Bacteriophages to control Vibrio alginolyticus in live feeds prior to their administration in larviculture.

AIMS: This study aimed to evaluate the efficiency of two phages [VB_VaC_TDDLMA (phage TDD) and VB_VaC_SRILMA (phage SRI)] alone and in a cocktail to control Vibrio alginolyticus in brine shrimp before their administration in larviculture. METHODS AND RESULTS: Phages were isolated from seawater samples and characterized by host spectrum, growth parameters, adsorption rate, genomic analysis, and inactivation efficiency. Both phages belong to the Caudoviricetes class and lack known virulence or antibiotic-resistance genes. They exhibit specificity, infecting only their host, V. alginolyticus CECT 521. Preliminary experiments in a culture medium showed that phage TDD (reduction of 5.8 log CFU ml-1 after 10 h) outperformed phage SRI (reduction of 4.6 log CFU ml-1 after 6 h) and the cocktail TDD/SRI (reduction of 5.2 log CFU ml-1 after 8 h). In artificial marine water experiments with Artemia franciscana, both single phage suspensions and the phage cocktail, effectively inactivated V. alginolyticus in culture water (reduction of 4.3, 2.1, and 1.9 log CFU ml-1 for phages TDD, SRI, and the phage cocktail, respectively, after 12 h) and in A. franciscana (reduction of 51.6%, 87.3%, and 85.3% for phages TDD, SRI, and the phage cocktail, respectively, after 24 h). The two phages and the phage cocktail did not affect A. franciscana natural microbiota or other Vibrio species in the brine shrimp. CONCLUSIONS: The results suggest that phages can safely and effectively control V. alginolyticus in A. franciscana prior to its administration in larviculture.

Feeding exposure and feeding behaviour as relevant approaches in the assessment of the effects of micro(nano)plastics to early life stages of amphibians.

The sedimentation of micro and nanosized plastics is of considerable environmental relevance and the need to assess its sublethal effects to biota increasingly recognized. In their majority, as bottom, non-selective grazers, independent-feeding young life stages of amphibians, an already severely endangered worldwide group, may be particularly vulnerable to sedimented plastics. Alongside, they may be good model organisms for the assessment of the effects of micro(nano)plastics (MNPs) through ingestion. However, to our knowledge, few studies have assessed amphibians' exposure to MNPs through contaminated food or its effects in feeding behaviour assays. The available studies reveal a lack of consistent methodology: organisms, food type, media of exposure, or exposure conditions (temperature and light) in the assessment of effects. This perspective article, will address major differences found in the available studies, identifying type, size and concentrations of the polymers tested, species, and observed effects, aiming to highlight the importance of feeding exposure assays when attempting to evaluate the effect of MNPs in amphibians.

The effects of nanoplastics on marine plankton: A case study with polymethylmethacrylate.

Marine biota is currently exposed to plastic pollution. The biological effects of plastics may vary according to polymer types (e.g. polystyrene, polyethylene, acrylate), size of particles (macro, micro or nanoparticles) and their shape. There is a considerable lack of knowledge in terms of effects of nanoplastics (NP) to marine biota particularly of polymers like polymethylmethacrylate (PMMA). Thus, this study aimed to assess its ecotoxicological effects using a battery of standard monospecific bioassays with four marine microalgae (Tetraselmis chuii, Nannochloropsis gaditana, Isochrysis galbana and Thalassiosira weissflogii) and a marine rotifer species (Brachionus plicatilis). The tested PMMA-NP concentrations allowed the estimation of median effect concentrations for all microalgae species. T. weissflogii and T. chuii were respectively the most sensitive (EC50,96h of 83.75 mg/L) and least sensitive species (EC50,96h of 132.52 mg/L). The PMMA-NP were also able to induce mortality in rotifers at concentrations higher than 4.69 mg/L with an estimated 48 h median lethal concentration of 13.27 mg/L. A species sensitivity distribution curve (SSD), constructed based on data available in the literature and the data obtained in this study, reveal that PMMA-NP appears as less harmful to marine biota than other polymers like polystyrene.

Short- and long-term toxicity of nano-sized polyhydroxybutyrate to the freshwater cnidarian Hydra viridissima.

The accumulation of increasingly smaller plastic particles in aquatic ecosystems is a prominent environmental issue and is causing a significant impact on aquatic biota. In response to this challenge, biodegradable plastics have emerged as a potential ecological alternative. Nevertheless, despite recent progress in polymer toxicology, there is still limited understanding of the ecological implications of biodegradable plastics in freshwater ecosystems. This study evaluated the toxicity of polyhydroxybutyrate nano-sized particles (PHB-NPLs) on the freshwater cnidarian Hydra viridissima assessing individual and population-level effects. Data revealed low toxicity of PHB-NPLs to H. viridissima in the short-term, as evidenced by the absence of significant malformations and mortality after the 96-h assays. In addition, hydras exhibited rapid and complete regeneration after 96 h of exposure to PHB-NPLs. Feeding assays revealed no significant alterations in prey consumption behavior in the 96-h mortality and malformations assay and the regeneration assay. However, significantly increased feeding rates were observed after long-term exposure, across all tested concentrations of PHB-NPLs. This increase may be attributed to the organisms' heightened energetic demand, stemming from prolonged activation of detoxification mechanisms. These changes may have a cascading effect within the food web, influencing community dynamics and ecosystem stability. Furthermore, a dose-dependent response on the hydras' populational growth was found, with an estimated 20 % effect concentration (EC20,8d) on this endpoint of 10.9 mg PHB-NPLs/L that suggests potential long-term impacts on the population's reproductive output and potential depression and local extinction upon long-term exposure to PHB-NPLs on H. viridissima. The obtained data emphasizes the importance of evaluating sublethal effects and supports the adoption of long-term assays when assessing the toxicity of novel polymers, providing crucial data for informed regulation to safeguard freshwater ecosystems. Future research should aim to unravel the underlying mechanisms behind these sublethal effects, as well as the impact of the generated degradation products.

Polyhydroxybutyrate (PHB) nanoparticles modulate metals toxicity in Hydra viridissima.

The use of bioplastics (e.g., polyhydroxybutyrate) emerged as a solution to help reduce plastic pollution caused by conventional plastics. Nevertheless, bioplastics share many characteristics with their conventional counterparts, such as degradation to nano-sized particles and the ability to sorb environmental pollutants, like metals. This study aimed to assess the potential impacts of the interaction of metals (cadmium - Cd, copper - Cu, and zinc - Zn) with polyhydroxybutyrate nanoplastics (PHB-NPLs; ~200 nm) on the freshwater cnidarian Hydra viridissima in terms of mortality rates, morphological alterations, and feeding behavior. The metal concentrations selected for the combined exposures corresponded to concentrations causing 20 %, 50 %, and 80 % of mortality (LC20, LC50, and LC80, respectively) and the PHB-NPLs concentrations ranged from 0.01 to 1000 µg/L. H. viridissima sensitivity to the metals, based on the LC50's, can be ordered as: Zn < Cd < Cu. Combined exposure to metals and PHB-NPLs yielded distinct outcomes concerning mortality, morphological changes, and feeding behavior, uncovering metal- and dose-specific responses. The interaction between Cd-LCx and PHB-NPLs progressed from no effect at LC20,96h to an ameliorative effect at Cd-LC50,96h. Cu-LCx revealed potential mitigation effects (LC20,96h and LC50,96h) but at Cu-LC80,96h the response shifts to a potentiating effect. For Zn-LCx, response patterns across the combinations with PHB-NPLs were like those induced by the metal alone. PHB-NPLs emerged as a key factor capable of modulating the toxicity of metals. This study highlights the context-dependent interactions between metals and PHB-NPLs in freshwater environments while supporting the need for further investigation of the underlying mechanisms and ecological consequences in forthcoming research.

Sub-chronic exposure to paroxetine disrupts ecologically relevant behaviours in fish.

The functional conservation of important selective serotonin reuptake inhibitor (SSRI) targets in non-target organisms raises concerns about their potential adverse effects on the ecosystems. Although the environmental levels of SSRIs like paroxetine (PAR) have risen, the knowledge regarding the effects of long-term exposure to PAR is limited. This study investigated the impact of sub-chronic exposure (21 days) to two sub-lethal concentrations of PAR (40 and 400 µg/L) on the behaviour of adult zebrafish in different scenarios: basal activity (under dark and light conditions), stress response (evoked by sudden light transitions) and stress response recovery. A new framework was employed for the integrative study of fish's swimming performance based on their innate ability to respond to light shifts. Several swimming-associated parameters (e.g., total swimming distance, time of inactivity, swimming angles) and thigmotaxis were monitored for an integrated analysis in each scenario. Data revealed reduced swimming activity, impaired behavioural response to stress and alterations in stress recovery of PAR-exposed fish. An anxiolytic effect was particularly noticeable in fish basal swimming activity in the dark at 400 µg/L and in the behavioural response to stress (from dark to light) and stress recovery (from light to dark) for organisms exposed to 40 µg/L. The detected PAR-induced behavioural modifications suggest a disruption of brain glucocorticoid signalling that may have implications at the individual level (e.g., changing behavioural responses to predators), with potential repercussions on the population and community levels. Therefore, the applied protocol proved sensitive in detecting behavioural changes induced by PAR.

Cation Composition Influences the Toxicity of Salinity to Freshwater Biota.

The effects of salinization on freshwater ecosystems have been estimated by testing sodium chloride (NaCl) since it is the most widely used salt as a deicing agent and Na+ and Cl- ions are the most representative in seawater composition. However, calcium, magnesium, and/or potassium are starting to be proposed as potential surrogates for NaCl, but for which ecotoxicological effects are less explored. This study aimed to identify (i) the less toxic salt to freshwater biota to be suggested as a safer alternative deicer and (ii) to contribute to the lower tiers of salinity risk assessment frameworks by identifying a more suitable surrogate salt than NaCl. The battery of ecotoxicity assays with five key trophic level species showed that among the tested salts (MgCl2, CaCl2, and KCl), KCl and CaCl2 seemed to induce the highest and lowest toxicity, respectively, compared with NaCl. CaCl2 is suggested as a safer alternative for use as a deicer and KCl as a surrogate for the risk assessment of seawater intrusion in coastal regions. These results enrich the salt toxicity database aiming to identify and propose more suitable surrogate salts to predict the effects of salinization to a broader extent.

Pre-exposure to seawater or chloride salts influences the avoidance-selection behavior of zebrafish larvae in a conductivity gradient.

The risk assessment of freshwater salinization is constructed around standard assays and using sodium chloride (NaCl), neglecting that the stressor is most likely a complex mixture of ions and the possibility of prior contact with it, triggering acclimation mechanisms in the freshwater biota. To date, as far as we are aware of, no information has been generated integrating both acclimation and avoidance behavior in the context of salinization, that may allow these risk assessments upgrading. Accordingly, 6-days-old Danio rerio larvae were selected to perform 12-h avoidance assays in a non-confined 6-compartment linear system to simulate conductivity gradients using seawater (SW) and the chloride salts MgCl2, KCl, and CaCl2. Salinity gradients were established from conductivities known to cause 50% egg mortality in a 96-h exposure (LC50,96h,embryo). The triggering of acclimation processes, which could influence organisms' avoidance-selection under the conductivity gradients, was also studied using larvae pre-exposed to lethal levels of each salt or SW. Median avoidance conductivities after a 12-h of exposure (AC50,12h), and the Population Immediate Decline (PID) were computed. All non-pre-exposed larvae were able to detect and flee from conductivities corresponding to the LC50,96h,embryo, selecting compartments with lower conductivities, except for KCl. The AC50,12h and LC50,96h overlapped for MgCl2 and CaCl2, though the former is considered as more sensitive as it was obtained in 12 h of exposure. The AC50,12h for SW was 1.83-fold lower than the LC50,96h, thus, reinforcing the higher sensitivity of the parameter ACx and its adequacy for risk assessment frameworks. The PID, at low conductivities, was solely explained by the avoidance behavior of non-pre-exposed larvae. Larvae pre-exposed to lethal levels of salt or SW were found to select higher conductivities, except for MgCl2. Results indicated that avoidance-selection assays are ecologically relevant and sensitive tools to be used in risk assessment processes. Stressor pre-exposure influenced organisms' avoidance-selection behavior under conductivity gradients, suggesting that under salinization events organisms may acclimate, remaining in altered habitats.

Contributions towards the hazard evaluation of two widely used cytostatic drugs.

Cytostatic drugs are one of the most important therapeutic options for cancer, a disease that is expected to affect 29 million individuals by 2040. After being excreted, cytostatics reach wastewater treatment plants (WWTPs), which are unable to efficiently remove them, and consequently, they will be released into the aquatic environment. Due to the highly toxic properties of cytostatics, it is particularly relevant to evaluate their potential ecological risk. Yet, cytostatics toxicity data is still not available for various species. In this work, the ecotoxicity of two widely consumed cytostatics, cyclophosphamide (CYP-as a model cytostatic) and mycophenolic acid (MPA-as a priority cytostatic), was evaluated on three freshwater species-Raphidocelis subcapitata, Brachionus calyciflorus, and Danio rerio, and the risk quotient (RQ) was assessed. Both drugs significantly affected the yield and growth inhibition of the microalgae, while for rotifers, the least sensitive species, only significant effects were registered for CYP. These drugs also caused significant effects on the mortality and morphological abnormalities on zebrafish. The estimation of the RQ discloses that CYP seems to pose a low risk to aquatic biota while MPA poses a very high risk. Altogether, these results emphasize the need for more complete environmental risk assessments, to properly prioritize and rank cytostatics according to their potentially toxic effects on the environment and aquatic biota.

Are early and young life stages of fish affected by paroxetine? A case study with Danio rerio.

Paroxetine (PAR) is a selective serotonin reuptake inhibitor (SSRI) antidepressant increasingly detected in surface waters worldwide. Its environmental presence raises concerns about the potential detrimental effects on non-target organisms. Thus, this study aimed to increase knowledge on PAR's potential environmental impacts, assessing the effects of commercial formulation (PAR-c) and active ingredient (PAR-a) on fish. Therefore, the short-term exposure effects of PAR-c and PAR-a were assessed on zebrafish (Danio rerio) embryos/larvae to determine the most toxic formulation [through median lethal (LC50) and effective concentrations (EC50)]. PAR-c and PAR-a induced morphological abnormalities (scoliosis) in a dose-dependent manner from 96 hours post-fertilization onwards, suggesting the involvement of a fully functional biotransformation system. As PAR-c exhibited higher toxicity, it was selected to be tested in the subsequent stage (juvenile stage), which was more sensitive (lower LC50). PAR-c significantly decreased fish swimming activity and disrupted fish stress response. Overall, the results highlight the ability of PAR-c to adversely affect fish swimming performance, an effect that persisted even after exposure ceases (21-day depuration), suggesting that PAR-c may impair individual fitness.

Adding knowledge to the design of safer hydrophobically modified poly(acrylic) acids: an ecotoxicological approach.

The architecture of hydrophobically modified polymers can be tailored to produce variants with different levels of functionality. This allows industry to apply rational design methods for the development of more environmentally friendly materials. In the present work, the ecotoxicity of six variants of hydrophobically modified poly(acrylic) acids (HMPAA), obtained by changing the crosslinked conformation, insertion position, and length of the hydrophobic groups, was assessed for the (i) bioluminescence production of Aliivibrio fischeri; (ii) population growth rate of Raphidocelis subcapitata and Chlorella vulgaris; (iii) mortality of Brachionus calyciflorus; (iv) feeding inhibition, somatic growth rate, reproduction, and mortality of Daphnia magna; and (iv) mortality and somatic growth rate of Pelophylax perezi tadpoles. The concentrations causing 50% and 20% of effects (L(E)C50 and 20, respectively) ranged from 9.64 up to > 2000 mg·L-1 for all six HMPAA and species. The bacterium A. fischeri and tadpoles of P. perezi were the most sensitive and most tolerant organisms to the six tested HMPAA, respectively. The computed 5% hazard concentrations (computed on the basis of L(E)C50 s) showed that HMPAA1 (13.0 mg·L-1) and HMPAA2 (26.1 mg·L-1) were the most toxic variants, while HMPAA6 (233 mg·L-1) the least one. These results suggest HMPAA6 (with low crosslink percentage modified by the addition of long and short hydrophobic groups at the surface) to be the most environmentally friendly variant and should be preferentially considered to be used in consumer products, compared to the other five studied variants.

Assessing the risks of capecitabine and its active metabolite 5-fluorouracil to freshwater biota.

Capecitabine (CAP, prodrug) and 5-fluorouracil (5-FU, its active metabolite) are two of the most prominent cytostatics, for which no clear picture can be drawn regarding potential concentrations of effect for freshwater biota, with CAP being grouped in the least studied cytostatic, whereas 5-FU has been classified as of no and of high environmental risk. Accordingly, the present work aimed to assess the ecotoxicity of CAP and 5-FU in three freshwater species, which included a 72-h assay with the producer Raphidocelis subcapitata; a 96-h assay with the invertebrate secondary consumer Hydra viridissima; and a 96-h assay with embryos of the vertebrate secondary consumer Danio rerio. The following endpoints were monitored: yield and population growth rate for the algae; mortality, morphological alterations, and post-exposure feeding rates for the cnidarian; and mortality, hatching, and malformations for the fish. Overall, organisms' sensitivity to CAP decreased in the following order: R. subcapitata > H. viridissima > D. rerio, whereas for 5-FU, it decreased in the following order: H. viridissima > D. rerio > R. subcapitata. For CAP, no median lethal effective concentrations (LC/EC50) were possible to compute for D. rerio, with no significant mortality or malformations registered in embryos exposed at concentrations up to 800 mg L-1. For R. subcapitata, the EC50s were 0.077 and 0.63 mg L-1 for yield and growth rate, respectively, and for H. viridissima, the EC50,30 min for feeding was 22.0 mg L-1. For 5-FU, no EC50s could be computed for R. subcapitata, whilst the EC50s for H. viridissima mortality and feeding were 55.4 and 67.9 mg L-1, respectively, and for D. rerio, the LC50,96 h and EC50,96 h (hatching and abnormalities) were 4546, 4100, and 2459 mg L-1, respectively. Assuming similar modes of action for both compounds and their co-occurrence, the combined risk quotient of the two chemicals was determined to be 7.97, which represents a risk for freshwater biota. Anticipating the increased consumption of these compounds and cancer development trends worldwide, these impacts may be further aggravated.

Ecotoxicity of non- and PEG-modified lanthanide-doped nanoparticles in aquatic organisms.

Various types of nanoparticles (NPs) have been widely investigated recently and applied in areas such as industry, the energy sector, and medicine, presenting the risk of their release into the environment. The ecotoxicity of NPs depends on several factors such as their shape and surface chemistry. Polyethylene glycol (PEG) is one of the most often used compounds for functionalisation of NP surfaces, and its presence on the surfaces of NPs may affect their ecotoxicity. Therefore, the present study aimed to assess the influence of PEG modification on the toxicity of NPs. As biological model, we chose freshwater microalgae, a macrophyte and invertebrates, which to a considerable extent enable the assessment of the harmfulness of NPs to freshwater biota. SrF2:Yb3+,Er3+ NPs were used to represent the broad group of up-converting NPs, which have been intensively investigated for medical applications. We quantified the effects of the NPs on five freshwater species representing three trophic levels: the green microalgae Raphidocelis subcapitata and Chlorella vulgaris, the macrophyte Lemna minor, the cladoceran Daphnia magna and the cnidarian Hydra viridissima. Overall, H. viridissima was the most sensitive species to NPs, which affected its survival and feeding rate. In this case, PEG-modified NPs were slightly more toxic than bare ones (non-significant results). No effects were observed on the other species exposed to the two NPs at the tested concentrations. The tested NPs were successfully imaged in the body of D. magna using confocal microscopy; both NPs were detected in the D. magna gut. The results obtained reveal that SrF2:Yb3+,Er3+ NPs can be toxic to some aquatic species; however, the structures have low toxicity effects for most of the tested species.

Effects of petroleum-based and biopolymer-based nanoplastics on aquatic organisms: A case study with mechanically degraded pristine polymers.

Mismanaged plastic litter submitted to environmental conditions may breakdown into smaller fragments, eventually reaching nano-scale particles (nanoplastics, NPLs). In this study, pristine beads of four different types of polymers, three oil-based (polypropylene, PP; polystyrene, PS; and low-density polyethylene, LDPE) and one bio-based (polylactic acid, PLA) were mechanically broken down to obtain more environmentally realistic NPLs and its toxicity to two freshwater secondary consumers was assessed. Thus, effects on the cnidarian Hydra viridissima (mortality, morphology, regeneration ability, and feeding behavior) and the fish Danio rerio (mortality, morphological alterations, and swimming behavior) were tested at NPLs concentrations in the 0.001 to 100 mg/L range. Mortality and several morphological alterations were observed on hydras exposed to 10 and 100 mg/L PP and 100 mg/L LDPE, whilst regeneration capacity was overall accelerated. The locomotory activity of D. rerio larvae was affected by NPLs (decreased swimming time, distance or turning frequency) at environmentally realistic concentrations (as low as 0.001 mg/L). Overall, petroleum- and bio-based NPLs elicited pernicious effects on tested model organisms, especially PP, LDPE and PLA. Data allowed the estimation of NPLs effective concentrations and showed that biopolymers may also induce relevant toxic effects.

Soil pH matters in the ecotoxicity of Basamid® to freshwater microalgae and macrophytes.

Intensive agriculture along with the use of agrochemicals has been associated with low soil fertility, soil erosion, and soil acidity. Management of soil pH through liming is a common practice in agriculture to increase soil fertility and nutrient availability. When altering soil pH, different chemical reactions occur depending on soil composition and agrochemicals presence. Basamid® is a fumigant used worldwide targeting soil nematodes, fungi, and weeds in diverse crops, that can reach freshwater ecosystems by leaching through the soil layers. The major goal of this work was to assess the influence of soil pH in the toxicity of Basamid® eluates to the microalgae Raphidocelis subcapitata and the duckweed Lemna minor. For this, eluates were prepared from soils with different pH (5.5, 6.5 and 7.5), contaminated with the recommended dose of Basamid® corresponding to 145.7 mg of dazomet/Kg soil. Soil was amended with calcium carbonate (CaCO3). Raphidocelis subcapitata and L. minor were exposed to the eluates during 72 h and 7 days respectively, and multiple endpoints were assessed: growth rate, biomass, pigment as chlorophyl content and cell damage. Results showed that soil pH can influence the performance of the tested species and also be a major factor in influencing Basamid®'s toxicity. However, a clear pattern of the influence of soil pH on Basamid®'s toxicity was not observed and was species dependent. For R. subcapitata lower soil pHs induced higher toxicity of Basamid®'s to the algae [ED50 for growth rate: 30 % (confidence limits-CL: 22.8-37.2) for soil pH 5.5; >100 % for soil pH 6.5 and pH 7.5], while for L. minor the opposite was observed [ED50 for number of fronds: 27.2 % (CL: 22.8-31.6) for pH 5.5; 20.3 % (CL: 10.0-30.6) for pH 6.5 and 10.7 % (CL: 6.3-15.1)]. Overall, these results showed that leachates of Basamid® through soils, at recommended doses, can have a severe impact on aquatic systems, with or without the influence of abiotic factors.

Ecotoxicity of eluates obtained from Basamid® contaminated soils is pH dependent: A study with Hydra viridissima, Xenopus laevis and Danio rerio.

Agrochemicals are mostly used to deplete pests and treat diseases in terrestrial agro-ecosystems. However, their transport through the soil, by leaching and/or runoff, may cause them to reach aquatic systems. Environmental parameters, such as soil pH, can affect this transport, by influencing the magnitude of agrochemicals degradation and chemical reaction. This work aimed at investigating the influence of soil pH on the toxicity of eluates obtained from Basamid® contaminated soils to Hydra viridissima, Xenopus laevis and Danio rerio. For this, a natural soil with pH amended to 5.5, 6.5 and 7.5, was spiked with the recommended dose (RD) of Basamid® (145 mg dazomet/kg soil) and eluates (Ba-E) were prepared with the respective species culture medium. Dilutions of the eluates (0.14-100%), obtained from the three soils (Ba-E 5.5, Ba-E 6.5 and Ba-E 7.5, corresponding to soil spiked with Basamid® RD at soil pH of 5.5, 6.5 and 7.5, respectively), were used to expose the organisms. Results showed that for H. viridissima increased soil alkalinity provoked less mortality comparatively to lower soil pH [LD50,96h of Ba-E 5.5: 10.6% and LD50,96h of Ba-E 7.5: 21.2%]. As for X. laevis and D. rerio Ba-E lethal ecotoxicity was similar across soil pH (LD50,96h varied from 5.7 to 6.9% and from 2.1 to 4.3%, respectively). For malformations, 20% effect dilution (ED) in H. viridissima was significantly higher at Ba-E 7.5 (ED20,96h: 17.4%), comparatively to Ba-E 5.5 and Ba-E 6.5 (ED20,96h: 7.9% and 7.7%, respectively). From the three tested organisms and based on both lethal and sublethal effects, H. viridissima presented the highest tolerance to Basamid® eluates and soil pH was a major factor determining the fumigant toxicity, with higher soil pH levels inducing, lower toxicity. The eluates obtained from soils contaminated with RD of Basamid® induced severe effects to the three aquatic species.

Soil pH influences the toxicity of Basamid® eluates to non-target species of primary consumers.

Basamid® is a fumigant nematicide and fungicide known to break down in several volatile compounds, mainly methyl isothiocyanate (MITC), when in contact with water. Soil abiotic parameters, such as pH, influences this breakdown process, and thus, the toxic effects of Basamid® to aquatic biota. This work studied the influence of soil pH (5.5, 6.5 and 7.5) on the toxicity of eluates (1:4, m:v), obtained from Basamid®-contaminated soils (with the recommended dose of 145 mg of dazomet/Kg of soil), on two primary consumers: Daphnia magna and Brachionus calyciflorus. For this, lethal and sublethal toxicity of eluates originated from soils at pH 5.5, 6.5 and 7.5, contaminated with Basamid® (Ba-E 5.5; 6.5 and 7.5, respectively), were assessed (dilutions between 0.096 - 100%). The LD50,24h of Basamid® eluates for D. magna varied from 3.07% to 7.82% (Ba-E 6.5 and Ba-E 5.5 respectively), while for B. calyciflorus varied from 18.1% to 84.7% (Ba-E 6.5 and Ba-E 7.5, respectively). Both species were less sensitive to Basamid® eluates originated from soils with pH 7.5 and more sensitive to those obtained from soils with pH 6.5. Regarding the sublethal effects, a lower soil pH was associated with a higher toxicity of Basamid® to D. magna reproduction (LOED: 0.125% Ba-E 5.5), while for B. calyciflorus such a higher toxicity was observed at the highest soil pH (ED20: 7.42% [5.10-9.74] at Ba-E 7.5). These results show a negative association between soil pH and the lethal toxicity of Basamid® contaminated eluates. However, such a pattern was not observed at sublethal level, at which a species dependency was observed regarding the influence of soil pH in the observed toxicity. Nevertheless, it is to highlight that very low concentrations of eluates (as 3.07%) caused significant mortality, indicating a high risk for freshwater biota. Considering that Basamid® is likely to reach the aquatic systems is real, for which reason the recommended dose must be reviewed at environmentally-relevant scenarios.

Bioplastics: known effects and potential consequences to marine and estuarine ecosystem services.

Bioplastics have been suggested as more sustainable alternatives to conventional, petroleum-based plastics. In this work, the available studies comparing effects of biopolymers and petroleum-based plastics were reviewed to improve the knowledge on the sustainability of biobased polymers, providing a benchmark regarding their ecotoxicological effects, as well as to highlight research priorities in this field. The literature review shows that, only a small number of the available biopolymers have been tested highlighting the need for more research diversifying the tested polymers. Overall, the available studies support the idea that bioplastics are likely to cause physiological impairments (feeding, reproduction, or locomotion) as well as cellular (proteome and enzyme activity) effects on biota. Furthermore, the studies on bioplastic degradation under realistic conditions report changes in water and sediment quality, which may also have consequences to biota. It is evident that some reservations must be kept regarding conventional plastics substitutions by bioplastics.

Seawater intrusion: an appraisal of taxa at most risk and safe salinity levels.

Seawater intrusion into low-lying coastal ecosystems carries environmental risks. Salinity levels at these coastal ecosystems may vary substantially, causing ecological effects from mortality to several sublethal endpoints, such as depression of rates of feeding, somatic growth, or reproduction. This review attempts to establish safe salinity levels for both terrestrial and freshwater temperate ecosystems by integrating data available in the literature. We have four specific objectives: (i) to identify the most sensitive ecological taxa to seawater intrusion; (ii) to establish maximum acceptable concentrations-environmental quality standards (MAC-EQSs) for sea water (SW) from species sensitivity distributions (SSDs); (iii) to compile from the literature examples of saline intrusion [to be used as predicted environmental concentrations (PECs)] and to compute risk quotients for the temperate zone; and (iv) to assess whether sodium chloride (NaCl) is an appropriate surrogate for SW in ecological risk assessments by comparing SSD-derived values for NaCl and SW and by comparing these with field data. Zooplankton, early life stages of amphibians and freshwater mussels were the most sensitive ecological receptors for the freshwater compartment, while soil invertebrates were the most sensitive ecological receptors for the terrestrial compartment. Hazard concentration 5% (HC5 ) values, defined as the concentration (herein measured as conductivity) that affects (causes lethal or sublethal effects) 5% of the species in a distribution, computed for SW were over 22 and 40 times lower than the conductivity of natural SW (≈ 52 mS/cm) for the freshwater and soil compartment, respectively. This sensitivity of both compartments means that small increments in salinity levels or small SW intrusions might represent severe risks for low-lying coastal ecosystems. Furthermore, the proximity between HC5 values for the soil and freshwater compartments suggests that salinized soils might represent an additional risk for nearby freshwater systems. This sensitivity was corroborated by the derivation of risk quotients using real saline intrusion examples (PECs) collected from the literature: risk was >1 in 34 out of 37 examples. By contrast, comparisons of HC5 values obtained from SSDs in field surveys or mesocosm studies suggest that natural communities are more resilient to salinization than expected. Finally, NaCl was found to be slightly more toxic than SW, at both lethal and sublethal levels, and, thus, is suggested to be an acceptable surrogate for use in risk assessment.

Ecotoxicity of cationic cellulose polymers to aquatic biota: The influence of charge density.

Better performances of cellulose-based polymers can be achieved by adjust their architecture including the density of cationic modifications. In this study, the influence of cationic substitution on the ecotoxicity of four quaternized hydroxyethyl cellulose polymers (SK-H, SK-L, SK-M, SK-MH) was studied, using an aquatic biota acute ecotoxicity classification, and rheological and physicochemical characterization. The ecotoxicity characterization was achieved by performing standard ecotoxicity assays with seven key trophic level species: Vibrio fischeri, Raphidocelis subcapitata, Chlorella vulgaris, Daphnia magna, Brachionus calyciflorus, Heterocypris incongruens, and Danio rerio. Median effective concentrations were used to compute hazard concentrations, through the species sensitive distribution curves method. The microalga C. vulgaris and rotifer B. calyciflorus were the most sensitive species to the studied polymers. The SK-H variant was highly toxic to the rotifer. Overall, variants with intermediate levels of cationic charge (SK-M, SK-MH) presented the lowest toxicity. The SK-M variant showed the lowest value of maximum acceptable concentration (0.00354 mg/L), thus being indicated as the least toxic variant. Therefore, the obtained results suggest that industry could direct the development of this type of polymers by tailoring its cationic substitution to moderate levels, in such a way that both functionality and environmental toxicity could be maximized.