne diseases (VBDs): standard deviation of altitude, mean daytime land surface temperature, mean amplitude and peak timing of the annual cycle of land surface temperature, mean and amplitude of the annual cycle of greenness value, and daily mean amount of rainfall. Principal Component Analysis followed by multivariate geographic clustering using the k-medoids technique were used to group the pixels with similar characteristics into different ecoregions, and at different spatial resolutions (250 m, 1 km and 2 km). We showed that the spatial structure of ecoregions is generally maintained at different spatial resolutions and we compared the resulting ecoregion maps with two datasets related to Bluetongue vectors and West Nile Disease (WND) outbreaks in Italy. The known characteristics of Culicoides imicola habitat were well captured by 2/22 specific ecoregions (at 250 m resolution). Culicoides obsoletus/scoticus occupy all sampled ecoregions, according to its known widespread distribution across the peninsula. WND outbreak locations strongly cluster in 4/22 ecoregions, dominated by human influenced landscape, with intense cultivations and complex irrigation network. This approach could be a supportive tool in case of VBDs, defining pixel-based areas that are conducive environment for VBD spread, indicating where surveillance and prevention measures could be prioritized in Italy. Also, ecoregions suitable to specific VBDs vectors could inform entomological surveillance strategies.

Crimean-Congo Hemorrhagic Fever Virus Genome in Tick from Migratory Bird, Italy.

We detected Crimean-Congo hemorrhagic fever virus in a Hyalomma rufipes nymph collected from a whinchat (Saxicola rubetra) on the island of Ventotene in April 2017. Partial genome sequences suggest the virus originated in Africa. Detection of the genome of this virus in Italy confirms its potential dispersion through migratory birds.

Experimental and numerical evaluation of Groundwater Circulation Wells as a remediation technology for persistent, low permeability contaminant source zones.

Contaminants removal stoked inside low permeability zones of aquifers is one of the most important challenge of groundwater remediation process today. Low permeability layers can be considered persistent secondary sources of contamination because they release pollutants by molecular diffusion after primary source of contamination is reduced, causing long plum tails (Back-Diffusion). In this study, the Groundwater Circulation Well (GCW) system was investigated as an alternative remediation technology to the low efficient traditional pumping technologies to restore contaminated low permeability layers of aquifers. The GCW system creates vertical groundwater circulation cells by drawing groundwater through a screen of a multi-screen well and discharging it through another screen. The suitability of this technology to remediate contaminated low permeability zones was investigated by laboratory test and numerical simulations. The collected data were used to calibrate a model created to simulate the Back-Diffusion process and to evaluate the effect of different pumping technologies on the depletion time of that process. Results show that the efficiency of the GCW is dependent on the position and on the geometry of the low permeability zones, however the GCW system appears more suitable to restore contaminated low permeability layers of aquifers than the traditional pumping technology.

Life Cycle Assessment of waste disposal from olive oil production: Anaerobic digestion and conventional disposal on soil.

Extra virgin olive-oil (EVO) production is an important economic activity for several countries, especially in the Mediterranean area such as Spain, Italy, Greece and Tunisia. The two major by-products from olive oil production, solid-liquid Olive Pomace (OP) and the Olive Mill Waste Waters (OMWW), are still mainly disposed on soil, in spite of the existence of legislation which already limits this practice. The present study compares the environmental impacts associated with two different scenarios for the management of waste from olive oil production through a comparative Life Cycle Assessment (LCA). The two alternative scenarios are: (I) Anaerobic Digestion and (II) Disposal on soil. The analysis was performed through SimaPro software and the assessment of the impact categories was based on International Life Cycle Data and Cumulative Energy Demand methods. Both the scenarios are mostly related to the cultivation and harvesting phase and are highly dependent on the irrigation practice and related energy demand. Results from the present study clearly show that the waste disposal on soil causes the worst environmental performance of all the impact categories considered here. Important environmental benefits have been identified when anaerobic digestion is chosen as the final treatment. It was consequently demonstrated that anaerobic digestion should be a feasible alternative for olive mills, to produce biogas from common olive oil residues, reducing the environmental burden and adding value to the olive oil production chain.

An empirical model for the evaluation of the dissolution rate from a DNAPL-contaminated area.

This paper investigates dynamic variation in the morphologic distribution of dense non-aqueous phase liquids (DNAPLs), which take into account the coupled mass transfer. Experiments were carried out in a 2D tank representing a reconstructed aquifer model. DNAPL dissolution rates were investigated over a wide range of DNAPL saturations, several source configurations, and different hydraulic conditions. Morphometric indexes are presented that take into consideration further factors affecting the dissolution process. Local information regarding transport parameters related to the characteristics of the medium was obtained through a neural network and an optimization algorithm applied to experimental tracer tests. The history of DNAPL source architecture, in terms of saturation, indentation grade, and orientation, was determined by image analysis. Dissolved concentrations were registered and mass transfer rate coefficients were obtained for a wide range of source-zone configurations. A statistical analysis was performed to develop a constitutive equation that is descriptive of the mass transfer rate as a function of source-zone metric characteristics. A new empirical dissolution model using the proposed morphometric parameters is presented and compared with other models. The mass transfer correlation reported incorporates morphometric parameters and considers the complex and variable architecture of non-miscible contaminants. The proposed correlation can be used for an initial assessment of non-aqueous phase liquid (NAPL) dissolution rates over a wide range of saturation (residual and non-residual) conditions and different aqueous phase velocities within the NAPL source zone.

Evaluation of biomarkers in Mytilus galloprovincialis as an integrated measure of biofilm-membrane bioreactor (BF-MBR) system efficiency in mitigating the impact of oily wastewater discharge to marine environment: a microcosm approach.

The large volumes of oily wastewater discharged to marine environment cause heavy impacts on the coastal marine ecosystem. The selection of an appropriate technology to reduce these impacts should be based on the respect of the discharge limits and on the effective assessment and monitoring of its effects on biological organism preservation. To this aim, we set up a controlled microcosm-scale system to compare the effects of a treated and untreated oily wastewater discharge in which the restore process is performed through a Membrane Bio-Reactor. The system is completed by other three microcosms to control and isolate any possible concurrent effect on the Mytilus galloprovincialis, used as sentinel organism. Mytilus galloprovincialis have been kept in all these microcosms, and then mRNA expression and morphology were evaluated on gills and digestive gland. The genes considered in this work are Heat Shock Protein 70 and Metallothionein 10, involved in response to physicochemical sublethal stressors, Superoxide dismutase 1, Catalase, and Cytochrome P450 involved in oxidative stress response. Our results evidenced a significant overexpression, both in gills and digestive gland, of HSP70 in samples maintained in the microcosm receiving the untreated effluent, and of MT10 in those animals kept in microcosm where the effluent was treated. Even though the mRNA modifications are considered "primary" and transient responses which do not always correspond to protein content, the study of these modifications can help to gain insights into the mechanisms of action of xenobiotic exposure. Morphological analysis suggested that, although different, depending on the microcosm, the most serious damages were found in the gill epithelium accompanied with severe haemocyte infiltration, whilst in digestive gland the tissue architecture alterations and the haemocyte infiltration were less pronounced. These observations suggest that the immune system was activated as a general response to stressful stimuli such as the presence of toxic compounds. Moreover, the results indicate that the treatment process is useful. In fact, samples derived from the microcosm receiving the treated effluent, even though presenting some signs of stress, seemed to partially recover the normal structure, although their mRNA expression indicated some cellular suffering.

Mosquito species involved in the circulation of West Nile and Usutu viruses in Italy.

Usutu (USUV) and West Nile (WNV) are mosquito-borne Flavivirus emerged in Italy in 1996 and 1998, respectively, and reappeared 10 years later. The aim of this work is to review the Italian mosquito species found positive for WNV and USUV between 2008 and 2014. Moreover, the role of mosquitoes in promoting the overwintering of these viruses is discussed, as a result of the mosquito collections performed in Molise region between September 2010 and April 2011. Overall 99,000 mosquitoes were collected: 337 and 457 mosquito pools tested positive by real time reverse transcriptase polymerase chain reaction (real time RT-PCR) for WNV and USUV, respectively. West Nile virus was detected in pools of Culex pipiens s.l. (329), Ochlerotatus caspius (4), Culex modestus (2), and Culex spp. (2). Positive USUV pools were from Cx. pipiens s.l. (435), Aedes albopictus (12), Oc. caspius (5), Culex spp. (2), Anopheles maculipennis s.l. (1), Culiseta annulata (1), and Ochlerotatus detritus (1). In Molise region, 1,694 mosquitoes were collected, and USUV was identi ed in Cx. pipiens s.l., Cs. annulata, and Oc. detritus pools. This paper shows that Cx. pipiens s.l. is the mosquito species most involved in the WNV and USUV circulation in Italy, although other species would also support the spread of both the viruses during Winter.

Shell fluctuating asymmetry in the sea-dwelling benthic bivalve Mytilus galloprovincialis (Lamarck, 1819) as morphological markers to detect environmental chemical contamination.

Investigations on asymmetries showed that deviations from perfect bilateral symmetry are interpreted as environmental changes inducing developmental instability. Since morphological abnormalities increase with pollution, deformations may be considered indicators of the organism exposition to pollution. Therefore, the onset of asymmetry in otherwise normally symmetrical traits has been used as a measure of some stresses as well. In this context, we studied how marine pollution affects the valve morphological alterations in the mussel Mytilus galloprovincialis. We used 180 specimens (30 per site) from the aquaculture area of Goro (River Po delta, northern Adriatic Sea), translocated, and released within 50 × 50 × 50 cm cages in five sites: two disturbed and one undisturbed near Naples (eastern Tyrrhenian Sea), and one disturbed and one undisturbed near Siracusa (western Ionian Sea). Disturbed sites were stressed by heavy industrialization and heavy tankers traffic of crude and refined oil, and were defined basing on sediment contamination. In particular, by the cone-beam computed tomography we obtained 3D virtual valve surfaces to be analyzed by the geometric morphometric techniques. Specifically, we focused the levels of the shell shape fluctuating asymmetry in relation to the degrees of marine pollution in different sites of the Tyrrhenian Sea. The Mahalanobis distances (interpreted as proxy of the individual shape asymmetry deviation from the mean asymmetry) significantly regressed with the sediment contamination gradient. Indeed, although the left-right differences were normally distributed in each studied site, the individual asymmetry scores (IAS) significantly varied amongst the investigated sites. IAS showed higher values in disturbed areas than those of undisturbed ones in both Tyrrhenian and Ionian Sea. Our results are consistent with past studies on molluscans and other taxa, demonstrating some detrimental effects of chemicals on organisms, although the investigated morphological marker did not discriminate the real disturbance source. Our findings indicate that the mussels act as a prognostic tool for sea pollution levels driving detrimental effects on benthic community.

Effects of petrochemical contamination on caged marine mussels using a multi-biomarker approach: Histological changes, neurotoxicity and hypoxic stress.

This work was designed to evaluate the biological effects of petrochemical contamination on marine mussels. Mytilus galloprovincialis, widely used as sentinel organisms in biomonitoring studies, were caged at the "Augusta-Melilli-Priolo" industrial site (eastern Sicily, Italy), chosen as one of the largest petrochemical areas in Europe, and Brucoli, chosen as reference site. Chemical analyses of sediments at the polluted site revealed high levels of PAHs and mercury, exceeding the national and international guideline limits. In mussels from the polluted site, severe morphological alterations were observed in gills, mainly involved in nutrient uptake and gas exchange. Changes in serotonergic and cholinergic systems, investigated through immunohistochemical, metabolomics and enzymatic approaches, were highlighted in gills, as well as onset of hypoxic adaptive responses with up-regulation of hypoxia-inducible factor transcript. Overall, the application of a multi-biomarker panel results effective in assessing the biological effects of petrochemical contamination on the health of aquatic organisms.