Microplastics in bulk atmospheric deposition along the coastal region of Victoria Land, Antarctica.

The increasing global concern over microplastic pollution has driven a surge in research efforts aimed at detecting microplastics across various ecosystems. Airborne microplastics (MPs) have been identified in remote environments worldwide, including Antarctica. However, data on bulk atmospheric deposition remain scarce. From January to December 2020, atmospheric deposition was directly collected using passive samplers placed in eight sites across Victoria Land. Using Raman Microspectroscopy, MPs were identified in six out of the seven samples collected (one sample was lost due to the extreme weather conditions). The average daily MP deposition for Victoria Land was 1.7 ± 1.1 MPs m-2 d-1, with values ranging from 0.76 to 3.44 MPs m-2 d-1. The majority (53 %) of MPs found in the atmospheric deposition were in the size class of 5-10 µm, and the main shape of MPs was fragments (95 %). The predominant plastic type was polypropylene (31 %), followed by polyethylene (19 %) and polycarbonate (12 %). Polystyrene, polyester, styrene and polyethylene terephthalate each accounted for ~6 %. Microplastics identified in the coastal sites may have local origins, potentially associated with scientific activities at research stations. Conversely, a backward trajectories analysis suggested a potential contribution of atmospheric transport to microplastic deposition at Larsen Glacier and Tourmaline Plateau, the two most remote sites of the study area, where the highest MP concentrations were detected. Our findings present the first evidence of microplastics in the Antarctic atmospheric deposition directly collected via passive samplers, highlighting the need for continued monitoring and research to assess the environmental impact of MPs, particularly in sensitive and remote ecosystems like Antarctica.

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.

A multidisciplinary approach to study the reproductive biology of wild prawns.

This work aims to provide deeper knowledge on reproductive biology of P. kerathurus in a multidisciplinary way. Upon 789 examined females, 285 were found inseminated. The logistic equation enabled to estimate the size at first maturity at 30.7 mm CL for female. The Gono-Somatic Index (GSI) showed a pronounced seasonality, ranged from 0.80 ± 0.34 to 11.24 ± 5.72. Histological analysis highlighted five stages of ovarian development. Gonadal fatty acids analysis performed with gas chromatograph evidenced a pronounced seasonal variation; total lipids varied from 1.7% dry weight (dw) in Winter, to 7.2% dw in Summer. For the first time, a chemometric approach (Principal Component Analysis) was applied to relate GSI with total lipid content and fatty acid composition of gonads. The first two components (PC1 and PC2) showed that seasonality explained about 84% of the variability of all data set. In particular, in the period February-May, lipids were characterized by high PUFAs content, that were probably utilized during embryogenesis as energy source and as constituent of the cell membranes. During the summer season, gonads accumulated saturated FAs, that will be used during embryogenesis and early larval stages, while in the cold season total lipids decreased drastically and the gonad reached a quiescent state.

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.

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.

Evidence that total extract of Hypericum perforatum affects exploratory behavior and exerts anxiolytic effects in rats.

Clinical trials have extensively reported the ability of Hypericum perforatum extracts to exert a significant antidepressant activity. Hypericin, the main constituent of H. perforatum extract, is no more regarded as the active principle of the antidepressant activity of the drug. Hence, the question of which constituents are involved in the basic activity of the total extract, is still waiting for an answer. In the present study we focused our attention on the potential anxiolytic activity of H. perforatum total extract, and of some pure components such as protohypericin and a fraction containing hypericin and pseudohypericin. Herein we report that the total extract of H. perforatum increases the locomotor activity in the open field and exerts anxiolytic activity in the light-dark test, whereas the single components did not show any effect. Interestingly, the anxiolytic activity of the total extract was blocked by pretreatment of rats with the benzodiazepine antagonist Flumazenil, hence suggesting an implication of benzodiazepine receptor activation in the anxiolytic effect of H. perforatum extract. Electrophysiological studies, performed to gain more information on the mechanism of action, showed that hypericin reduced the GABA-activated chloride currents, while pseudohypericin did an opposite effect. Furthermore, both hypericin and pseudohypericin inhibited the activation of NMDA receptors.

Prenatal exposure to methylmercury during late gestation affects cerebral opiatergic system in rat offspring.

Pregnant female rats were orally administered a single dose (8 mg/kg) of methylmercury chloride (MMC) on Day 15 of gestation. The binding characteristics of opioid receptors were studied in the brain of developing rats at different stages of age. An increased density of opioid receptors was found in whole brain of MMC-exposed rats at 21 days (delta receptors) and 60 days (mu and delta receptors) of age, in comparison with matched controls. An enhanced response to morphine administration was detected in MMC-exposed rat offspring at Day 60 of postnatal life, which, however, was not apparently due to an impaired liver metabolization or renal excretion. Hence, it is reasonable to surmise a possible correlation between receptor up-regulation and increased response to pharmacological challenge. These data seem to indicate that neurochemical alterations produced in the rat developing organism by prenatal exposure to methylmercury involve the opiatergic system which undergoes a supersensitivity phenomenon. This effect, which is not detectable in the first postnatal period, shows a delayed onset, being detectable only at the adult stage. These findings seem to indicate that pre- and postnatal methylmercury exposure induces latent neurochemical and behavioral alterations which could last even after the clearance of the metal from the brain.

Effect of 2-methyltaurine and its enantiomers on arterial blood pressure.

Taurine has been shown to exert many biological effects. Among them, the ability of this amino acid to induce a reduction of arterial blood pressure, when administered intracerebroventricularly (i.c.v.) to mammals, has been established since a long time ago. To gain further information on the structure-activity requirements for taurine's hypotensive effect, the synthesis of 2-methyltaurine (2-aminopropanesulphonic acid) and the asymmetric synthesis of its enantiomers were performed; the enantiomeric purity of (R)- and (S)-2-methyltaurine was assayed by differential scanning calorimetry (DSC) and HPLC. The findings here reported show that the hypotensive effect elicited by the i.c.v. administration of racemate is mainly due to the (S)-enantiomer. The specificity of this effect was proved by the ability of the taurine antagonist TAG (6-aminomethyl-4H-3-methyl-1,2,4-benzothiadiazine 1,1-dioxide) to prevent it.

Increased brain concentrations of polyamines in rats with encephalopathy due to a galactosamine-induced fulminant hepatic failure.

Polyamine concentrations including putrescine, spermidine and spermine were documented in two brain areas of rats with mild and severe stages of hepatic encephalopathy (HE) due to fulminant hepatic failure induced by galactosamine HC1 injection (3 g kg-1 i.p.). In the mild stage of HE putrescine increased by 3-4 times whereas spermidine and spermine showed a slight increase. The scenario, however, was found to be changed going from the mild to the severe stage of HE, since in this last stage spermidine and spermine showed a further rise while putrescine was found to be significantly lower than in the mild stage of HE in both the brain areas studied. The changes in the ratio among the three polyamines with an enhanced prevalence in the severe stage of HE of spermidine and spermine are likely to be related to the exhaustion of the synthetic pathway of putrescine or to a reduction of the interconversion to this polyamine from spermidine and spermine. Considering that these last two polyamines potentiate the N-methyl-D-aspartate glutamate receptor mediated toxicity and that they might exert neurotoxic effects per se, there are clear reasons for suspecting an implication of the described changes of polyamines in the neurochemical mechanism which sustain HE and to surmise a potential therapeutic effect in this pathology of non-competitive antagonists of polyamine-site on N-methyl-D-aspartate glutamate receptors.

Methyl mercury during late gestation affects temporarily the development of cortical muscarinic receptors in rat offspring.

Pregnant Sprague-Dawley rats were treated by gavage with a single dose of 8 mg/kg of methyl mercury on gestational day 15. Offspring of control and treated rats were killed at 14, 21 and 60 days of age. The binding characteristics of muscarinic receptors labelled in cortical membrane preparation by 3H-L-quinuclidinyl benzilate were studied together with the assessment of mercury level in the same brain area. Furthermore, the performance in passive avoidance tasks was evaluated in 8 weeks old rats. Perinatal exposure to methyl mercury significantly reduced the maximum number of muscarinic receptors (Bmax) in the brain of 14 (53%) and 21 day old rats (21%), while this change was no more present in 60 day old rats. This phenomenon seems to be strictly related to the presence of mercury in the cortex since it disappeared with the normalization of mercury levels in the brain. Despite the recovery of muscarinic receptor densities in methyl mercury exposed rats at 8 weeks of age, the avoidance latency was reduced in passive avoidance test as an indication of learning and memory deficits in these animals. Results from this study indicate that prenatal methyl mercury exposure induces latent cognitive dysfunction which does not seem to be related to transient muscarinic receptor alteration found in the early period of postnatal life.

Synthesis and antiinflammatory activity of 2,6-bis(1,1-dimethylethyl)phenol derivatives.

The influence of 11 newly synthesized 2,6-bis(1,1-dimethylethyl)phenol derivatives substituted in the 4 position as measured on the carrageenan paw edema assay in Sprague-Dawley rats, was studied using indomethacin as a reference drug. Furthermore we studied the possible interference of few of these compounds on the calcium binding sites by using the specific ligand [3H]-PN 200-110 in "in vitro" experiments. As far as regard the antiinflammatory activity only the compounds 2b, 2j and 2k, dosed at 20 mg/Kg/os, exerted an inhibitory effect on paw edema which was practically equal, after 6 h, to that of indomethacin (approximately 30%) dosed at 2.5 mg/Kg. The compound 2k, however, showed, in comparison with indomethacin and the other new tested compounds, a longer lasting effect, reaching, after 8 h, a 56.7% inhibition of the edema. Finally the above mentioned compounds, when tested alone or in combination with nitrendipine, did not exert any displacing activity on [3H]-PN 200-110 binding to synaptosomal membranes. It is noteworthy however that compound 2e, which incidentally was inactive as antiinflammatory agent, showed a negative allosteric modulatory activity on the ability of nitrendipine to displace [3H]-PN 200-110 binding.

Urine retention due to intra-spinal cord injection of colchicine in rats: improved recovery of bladder function by monosialoganglioside GM1 and nerve growth factor administration.

Intra-spinal cord injection of a low dose of colchicine (2 micrograms/rat) at the lumbar level affects the micturition reflex leading to voiding suppression, bladder hypertrophy and overflow incontinence which lasts about four weeks. The administration of nerve growth factor and monosialoganglioside GM1 normalizes urine output within 3 days and improves recovery of the bladder contraction tested by a cystometric analysis.

Rigid analogs of taurine as potential taurine antagonists.

In this study we tested the potential taurine-antagonistic properties of three rigid analogs of taurine, 3-amino- (1), 3-hydrazino- (2) and 3-aminomethyl-1,2,4-benzothiadiazine-1,1-dioxide (3), which were prepared in our laboratory, using TAG (6-aminomethyl-3-methyl-1,2,4-benzothiadiazine-1,1- dioxide) (4), the only antagonist of taurine so far available, as reference compound in "in vivo" and "in vitro" experiments. Some physicochemical properties of (1), (2) and (3) were studied and the synthesis of TAG (4) was improved with a new preparative method. A dose-effect study performed by injecting intracerebroventricularly (1), (2) and (3) showed that these compounds have none of exciting effects exerted by the high doses of TAG (4). (1) and (3) as well as TAG (4), were found to antagonize the controlateral turning induced by the intracerebro injection of taurine and to potentiate the sedative effect of diazepam. We failed to find specific binding for taurine in different brain synaptic membrane preparations using 3H-taurine as radioligand and taurine, (1) and (3) as binding displacer. (1), (3) and TAG (4) however were found to antagonize the inhibitory effect of taurine on 3H-diazepam binding. These results seem to indicative that at least (1) and (3), which were more extensively studied than (2) because of their better solubility, are taurine antagonists with an apparent better selectivity than TAG (4).