A Practical Tool for the Assessment of Polymer Biodegradability in Marine Environments Guides the Development of Truly Biodegradable Plastics.

Environmental persistence is one of the few shortcomings of plastic materials. As a consequence, alternative plastics labeled as compostable are replacing polyolefins in some commercial applications, such as food bags and trash bags. A rapid, high-throughput, and environmentally relevant method to assess the potential biodegradability in marine conditions is used to assess these materials already on the market, as well as novel bio-based polymers still in development. By fitting experimental data to a non-linear logistic model, ultimate biodegradability can be calculated without regard for incubation time. Whereas the commercial products show negligible or very low marine biodegradability, one of the novel materials exceeds the 20% biodegradation threshold relative to fully marine biodegradable PHB after 28 days. In addition, the sensitivity of the method can be enhanced and its duration reduced, at the expense of labor-demanding preconditioning of the microbial inoculum, by increasing the bacterial density in the incubation vessels. In contrast, pre-exposure of the inoculum to plastic, either in laboratory or field conditions, does not enhance the performance of the test.

Assessment of Toxicity and Biodegradability of Poly(vinyl alcohol)-Based Materials in Marine Water.

Due to the continuous rise in conventional plastic production and the deficient management of plastic waste, industry is developing alternative plastic products made of biodegradable or biobased polymers. The challenge nowadays is to create a new product that combines the advantages of conventional plastics with environmentally friendly properties. This study focuses on the assessment of the potential impact that polyvinyl alcohol (PVA)-based polymers may have once they are released into the marine environment, in terms of biodegradation in seawater (assessed by the percentage of the Theoretical Oxygen Demand, or % ThOD, of each compound) and aquatic toxicity, according to the standard toxicity test using Paracentrotus lividus larvae. We have tested three different materials: two glycerol-containing PVA based ones, and another made from pure PVA. Biodegradation of PVA under marine conditions without an acclimated inoculum seems to be negligible, and it slightly improves when the polymer is combined with glycerol, with a 5.3 and 8.4% ThOD achieved after a period of 28 days. Toxicity of pure PVA was also negligible (<1 toxic units, TU), but slightly increases when the material included glycerol (2.2 and 2.3 TU). These results may contribute to a better assessment of the behavior of PVA-based polymers in marine environments. Given the low biodegradation rates obtained for the tested compounds, PVA polymers still require further study in order to develop materials that are truly degradable in real marine scenarios.

Integrative assessment of ecological responses and chemical contamination of urban wastewater outfalls on soft bottom sediments of an estuarine system.

Sewage outfalls are one of the main anthropogenic impacts to coastal systems. Linking the chemical analysis in sediment, and changes in the ecological responses is one of the main tasks in ecosystem assessment for the protection of the marine environment. A three-tiered approach to evaluate the impact of the sewage of a recently built and obsolete Urban Waste Water Treatment Plants (UWWTPs) was performed. The sewage from each UWWTP differentially affected surrounding sediments. Macrobenthic community changes were influenced by the chemical composition in the sediments but also by grain size and organic matter. Abundance of the different families of the macrobenthic communities were correlated with most of the chemicals analyzed. On the contrary, toxicity bioassays were correlated with Zn and PCB concentrations but not with environmental variables. The toxicity tests linked the effect of chemical contamination on macrobenthic communities. A joint assessment of both chemical concentrations and their ecological effects is recommended.

Combined use of chemical, biochemical and physiological variables in mussels for the assessment of marine pollution along the N-NW Spanish coast.

This study undertakes an overall assessment of pollution in a large region (over 2500 km of coastline) of the N-NW Spanish coast, by combining the use of biochemical (AChE, GST, GPx) and physiological (SFG) responses to pollution, with chemical analyses in wild mussel populations (Mytilus galloprovincialis). The application of chemical analysis and biological techniques identified polluted sites and quantified the level of toxicity. High levels of pollutants were found in mussel populations located close to major cities and industrialized areas and, in general, average concentrations were higher in the Cantabrian than in the Iberian Atlantic coast. AChE activities ranged between 5.8 and 27.1 nmol/min/mg prot, showing inhibition in 12 sampling sites, according to available ecotoxicological criteria. GST activities ranged between 29.5 and 112.7 nmol/min/mg prot, and extreme variability was observed in GPx, showing activities between 2.6 and 64.5 nmol/min/mg prot. Regarding SFG, only 5 sites showed 'moderate stress' (SFG value below 20 J/g/h), and most sites presented a 'high potential growth' (>35 J/g/h) corresponding to a 'healthy state'. Multivariate statistical techniques applied to the chemical and biological data identified PCBs, organochlorine pesticides and BDEs as the main responsible of the observed toxicity. However, the alteration of biological responses caused by pollutants seems to be, in general, masked by biological variables, namely age and mussel condition, which have an effect on the mussels' response to pollutant exposure.

Prospective ecological risk assessment of sediment resuspension in an estuary.

This study assesses potential ecological risk of resuspended sediment in the water column during the construction of a viaduct in the estuary of the Ulla river (Galicia, NW Iberian Peninsula), a shellfish production area. Chemical analyses and toxicity bioassays with elutriates were performed with sediments from the area where the three pillars of the viaduct will be located (CT1, CT2 and CT3) and a reference sediment (A2). Acute toxicity of the elutriate was evaluated in five species of three trophic levels (Isochrysis galbana, Paracentrotus lividus, Mytilus galloprovincialis, Venerupis pullastra and Siriella armata). The sediments of the pillars showed moderate levels of contamination by trace elements (Cu, Cr). Clam and sea urchin embryo-larval toxicity tests showed slightly higher sensitivity than mussel embryo tests, and toxicity was not detected for phytoplankton and mysid bioassays. The predicted no-effect environmental concentration (PNEC) was calculated from the arithmetic mean of the lowest calculated EC(50)s for each sampling site. The predicted environmental concentration (PEC) was estimated from a simple dilution model and the PEC/PNEC ratio was calculated according to different scenarios of resuspension. Negligible ecological risk in the water column is expected during construction of the pillars.

Ecological risk assessment of perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) in marine environment using Isochrysis galbana, Paracentrotus lividus, Siriella armata and Psetta maxima.

Perfluorooctanoic acid (PFOA) and perfluorooctane sulfonic acid (PFOS) are anthropogenic substances classified as persistent bioaccumulative compounds and are found in various environmental compartments throughout the world, from industrialized regions to remote zones far from areas of production. In this study, we assessed the effects of PFOA and PFOS on early life stages of marine test species belonging to three different trophic levels: one microalga (Isochrysis galbana), a primary consumer (Paracentrotus lividus) and two secondary consumers (Siriella armata and Psetta maxima). Acute EC(50) values for PFOS were 0.11 mg L(-1) in P. maxima, 6.9 mg L(-1) in S. armata, 20 mg L(-1) in P. lividus and 37.5 mg L(-1) in I. galbana. In the case of PFOA, the toxicity was lower but the ranking was the same; 11.9 mg L(-1) in P. maxima, 15.5 mg L(-1) in S. armata, 110 mg L(-1) in P. lividus and 163.6 mg L(-1) in I. galbana. The Predicted No Effect Concentration (PNEC) for PFOS and PFOA in marine water derived from these acute toxicity values are 1.1 µg L(-1) for PFOS and 119 µg L(-1) for PFOA. This study established a baseline dataset of toxicity of PFOS and PFOA on saltwater organisms. The data obtained suggest that PFOA pose a minor risk to these organisms through direct exposure. In the perspective of risk assessment, early life stage (ELS) endpoints provide rapid, cost-effective and ecologically relevant information, and links should be sought between these short-term tests and effects of long-term exposures in more realistic scenarios.

Assessment criteria for using the sea-urchin embryo test with sediment elutriates as a tool to classify the ecotoxicological status of marine water bodies.

A large, multiyear data set was generated by pooling the sediment elutriate (SET) results collected during previous studies conducted in the Galician Rias (northwest Iberian Peninsula) that met the acceptability criteria in the controls, to ensure optimum quality of data (n=162). Two subsets of equal to reference and lower than reference sites were identified by comparing the Percentage Net Response (PNR) value from each sampling site with nontoxic, cruise-specific, reference sites by using the t test with the unequal variance assumption. Ecotoxicological Assessment Criteria (EAC0, EAC1, EAC2, and EAC3) were then derived from those two subsets to classify the SET results into five categories of ecotoxicological status: high, good, moderate, poor, and bad, in line with the European legislation. The 50th and 5th percentiles of the PNR distribution of the equal to reference sites subset were EAC0=0.879 and EAC1=0.694. An EAC2=0.508 was obtained from the 50th percentile of the lower than reference sites subset. Because the PNR values of the entire database showed a distribution that can be adjusted to two normal populations, the EAC3=0.240 PNR was calculated as the intersection between the first and second normal distributions identified. Power analysis proved that the limit between acceptable and unacceptable status (EAC1) corresponded to a detectable PNR difference to control with a confidence level>99% and a power of 95%.

Acute toxicity of a shoreline cleaner, CytoSol, mixed with oil and ecological risk assessment of its use on the Galician Coast.

The application of embryo-larval bioassay with the purple sea urchin Paracentrotus lividus and the mussel Mytilus galloprovincialis at 48 hours, and with neonates of the mysid Siriella armata at 96 hours, was used to evaluate the acute toxicities of the following preparations: (1) the shoreline cleaning agent CytoSol; (2) the water-accommodated fraction of CytoSol plus a light crude oil; and (3) the runoff from a pilot-scale treatment with CytoSol of a rocky coastal substrate impregnated with residues from the Prestige oil spill (which occurred on November 19, 2002). The mussel was the most sensitive organism to CytoSol and runoff effects (EC(50) = 8.0 microL/L and 64.3 mL/L, respectively), and the mysid was the least sensitive to the runoff (EC(50) > 200 mL/L). The predicted no-effect environmental concentration (PNEC) was calculated from the no observed-effect concentration of the species most sensitive to the runoff. The predicted environmental concentration (PEC) was estimated from a simple and reasonable dilution model, and the PEC/PNEC ratio was calculated according to the area treated and the values of the variables considered in the model. Implications for the management of the treatment operations are discussed.

Methodological basis for the optimization of a marine sea-urchin embryo test (SET) for the ecological assessment of coastal water quality.

The sea-urchin embryo test (SET) has been frequently used as a rapid, sensitive, and cost-effective biological tool for marine monitoring worldwide, but the selection of a sensitive, objective, and automatically readable endpoint, a stricter quality control to guarantee optimum handling and biological material, and the identification of confounding factors that interfere with the response have hampered its widespread routine use. Size increase in a minimum of n=30 individuals per replicate, either normal larvae or earlier developmental stages, was preferred to observer-dependent, discontinuous responses as test endpoint. Control size increase after 48 h incubation at 20 degrees C must meet an acceptability criterion of 218 microm. In order to avoid false positives minimums of 32 per thousand salinity, 7 pH and 2mg/L oxygen, and a maximum of 40 microg/L NH(3) (NOEC) are required in the incubation media. For in situ testing size increase rates must be corrected on a degree-day basis using 12 degrees C as the developmental threshold.

Integrative assessment of coastal pollution in a Ría coastal system (Galicia, NW Spain): correspondence between sediment chemistry and toxicity.

Elutriate embryo-larval bioassays with sea-urchins (Paracentrotus lividus) and ascidians (Ciona intestinalis) were conducted concurrently with trace metal analyses as part of an integrative assessment of sediment pollution at Ría de Pontevedra (Galicia, NW Spain). High metal contents in sediments were found in localised areas from the inner part of the estuary indicating a clear anthropogenic influence. In particular, very high Cu, Zn and Pb levels were found at sites P2 and P3, which were also the most toxic to the embryo-larval bioassays. Sediment quality guidelines were used to help in the ecological interpretation of sediment chemistry data and to identify pollutants of concern. Cu and Zn in P3 were consistently above the effects range median (ERM) values, which seem to be good predictors of toxicity to sea-urchin and ascidian embryos. A toxic unit approach, based on published EC(50) values and metal levels in elutriates, was used to assess the harmful ecological effects associated to sediment chemistry. Toxicity detected in P3 may be explained on the basis of the toxic unit model; however, the high toxicity detected at P2 may be attributable not only to the metals quantified in the analyses but also to unmeasured organic pollutants. Multidimensional scaling applied independently to the toxicology and chemistry data resulted in a good agreement between both types of configurations. Moreover, the Mantel test revealed a significant correlation (r(M)=0.481; p=0.019) between metal concentrations and toxicity data profiles, supporting the correspondence between configurations.

Toxicity assessment of crude and partially purified extracts of marine Synechocystis and Synechococcus cyanobacterial strains in marine invertebrates.

Among the Cyanoprokaryota, the genera Synechocystis and Synechococcus have rarely been studied with respect to potential toxicity. This is particularly true with marine environments where studies about the toxicity of cyanobacteria are restricted to filamentous forms at the warmer temperate and tropical regions and also to filamentous forms at cold seas such as the Baltic Sea. In this study, we describe the effects of cyanobacterial strains of the Synechocystis and Synechococcus genera isolated from the marine coast of Portugal, on marine invertebrates. Crude and partially purified extracts at a concentration of 100 mg/ml of freeze-dried material of the marine strains were tested for acute toxicity in nauplii of the brine shrimp Artemia salina, in the rotifer Brachionus plicatillis and in embryos of the sea urchin Paracentrotus lividus and the mussel Mytilus galloprovincialis. The cyanobacterial extracts, especially the crude extract, had an impact on A. salina nauplii. No significant toxic effects were registered against the rotifer. A negative impact of all strains was recorded on the embryonic development of the sea urchin, with toxic effects resulting in an inhibition of embryogenesis or development of smaller larvae. To the mussel embryos, the effects of cyanobacterial extracts resulted in a complete inhibition of embryogenesis. The results of all assays indicate that Synechocystis and Synechococcus marine strains contained toxic compounds to marine invertebrates.