The role of warming in modulating neodymium effects on adults and sperm of Mytilus galloprovincialis.

The use of rare earth elements (REEs) has been increasing and one of the most used is neodymium (Nd). Being an emergent contaminant, its negative impacts are poorly understood. Aquatic organisms are also threatened by climate change-related factors, as is the case of warming, which can change the effects of REEs. Thus, the impacts of Nd, warming, and the combination of both stressors were studied in adult mussels and sperm of the species Mytilus galloprovincialis, after an exposure period of 28 days (adults) and 30 min (sperm). The effects were evaluated through the analysis of biochemical and histopathological alterations in adults and biochemical and physiological responses given by sperm. The results showed that mussels only activated their biotransformation capacity when exposed to the stressors acting alone, which was insufficient to avoid lipid peroxidation. Furthermore, warming (alone and combined with Nd) also produces damage to proteins. The digestive gland was the most sensitive organ to Nd, presenting several histopathological alterations. In the case of sperm, all stressors induced lipid peroxidation, a higher oxygen demand, and a decrease in velocity, even if the sperm viability was maintained. It seems that warming influenced the effects of Nd to some extent. The present findings contribute significantly to the field of REEs environmental toxicology by offering valuable insights into the impacts of Nd on various biological levels of mussels. Additionally, within the context of climate change, this study sheds light on how temperature influences the effects of Nd. The obtained results indicate that both stressors can potentially compromise the overall health of mussel populations, thereby affecting other species reliant on them for food and habitat. Moreover, this study highlights impaired sperm health, which could adversely affect their reproductive capacity and ultimately lead to population decline.

Toxic effects of a mixture of pharmaceuticals in Mytilus galloprovincialis: The case of 17α-ethinylestradiol and salicylic acid.

The impact of pharmaceuticals on marine invertebrates has been a topic of rising concern, with an increasing number of studies regarding the impacts on bivalves. However, very few investigated the toxicity of mixtures of pharmaceuticals. This knowledge gap was investigated in the present study, where the toxicity of 17α-ethinylestradiol (EE2) and salicylic acid (SA) mixture was evaluated. To this end, Mytilus galloprovincialis mussels were chronically subjected to both pharmaceuticals, acting alone and in combination, and the effects at the cellular level were measured. The Independent Action (IA) model was performed aiming to compare obtained with predicted responses. The integrated biomarker response (IBR) index was used to assess the overall biochemical response given by mussels. The results obtained revealed that the most stressful condition was caused by the combined effect of EE2 and SA, with the highest metabolic capacity, antioxidant (catalase activity) and biotransformation (carboxylesterases activity) activation and cellular damage in organisms exposed to the mixture of both drugs in comparison to responses observed when each drug was acting alone. Predicted responses obtained from the IA model indicate that caution should be paid as frequent deviations to observed responses were found. This study highlights the need for future studies considering the mixture of pollutants, mimicking the actual environmental conditions.

Metabolic and oxidative status alterations induced in Ruditapes philippinarum exposed chronically to estrogen 17α-ethinylestradiol under a warming scenario.

The presence of pharmaceuticals in the aquatic environment is an ongoing concern. However, the information regarding their effects under different climate change scenarios is still scarce. 17α-ethinylestradiol (EE2) is widely present in different aquatic systems showing negative impacts on aquatic organisms even when present at trace concentrations (≈1 ng/L). Nevertheless, its impact on bivalves is poorly understood, especially considering the influence of climate change factors. This study aimed to assess the toxicological impacts of EE2 under current and predicted warming scenarios, in the edible clam Ruditapes philippinarum. For this, clams were exposed for 28 days to different EE2 concentrations (5, 25, 125, 625 ng/L), under two temperatures (17 °C (control) and 21 °C). Drug concentrations, bioconcentration factors and biochemical parameters, related to oxidative stress and energy metabolism, were evaluated. Results showed that under actual and predicted temperature scenarios EE2 concentrations led to a disturbance in redox homeostasis of the clams, characterized by an increase in oxidized glutathione in contaminated organisms compared to control ones. Nevertheless, clams were capable to cope with the stressful conditions, activating their defence mechanisms (especially at the highest exposure concentration and in particular at increased temperature), and no oxidative damage occured. Although limited effects were observed, the present findings indicate that under both temperatures contaminated clams altered their biochemical performance, which can impair their sensitivity and protection capacity to respond to other environmental changes and/or affect their capacity to grow and reproduce. The results presented here highlight the need for further research on this thematic, considering that climate change is an ongoing problem, and the levels of some pharmaceutical drugs will continue to increase in marine/estuarine environments.

Cell death and changes in primary metabolism: the onset of defence in Eucalyptus in the war against Leptocybe invasa.

BACKGROUND: Here, we investigated changes in primary metabolism and cell death around oviposition sites in two hybrid clones of Eucalyptus with different degrees of resistance to Leptocybe invasa Fisher & La Salle (Hymenoptera: Eulophidae), as well as tolerance to water deficiency. RESULTS: We showed that apices of the resistant clone with oviposition had a higher content of amino acids, organic acids and the compound putrescine compared with those of the susceptible clone with oviposition. By contrast, apices of the resistant clone with oviposition had lower sugar and pyruvate organic acid content than those of the susceptible clone with oviposition. Small areas of necrosis were induced around the oviposition sites in the stem apices of Eucalyptus 24 h after infestation. The resistant clone developed larger necrotic areas that showed progressive increases 24-72 h after infestation compared with the susceptible clone, in which cell death was significantly lower and no changes were observed in necrotic area over time. Thus, the programmed death of cells around the egg, modulated by several amino acids, is likely the first defence response of Eucalyptus against L. invasa. CONCLUSION: Our results serve as the basis for the early identification of key metabolites produced in plants in defence against galling insects. © 2022 Society of Chemical Industry.

Influence of salinity on the toxicity of copper and cadmium to Zebrafish embryos.

Salinization has become a serious worldwide environmental perturbation in freshwater ecosystems. Concomitantly, many of such ecosystems are already impacted by other toxicants, which together with increased salinity may result in synergistic, antagonistic or additive toxic effects to biota. This work intended to assess the influence of increasing salinity (by using NaCl) on the lethal and sublethal toxicity of two metallic elements (copper and cadmium) in embryos of the fish species Danio rerio. This goal was achieved by exposing zebrafish embryos to seven concentrations of NaCl, individually or combined with each metal, using a full factorial design. The following endpoints were monitored in the test organisms: mortality, hatching, malformations and the enzymatic activity of glutathione S-transferase (GST) and cholinesterase (ChE). Overall, moderate salinity levels alleviated the lethal toxicity of both copper and cadmium although this effect was stronger in the copper assay. This effect was also influenced, as expected, by the concentrations of the metals indicating that the protective effect of salt only reaches some levels, after what is overwhelmed by the high metal toxicity, especially with the non-essential metal cadmium. At sub-lethal concentrations, the interactive effect resulting from NaCl and metals was not consistent and varied with the endpoint analyzed and the metal tested. Overall, the interactions between the salt and metals seem complex and with more drastic effects (positive or negative) on lethal endpoints than sub-lethal.

Differential development times of galls induced by Leptocybe invasa (Hymenoptera: Eulophidae) reveal differences in susceptibility between two Eucalyptus clones.

BACKGROUND: The significance of morphological responses of hosts on susceptibility against gall-inducing insects is relatively unknown, especially in planted forests. Here, we investigate the temporal morphological responses (gall development) induced by the invasive gall wasp Leptocybe invasa and the subsequent insect development in two Eucalyptus clones. RESULTS: Our results identified a novel stage of gall development, not previously reported, termed here 'brownish ring'. In both hosts similar gall development stages were observed. Although L. invasa oviposited in both clones, comparison of external morphological traits of galls over time revealed a differential response in the number of galls between clones. Comparison of the developmental time of each gall and insect stage between clones suggests that plant defense mechanisms against L. invasa are activated shortly after oviposition by the wasp, yet before gall formation. CONCLUSION: Gall number is an important parameter that should be used to measure host susceptibility among Eucalyptus clones. To the best of our knowledge, this is the first study showing differential morphological responses induced by a galling insect, even before gall formation, revealing differences in susceptibility between different plant hosts. These findings provide insight into the use of early stages of gall formation by L. invasa to prevent invasion and establishment of this pest.

Lumbriculus variegatus (oligochaeta) exposed to polyethylene microplastics: biochemical, physiological and reproductive responses.

Freshwater sediments are a repository of microplastics (MPs) resulting from inland anthropogenic activities. Benthic invertebrates, particularly endobenthic sediment-ingesting species such as the annelid Lumbriculus variegatus (blackworm), are commonly found in contaminated sediments where they likely find and ingest MPs. In the present study, L. variegatus was exposed to concentrations between 0.51 and 20 g kg-1 dry sediment of four size-classes of irregularly-shaped polyethylene MPs (PE-MPs; size-class A: 32-63, B: 63-125, C: 125-250 and D: 250-500 µm) for 48 h to assess their sub-cellular responses to particles ingested, and for 28 days to determine chronic effects on worm's reproduction and biomass. After the short-term exposure (48 h), number of PE-MPs in blackworms' gut were related to MPs concentration in the sediment. In general, PE-MPs ingestion by blackworms induced depletion of their energy reserves (e.g., sugars in all size classes and lipids in the size-classes of PE-MPs > 125 µm), concomitant with the activation of antioxidant and detoxification mechanisms (increased level of total glutathione in all size-classes, and increased glutathione-S-transferase activity in PE-MPs > 250 µm), preventing lipid peroxidation. In addition, it was observed a reduction of aerobic energy production (decreased activity of the electron transport system) and a slight increase in neurotransmission (cholinesterase activity). After a long-term exposure (28 d), the presence and ingestion of PE-MPs did not affect reproduction and biomass of L. variegatus. The activation and efficiency of the antioxidant and detoxification mechanisms allied with the anatomy and physiology of L. variegatus, its feeding strategy and potentially dynamic ingestion/egestion capacity seem to be key features preventing MP deleterious effects under short- and chronic-exposures. Considering the MPs levels reported for freshwater sediments, and despite evidence of MPs ingestion and some sub-organismal effects, our results suggest no adverse impacts of PE-MPs contamination on L. variegatus populations fitness. This study applies an integrative approach in which data concerning the ingestion of different sized MPs and subsequent sub-cellular and apical responses are delivered, raising knowledge on endobenthic invertebrates' strategies to potentially overcome MP toxicity in field contaminated sites.

Daphnia magna responses to fish kairomone and chlorpromazine exposures.

To avoid being preyed, organisms must be able to identify predatory threats by sensing molecules released by predators (kairomones), and to employ effective strategies to prevent detection by predators. Furthermore, in the wild, organisms are also exposed to chemicals that may alter their behavioral traits, such as neuroactive pharmaceuticals. Considering the co-occurrence of both types of chemicals, their possible interaction needs to be studied. To address this topic, the aim of this study was to verify the effects of fish kairomone (FK - a chemical associated to putative predation by fish) and chlorpromazine (CPZ - neuroactive pharmaceutical drug, environmental contaminant), isolated and in combination, in different functional endpoints of Daphnia magna, such as oxygen consumption, feeding rate, behavior and reproduction. Among these endpoints, oxygen consumption was only affected by the combination of compounds (FK + CPZ). On the other hand, feeding rate was affected by all treatments, being lower than control. For life history traits and phototactic behavior, the effects of FK predominated over the ones caused by CPZ exposure, incrementing the reproductive output of females, leading to greater population growth rates and increasing negative phototactic behaviour.