Investigation of Metal Toxicity on Microalgae Phaeodactylum tricornutum, Hipersaline Zooplankter Artemia salina, and Jellyfish Aurelia aurita.

The escalating global anthropogenic activities associated with industrial development have led to the increased introduction of heavy metals (HMs) into marine environments through effluents. This study aimed to assess the toxicity of three HMs (Cr, Cu, and Cd) on organisms spanning different trophic levels: Phaeodactylum tricornutum (a primary producer), Artemia salina (a primary consumer), and Aurelia aurita (a secondary consumer). The EC50 values obtained revealed varying relative toxicities for the tested organisms. Phaeodactylum tricornutum exhibited the highest sensitivity to Cu, followed by Cd and Cr, while Artemia salina displayed the highest sensitivity to Cr, followed by Cu and Cd. A. aurita, on the other hand, demonstrated the highest sensitivity to Cu, followed by Cr and Cd. This experimental investigation further supported previous studies that have suggested A. aurita as a suitable model organism for ecotoxicity testing. Our experiments encompassed sublethal endpoints, such as pulsation frequency, acute effects, and mortality, highlighting different levels of sensitivity among the organisms.

Effects of life cycle exposure to polystyrene microplastics on medaka fish (Oryzias latipes).

The number of published studies evaluating the effects of microplastics (MPs) in fish has increased in the last decade. However, of the available studies, few have explored the long-term effects of MPs on fish growth and reproduction and have resorted to MPs in the form of µm-sized beads/microspheres. In this study, 6-10 day-old post-hatch medaka (Oryzias latipes) fish were exposed to 50 (i.e. 1X) and 500 (i.e. 10X) µg of heterogeneously sized and irregularly shaped virgin polystyrene (PS) MP particles (200-µm range)/L for 150 days. These concentrations corresponded to respective daily mean values of 247 and 3087 particles/L administered through the diet. The PS MPs dietary exposure resulted in body burdens of 114 and 440 particles/g fish on day 50, and of 78 and 173 particles/g fish on day 100 since the respective exposures to the 1X and the 10X treatments started. The biometric analyses found no incidence of PS MPs ingestion on overall fish growth and development. The histological survey in the 10X group did not reveal alterations in gills or in the digestive tract. Mild alterations in other organs were seen and included increased fluid material in the peritoneal cavity, glomerular and tubular alterations in kidneys, and differences in the diameter of the thyroid follicles and thickness of the follicular epithelial cells. The initial days of the reproductive phase revealed MP-related differences in the number of gravid females, fecundity, and fertilization rates. Overall, these values reverted to normal rates throughout the succeeding days. No significant effects of PS MPs exposure were evidenced on offspring success. The 150-day PS MPs dietary exposure used in this study provided clues of histological effects and a reproduction delay. However, it did not seem to compromise overall growth/thriving and the ongoing reproduction.

Soil and Freshwater Bioassays to Assess Ecotoxicological Impact on Soils Affected by Mining Activities in the Iberian Pyrite Belt.

This study aims to use geochemical, mineralogical, ecotoxicological and biological indicators for a comprehensive assessment of the ecological risks related to the mobility, ecotoxicity and bioavailability of potentially harmful elements in the Lousal mining district. Particularly, toxicity was evaluated using four assays: algae, cytotoxicity assays with HaCaT cell line (dermal), earthworms and Daphnia magna. The geochemical and mineralogical characterization of the studied area shows that the mine wastes underwent intense weathering processes, producing important contamination of the adjacent soils, which also led to the release and mobilization of PHEs into nearby water courses. Total PTE results indicate that the soils affected by mining activities were highly contaminated with As and Cu, while Zn and Pb content ranged from low to very high, depending on the analyzed samples. Cadmium levels were found to be very low in most of the soil samples. The test using Daphnia magna was the most sensitive bioassay, while the Eisenia foetida test was the least sensitive. Except for the LOS07 soil sample, the rest of the soils were classified as "High acute toxicity" and "Very high acute toxicity" for aquatic systems. The results in HACaT cells showed results similar to the ecotoxicological bioassays. The application of biotests, together with geochemical and mineralogical characterization, is a very useful tool to establish the degree of contamination and the environmental risk of potentially harmful elements.

Application of an Ecotoxicological Battery Test to the Paddy Field Soils of the Albufera Natural Park.

Albufera Natural Park (ANP) (Valencia, Spain) is one of the most important wetland areas of the Mediterranean coast subject to high anthropogenic pressure, on whose soils a battery of bioassays has never been applied to evaluate the ecotoxicological risk. The present study determined available and water-soluble heavy metal content in four paddy soils used in the ANP, and the ecotoxicological effect on these soils was evaluated by performing the bioassays regulated in Spanish Royal Decree 9/2005. Soil properties and extractable Co, Cr, Cu, Ni, Pb and Zn (EDTA pH = 7) were analyzed in soils. These elements and macro- and micronutrients were also assessed in soil leachate. A test battery covering the following was needed: acute toxicity test in Eisenia foetida (OECD TG 207); mineralization tests of nitrogen (OECD TG 2016) and carbon (OECD TG 217); growth inhibition test in Raphidocelis subcapitata (OECD TG 201); mobility inhibition test in Daphnia magna (OECD TG 202). The soils found in the most anthropized areas to the north of the ANP (Massanassa and Alfafar) demonstrated a higher concentration of available heavy metals than in the southern ones (Sueca and Sollana). The aqueous leachate of the studied soils contained very low concentrations, which would be related to soil properties. Despite the high concentration of available potentially toxic elements (PTEs) in the Massanassa and Alfafar soils, the studied soils showed no toxicity during the performed battery bioassays. Therefore, soils can be considered non-toxic despite the obtained PTEs available concentration.

Comparing in vivo data and in silico predictions for acute effects assessment of biocidal active substances and metabolites for aquatic organisms.

The purpose of this study was to determine the acute toxicity in aquatic organisms of one biocidal active substance and six metabolites derived from biocidal active substances and to assess the suitability of available QSAR models to predict the obtained values. We have reported the acute toxicity in sewage treatment plant (STP) microorganisms, in the freshwater microalgae Pseudokirchneriella subcapitata and in Daphnia magna following OECD test methods. We have also identified in silico models for acute toxicity of these trophic levels currently available in widely recognized platforms such as VEGA and the OECD QSAR ToolBox. A total of six, four and two models have been selected for Daphnia, algae and microorganisms, respectively. Finally, we have compared the in silico and in vivo data for the seven compounds plus two previously assayed biocidal substances. None of the compounds tested were toxic for Daphnia and STP microorganisms. For microalgae, CGA71019 (1,2,4 triazole) presented an ErC50 value of 38.3 mg/L. The selected in silico models have provided lower EC50 values and are therefore more conservative. Models from the OECD QSAR ToolBox predicted values for 7 out of 9 and for 4 out of 9 chemicals for Daphnia and P. subcapitata, respectively. No predictive models were identified in such platform for STP microorganism's acute effects. In terms of models's specificity, biocide-specific models, developed from curated datasets integrated by biocidal active substances and implemented in VEGA, perform better in the case of microalgae but for Daphnia an alternative, non biocide-specific has revealed a better performance. For STP microorganisms only biocide-specific models have been identified.

Acute hazard of biocides for the aquatic environmental compartment from a life-cycle perspective.

One of the aims of the European project LIFE-COMBASE is to build a computational tool to predict the acute toxicity for aquatic organisms of biocidal active substances and its environmental degradation products. A database was implemented compiling toxicity data for these substances in organisms of the freshwater/marine and sewage treatment plant compartments. The goal of this study is to analyze the compiled data to identify the possible hazard of these compounds for the aquatic compartments. Several official and scientific databases were consulted. Data from 196 biocidal substances and 206 environmental metabolites were collected for the taxonomic groups, including fish, invertebrates, algae and sewage treatment plant (STP) microorganisms. Substances were categorized for their toxicity in four groups, considering values of L(E)C50, according to EU Regulation (EC) No 1272/2008. >50% of the parent were located in category 1 (L(E)C50 ≤ 1 mg/L) for fish, invertebrates and algae, indicating a high toxicity for the freshwater/marine compartments. However >60% were not toxic for STP microorganisms. Metabolites were mainly less toxic than the parent compounds, but 22-36% presented the same toxicity and ~6% were more toxic. No toxicological information was found for ~50% of the metabolites for fish, invertebrates and algae, reaching the 96% for the microorganisms. In addition, information on toxicity to the STP microorganisms was only found for 40% of the parent compounds. The high percentage of toxic metabolites and the scarcity of data for these compounds indicate the need to further study their impact in the aquatic compartments.

Acute toxicity and bioconcentration of fungicide tebuconazole in zebrafish (Danio rerio).

This research work investigated the bioconcentration of tebuconazole [(±)-α-[2-(4-chlorophenyl)ethyl]-α-(1,1-dimethylethyl)-1H-1,2,4-triazole-1-ethanol] fungicide in zebrafish (Danio rerio) under laboratory conditions and a first-order kinetic pesticide dissipation in the water. The concentrations of tebuconazole fitted to an equivalent nonlinear kinetic type model which allowed the calculation of the following parameters: bioconcentration factor (38.80 L kg(-1) ), time to reach maximum fish concentration (6 days), maximum concentration in fish (0.0075 µg mg(-1) ), half-life in fish (24 days) and time needed for the fish to eliminate 95% of the maximum concentration (105 days). These calculations permitted the establishment of theoretical reference limit values for human consumption of fish and the establishment of safe limits for the water pesticide concentration. The data would also be useful in safe strategies associated with fishery activities that are conducted in aquatic regions close to crops using tebuconazole. The information will contribute to enlarge the tebuconazole toxicokinetics database of aquatic organisms.

Assessment of pollution along the Northern Iberian shelf by the combined use of chemical and biochemical markers in two representative fish species.

Muscle concentrations of organochlorinated compounds as well as biliary levels of polycyclic aromatic hydrocarbons (PAHs) and alkylphenols (APEs) were determined in two different fish species, the four-spotted megrim (Lepidorhombus boscii) and the pouting (Trisopterus luscus) collected along the Northern Iberian coast. Additionally, a set of biochemical markers namely, 7-ethoxyresorufin O-deethylase (EROD), UDP-glucuronosyltransferase (UGT) and catalase (CAT) were measured in liver subcellular fractions. Chemical analysis indicated geographical differences in pollutant loads that were further reinforced by biomarker responses. Thus, EROD activity showed a good correlation with the amount of PCBs bioaccumulated in muscle tissue of both fish species. Elevated UGT activity was observed in those individuals highly exposed to APEs and 1-naphthol. The study reinforces the need to select representative sentinel species from different habitats for biomonitoring purposes and provides further support for the use of biomarkers in assessing the health of coastal areas.