Evidence of small microplastics (<100 µm) ingestion by Pacific oysters (Crassostrea gigas): A novel method of extraction, purification, and analysis using Micro-FTIR.

Microplastics (MPs) are present in fresh, brackish, or marine waters. Micro- and macroinvertebrates can mistake MPs or small microplastics (SMPs, <100 µm) to be food particles and easily ingest them according to the size of their mouthparts. SMPs may then block the passage of food through the intestinal tract (i.e. hepatopancreas), accumulate within the organism, and enter the food web. Pacific oysters (Crassostrea gigas) are allochthonous filter-feeding bivalve mollusks, which have been introduced in coastal seas around the world in both natural banks and farms. Considering their economic and ecological value, these bivalves have been chosen as a model to study the ingestion of SMPs. A novel method for the extraction and purification of SMPs in bivalves was developed. Quantification and simultaneous polymer identification of SMPs using Micro-FTIR (Fourier Transform Infrared Spectroscopy) were performed, with a limit of detection for the particle size of 5 µm.

Domoic acid at trace levels in lagoon waters: assessment of a method using internal standard quantification.

Domoic acid (DA) is a neurotoxin produced by different algae, including pennate diatoms, principally from the genus Pseudo-nitzschia, and it is the main cause of amnesic shellfish poisoning. Determination of this toxin in seawater samples is fundamental to define the real contamination risks for aquatic species. We have developed two very sensitive instrumental methods using hydrophilic interaction liquid chromatography coupled using tandem mass spectrometry in positive and negative polarity modes. Instrumental detection limits were 9 pg mL(-1) for positive and 19 pg mL(-1) for negative ionisation. A procedural method based on solid-phase extraction for the determination of dissolved DA present in seawater has been developed, and an extraction procedure was employed for the determination of the toxin in the particulate fraction. DA quantification was performed using the internal standard method to account for signals fluctuations and random errors during sample treatment. To our knowledge, this is the first study to use this quantification method for DA determination. Trueness, extraction yield, matrix effects, repeatability and procedural detection limits were evaluated during method validation. Procedural detection limits of 0.3 pg mL(-1) (positive mode) and 0.6 pg mL(-1) (negative mode) were found for the dissolved fraction, and absolute limits of 0.4 pg (positive mode) and 6.0 pg (negative mode) for particulate samples were obtained. The most sensitive method in positive mode was applied to define DA occurrence in the Venice Lagoon. Trace concentrations of domoic acid ranging from 1.5 to 16.2 pg mL(-1) were found for the first time in the Venetian environment.

Complexation of cadmium and copper by fluvial humic matter and effects on their toxicity.

The effects of humic acids and fulvic acids isolated from the River Arno (Italy) on the bioavailability and toxicity of cadmium and copper were assessed in relation to changes in their speciation. Measurements of the complexing capacity of solutions containing these organic ligands were carried out by a titration procedure followed by DPASV and toxicity tests were carried out using lysosomes isolated from rat liver. The complexing capacity of the physiological medium containing about 13 mg/L of humic acids, expressed as ligand concentrations, was 0.30 and 0.072 micromol/L for cadmium and copper respectively; the corresponding conditional stability constants were 4.2 x 10(11) and 1.3 x 10(8) (mol/L)-1. The complexing capacities of the solution containing the same amount of fulvic acids were 0.33 and 0.164 micromol/L for cadmium and copper respectively, the conditional stability constants were 3.2 x 10(11) and 2.4 x 10(7) (mol/L)-1. The humic acids reduced the toxicity of cadmium by about 5 times: the EC50 changed from 4.4 to 20.4 micromol/L. The dose effect curve of copper presented a bi-sigmoid trend and two EC50 values can be determined: The EC50(1) in the presence of humic acids changed from 2.0 to 3.1 micromol/L, while the EC50(2) increased from 22.3 to 45.3 micromol/L. The fulvic acids reduced the cadmium toxicity by about the same amount as humic acids, from 4.4 to 18.6 micromol/L, but they had no effect on copper toxicity. Analysing the chemical speciation of cadmium and copper in the presence of humic components and under toxicity test conditions we can say that the appreciable decrease of EC50 is not related to changes in their speciation; we can hypothesize that this is due to different processes, as well as to blocking of the lysosomal membrane. On the basis of the shape of the dose-effect curves obtained for cadmium and copper respectively, we can say that the toxic effects of the two metals are different and we can hypothesize that copper could exercise its toxic activity by inhibiting the ATP-driven proton pump and the function of the Cl- selective channel.

Summer distribution of trace metals in the western sector of the Ross Sea, Antarctica.

The more important water masses generated by the interaction of Circumpolar Deep Water and the shelf waters in the western sector of the Ross Sea are characterized for trace element contents. The distribution of cadmium, lead, copper, zinc, iron, manganese and chromium during the austral summer is analysed and discussed according to the physical, chemical and biological processes which affect the composition of the water masses. The Cd concentration is found to have a relative high variability that can be related to biological activity and the water mass age, MCDW presents a mean dissolved concentration (SD) of 0.77 (0.07) nmol kg(-1), while the HSSW and AASW have a mean concentration of 0.63 (0.06) and 0.61 (0.16) nmol kg(-1), respectively. Lead features the typical distribution of a scavenged element with a surface maximum ranging between 22 and 130 pmol kg(-1) decreasing to 11 pmol kg(-1) in deep waters. However, the vertical distribution in the shelf area features a maximum concentration in intermediate/deep waters and we can hypothesize that the distribution may be influenced by more than one source. The surface dissolved concentration of zinc and copper were un-homogeneously distributed, the mean (SD) values were 5.25 (2.92) and 1.99 (1.49) nmol kg(-1) for zinc and copper, respectively and increased with depth for both the elements. We may therefore hypothesize enrichment in the dissolved phase deriving from recycling in deep waters. 95% of the chromium was in dissolved form and showed a superficial depletion; the mean concentrations were 1.6 +/- 0.2 and 2.6 +/- 0.8 nmol kg(-1) for surface and deep waters respectively. The vertical distribution of dissolved manganese was quite homogeneous with a mean concentration 0.96 +/- 0.7 nmol kg(-1). The particulate iron and manganese concentration trends are similar and feature a significant bottom increase implying a significant input from resuspension; the mean concentration of particulate ranged between 1.4 and 7.4 nmol kg(-1) for iron and ranged between 0.072 and 0.29 nmol kg(-1) for manganese.