The aim of this study was to investigate the ecotoxicity of polyvinylidene difluoride (PVDF) and polylactic acid (PLA) microplastics (MPs) in two marine zooplankton: the crustacean Artemia franciscana and the cnidarian Aurelia sp. (common jellyfish). To achieve this goal, (i) MP uptake, (ii) immobility, and (iii) behavior (swimming speed, pulsation mode) of crustacean larval stages and jellyfish ephyrae exposed to MPs concentrations (1, 10, 100 mg/L) were assessed for 24 h. Using traditional and novel techniques, i.e., epifluorescence microscopy and 3D holotomography (HT), PVDF and PLA MPs were found in the digestive systems of the crustaceans and in the gelatinous tissue of jellyfish. Immobility was not affected in either organism, while a significant behavioral alteration in terms of pulsation mode was found in jellyfish after exposure to both PVDF and PLA MPs. Moreover, PLA MPs exposure in jellyfish induced a toxic effect (EC50: 77.43 mg/L) on the behavioral response. This study provides new insights into PLA and PVDF toxicity with the potential for a large impact on the marine ecosystem, since jellyfish play a key role in the marine food chain. However, further investigations incorporating additional species belonging to other trophic levels are paramount to better understand and clarify the impact of such polymers at micro scale in the marine environment. These findings suggest that although PVDF and PLA have been recently proposed as innovative and, in the case of PLA, biodegradable polymers, their effects on marine biota should not be underestimated.

Fluoxetine (FLX), one of the most widely prescribed selective serotonin reuptake inhibitors, is frequently detected in the aquatic environment. In this study we assessed the ecotoxicological effects of FLX on the early life-stages of the sea urchin Paracentrotus lividus, a key species in the Mediterranean Sea. Fertilization rate, developmental anomalies and behavioural alterations were evaluated up to 72 h by exposing gametes, zygotes, and embryos (gastrula) to environmental (0.001, 0.01 mg/L) and high concentrations (0.1, 1, 10 mg/L). Further, the different types and frequency of morphological anomalies at larval level were classified to estimate the Index of Contaminant Impact (ICI) at relevant and high concentrations. The ICI was applied to predict which FLX concentrations may pose a risk to sea urchins. Although FLX did not affect fertilization, significant skeletal anomalies and behavioural alterations were found in plutei from each exposed stage. Based on EC50 values, the sensitivity level ranks as follows: zygote > gastrula > sperm. The ICI values indicated high and moderate impacts only at high concentrations. However, a slight impact was also found in plutei from zygote exposure at relevant environmental concentrations, highlighting a potential risk for sea urchin early development. Considering increasing FLX consumption, we suggest to include this PC in monitoring plans, to not exceed levels that may impair and severely affect the early developmental stages of echinoderms. In addition, our findings promote the use of ICI as a novel tool for FLX impact assessment.

The sea urchin Paracentrotus lividus is a good model in ecotoxicology, but adults living along the Italian coasts have a limited reproductive period. In this species, natural or human-driven pressures may lead to limited gamete availability for ecotoxicological surveys. This study investigates the quality of early developmental stages of wild and cultured sea urchins to be used in ecotoxicology, avoiding field collection of mature specimens. Adult sea urchins were cultured in the laboratory for 2 years. Every 45 days, fertilization and larval quality were checked and compared to those from adults sampled in the wild. Fertilization was never affected, differently from development, which was impaired in the larvae obtained from sea urchins reared for more than one year. Fertilization and embryotoxicity were performed using copper nitrate in wild and cultured sea urchins. Fertilization did not differ up to ten months, while similar embryotoxicity was only found up to 5 months. This study promotes rearing sea urchins in ‘ready-to-spawn’ conditions for ecotoxicology surveys by recommending 10- and 5-month rearing times to assess fertilization and embryo toxicity, respectively. Here, we provided a baseline in marine ecotoxicology to obtain gametes on demand, irrespective of reproductive period and other pressures that may impact gamete availability.

The Mediterranean Basin is facing an increasing frequency and intensity of wildfires driven by climate change, raising concern about their off-site impacts on marine ecosystems. This study provides for the first time the effects of wildfire ash on the early developmental stages of the sea urchin Paracentrotus lividus, a keystone Mediterranean species. Environmentally relevant aqueous ash extracts (AEAs; 5 g/L) and their dilutions were tested through spermio- and embryotoxicity assays, assessing responses across multiple levels of biological organization. Fertilization success, larval development and morphology, apoptosis and enzymatic activity were evaluated up to 72 h of exposure. In addition, the Index of Contaminant Impact (ICI) was estimated by classifying the type and frequency of larval morphological anomalies, allowing the prediction of ash concentrations that may pose an ecotoxicological risk to sea urchins. Although fertilization was not affected, early development was significantly impaired, revealing a higher sensitivity of fertilized egg exposure rather than sperm. Choline acetyltransferase (ChAT) inhibition, increased acetylcholinesterase (AChE) activity, and enhanced apoptosis were observed, indicating the activation of inflammatory and stress-related pathways. Alterations in larval morphology and development were found from 25% AEA dilution upwards, being responsible from slight to moderate impact. Overall, these results suggest that wildfire ash represents an emerging threat to the early life stages of coastal marine species such as echinoderms. Furthermore, they support the use of sea urchins as sensitive bioindicators of wildfire-related marine pollution.