There is limited information regarding the toxicity of the trace element thallium (Tl) to aquatic biota, most of which assesses acute toxicity and bioaccumulation. The relative lack of chronic Tl toxicity data compromises the establishment of water quality criteria for this trace metal. In the presented work, chronic toxicity endpoints (final body weight (a proxy measure of growth), survival, and reproduction) and Tl body burden were measured in the freshwater crustacean Daphnia magna during a 21-day exposure to dissolved Tl. Thallium caused complete mortality in daphnids between exposure concentrations of 424 and 702 μg L⁻¹. In contrast with previously published work examining acute Tl toxicity, exposure to Tl for 21 days was not associated with changes in whole-body potassium concentration. This was despite a 710-fold increase in Tl body burden in animals exposed to 424 μg L⁻¹ relative to the control. Median effect concentrations (EC₅₀’s) for growth and reproduction (total neonates produced), were 1.6 (95% confidence interval: 1.0–3.1) and 11.1 (95% confidence interval: 5.5–21.8) μg Tl L⁻¹, respectively. A no observable effect concentration (NOEC) of 0.9 μg Tl L⁻¹ for growth, and a NOEC range of 0.9–83 μg Tl L⁻¹ for a variety of reproductive metrics, was measured. A lowest observable effect concentration (LOEC) of 8.8 μg Tl L⁻¹ was determined for the effects of Tl on growth and most of the reproductive endpoints examined. These data indicate that under controlled laboratory conditions D. magna is significantly less sensitive to Tl than the species on which the current Canadian Council of Ministers of the Environment regulatory guideline value of 0.8 μg L⁻¹ is based.

Hyperbranched polymers are nanomaterials belonging to the class of dendritic architectures with increasing applications in many diverse fields. We studied the toxicity of two hyperbranched polymers to the freshwater crustacean Daphnia magna. A hyperbranched hydroxyl-terminated polyester and a commercial hyperbranched polyamidoamine, Helux-3316 were tested for the acute immobilization of daphnids, the overproduction of reactive oxygen species and the activity of the antioxidant enzymes catalase and glutathione S-transferase. The effect for D. magna immobilization was higher for the hyperbranched polyamidoamine Helux-3316, which was attributed to the presence of primary amino groups on its surface. Following exposure to both hyperbranched polymers, a clear overproduction of reactive oxygen species took place accompanied by concentration-dependent enzymatic antioxidant response. Our results showed that the overproduction of reactive oxygen species activated antioxidant defence mechanisms and was responsible for the immobilization of daphnids exposed to both hyperbranched polymers. We showed evidence of the uptake of fluorescently labelled Helux-3316 that accumulated into the gastrointestinal tract of D. magna, and its removal via excretion within fecal pellets. This is the first work reporting the internalization of hyperbranched polymers in aquatic organisms.

Plastic waste is continuously introduced not only into marine, but also freshwater environments, where it fragments into microplastics. Organisms may be affected by the particles themselves due to ingestion and indirectly via incorporated additives such as plasticizers, since these substances have the ability to leach out of the polymer matrix. Although it has been indicated that the likelihood of additives leaching out into the gut lumen of organisms exposed to microplastics is low, studies distinguishing between the effects of the synthetic polymer itself and incorporated additives of the same polymer are scarce. Since this is obligatory for risk assessment, we analyzed the chronic effects of flexible polyvinylchloride (PVC), a widely used polymer, containing the plasticizer diisononylphthalate (DiNP) on morphology and life history of the freshwater crustacean Daphnia magna and compared these effects with the effects of rigid PVC, lacking DiNP, as well as a glass bead control. After up to 31 days of exposure, rigid PVC and glass beads did not affect body length and relative tail spine length of D. magna, whereas flexible PVC led to an increased body length and a reduced number of offspring. None of the treatments increased the mortality significantly. We were able to show that 2.67μg/L DiNP leached out of the flexible PVC into the surrounding medium using GC-MS. Yet, we were not able to measure leachate inside the gut lumen of D. magna. The effects emerged towards the end of the experiment, due to the time dependent process of leaching. Therefore, the results highlight the relevance of long-term chronic exposure experiments, especially as leaching of additives takes time. Further, our study shows the importance to distinguish between microplastics containing leachable additives and the raw polymer in ecotoxicological testing.

Rare earth elements (REE) have become essential in high- and green-technologies. Their increasing use lead to the release of anthropogenic REE into the environment including aquatic systems. The limited data available on the aquatic ecotoxicology of REE indicate their biological effects are highly dependent on their speciation, posing challenges for a reliable environmental risk assessment (ERA). The current study assessed the influence of speciation on the toxicity of neodymium (Nd), gadolinium (Gd) and ytterbium (Yb) in the Daphnia magna mobility inhibition test (ISO 6341:2012). REE toxicity was assessed individually and in ternary mixture, in the absence and presence of dissolved organic matter (DOM). Speciation was predicted by modeling and REE bioaccumulation by D. magna was measured to better understand the relationship between REE speciation and toxicity. DOM decreased significantly the toxicity of Nd, Gd and the mixture towards this freshwater crustacean. This was explained by a lower REE bioaccumulation in the presence of DOM due to REE-DOM complexation, which reduced REE bioavailability. DOM effects on Yb toxicity and bioaccumulation were limited because of Yb precipitation. We show that the way of expressing EC50 values (based on nominal, measured or predicted REE concentrations in solution) drastically changed REE toxicity assessment and that these changes were influenced by REE speciation. This study demonstrates for the first time that REE speciation, and especially REE-DOM complexation, significantly influences REE bioaccumulation and toxicity towards D. magna. Our results have implications for the subsequent ERA of REE.

The ongoing development of nanotechnology has raised concerns regarding the potential risk of nanoparticles (NPs) to the environment, particularly aquatic ecosystems. A relevant aspect that drives NP toxicity is represented by the abiotic and biotic processes occurring in natural matrices that modify NP properties, ultimately affecting their interactions with biological targets. Therefore, the objective of this study was to perform an ecotoxicological evaluation of CeO₂NPs with different surface modifications representative of NP bio-interactions with molecules naturally occurring in the water environment, to identify the role of biomolecule coatings on nanoceria toxicity to aquatic organisms. Ad hoc synthesis of CeO₂NPs with different coating agents, such as Alginate and Chitosan, was performed. The ecotoxicity of the coated CeO₂NPs was assessed on the marine bacteria Aliivibrio fischeri, through the Microtox® assay, and with the freshwater crustacean Daphnia magna. Daphnids at the age of 8 days were exposed for 48 h, and several toxicity endpoints were evaluated, from the molecular level to the entire organism. Specifically, we applied a suite of biomarkers of oxidative stress and neurotoxicity and assessed the effects on behaviour through the evaluation of swimming performance. The different coatings affected the hydrodynamic behaviour and colloidal stability of the CeO₂NPs in exposure media. In tap water, NPs coated with Chitosan derivative were more stable, while the coating with Alginate enhanced the aggregation and sedimentation rate. The coatings also significantly influenced the toxic effects of CeO₂NPs. Specifically, in D. magna the CeO₂NPs coated with Alginate triggered oxidative stress, while behavioural assays showed that CeO₂NPs coated with Chitosan induced hyperactivity. Our findings emphasize the role of environmental modification in determining the NP effects on aquatic organisms.

Juvenile hormone analogue (JHA) insecticides are endocrine disrupters that interfere with hormonal action in insects by mimicking their juvenile hormones (JH). As the structure and functions of methyl farnesoate in crustaceans are similar to those of JH in insects, exogenous JHA insecticides could have adverse effects on the development and reproduction of crustaceans. This study examined the toxic effects of two JHA insecticides, fenoxycarb and methoprene, on a freshwater shrimp model of cherry shrimp, Neocaridina davidi. Both insecticides had detrimental effects on cherry shrimp, but fenoxycarb was more toxic than methoprene. Chronic exposure to these insecticides reduced the shrimp's body length and molting frequency. Based on transcriptome annotations for N. davidi, we identified important gene homologues that were active in both insect JH biosynthetic and degradative pathways as well as JH and ecdysteroid signaling pathways. Chronic treatments with JHAs had significant effects on these genes in N. davidi. Our transcriptomic analysis showed that genes involved in the pathways related to cuticle development, serine protease activity, and carbohydrate, peptide and lipid metabolic processes were differentially expressed in shrimp exposed to JHAs. These results demonstrate the toxicity of fenoxycarb and methoprene to freshwater crustaceans and indicate the need to monitor the use of JHA insecticides.

Toxicity of Cu and Cu-based nanoparticles (NPs) to aquatic biota is usually mitigated in natural freshwater compared to organics-free artificial freshwater. The main aim of this study was to evaluate whether mitigated toxicity is accompanied by lower total copper body burden in the freshwater crustacean Daphnia magna and whether CuO NPs are more hazardous in this aspect than soluble Cu salts.Total copper body burden in different media (OECD202 artificial freshwater and two natural freshwaters) was measured by a relatively novel technique - total reflection X-ray fluorescence (TXRF) spectroscopy - which proved suitable for the analysis of individual juvenile daphnids. Mean copper body burden was 2.8–42 times higher in daphnids exposed to CuO NPs (0.05 mg Cu/L and 1 mg Cu/L) than in daphnids exposed to equal or equitoxic concentrations (0.025 mg Cu/L and 0.05 mg Cu/L) of CuSO4. Using natural freshwater instead of artificial one resulted in increased copper burden after exposure to CuO NPs but not after exposure to Cu salt. After 24 h post-exposure depuration in the presence of algae Raphidocelis subcapitata, total copper body burden in daphnids exposed to CuO NPs sharply decreased while in daphnids exposed to Cu salt it did not. Despite the CuO NP toxicity mitigating effect of natural freshwater, total copper body burden of aquatic crustaceans in natural waterbodies may be greater than could be predicted based on the results obtained using artificial freshwater as the test medium.

With the ever-increasing consumption of pharmaceutical compounds, their presence in the environment is now an undisputable reality. The majority of these compounds are released into the wild after their therapeutic use, as biotransformation products or in their original form. The presence of this class of compounds in the environment, due to their biological properties, can exert effects on non-target organisms, with adverse consequences. In addition, some bioactive substances, such as stimulants of the central nervous system, are also used by humans as part of their diet. The adverse consequences posed by such chemicals may be permanent or transient, if the exposure to xenobiotics is halted; it is thus of the paramount importance to study effects that result from long-term exposure to toxicants, but also the recovery of organisms previously exposed to such substances, especially if such chemicals may cause some type of addiction. Caffeine (1,3,7-trimethylxanthine) is a naturally occurring alkaloid found in many plants, being one of the most common stimulant/pharmaceutical compounds found in the environment. In addition, it is addictive, and strongly consumed by humans, a factor that contributes also for its continuous presence in the aquatic environment. The aim of this study was to evaluate the effects of environmentally relevant concentrations (0.08; 0.4; 2; 10; and 50 μg/L) of caffeine on behavior and physiological parameters (that are proxies of metabolic traits, such as oxygen uptake and glycogen content), in individuals of the freshwater crustacean species Daphnia magna, of distinct ages, and with or without a recovery period in the absence of caffeine. Regarding behavior, the results indicated that caffeine exposure altered the moved distance of the test organisms, but not according to a coherent pattern; low concentrations of caffeine reduced the movement of exposed daphnids, while higher levels did not have any measurable effect on this parameter. In addition, it was possible to identify subtle withdrawal effects (animals exposed to caffeine during 21 days and kept in uncontaminated media for 2 days). Regarding the other two studied parameters, caffeine exposure did not result in any significant modification in oxygen uptake and glycogen stores/reserves of the test organisms, in animals continuously exposed, or in those subjected to a recovery period, suggesting that despite a behavioral stimulatory effect, this was not followed by any metabolic change, and no addictive effect was possible to infer. The results showed that the presence of caffeine in environmental concentrations can induce mild behavioral effects at low, albeit realistic levels, but not capable of establishing clear biochemical changes.

Hospital wastewater (HWW) contains different hazardous substances resulting from a combination of medical and non-medical activities of hospitals, including pharmaceutical residues. These substances may represent a threat to the aquatic environment if they do not follow specific treatment processes. Therefore, we aimed to investigate the effects of the untreated effluent collected from a general hospital in Mahdia City (Tunisia) on neonatal stages of the freshwater crustacean Daphnia magna. Test organisms were exposed to three proportions (3.12%, 6.25%, and 12.5% v/v) of HWW. After 48 h of exposure, a battery of biomarkers was measured, including the quantification of antioxidant enzymes [catalase (CAT) and total and selenium-dependent glutathione peroxidase (total GPx; Se-GPx)], phase II biotransformation isoenzymes glutathione-S-transferases (GSTs), cyclooxygenases (COX) involved in the regulation of the inflammatory process, and total cholinesterases (ChEs) activities. Lipid peroxidation (LPO) was measured to estimate oxidative damage. The here-obtained results showed significant decreases of CAT and GSTs activities and also on LPO content in daphnids, whereas Se-GPx activity was significantly increased in a dose-dependent manner. Impairment of cholinesterasic and COX activities were also observed, with a significant decrease of ChEs and an increase of COX enzymatic activities. Considering these findings, HWW was capable of inducing an imbalance of the antioxidant defense system, but without resulting in oxidative damage in test organisms, suggesting that peroxidases and alternative detoxifying pathways were able to prevent the oxidant potential of several drugs, which were found in the tested effluents. In general, this study demonstrated the toxicity of hospital effluents, measured in terms of the potential impairment of key pathways, namely neurotransmission, antioxidant defense, and inflammatory homeostasis of crustaceans.

Understanding how temperature alone affects biomarkers commonly used in ecotoxicology studies and biomonitoring programs is important to obtain a more real response in field studies, especially in freshwater. Thus, we analyzed the behavioral responses, the lethality, and the biochemical biomarkers in the freshwater crustacean Aegla longirostri at different water temperatures. Animals were exposed under laboratory conditions, to 18 °C, 21 °C, 24 °C, and 26 °C for 48 h. There were significant changes in biochemical parameters in different tissues (hepatopancreas, gills, and muscle) and in the behavioral tests in A. longirostri. Hepatopancreas was especially affected by the elevation of temperature, as showed by the high levels of carbonyl proteins. The activity of acetylcholinesterase increased in a temperature-dependent manner in muscle. Glutathione S-transferase activity decreased with the elevation of temperature in all tissues sampled. The results obtained in this study indicate that when assessing the health of polluted limnic ecosystems through the use of organisms in situ, the intrinsic effect of abiotic factors, such as temperature, on biomarkers must be considered.