Effects of biological factors and seasonality on the level of polycyclic aromatic hydrocarbons in red mullet (Mullus barbatus).

This study evaluates the effects of biological factors of fish and seasonality on Polycyclic Aromatic Hydrocarbon (PAH) accumulation in red mullet (Mullus barbatus) tissue. Specimens were collected monthly with a bottom trawl net in an offshore fishing ground in the Northern and Central Adriatic Sea (Geographical Sub Area 17) throughout 2016. The edible fillets of 380 individuals were analyzed for the concentrations of individual PAH, total PAH, and low, medium and high molecular weight (MW) PAHs. PAH bioaccumulation was related to their physicochemical characteristics (MW, and logarithm of the octanol-water partition coefficient, log Kow), some biological parameters of fish (body size, age, sex, reproductive stage and total lipid content), and catch season. The PAH bioaccumulation pattern and the effects of the different factors varied according to PAH MW. The heavier (medium and high MW) PAHs showed higher levels in winter-autumn and in pre-spawners compared with spawners and post-spawners. Our findings suggest that an important detoxification mechanism, albeit limited to the heavier PAHs, acts in the spawning and post-spawning stage. Low MW PAHs appeared to be unaffected by reproductive stage, lipid content and seasonality. Reproductive stage and seasonality seem to play an important role in the accumulation of heavier PAH, whereas total lipid content and age seem to exert a limited influence, and body size no effect at all.

Heat shock influences the fatty acid composition of the muscle of the Antarctic fish Trematomus bernacchii.

In the Ross Sea region (average temperature of -1.87 °C), shelf water warming up to +0.8-+1.4 °C is predicted by 2200, so there is an urgent need to understand how organisms can respond to rising temperatures. In this study, we analyzed the effect of a heat shock on the fatty acid (FAs) composition of muscle of the Antarctic teleost Trematomus bernacchii, caught in Terra Nova Bay (Ross Sea), and held in fish tanks at 0, +1 or +2 °C, for 1, 5 and 10 days. In general, heat shock produced, beyond a reduction in total lipid content correlated to the temperature, an increase in the percentage of saturated FAs, and a decrease in mono-unsaturated FAs; however, the level of poly-unsaturated FAs did not seem to directly correlate with temperature. Principal component analysis indicated that both temperature and exposure time affect the composition of FAs in the muscle probably through an alteration of the metabolic pathways of FAs. In this study, we demonstrated that T. bernacchii was capable to rapidly acclimatize to a heat shock. This study contributes to increasing knowledge on the effect of temperature on the lipid composition of T. bernacchii and is complementary to previous studies on the gene expression and biochemistry of this species face multiple stressors.

Distribution of Cd, Pb and Cu between dissolved fraction, inorganic particulate and phytoplankton in seawater of Terra Nova Bay (Ross Sea, Antarctica) during austral summer 2011-12.

During the austral summer 2011-2012, the metal quotas of Cd, Pb and Cu in the phytoplankton of Terra Nova Bay (TNB, Antarctica) were measured for the first time. Evolution of all the three metal distributions between dissolved and particulate fractions during the season was also evaluated. Metal concentrations were mainly affected by the dynamic of the pack ice melting and phytoplankton activity. In mid-December when TNB area was covered by a thick pack ice layer and phytoplankton activity was very low, all the three metals were present mainly in their dissolved species. When the pack ice started to melt and the water column characteristics became ideal (i.e. moderate stratification, ice free area), the phytoplankton bloom occurred. Cd showed a nutrient-type behaviour with dissolved and particulate fractions mainly influenced by phytoplankton activity. Cd quota showed a mean value of 0.12 ± 0.07 nmol L-1 (30-100% of the total particulate). Also Cu showed a nutrient-type behaviour, with its quota in phytoplankton varying between 0.08 and 2.1 nmol L-1 (20-100% of the total particulate). Pb features the typical distribution of a scavenged element with very low algal content (0.03 ± 0.02 nmol L-1, representing 20-50% of the total particulate). The vertical distribution of this element was influenced by several factors (e.g. pack ice melting, atmospheric inputs), the phytoplankton activity affecting Pb behaviour only partially. Metal:C ratios provide valuable information on the biological requirements for Cd, Pb and Cu, leading us to better understand their biogeochemical cycles.

Quantification of fatty acids in the muscle of Antarctic fish Trematomus bernacchii by gas chromatography-mass spectrometry: Optimization of the analytical methodology.

This work presents data on the quantification of fatty acids (FAs, in terms of mass unit per tissue weight) in the muscle of Trematomus bernacchii, a key species in Antarctica, often used as bioindicator for contamination studies. Modifications in fatty acids content should be considered a useful biomarker to study how contaminants affect Antarctic biota. Until now, very few studies quantified fatty acids of muscle of T. bernacchii, and only as percentage of a single fatty acid on total lipids. To perform the quantification of fatty acids, we used an analytical method based on a fast microwave-assisted extraction of lipids from a lyophilized sample, a base-catalyzed trans-esterification of lipid extract to obtain Fatty Acids Methyl Esters (FAMEs), and a separation and identification of FAMEs by gas chromatography-mass spectrometry. With the optimized and validated method, a fast and accurate separation of Fatty Acids Methyl Esters was performed in 43 min. The linearity was checked up to about 320 µg mL-1; limit of detection and limit of quantification are in the range 4-22 µg mL-1 and 13-66 µg mL-1, respectively. The optimized method showed a good accuracy and precision. Major fatty acids were 14:0, 16:0, 16:1n7, 18:1n9, 18:1n7, 20:1n9, 20:5n3 and 22:6n3. Quantified FAs compute for about 47 mg g-1 tissue dry weight (dw), with 9.1 ± 0.1 mg g-1 dw of saturated FAs, 25.5 ± 0.1 mg g-1 dw of mono-unsaturated FAs, and 12.2 ± 0.1 mg g-1 dw of poly-unsaturated FAs.

Heavy metal distribution in organic and siliceous marine sponge tissues measured by square wave anodic stripping voltammetry.

May sponge spicules represent a "tank" to accumulate heavy metals? In this study we test this hypothesis determining the distribution of Cd, Pb and Cu concentrations between organic and siliceous tissues in Antarctic Demospongia (Sphaerotylus antarcticus, Kirkpatrikia coulmani and Haliclona sp.) and in the Mediterranean species Petrosia ficiformis. Results show that although, in these sponges, spicules represent about 80% of the mass content, the accumulation of pollutant is lower in the spicules than in the corresponding organic fraction. The contribution of tissues to the total sponge content of Cd, Pb and Cu is respectively 99%, 82% and 97% for Antarctic sponges and 96%, 95% and 96% for P. ficiformis, similar in polar and temperate organisms. These results pave the way to a better understanding of the role of marine sponges in uptaking heavy metals and to their possible use as monitor of marine ecosystems, recommend by the Water Framework Directive.

Square-wave anodic-stripping voltammetric determination of Cd, Pb, and Cu in a hydrofluoric acid solution of siliceous spicules of marine sponges (from the Ligurian Sea, Italy, and the Ross Sea, Antarctica).

Square-wave anodic-stripping voltammetry (SWASV) was set up and optimized for simultaneous determination of cadmium, lead, and copper in siliceous spicules of marine sponges, directly in the hydrofluoric acid solution (approximately 0.55 mol L(-1) HF, pH approximately 1.9). A thin mercury-film electrode (TMFE) plated on to an HF-resistant epoxy-impregnated graphite rotating-disc support was used. The optimum experimental conditions, evaluated also in terms of the signal-to-noise ratio, were as follows: deposition potential -1100 mV vs. Ag/AgCl, KCl 3 mol L(-1), deposition time 3-10 min, electrode rotation 3000 rpm, SW scan from -1100 mV to +100 mV, SW pulse amplitude 25 mV, frequency 100 Hz, DeltaE(step) 8 mV, t(step) 100 ms, t(wait) 60 ms, t(delay) 2 ms, t(meas) 3 ms. Under these conditions the metal peak potentials were Cd -654 +/- 1 mV, Pb -458 +/- 1 mV, Cu -198 +/- 1 mV. The electrochemical behaviour was reversible for Pb, quasi-reversible for Cd, and kinetically controlled (possibly following chemical reaction) for Cu. The linearity of the response with concentration was verified up to approximately 4 microg L(-1) for Cd and Pb and approximately 20 microg L(-1) for Cu. The detection limits were 5.8 ng L(-1), 3.6 ng L(-1), and 4.3 ng L(-1) for Cd, Pb, and Cu, respectively, with t(d) = 5 min. The method was applied for determination of the metals in spicules of two specimens of marine sponges (Demosponges) from the Portofino natural reserve (Ligurian Sea, Italy, Petrosia ficiformis) and Terra Nova Bay (Ross Sea, Antarctica, Sphaerotylus antarcticus). The metal contents varied from tens of ng g(-1) to approximately 1 microg g(-1), depending on the metal considered and with significant differences between the two sponge species.

RelA/NF-kappaB recruitment on the bax gene promoter antagonizes p73-dependent apoptosis in costimulated T cells.

The balance between antiapoptotic and proapoptotic proteins of the Bcl-2 family is critical in determining the fate of T cells in response to death stimuli. Proapoptotic genes, such as bax, are generally regulated by the p53 family of transcription factors, whereas NF-kappaB subunits can activate the transcription of antiapoptotic Bcl-2 members. Here, we show that CD28 activation protects memory T cells from irradiation-induced apoptosis by both upregulating bcl-xL and inhibiting bax gene expression. We found that p73, but not p53, binds to and trans-activates the bax gene promoter in irradiated T cells. The activation of RelA/NF-kappaB subunit in CD28 costimulated T cells and its binding onto the bax gene promoter results in suppression of bax transcription and decrease in both p73 and RNA polymerase II recruitment in vivo. RelA recruitment on the bax gene promoter is also accompanied by the lost of p300 binding and the parallel appearance of histone deacetylase-1-containing complexes. These findings identify RelA/NF-kappaB as a critical regulator of T-cell survival by affecting the balance of Bcl-2 family members.

Determination of water-soluble and insoluble (dilute-HCl-extractable) fractions of Cd, Pb and Cu in Antarctic aerosol by square wave anodic stripping voltammetry: distribution and summer seasonal evolution at Terra Nova Bay (Victoria Land).

Eight PM10 aerosol samples were collected in the vicinity of the "Mario Zucchelli" Italian Antarctic Station (formerly Terra Nova Bay Station) during the 2000-2001 austral summer using a high-volume sampler and precleaned cellulose filters. The aerosol mass was determined by differential weighing of filters carried out in a clean chemistry laboratory under controlled temperature and humidity. A two-step sequential extraction procedure was used to separate the water-soluble and the insoluble (dilute-HCl-extractable) fractions. Cd, Pb and Cu were determined in the two fractions using an ultrasensitive square wave anodic stripping voltammetric (SWASV) procedure set up for and applied to aerosol samples for the first time. Total extractable metals showed maxima at midsummer for Cd and Pb and a less clear trend for Cu. In particular, particulate metal concentrations ranged as follows: Cd 0.84-9.2 microg g(-1) (average 4.7 microg g(-1)), Pb 13.2-81 microg g(-1) (average 33 microg g(-1)), Cu 126-628 microg g(-1) (average 378 microg g(-1)). In terms of atmospheric concentration, the values were: Cd 0.55-6.3 pg m(-3) (average 3.4 pg m(-3)), Pb 8.7-48 pg m(-3) (average 24 pg m(-3)), Cu 75-365 pg m(-3) (average 266 pg m(-3)). At the beginning of the season the three metals appear widely distributed in the insoluble (HCl-extractable) fraction (higher proportions for Cd and Pb, 90-100%, and lower for Cu, 70-90%) with maxima in the second half of December. The soluble fraction then increases, and at the end of the season Cd and Pb are approximately equidistributed between the two fractions, while for Cu the soluble fraction reaches its maximum level of 36%. Practically negligible contributions are estimated for crustal and sea-spray sources. Low but significant volcanic contributions are estimated for Cd and Pb (approximately 10% and approximately 5%, respectively), while there is an evident although not quantified marine biogenic source, at least for Cd. The estimated natural contributions (possibly including the marine biogenic source) cannot account for the high fractions of the metal contents, particularly for Pb and Cu, and this suggests that pollution from long-range transport is the dominant source.

Effects of cadmium exposure on testis apoptosis in the marine teleost Gobius niger.

It is known that heavy metals can accumulate in tissues during aquatic organism growth (bioaccumulation) and often biomagnify up the food chain interfering with the health and reproduction of both wildlife and humans. Recently, cadmium (Cd) was included in the endocrine disruptors list, exerting its effect on gametes quality and reproductive functions; in addition, its role as apoptotic factor was evidenced in different cell types and tissues. In the present study, the effects of two different Cd doses on testis and liver of the black goby Gobius niger were analyzed. Cd concentration in the water and its uptake by the gills were measured by differential pulse anodic stripping voltammetry. Toxic, apoptotic, and stressor Cd effects were analyzed using metallothionein (MTT), caspase 3 and heath shock protein 70 (HSP70), respectively, as bioindicators. The results of the present study suggested that, in the gills, the saturation of all specific metal sites was reached only with the highest Cd dose exposure. Either testis and liver showed an increase of MTT gene expression and protein synthesis in addition to HSP70 gene expression, related with Cd concentration in the water indicating that both tissues were affected by Cd exposure. In conclusion, the present study, not only shows the toxic effect of Cd on hepatic tissue, but also indicates its potency as apoptotic factor in the testis. This is supported by the increase of caspase 3 gene expression and the presence of its active form in testis of exposed fish.