Effects of Bacterioruberin-Rich Haloarchaeal Carotenoid Extract on the Thermal and Oxidative Stabilities of Fish Oil.

This study aimed to assess the efficacy of a bacterioruberin-rich carotenoid extract (HAE) derived from the halophilic archaea Halorubrum ezzemoulense DSM 19316 in protecting crude fish oil against thermal oxidation. The research used fish oil derived from anchovies, which had a peroxide value (PV) of 6.44 ± 0.81 meq O2 kg-1. To assess the impact of HAE on the thermal stability and post-oxidation characteristics of fish oil, several concentrations of HAE were added to the fish oil samples: 0 ppm (no additive) (HAE0), 50 ppm (HAE50), 100 ppm (HAE100), 500 ppm (HAE500), and 1000 ppm (HAE1000). Furthermore, a control group was established with the addition of 100 ppm butylated hydroxytoluene (BHT100) in order to evaluate the effectiveness of HAE with a synthetic antioxidant that is commercially available. Prior to the fast oxidation experiment, thermogravimetric analysis was conducted on samples from all experimental groups. At the conclusion of the examination, it was seen that the HAE500 and HAE1000 groups exhibited a delay in the degradation temperature. The experimental groups underwent oxidation at a temperature of 55.0 ± 0.5 °C for a duration of 96 h. The measurement of PV was conducted every 24 h during this time. PV in all experimental groups exhibited a time-dependent rise (p < 0.05). However, the HAE500 group had the lowest PV measurement at the conclusion of the 96 h period (p < 0.05). Significant disparities were detected in the fatty acid compositions of the experimental groups at the completion of the oxidation experiment. The HAE500 group exhibited the highest levels of EPA, DHA, and ΣPUFA at the end of oxidation, with statistical significance (p < 0.05). Through the examination of volatile component analysis, specifically an oxidation marker, it was shown that the HAE500 group exhibited the lowest level of volatile components (p < 0.05). Consequently, it was concluded that the addition of HAE to fish oil provided superior protection compared to BHT at an equivalent rate. Moreover, the group that used 500 ppm HAE demonstrated the highest level of performance in the investigation.

Flow cytometry and micro-Raman spectroscopy: Identification of hemocyte populations in the mussel Mytilus galloprovincialis (Bivalvia: Mytilidae) from Faro Lake and Tyrrhenian Sea (Sicily, Italy).

Immunological and structural characteristics of hemocyte populations in the mussel Mytilus galloprovincialis (Bivalvia: Mytilidae), going from two different Sicilian habitats (Faro Lake and Tyrrhenian sea), was investigated by means of two different techniques (flow cytometric and micro-Raman spectroscopy analyses). For this purpose, three hundred and sixty mussels Mytilus galloprovincialis were analyzed during November 2017. They were divided into two equal groups (triplicate sample) on the basis of the site of collection (n = 60 caught in Faro Lake - group A, and n = 60 caught in Tyrrhenian Sea - group B). Some several differences between the species of Faro Lake and Tyrrhenian Sea are observed and ascribed to the disruption of immune parameters induced by the variations of some qualitative water parameters (temperature, salinity, dissolved oxygen, pH, ammonium 10, free chlorine, total chlorine, total phosphate, orthofhosphate) recorded in the two habitats. This study is relevant for monitoring the conditions of the sea and Faro Lake, which is strongly influenced by the currents of the Tyrrhenian Sea. Faro lake is well known for the cultivation of mussels and this is part of a coastal habitat of particular interest, consisted of a peculiar biocenotic complex. Further, for the first time, significant different arrangement in the mussels cell structural organization was evidenced by simply following their highly reproducible Raman biomolecular signatures.

Automated Hematological Approach and Protein Electrophoretic Pattern in Tilapia (Oreochromis niloticus): An Innovative and Experimental Model for Aquaculture.

This study aimed to assess the usefulness of two innovative automated methods (automated blood count counters and flow cytometry) for hematological investigation in Tilapia to make a contribution to the clinical diagnostics of this farmed species. Moreover, serum total proteins and their electrophoretic fractions (prealbumin, albumin, α-, ß-, and γ-fraction), as health condition indicators, were assessed. The analysis of serum total proteins and electrophoretic fraction showed a normal and typical electrophoretic pattern of healthy fish (serum total proteins, 3.70 ± 0.62 g/dL; prealbumin, 0.44 ± 0.20 g/dL; albumin, 1.17 ± 0.66 g/dL; α-fraction, 1.49 ± 0.64 g/dL; ß-fraction, 0.32 ± 0.16 g/dL; and γ-fraction, 0.29 ± 0.13 g/dL). The relationships between the values of red blood cells (RBCs), white blood cells (WBCs), and thrombocytes (TCs) obtained with the two automated methods were determined using Pearson correlation analysis. The results showed a significant positive correlation between automatic blood cell counting and flow cytometry analysis for RBCs (r = 0.97, p < 0.0001) and WBCs (r = 0.91, p < 0.0001), whereas no correlation was found for TCs (r = -0.11, p = 0.66). The preliminary results gathered in this study seem to highlight the usefulness of the new analytical techniques herein investigated in tilapia, suggesting their application in the hematological investigation of farmed fish species and their usefulness for monitoring the health and well-being of fish reared in aquaculture.

Heavy Metals Bioaccumulation in Mytilus galloprovincialis and Tapes decussatus from Faro Lake (Messina), Italy.

The aim of this study was to evaluate the potential bioaccumulation of arsenic (As), copper (Cu), zinc (Zn), cadmium (Cd), lead (Pb) and mercury (Hg) in the haemolymph and corpus of Mytilus galloprovincialis and Tapes decussatus from Lake Faro. The lake is particularly prone to the accumulation of substances that are potentially toxic to aquatic organisms, due to the input of pollutants from urban and agricultural sources and the low rate of water exchange. The combination of saltwater from the Tyrrhenian Sea, the Strait of Messina and freshwater from hilly aquifers has created brackish conditions in the lake, resulting in an area of high commercial shellfish productivity. As, Cd, Cu, Pb and Zn were determined using a single quadrupole inductively coupled plasma mass spectrometer; Hg was determined using a direct mercury analyser (DMA-80). Physicochemical parameters of the water from Lake Faro were also performed. Statistical analysis was carried out using GraphPad Prism 9.0 (GraphPad Software, Inc., Boston, MA, USA) and Shapiro-Wilk normality was applied. Concentrations of Cd, Hg and Pb below the permitted MRLs in Mytilus galloprovincialis and Tapes decussatus used as ''biological indicators'' show that Lake Faro is not at risk of contamination by these pollutants and, moreover, is free of health problems for the consumer based on regulatory limits.

Biological responses of juvenile European sea bass (Dicentrarchus labrax) exposed to contaminated sediments.

Multiple anthropogenic activities present along coastal environments may affect the health status of aquatic ecosystems. In this study, specimens of European sea bass (Dicentrarchus labrax) were exposed for 30 days to highly contaminated sediment collected from the industrial area between Augusta and Priolo (Syracuse, Italy), defined as the most mercury polluted site in the Mediterranean. The aim was to evaluate the responses of juvenile D. labrax to highly contaminated sediments, particularly enriched in Hg, in order to enhance the scarce knowledge on the potential compensatory mechanisms developed by organisms under severe stress conditions. Apoptotic and proliferative activities [cell turnover: Proliferating Cell Nuclear Antigen (PCNA) and FAS Ligand (FasL)], onset of hypoxic condition [hypoxia: Hypoxia Inducibile Factor-1α (HIF-1α)], and changes in the neuroendocrine control mechanisms [neurotransmission: Tyrosine Hydroxylase (TH), Choline Acetyltransferase (ChAT), Acetylcholinesterase (AChE), 5-Hydroxytryptamine (5-HT) and 5-Hydroxytryptamine receptor 3 (5-HT3)] were investigated in sea bass gill tissues. In the specimens exposed to the polluted sediment, the occurrence of altered cell turnover may result in impaired gas exchange that leads to a condition of "functional hypoxia". Changes in neurotransmission pathways were also observed, suggesting a remodeling process as an adaptive response to increase the O2-carrying capacity and restore the normal physiological conditions of the gills. Overall, these findings demonstrated that although chronic exposure to heavy metal polluted sediments alters the functioning of both the nervous and endocrine systems, as well as plasticity of the gill epithelium, fish are able to trigger a series of physiological adjustments or adaptations interfering with specific neuroendocrine control mechanisms that enable their long-term survival.

Effectiveness and safety of secukinumab in ankylosing spondylitis and psoriatic arthritis: a 52-week real-life study in an Italian cohort.

BACKGROUND: Secukinumab has shown high efficacy in randomized controlled trials in both ankylosing spondylitis (AS) and psoriatic arthritis (PsA). Here, we investigated its real-life effectiveness and tolerability in a cohort of AS and PsA patients. METHODS: We retrospectively analyzed medical records of outpatients with AS or PsA treated with secukinumab between December 2017 and December 2019. ASDAS-CRP and DAS28-CRP scores were used to measure axial and peripheral disease activity in AS and PsA, respectively. Data were collected at baseline and after 8, 24, and 52 weeks of treatment. RESULTS: Eighty-five adult patients with active disease (29 with AS and 56 with PsA; 23 males and 62 females) were treated. Overall, mean disease duration was 6.7 years and biologic-naïve patients were 85%. Significant reductions in ASDAS-CRP and DAS28-CRP were observed at all time-points. Body weight (in AS) and disease activity status at baseline (particularly in PsA) significantly affected disease activity changes. ASDAS-defined inactive disease and DAS28-defined remission were achieved in comparable proportions between AS and PsA patients, at both 24 weeks (45% and 46%) and 52 weeks (65.5% and 68%, respectively); male sex was found an independent predictor of positive response (OR 5.16, P = 0.027). After 52 weeks, achievement of at least low disease activity and drug retention were observed in 75% of patients. Secukinumab was well-tolerated and only mild injection-site reactions were recorded in 4 patients. CONCLUSION: In a real-world setting, secukinumab confirmed great effectiveness and safety in both AS and PsA patients. The influence of gender on treatment response deserves further attention.

Seasonal Variations in the Trace Elements and Mineral Profiles of the Bivalve Species, Mytilus galloprovincialis, Chamelea gallina and Donax trunculus, and Human Health Risk Assessment.

This study aimed to provide data on selected toxic (Cd, Pb and Ni), essential (Cr, Cu, Fe, Mn and Zn) and microelement (Na, K, Ca and Mg) concentrations in edible tissues of the Mediterranean mussel (Mytilus galloprovincialis), striped venus clam (Chamelea gallina) and the wedge clam (Donax trunculus). Samples were collected from the Black Sea (Bulgaria) four times over, a period of one year (2022). In comparison with the maximum permissible levels set by the EU and USFDA, all elemental concentration found in the bivalve species were lower than the prescribed limits. An estimation of the dietary metal intake through calculation of the target hazard quotients (THQ), hazard index (HI) and target risk (TR) was performed. The target hazard quotient (THQ) for individual metal and HI for combined metals were lower than 1, indicating no health risk for consumers due to the intake of either individual element or combined ones. The target risk value for toxic inorganic Pb and Cr was below 10-6, indicating no carcinogenic risk. According to these results, the consumption of these bivalve species is completely safe for human health.

Toxic metals and total lipids comparison between wild and farmed fish of South mediterranean.

A total of 151 wild and farmed fish samples of three species (S. aurata, D. labrax and U. cirrosa) from south Mediterranean were examined for cadmium, lead and mercury and total lipids detection to verify possible differences between wild and farmed fish. Mercury was detected only in S. aurata samples, with mean values of 0.056 ± 0.128 mg/kg and 0.031 ± 0.033 mg/kg for farmed and wild samples, respectively. The results verified no significant differences in Pb, Cd and Hg levels between wild and farmed fish samples (p < 0.05). The farmed fish samples showed higher total lipids contents than wild (p > 0.05). As far as we know, this work report first findings on the toxic metal presence in farmed Umbrina cirrosa, showing no detectable Hg, Cd and Pb values. The results of this work seems to confirm that the presence of toxic metals could be more related to the pollution the site than the type of production.

Metabolomic investigation of Mytilus galloprovincialis (Lamarck 1819) caged in aquatic environments.

Environmental metabolomics was applied to assess the metabolic responses in transplanted mussels to environmental pollution. Specimens of Mytilus galloprovincialis, sedentary filter-feeders, were caged in anthropogenic-impacted and reference sites along the Augusta coastline (Sicily, Italy). Chemical analysis revealed increased levels of PAHs in the digestive gland of mussels from the industrial area compared with control, and marked morphological changes were also observed. Digestive gland metabolic profiles, obtained by 1H NMR spectroscopy and analyzed by multivariate statistics, showed changes in metabolites involved in energy metabolism. Specifically, changes in lactate and acetoacetate could indicate increased anaerobic fermentation and alteration in lipid metabolism, respectively, suggesting that the mussels transplanted to the contaminated field site were suffering from adverse environmental condition. The NMR-based environmental metabolomics applied in this study results thus in it being a useful and effective tool for assessing environmental influences on the health status of aquatic organisms.

Intra-Decadal (2012-2021) Dynamics of Spatial Ichthyoplankton Distribution in Sevastopol Bay (Black Sea) Affected by Hydrometeorological Factors.

High mortality of fish in the early stages of ontogenesis requires the detection of factors affecting it and is an extremely important problem. For this reason, we tried to consider the influence of some hydrometeorological factors on the intra-decadal dynamics and spatial distribution of ichthyoplankton abundance in Sevastopol Bay. To this end, we analyzed ichthyoplankton samples collected in 2012−2021 in Sevastopol Bay, and the four districts were identified. The ichthyoplankton distribution by areas was associated with the peculiarities of the pollution distribution. The maximum numbers in eggs (34.7 ± 7.5 ind. m−2) and larvae (10.0 ± 2.4 ind. m−2) were recorded in July−August. For the first time, a positive effect of temperature and repeatability of north and northeast winds on the ichthyoplankton number during the summer spawning in all identified areas of Sevastopol Bay using principal component analysis (PCA) was reported. In the southern region, positive influence of temperature and northerly winds on the ichthyoplankton abundance was confirmed by ANOVA (p = 0.02), and Pearson's correlation analysis revealed significant correlations between these parameters on inter-annual scale (r > 0.5). In the eastern area, the discharge of rivers is an additional factor affecting the abundance of ichthyoplankton, while in the western and central areas, these factors were the wind characteristics caused by the orography peculiarities. The pollution spread in Sevastopol Bay is also largely due to the wind regime. The importance of comprehensive monitoring studies using intra-decadal data on ichthyoplankton is emphasized.

Comparison of cellular mechanisms induced by pharmaceutical exposure to caffeine and its combination with salicylic acid in mussel Mytilus galloprovincialis.

Urban and hospital-sourced pharmaceuticals are continuously discharged into aquatic environments, threatening biota. To date, their impact as single compounds has been widely investigated, whereas few information exists on their effects as mixtures. We assessed the time-dependent biological impact induced by environmental concentrations of caffeine alone (CAF; 5 ng/L to 10 µg/L) and its combination with salicylic acid (CAF+SA; 5 ng/L+0.05 µg/L to 10 µg/L+100 µg/L) on gills of mussel Mytilus galloprovincialis during a 12-day exposure. Although no histological alteration was observed in mussel gills, haemocyte infiltration was noticed at T12 following CAF+SA exposure, as confirmed by flow cytometry with increased hyalinocytes. Both the treatments induced lipid peroxidation and cholinergic neurotoxicity, which the antioxidant system was unable to counteract. We have highlighted the biological risks posed by pharmaceuticals on biota under environmental scenarios, contributing to the enhancement of ecopharmacovigilance programmes and amelioration of the efficacy of wastewater treatment plants.

Histological endpoints and oxidative stress transcriptional responses in the Mediterranean mussel Mytilus galloprovincialis exposed to realistic doses of salicylic acid.

Despite the availability of analytic data, little is known about the toxicity of salicylic acid (SA) on aquatic non-target organisms. The present study aimed at evaluating the impact of SA through a short-term exposure of the Mediterranean mussel Mytilus galloprovincialis to five environmentally relevant concentrations of SA. A set of suitable biomarkers was applied at selected time-points on mussel digestive glands, including histological observations and expression of oxidative stress related genes. The obtained results showed a conspicuous hemocytic infiltration among mussel digestive tubules, as confirmed also by a flow cytometric approach that revealed an increase of halinocytes and granulocytes. Interestingly, a significant dose and time dependent decrease in the expression levels of oxidative stress related genes was found in mussels exposed to SA except for the glutathione S-transferase gene that was significantly up-regulated in a time-dependent manner confirming its important role against oxidant species and in the metabolism of pharmaceuticals.

Determination of the Heavy Metal Bioaccumulation Patterns in Muscles of Two Species of Mullets from the Southern Caspian Sea.

Although fish is a food that supplies nutrients of a high biological value, they can also be a source of some harmful substances, such as heavy metals. In the same context, some human activities in the Caspian Sea have contaminated this ecosystem during the past few years. For those reasons, our objective consisted of determining the concentrations of heavy metals and evaluating their bioaccumulation patterns in the different types of musculature in two species of mullets of commercial interest, Chelon auratus and Chelon saliens, from the southern coast of this sea. For this purpose, 20 C. auratus and 29 C. saliens were caught off this coastline and the metal concentrations in 3 different muscle locations were analyzed: the ventral, dorsal and caudal muscles of each fish. The caudal muscle had higher concentrations of Cd, Cu, Pb, and Zn, whereas As, Hg and Ni accumulation seemed to be independent of the musculature type. Overall, the Cd, Hg, and Pb concentrations exceeded the maximum levels permitted in fish by the European Union. In addition, the relationships between pairs of metals were positive and elevated in all the cases, which could be a sign of heavy metal pollution in the region sampled. Therefore, it will be necessary to continue monitoring and evaluating the degree of pollution in the Caspian Sea.

Risk Assessment of Essential and Toxic Elements in Freshwater Fish Species from Lakes near Black Sea, Bulgaria.

The aims of this study were to measure the concentrations of selected toxic and essential elements in the muscle tissue of five common freshwater fish species ((roach (Rutilus rutilus), freshwater bream (Abramis brama), prussian carp (Carassius gibelio), crucian carp (Carassius carassius) and common carp (Cyprinus carpio)) from Lake Burgas and Lake Mandra (Bulgaria). In all samples the levels of Cd, Cr, Cu, Mn, Ni, Pb, Fe and Zn were under the maximum allowed concentrations for safe human consumption in Bulgaria and ranged as follows: Cd 0.02-0.05; Cr 0.03-0.06; Cu 0.11-0.20; Mn 0.05-0.71; Ni 0.06-0.11; Pb 0.15-0.27, Fe 1.68-5.86 and Zn 1.94-9.06 mg/kg wet weight. The concentration of As was under detection limit. An assessment of the human risk by calculation of the target hazard quotients (THQ), hazard index (HI) and target risk (TR) was performed. The target hazard quotient (THQ) for individual elements and HI for combined metals were lower than 1, indicating no health risk for consumers due to the intake of either individual or combined metals. The target risk for iAs, Pb and Ni was below 10-6, indicating no carcinogenic risk. According to these results, the consumption of these freshwater fish species is safe for human health.

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.

Trace Elements and Omega-3 Fatty Acids of Wild and Farmed Mussels (Mytilus galloprovincialis) Consumed in Bulgaria: Human Health Risks.

The unique, closed ecosystem of the Black Sea is of significant global importance. The levels and health risk of some trace elements (As, Cd, Cr, Cu, Fe, Ni, Pb and Zn) in wild and farmed mussels (Mytilus galloprovincialis) collected from the Bulgarian part of the Black Sea were determined and using different approaches such as Estimated Daily Intake (EDI), Target Hazard Quotient (THQ), Hazard Index (HI), Target risk (TR), human health risk levels were assessed. The mean maximum concentrations of the elements Cd, Cr, Cu, Fe, Ni, Pb and Zn in all mussel samples were below the maximum permissible limits (MPLs) except that which exceeded the limit of 2.00 mg/kg ww. Eicosapentaenoic (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) were the major polyunsaturated fatty acids. The fatty acids profile studied mussels showed that the farmed mussels had higher PUFA/SFA ratios, DHA and EPA + DHA content and lower SFA, AI and TI values. The target risk (TR) values for Pb, Cr, Ni and As were calculated, evaluated and showed acceptable or negligible levels. Target hazard quotients (THQs) and hazard index (HI) from elemental intake were below 1 indicated no hazard from consumption. The benefit-risk ratio indicated that wild and cultured M. galloprovincialis are safe for human consumption.

Effects of Essential Oil Derived from the Bitter Orange (Citrus aurantium) on Growth Performance, Histology and Gene Expression Levels in Common Carp Juveniles (Cyprinus carpio).

The aim of this study was to detect effects of bitter orange (Citrus aurantium) essential oil, commonly called neroli oil (NO) (0, 0.25, 0.50, 1, and 1.5% referred to as NO0 NO0.25, NO0. 05, NO1 and NO1.5, respectively) on growth performance output and expression levels of some growth-related genes in the muscle tissue and some immune-related genes in the head kidney and pathological differences in digestive system organs of common carp Cyprinus carpio. The NO0.25 group had a large improvement in growth efficiency at the end of the 60-day feeding cycle. Real-time PCR (Bio RAD, USA) system was used to detect variations in gene expression levels. Furthermore, NO supplementation of up to 0.25% in muscle tissue controlled the release of growth hormone (GH) and insulin-like growth factor I (IGF-I). Furthermore, in the NO0.25 treatment category, immune response gene levels TNF-α, IL-8 and IL-1ß increased in head kidney tissue. In the histological examination of the liver and intestine, there were significant differences between fish fed with N1 and N1.5 diets. This study confirms that dietary supplementation of NO up to 0.25% can improve common carp growth efficiency and increase the expression of genes (GH and IGF-I) related to muscle growth, TNF-α, IL-8 and IL-1ß genes related to immune status, and liver and intestine histological status of common carp.

Trace elements (Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) in Mytilus galloprovincialis and Tapes decussatus from Faro and Ganzirri Lakes (Sicily, Italy): Flow cytometry applied for hemocytes analysis.

BACKGROUND: Trace elements present in sessile molluscs, are important because they are used in human consumption and it has significantly increased in recent years. While their filtering of the water can lead to their build-up of organic and inorganic materials that can be sampled and analyzed, this can also lead to bioaccumulation of harmful substances, such as essential and non-essential elements, that can harm the human health if in taken in high concentrations or for a long period of time. METHODS: In the present study, the trace metal content (Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) of two sessile crustaceans, 20 Mytilus galloprovincialis (mussel) and 20 Tapes decussatus (clam) in Faro and Ganzirri Lakes (Messina, Sicily, Italy) were analyzed. Haemolymph samples were taken on both molluscs in order to analyze the haemocyte population by flow cytometric analysis. Unpaired t-tests were used to determine significant differences for the essential and non-essential metallic elements concentrations in the lake waters and in the tissues of M. galloprovincialis and T. decussatus and for hemocyte populations R1 (halinocytes) and R2 (granulocytes). RESULTS: The results suggested that that in Faro Lake, the tissue Al, Cr and Pb levels in M. galloprovincialis were higher than those for T. decussatus, in contrast to Mn, Fe, Ni, Cu, Zn and Cd, which were higher in T. decussatus. Unpaired t-tests showed that there were significantly higher proportions of halinocytes in M. galloprovincialis versus T. decussatus for both Faro Lake (41.8 % vs. 24.3 %; P < 0.001) and Ganzirri Lake (43.0 % vs. 22.4 %; P < 0.001). In contrast, while there were significantly higher proportions of granulocytes in Faro Lake (21.2 % vs. 9.1 %; P < 0.001), this difference was not seen for the granulocytes of M. galloprovincialis versus T. decussatus in Ganzirri Lake (9.6 % vs. 13.0 %). CONCLUSION: This study shows that M. galloprovincialis and T. decussatus can indeed bioaccumulate some of these metal, such that activation of the immune responses is specific to certain cell types. Future research must focus on the balance of trace elements in the consumption of these shellfish, and analyzes with more sophisticated tools can be used to diagnose the increased concentration of trace elements and the quantification of trace metals from shellfish to clams.

Immunohistochemical and molecular biomarkers in Coris julis exposed to environmental contaminants.

When a contaminant interacts with biotic components of a marine ecosystem, it causes a series of changes that can compromise an entire community (Stebbing, 1985). This present study wants to focus on changes in the gills of a bioindicator benthic organism, Coris julis, collected in Milazzo (Messina, Italy), characterized by a strong anthropical impact), compared with individuals from the control site (Marinello, Messina). RT-PCR has been used for both MT and HSP70, and the respective mRNAs have been visualized by FISH. MT and HSP70 expression levels increased in individuals collected in Milazzo. The presence of numerous apoptotic and proliferating cells and the analysis of several neuronal markers by immunohistochemical method give information about the adaptation to a heavy metal mixture. The obtained results show that, in stressed fishes, defensive processes increase to maintain the normal functions of the organs more exposed to the action of polluted substances.

A multibiomarker approach in Coris julis living in a natural environment.

To monitor the health of aquatic organisms, biomarkers have been used as effective tools in assessing environmental risk. In this study was examined the teleost Coris julis, sampled in two marine sites in Messina (Italy) at different pollution degree, Milazzo, characterized by a strong anthropogenic impact, and Marinello, the natural reserve. C. julis is a species particularly suitable to biomonitoring because its feeding habits favor bio-accumulation of xenobiotics. The following biomarkers were used to estimate the impact of highly persistent pollutants: cellular localization of cytochrome P4501A (CYP1A) and glutathione-S-transferase (GST) in the liver, their hepatic expression at the mRNA level, the enzymatic activity (EROD and BPMO), the micronucleus and comet assays in the blood, esterases (AChE in the brain and BChE in the blood) activity and evaluation of PAH metabolites in the bile. The present findings provide evidence of statistically significant differences in parameters between individuals collected in two sites.