Microplastics from agricultural mulch films: Biodegradation and ecotoxicity in freshwater systems.

The application of bio-based biodegradable mulch films in agriculture has raised environmental concerns regarding their potential impacts on adjacent freshwater ecosystems. This study investigated the biodegradation of microplastics derived from a bio-based biodegradable mulch (bio-MPs) and its acute and chronic ecotoxicity considering relevant scenarios (up to 200 and 250 mg/kg of sediment, using pristine and/or UV-aged particles), using the fungus Penicillium brevicompactum and the dipteran Chironomus riparius as model organisms, respectively, due to their ecological relevance in freshwater environments. Fourier-transform infrared spectroscopy analysis suggested changes in the fungus's carbohydrate reserves and bio-MP degradation through the appearance of low molecular weight esters throughout a 28 day biodegradation test. In a short-term exposure (48 h), C. riparius larvae exposed to pristine or UV-aged bio-MPs had up to 2 particles in their gut. Exposure to pristine bio-MPs decreased larval aerobic metabolism (<20 %) and increased neurotransmission (>15 %), whereas exposure to UV-aged bio-MPs activated larval aerobic metabolism (>20 %) and increased antioxidant defences (catalase activity by >30 % and glutathione-s-transferase by >20 %) and neurotransmission (>30 %). Longer-term (28-d) exposure to UV-aged bio-MPs did not affect larval survival and growth nor the dipteran's emergence but increased male numbers (>30 %) at higher concentrations. This study suggests that the selected agricultural bio-based mulch film is prone to biodegradation by a naturally occurring fungus. However, there is a potential for endocrine disruption in the case of prolonged exposures to UV-aged microplastics. This study emphasises the importance of further research to elucidate the potential ecological effects of these plastic products, to ensure effective management practices, and to establish new regulations governing their use.

PPCPs abatement using TiO2-based catalysts by photocatalytic oxidation and ozonation: The effect of nitrogen and cerium loads on the degradation performance and toxicity impact.

Pharmaceutical and personal care products (PPCPs) have been consumed in great extension and most of these are found in water bodies, owing to the inefficiency of conventional wastewater treatments. To face against these recalcitrant contaminants, advanced oxidation processes such as photocatalysis and ozonation have been studied. Moreover, the combination of these technologies can improve the degradation of PPCPs, reducing the ozone consumption and the effluent toxicity with the presence of photocatalysts. In particular, this study aimed to evaluate the effects of different N and Ce loads in co-doping TiO2 catalysts on the efficiency of photocatalytic oxidation and photocatalytic ozonation for PPCPs abatement, as well as on the resultant toxicity to aquatic species. Different radiation sources (UVA and solar radiation) were considered for the photocatalytic oxidation. A mixture of 5 PPCPs: paracetamol, sulfamethoxazole, carbamazepine, methylparaben and propylparaben was used as a model synthetic effluent. Photocatalysis showed a low efficiency on the PPCPs removal (<20 %), which was not affected by the radiation source. In general, the tested catalysts showed no or low added-value for reducing the toxicity of the synthetic effluent. Concerning photocatalytic ozonation, the lowest N amount (2.5 % w/w) promoted the best results for PPCPs removal, achieving values up to 100 % with significant reduction of ozone dose compared to photolytic ozonation. In general, photocatalytic ozonation showed better ecotoxicological performance than single photocatalysis. Compared to single photolytic ozonation, a benefitial effect was observed for two aquatic species, using a specific catalyst. This catalyst, prepared by doping TiO2 with 2.5 % w/w N and 1.2 % w/w Ce, showed to be the most promisong one, with potential to be used in photocatalytic ozonation. Hence, this work highlights the potential role of N and Ce co-doped TiO2-based catalysts in photocatalytic ozonation for wastewater treatment.

Ecotoxicological and biochemical effects of a binary mixture of pesticides on the marine diatom Thalassiosira weissflogii in a scenario of global warming.

Under the current scenario of global warming, it is ecologically relevant to understand how increased temperature influences the combined toxicity of pesticides to aquatic species. Hence, this work aims to: a) determine the temperature effect (15 °C, 20 °C and 25 °C) on the toxicity of two pesticides (oxyfluorfen and Copper (Cu)), on the growth of Thalassiosira weissflogii; b) assess whether temperature affects the type of toxicity interaction between these chemicals; and c) assess the temperature effect on biochemical responses (fatty acids (FA) and sugar profiles) of the pesticides on T. weissflogii. Temperature increased the tolerance of the diatoms to the pesticides with EC50 values between 3.176 and 9.929 µg L-1 for oxyfluorfen and 42.50-230.75 µg L-1 for Cu, respectively, at 15 °C and 25 °C. The mixtures toxicity was better described by the IA model, but temperature altered the type of deviation from dose ratio (15 °C and 20 °C) to antagonism (25 °C). Temperature, as well as the pesticide concentrations, affected the FA and sugar profiles. Increased temperature increased saturated FA and decreased unsaturated FA; it also affected the sugar profiles with a pronounced minimum at 20 °C. Results highlight effects on the nutritional value of these diatoms, with potential repercussion on food webs.

Do Freshwater and Marine Bivalves Differ in Their Response to Wildfire Ash? Effects on the Antioxidant Defense System and Metal Body Burden.

Wildfires constitute a source of contamination to both freshwater and marine ecosystems. This study aimed to compare the antioxidant defense response of the freshwater clam Corbicula fluminea and the marine cockle (Cerastoderma edule) to wildfire ash exposure and the concomitant metal body burden. Organisms were exposed to different concentrations (0%, 12.5%, 25%, 50%, and 100%) of aqueous extracts of Eucalypt ash (AEAs) from a moderate-to-high severity wildfire. The activity of various enzymes, as well as lipid peroxidation, protein content, and metal body burden, were determined after 96 h of exposure. A significant increase in the protein content of soft tissues was observed for C. edule at AEA concentrations ≥ 25%, unlike for C. fluminea. Similarly, significant effects on lipid peroxidation were observed for cockles, but not for clams. For both species, a significant effect in the total glutathione peroxidase activity was observed at AEA concentrations ≥ 25%. Relative to the control, AEAs-exposed clams showed higher Cd content, whereas AEAs-exposed cockles showed higher Cu content, thus exhibiting different responses to the exposure to wildfire ash. The susceptibility of bivalves to ashes, at environmentally relevant concentrations, raises concern about the effects of post-fire runoff to bivalve species.

Ozonation of Selected Pharmaceutical and Personal Care Products in Secondary Effluent-Degradation Kinetics and Environmental Assessment.

The efficiency of ozonation depends on the water matrix and the reaction time. Herein, these factors were addressed by assessing the removal of five pharmaceutical and personal care products (PPCPs) by ozonation. The main aims were: (i) to assess the effects of the water matrix on the degradation kinetics of PPCPs, individually and in mixture, following ozonation; and (ii) to assess the ecotoxicological impact of the ozone reaction time on the treatment of a spiked municipal wastewater (MW) added the five PPCPs over several species. The degradation of the PPCPs was faster in ultrapure water, with all PPCPs being removed in 20 min, whereas in the MW, a 30 min ozonation period was required to achieve a removal close to 100%. Increasing the number of PPCPs in the water matrix did not affect the time required for their removal in the MW. Regarding the ecotoxicity assessment, Raphidocelis subcapitata and Daphnia magna were the least sensitive species, whereas Lemna minor was the most sensitive. The temporal variation of the observed effects corroborates the degradation of the added PPCPs and the formation of toxic degradation by-products. The removal of the parent compounds did not guarantee decreased hazardous potential to biological species.

Do sediment-bound nickel and lead affect chironomids life-history? Toxicity assessment under environmentally relevant conditions.

Metal pollution in aquatic ecosystems translates into increased concentrations of sediment-bound metals, representing a risk for benthic species. This risk might be enhanced in soft and moderately hard waters, world widely distributed, due to the protective role of hardness against metal toxicity. As lead (Pb) and nickel (Ni) are amongst the more abundant metals in aquatic systems, and since their combined effects to benthic species have been overlooked, in this study we aimed to investigate the life-cycle toxicity of Pb and Ni (using spiked sediment) to the benthic species Chironomus riparius, considering both single and mixture exposures, in moderately hard water. Environmentally relevant concentrations of each metal were used (25 and 75 mg kg-1, based on a scenario of pollution by runoff waters from burnt forests), following a full factorial design. Effects of the mixture with the highest metal concentrations (Pb 75 mg kg-1 dw + Ni 75 mg kg-1 dw) were also assessed in the second generation. In the first generation, exposure to Pb increased emergence and the weight of males, and decreased time to emergence of both males and females. Conversely, exposure to Ni delayed female emergence and decreased the weight of imagoes. Summarizing, Pb affected more endpoints but showed an apparent positive effect, whereas Ni affected less endpoints but exhibited adverse effects. Reproduction was not affected by these metals. In the second generation, the mixture Pb 75 mg kg-1 + Ni 75 mg kg-1 dw delayed emergence and reduced the emerged female fraction and their weight. These results highlight that Pb and Ni can alter the structure of C. riparius populations at environmentally relevant concentrations, which signals potential repercussions in the dynamics and functioning of freshwater ecosystems under these contamination scenarios. The findings of the present study are relevant not only for metal-polluted environments, in general, but also for fire-affected ecosystems.

Effects of Eucalypt ashes from moderate and high severity wildfires on the skin microbiome of the Iberian frog (Rana iberica).

Forest fires can threaten amphibians because ash-associated contaminants transported by post-fire runoff impact both terrestrial and aquatic ecosystems. Still, the effects of these contaminants on the skin microbiome of amphibians have been overlooked. Thus, the main objective of this study was to assess the effects of ash from different severity wildfires (moderate and high) on the skin microbiome of the Iberian frog (Rana iberica). Bacterial isolates sampled from R. iberica skin microbiome were tested for their antimicrobial activity against the pathogen Aeromonas salmonicida. The isolates with antimicrobial activity were identified and further exposed to several concentrations (0, 6.25, 12.5, 25, 50, 75, and 100%) of Eucalypt (Eucalyptus globulus) aqueous extracts (AAEs) of ash from both a moderate and a high severity wildfire. The results showed that 53% of the bacterial isolates presented antimicrobial activity, with Pseudomonas being the most common genus. Exposure to AAEs had diverse effects on bacterial growth since a decrease, an increase or no effects on growth were observed. For both ash types, increasing AAEs concentrations led to an increase in the number of bacteria whose growth was negatively affected. Ash from the high severity fire showed more adverse effects on bacterial growth than those from moderate severity, likely due to the higher metal concentrations of the former. This study revealed that bacteria living in Iberian frogs' skin could be impaired by ash-related contaminants, potentially weakening the individual's immune system. Given the foreseen increase in wildfires' frequency and severity under climate change, this work raises awareness of the risks faced by amphibian communities in fire-prone regions, emphasising the importance of a rapid implementation of post-fire emergency measures for the preservation and conservation of this group of animals.

Effects of Pine and Eucalypt ashes on bacterial isolates from the skin microbiome of the fire salamander (Salamandra salamandra).

Environmental contamination influences the diversity of the resident skin microbial community of amphibians, ultimately affecting the individual's immune system. Wildfires are expected to impact the skin microbiome, since post-fire runoff typically transports hazardous substances, that can affect terrestrial and aquatic ecosystems. The present study is the first to assess the effects of Eucalypt and Pine wildfire ash on cultivable bacterial isolates from the skin microbiome of amphibians, in particular the fire salamander (Salamandra salamandra), a common species in fire-prone Mediterranean ecosystems. To achieve this goal, samples of skin bacteria of adult individuals of S. salamandra were collected at a site without influence of wildfires. The bacterial isolates were tested against the pathogenic agent Aeromonas salmonicida for assessing their antimicrobial activity, before exposing them to a series of dilutions of aqueous extracts of Eucalypt and Pine ashes (AAEs) from high severity wildfires. From the 80 bacterial isolates collected, 48 (mostly Pseudomonas spp.) showed antimicrobial activity. Exposure of bacteria with antimicrobial activity to the Eucalypt and Pine AAEs at concentrations of 0, 6.25, 12.5, 25, 50, 75, and 100%, revealed that bacterial growth could be significantly inhibited, stimulated or unaffected by ash. Growth inhibition was found for Pine and Eucalypt AAEs at concentrations as low as 6.25% and 12.5%, respectively, but were more expressive at concentrations equal or above 50%. Eucalypt AAEs had a higher negative impact on bacterial growth than Pine AAEs, likely due to differences in metal concentrations between ash types. These findings raise concern about the future of amphibians in fire-prone regions since the foreseen increase in fire frequency and severity owing to climate changes are likely to alter the skin microbiome of amphibians, weaken the immune system and consequently increasing the incidence of infections or diseases, further contributing to the decline of the populations.

Key Challenges to the Effective Management of Pollutants in Water and Sediment.

The intensification of human activities is placing increasing pressure on the ecosystems of riverine, estuarine, and coastal waters, as these compartments are sinks for many anthropogenic contaminants [...].

Automated Counting of Daphnid Neonates, Artemia Nauplii, and Zebrafish Eggs: A Proof of Concept.

In aquatic invertebrate (e.g., daphnids and Artemia sp.) and zebrafish cultures, in ecotoxicological bioassays, or when addressing complex population-level experimental designs, the counting of an organism's progeny is often required. This counting process is laborious, repetitive, and time-consuming, potentially posing health hazards to the operators, and necessarily entailing a higher likelihood of human error. We present an experimental evaluation of a computer-based device for counting neonates (Daphnia magna, Daphnia longispina, and Ceriodaphnia sp.), nauplii (Artemia salina and Artemia franciscana), and zebrafish (Danio rerio) eggs. Manual counts by an experienced technician were compared with the corresponding automated counts achieved by the computer-based counting device. A minimum of 55 counts/species was performed, with the number of counted organisms being up to a maximum of 150 neonates of Ceriodaphnia dubia, 200 neonates of D. magna and D. longispina, 200 nauplii of A. franciscana and A. salina, and 500 zebrafish eggs. Manual and automated counts were both performed in culture medium solutions of 50 ml of volume. Automated counts showed a mean relative acccuracy of 98.9% (97.9%-99.4%) and a relative standard deviation of 1.72%. The results demonstrate that the computer-based device can be used for accurately counting these aquatic organisms. This computer-based counting might be extended to other organisms of similar size, thus facilitating reproduction and life-cycle ecotoxicity tests. Environ Toxicol Chem 2022;41:1451-1458. © 2022 SETAC.

A review on polycyclic aromatic hydrocarbons distribution in freshwater ecosystems and their toxicity to benthic fauna.

Polycyclic aromatic hydrocarbons (PAHs) are a group of organic compounds, found ubiquitously in all environmental compartments. PAHs are considered hazardous pollutants, being of concern to both the environmental and human health. In the aquatic environment, PAHs tend to accumulate in the sediment due to their high hydrophobicity, and thus sediments can be considered their ultimate sink. Concurrently, sediments comprise important habitats for benthic species. This raises concern over the toxic effects of PAHs to benthic communities. Despite PAHs have been the subject of several reviews, their toxicity to freshwater benthic species has not been comprehensively discussed. This review aimed to provide an overview on PAHs distribution in freshwater environments and on their toxicity to benthic fauna species. The distribution of PAHs between sediments and the overlying water column, given by the sediment-water partition coefficient, revealed that PAHs concentrations were 2 to 4 orders of magnitude higher in sediments than in water. The sediment-water partition coefficient was positively correlated to PAHs hydrophobicity. Toxicity of PAHs to benthic fauna was addressed through Species Sensitivity Distributions. The derived hazardous concentration for 5% of the species (HC5) decreased as follows: NAP (376 µg L-1) > PHE > PYR > FLT > ANT (0.854 µg L-1), varying by 3 orders of magnitude. The hazardous concentrations (HC5) to benthic species were inversely correlated to the hydrophobicity of the individual PAHs. These findings are pertinent for environmental risk assessment of these compounds. This review also identified future challenges regarding the environmental toxicity of PAHs to freshwater benthic communities, namely the need for updating the PAHs priority list and the importance of comprehensively and more realistically assess the toxicity of PAHs in combination with other stressors, both chemical and climate-related.

New insights on the effects of ionic liquid structural changes at the gene expression level: Molecular mechanisms of toxicity in Daphnia magna.

Knowledge on the molecular basis of ionic liquids' (ILs) ecotoxicity is critical for the development of these designer solvents as their structure can be engineered to simultaneously meet functionality performance and environmental safety. The molecular effects of ILs were investigated by using RNA-sequencing following Daphnia magna exposure to imidazolium- and cholinium-based ILs: 1-ethyl-3-methylimidazolium chloride ([C2mim]Cl), 1-dodecyl-3-methylimidazolium chloride ([C12mim]Cl) and cholinium chloride ([Chol]Cl)-; the selection allowing to compare different families and cation alkyl chains. ILs shared mechanisms of toxicity focusing e.g. cellular membrane and cytoskeleton, oxidative stress, energy production, protein biosynthesis, DNA damage, disease initiation. [C2mim]Cl and [C12mim]Cl were the least and the most toxic ILs at the transcriptional level, denoting the role of the alkyl chain as a driver of ILs toxicity. Also, it was reinforced that [Chol]Cl is not devoid of environmental hazardous potential regardless of its argued biological compatibility. Unique gene expression signatures could also be identified for each IL, enlightening specific mechanisms of toxicity.

Effects of pH and nitrites on the toxicity of a cypermetrin-based pesticide to shrimps.

The Pantanal (Brazil) is a wetland region characterized by seasonal flooding. Hydrological cycles influence the water physicochemical parameters, causing seasonal variations in pH and nitrites. Thus, the objective of this study was to evaluate the effects of varying pH and nitrite concentrations on the toxicity of the cypermethrin-based pesticide Barrage®, considering both lethal (mortality) and sublethal endpoints (growth and development). Larvae of the endemic shrimp Macrobrachium pantanalense and of the estuarine Amazonian congener Macrobrachium amazonicum were exposed to cypermethrin (through Barrage®) under several pH levels (6.5, 7.5 and 8.5) or nitrite concentrations (0.1, 0.2 and 0.4 mg/L). The pH had direct effects on all the tested endpoints for both species. For M. pantanalense, the lethal effects of the cypermethrin formulation were more pronounced at low pH (96-h LC50 = 0.004 µg/L at pH 6.5, and 0.146 µg/L at pH 8.5). For M. amazonicum, an opposite response was observed, with increased toxicity of the formulation at high pH (96-h LC50 = 0.110 µg/L at pH 6.5 and 0.044 µg/L at pH 8.5). Variations in pH also seemed to modify the sublethal effects of the formulation on larval growth and development of M. pantanalense. Nitrite concentrations affected larval growth of both species, modifying also the effects of the cypermethrin formulation on the larval development of M. amazonicum. This work shows the importance of considering abiotic factors for risk assessment either due to possible direct effects on the physiology of organisms and/or due to interactions with other stressors, particularly in fragile biomes such as Pantanal.

The role of humic acids on gemfibrozil toxicity to zebrafish embryos.

Climate change is expected to alter the dynamics of water masses, with consequent changes in water quality parameters such as dissolved organic carbon (DOC) concentration. DOC levels play a critical role in the fate of organic chemicals, influencing their bioavailability and toxicity to aquatic organisms. This study aimed to evaluate the effects of DOC, particularly humic acids (HA), in the toxicity of gemfibrozil (GEM) - a human pharmaceutical frequently detected in surface waters. Lethal and sublethal effects (genotoxic, biochemical and behavioural alterations) were evaluated in zebrafish embryos exposed to several concentrations of GEM and three HA levels, in a full factorial design. HA significantly increased GEM LC50 values, mainly in the first 72 h of exposure, showing a protective effect. At sublethal levels, however, such protection was not observed since HA per se elicited adverse effects. At a biochemical level, individual exposure to HA (20 mg/L) elicited significant decreases in cholinesterase and glutathione S-transferase activities. Regarding behaviour, effects of individual exposure to HA appear to surpass the GEM effects, reducing the total distance moved by larvae. Both GEM and HA significantly increased DNA damage. Hence, this study demonstrated that abiotic factors, namely HA, should be considered in the assessment of pharmaceuticals toxicity. Moreover, it showed that lethality may not be enough to characterize combined effects since different patterns of response may occur at different levels of biological organization. Testing sublethal relevant endpoints is thus recommended to achieve a robust risk assessment in realistic scenarios.

Unraveling the ecotoxicity of deep eutectic solvents using the mixture toxicity theory.

The interest on deep eutectic solvents (DES) has been increasing. However, the ecotoxicological profile of DES is scarcely known. Also, despite previous studies showed that DES components dissociate in water, none assessed DES toxicity using the classical and adequate models for mixture toxicity prediction - concentration addition (CA) and independent action (IA). This study evaluates the ecotoxicological profile of DES based on [N1111]Cl, [N2222]Cl and [N3333]Cl as hydrogen bond acceptors (HBA) combined with hydrogen-bond donors (HBD) vis. ethylene glycol and 1-propanol, through the Microtox® Acute Toxicity Test. CA and IA with deviations describing synergism/antagonism, dose-ratio and dose-level effects were fitted to the toxicity data. Neither the starting materials nor DES were found hazardous to Aliivibrio fischeri, in this specific case agreeing with the claimed "green character" of DES. Among the starting materials, ethylene glycol was the least toxic, whereas [N3333]Cl was the most toxic (30 min-EC50 = 96.49 g L-1 and 0.5456 g L-1, respectively). DES toxicity followed the same trend as observed for the salts: [N1111]Cl-based DES < [N2222]Cl-based DES < [N3333]Cl-based DES. The IA model, with specific deviations, adjusted better in 5 out of 6 DES. Antagonism was observed for [N1111]Cl-based DES, and synergism for [N3333]Cl-based DES and for 1-propanol:[N2222]Cl. The application of the mixture toxicity models represents a breakthrough in the problematic of assessing the toxicity of the countless number of DES that can be created with the same starting materials, since they provide the expected toxicity of any virtual combination between HBA and HBD.

Phytotoxicity of silver nanoparticles to Lemna minor: Surface coating and exposure period-related effects.

Silver nanoparticles (Ag NPs) exponential production raises concern about their environmental impact. The effects of Ag NPs to aquatic plants remain scarcely studied, especially in extended exposures. This paper aims to evaluate Ag NPs effects in Lemna minor at individual and sub-individual levels, focusing on three variables: Ag form (NPs versus ions - Ag+), NPs surface coating (citrate vs polyvinylpyrrolidone - PVP) and exposure period (7 vs 14days). Endpoints were assessed at individual level (specific growth rate, chlorosis incidence and number of fronds per colony) and sub-individual level (enzymatic activities of catalase (CAT), guaiacol peroxidase (GPx) and glutathione-S-transferase (GST)). Generally, plants exposed to all Ag forms underwent decays on growth rate and fronds per colony, and increases on chlorosis, GPX and GST, but no effects on CAT. The most sensitive endpoints were specific growth rate and GPx activity, showing significant effects down to 0.05mg/L for Ag NPs and 3µg/L for Ag+, after 14days. Ag+ showed higher toxicity with a 14d-EC50 of 0.0037mg Ag/L. Concerning surface coating, PVP-Ag NPs were more deleterious on growth rate and fronds per colony, whereas citrate-Ag NPs affected more the chlorosis incidence and GPx and GST activities. The exposure period significantly affected chlorosis: 14days triggered a chlorosis increase in Ag+-exposed plants and a decrease in Ag NPs-exposed plants when compared to 7days. Ag NPs induced an oxidative stress status in cells, thus ensuing upregulated enzymatic activity as a self-defense mechanism. Since Ag NPs dissolution might occur on a steady and continuous mode along time, and the average longevity of fronds, we propose longer exposures periods than the recommended by the OECD guideline. This approach would provide more relevant and holistic evidences on the overall response of freshwater plants to Ag NPs in an ecological relevant scenario.

The effects of Copper and Zinc on survival, growth and reproduction of the cladoceran Daphnia longispina: introducing new data in an "old" issue.

Metal contamination is still a major environmental issue due to their continuous deposition and persistence. In this work we intended to assess the impact that Copper (Cu) and Zinc (Zn) exert in life-history parameters of Daphnia longispina, a common cladoceran in freshwater environments. Thus, we studied the effects of Cu (20-300 µg/L) and Zn (500-4000 µg/L) on the survival, growth, reproduction, feeding rate and population growth rate of D. longispina. Though survival was only reduced for the highest concentration of each metal, other endpoints were strongly affected by lower concentrations. Growth was affected by both metals, especially in the period 0-7 d, being significant for Cu ≥ 40 µg/L and Zn ≥ 500 µg/L. Indeed, growth endpoints at day 7 (body length and growth rate) were equally or more sensitive than the corresponding endpoints at day 21. The size at first reproduction decreased (significant for Cu ≥ 40 µg/L and Zn ≥ 500 µg/L). Reproduction decreased, which was partially explained by the reduced growth, but additional effects were supported by a direct effect of metals on reproduction (based on the relationship body length-brood size). Reduced growth and reproduction are likely a food mediated effect due to feeding inhibition caused by the metals. Globally, the studied endpoints were affected by Cu and Zn differently, supporting a chemical-specificity of the effects, with Zn causing more pronounced effects than Cu. This work presents an innovative approach to the effects of Cu and Zn to D. longispina, giving a general and comprehensive overview of those effects.

Endemic shrimp Macrobrachium pantanalense as a test species to assess potential contamination by pesticides in Pantanal (Brazil).

Pantanal is a biome characterized by an extraordinary diversity and abundance of wildlife and houses several endemic species such as the freshwater shrimp Macrobrachium pantanalense. However, the increase in agriculture and husbandry activities in the region has contributed with residues of pesticides reaching aquatic systems. The main objective of this study is to assess the sensitivity of the endemic shrimp M. pantanalense compared with other freshwater species: the shrimp M. amazonicum, the crustacean Daphnia similis and the fish Danio rerio. The sensitivity of these organisms was assessed through acute exposure to copper and cypermethrin (through the formulation Barrage®, widely used in Pantanal). For copper the species sensitivity decreased in the following order: D. similis (48 h-EC50 0.051 mg/L) > M. pantanalense > D. rerio > M. amazonicum (48 h-LC50 26.34 mg/L). Copper caused reduced length of shrimps and zebrafish and reduced heartbeat of zebrafish embryos. For cypermethrin the species sensitivity decreased in the following order: M. pantanalense (96 h-LC50 0.05 µg/L) > M. amazonicum > D. similis > D. rerio (144 h-LC50 1680 µg/L). Major effects of cypermethrin included reduced length of shrimps and zebrafish, as well as early hatching and increased incidence of developmental deformities in zebrafish embryos. This study highlights the importance of using endemic species for risk evaluations in sensitive biomes such as Pantanal. Moreover, it emphasizes the importance of testing pesticides toxicity as commercial formulations. Furthermore, we suggest that the endemic shrimp species M. pantanalense can be successfully used as a test species in ecotoxicology.

Lethal and sub lethal effects of the biocide chlorhexidine on aquatic organisms.

Chlorhexidine is among the most used biocides in Europe, however its toxicity to aquatic organisms is scarcely known. The main objective of this study was to assess the lethal and sub lethal effects of chlorhexidine digluconate (ChD) on four aquatic model organisms: the bacteria Vibrio fischeri, the algae Pseudokirchneriella subcapitata, the crustacean Daphnia magna and the embryos of the fish Danio rerio. ChD was very toxic to algae and crustaceans, with a 72 h-EC50 of 62.2 µg/l and a 48 h-EC50 of 45.0 µg/l, respectively. Toxicity to fish embryos and the bacteria was lower, with a 96 h-EC50 of 804.0 µg/l and a 15 min-EC50 of 1,694.0 µg/l, respectively. Concerning sub lethal effects on D. magna (feeding inhibition) a 6 h-EC50 of 503.7 µg/l was obtained. In fish, ChD caused developmental abnormalities, namely alterations in the amniotic fluid (48 h-EC20 of 753.6 µg/l) and early hatching. Moreover, enzymatic biomarkers on fish embryos showed an induction of cholinesterase activity in all ChD tested concentrations (80-900 µg/l). The catalase activity was also induced at the highest concentration tested (900 µg/l) whereas no changes were observed for glutathione-S-transferase and lactate dehydrogenase activities. The toxicity of ChD to the algae and crustacean raises concerns about its potential effects in aquatic food webs, since these organisms are in the base of trophic chains, and highlights the need for further studies on ChD toxicity to aquatic organisms.

Mercury toxicity to freshwater organisms: extrapolation using species sensitivity distribution.

Mercury toxicity to aquatic organisms was evaluated in different taxonomic groups showing the following species sensitivity gradient: Daphnia magna > Daphnia longispina > Pseudokirchneriella subcapitata > Chlorella vulgaris > Lemna minor > Chironomus riparius. Toxicity values ranged from 3.49 µg/L (48 h-EC50 of D. magna) to 1.58 mg/L (48 h-EC50 of C. riparius). A species sensitivity distribution was used to estimate hazardous mercury concentration at 5 % level (HC5) and the predicted no effect concentration (PNEC). The HC5 was 3.18 µg Hg/L and the PNEC varied between 0.636 and 3.18 µg Hg/L, suggesting no risk of acute toxicity to algae, plants, crustaceans and insects in most freshwaters.