Recommended rates of azoxystrobin and tebuconazole seem to be environmentally safe but ineffective against target fungi.

The use of fungicides in agriculture has been playing a role in the enhancement of agricultural yields through the control of pathogens causing serious diseases in crops. Still, adverse environmental and human health effects resulting from its application have been reported. In this study, the possibility of readjusting the formulation of a commercial product combining azoxystrobin and tebuconazole (active ingredients - AIs; Custodia®) towards environmentally safer alternative(s) was investigated. Specifically, the sensitivity of non-target aquatic communities to each AI was first evaluated by applying the Species Sensitivity Distributions (SSDs) approach. Then, mixtures of these AIs were tested in a non-target organism (Raphidocelis subcapitata) denoting sensitivity to both AIs as assessed from SSDs. The resulting data supported the design of the last stage of this study, where mixtures of those AIs at equivalent vs. alternative ratios and rates as in the commercial formulation were tested against two target fungal species: Pyrenophora teres CBS 123929 and Rhynchosporium secalis CBS 110524. The comparison between the sensitivity of non-target aquatic species and the corresponding efficacy towards target fungi revealed that currently applied mixture and rates of these AIs are generally environmentally safe (antagonistic interaction; concentrations below the EC1 for R. subcapitata and generally below the HC5 for aquatic non-target communities), but ineffective against target organisms (maximum levels of inhibition of 70 and 50% in P. teres CBS 123929 and R. secalis CBS 110524, respectively). Results additionally suggest a potentiation of the effects of the AIs by the other formulants added to the commercial product at tested rates. Overall, this study corroborates that commercial products can be optimized during design stages based on a systematic ecotoxicological testing for ingredient interactions and actual efficacy against targets. This could be a valuable pathway to reduce environmental contamination during transition to a more sustainable agricultural production.

Wildfire effects on two freshwater producers: Combining in-situ and laboratory bioassays.

Mediterranean forests are highly susceptible to wildfires, which can cause several impacts not only within burnt areas but also on downstream aquatic ecosystems. The ashes' washout from burnt areas by surface runoff can be a diffuse source of toxic substances, such as metals, when reaching the nearby aquatic systems, and can be noxious to aquatic organisms. The present work aimed at assessing the ecotoxicological effects of post-fire contamination on two aquatic producers (the microalgae Raphidocelis subcapitata and the macrophyte Lemna minor) through in-situ bioassays, validating the obtained results with the outcomes of laboratory bioassays with surface water collected simultaneously. Four distinct sites were selected in a basin partially burnt (Ceira river basin; Coimbra district, Portugal) for bioassay deployment: one site upstream the burnt area in the Ceira river (RUS); three sites located under the influence of the burnt area, one immediately downstream of the burnt area in the Ceira river (RDS) and the other two in tributary streams within the burnt area (BS1 and BS2). The in-situ bioassays lasted for 13 days and began following the first post-fire major rain events. Results showed that the microalgae growth rate was able to distinguish the three sites within and downstream of the burnt area (BS1, BS2, RDS) from the site upstream (RUS). By contrast, the macrophytes growth rate only allowed to differentiate between the sites within the burnt area (BS1 and BS2) and those up- and downstream of the burnt area (RUS and RDS). The in-situ results for both species were corroborated with the results of the laboratory experiments, supporting the use of laboratory surrogates for a screening assessment of wildfire impacts in aquatic ecosystems. Direct causal relationships between the observed ecotoxicological effects on R. subcapitata and L. minor and the physical-chemical parameters of the water samples were difficult to establish, although the results suggest (i) a role of differential major and trace metal load in explaining species growth variation; (ii) interaction between metals and/or between metals and other field parameters are likely to modulate the biological responses to the challenges deriving from wildfire runoff.

Synthesis and Characterization of Surface-Active Ionic Liquids Used in the Disruption of Escherichia Coli Cells.

Twelve surface-active ionic liquids (SAILs) and surface-active derivatives, based on imidazolium, ammonium, and phosphonium cations and containing one, or more, long alkyl chains in the cation and/or the anion, were synthetized and characterized. The aggregation behavior of these SAILs in water, as well as their adsorption at solution/air interface, were studied by assessing surface tension and conductivity. The CMC values obtained (0.03-6.0 mM) show a high propensity of these compounds to self-aggregate in aqueous media. Their thermal properties were also characterized, namely the melting point and decomposition temperature by using DSC and TGA, respectively. Furthermore, the toxicity of these SAILs was evaluated using the marine bacteria Aliivibrio fischeri (Gram-negative). According to the EC50 values obtained (0.3-2.7 mg L-1 ), the surface-active compounds tested should be considered "toxic" or "highly toxic". Their ability to induce cell disruption of Escherichia coli cells (also Gram-negative), releasing the intracellular green fluorescent protein (GFP) produced, was investigated. The results clearly evidence the capability of these SAILs to act as cell disruption agents.

Glycine-betaine-derived ionic liquids: Synthesis, characterization and ecotoxicological evaluation.

Ionic Liquids (ILs) are generically regarded as environmentally "harmless" and thus, assumed as "non-toxic". However, due to the endless design possibilities, their ecotoxicological profile is still poorly known. An accurate knowledge on the toxicity of a substance is required, under the scope of environmental regulation worldwide, before their application and commercialization. Knowledge on the relationship between the chemical structure and toxic effects is essential for the future design of more biocompatible solvents. Focusing on the use of ILs as base lubricants, lubricant additives, or even as potential working fluids for absorption heat pumps, the knowledge on its environmental impact is of great importance, due to the possibility of spills. In this specific context, four analogues of glycine-betaine-based ILs (AGB-ILs) and four glycine-betaine based ILs (GB-ILs) were synthesized and characterized. Their ecotoxicity was assessed using representatives of two trophic levels in aquatic ecosystems, the bacteria Allivibrio fischeri (commonly used as a screening test organism) and the microalgae Raphidocelis subcapitata (as an alternative test organism that has been proven very sensitive to several IL families). The microalgae were more sensitive than the bacteria, hence, following a precautionary principle, we recommend considering the toxicity towards microalgae as an indicator in future studies regarding the focused ILs. Although four of the studied ILs were derived from a natural amino acid, all were considered hazardous for the aquatic environment, disproving the primary theory that all ILs derived from natural compounds are benign. Furthermore, the modification in the structure of anion and the cation can lead to the increase of toxicity.

Impacts of S-metolachlor and terbuthylazine in fatty acid and carbohydrate composition of the benthic clam Scrobicularia plana.

S-metolachlor (SMOC) and terbuthylazine (TBA) are herbicides that commonly appear as active ingredients (a.i.) in the composition of plant protection products. In a previous work, experimental bioassays were performed using those chemicals to find suitable molecular biomarkers to assess its toxicity to the non-target species Scrobicularia plana. The results obtained showed that the pollutants produce mortality and biochemical changes at the species, namely in protein contents and enzymatic activity levels. Thus, for a better understanding of the total biochemical impacts of those pollutants in S. plana, the composition of fatty acids (FA) and carbohydrates (CH) of the survival organisms are investigated here. In addition, since this species is edible its biochemical profile is directly related to its nutritious quality, which is analysed in this study. Furthermore, the analyses were performed in two types of tissue - the muscle and visceral mass of each survival organism. The greatest changes in FA composition are observable in small size class, being the most sensitive size class both at the toxicological and biochemical level. FA contents are higher in small organisms, both at the field and under laboratory conditions, being the disparity between size classes higher in visceral masses than in muscles. Indeed, muscles adequately represent the FA profile since those molecules appear in higher content in this tissue compared to visceral masses, becoming the better indicator tissue of biochemical changes. Besides, using muscles, less amount of biomass is needed, so it turns out to be the most cost-effective tissue to be used as endpoint in future studies. FA profiles observed at SMOC and TBA exposure are different, organisms from TBA exposure presenting a lower nutritious quality, in terms of FA abundance and diversity, than the organisms exposed to SMOC. Still, SMOC produces reductions of HUFA, essential fatty acids that cannot be synthesized by the species. Moreover, HUFA (mostly EPA and DHA) occupied the greatest part of the FA composition of organisms exposed to the control treatments and to TBA; however, the decreases of HUFA caused by the SMOC exposure change the profiles and make SFA the most dominant group. These findings represent a risk of low occurrence of essential fatty acids in entire aquatic environments exposed to the chemicals studied. Regarding CH, glucose is the only monosaccharide found in S. plana which was expected since glycogen is the main polysaccharide in animal tissues. In general, the glucose content increases with a concentration of pollutants, whereas the glycogen concentration decreases, suggesting that the glucose is being released as a response to chemical stress. Thus, this work presents tools to assess biochemical impacts of S-metolachlor and terbuthylazine in aquatic systems and to goes deeper in the knowledge of these pollutants' toxicity to non-target species to predict its propagation through aquatic trophic webs.

Synthesizing the role of epigenetics in the response and adaptation of species to climate change in freshwater ecosystems.

Freshwater ecosystems are amongst the most threatened ecosystems on Earth. Currently, climate change is one of the most important drivers of freshwater transformation and its effects include changes in the composition, biodiversity and functioning of freshwater ecosystems. Understanding the capacity of freshwater species to tolerate the environmental fluctuations induced by climate change is critical to the development of effective conservation strategies. In the last few years, epigenetic mechanisms were increasingly put forward in this context because of their pivotal role in gene-environment interactions. In addition, the evolutionary role of epigenetically inherited phenotypes is a relatively recent but promising field. Here, we examine and synthesize the impacts of climate change on freshwater ecosystems, exploring the potential role of epigenetic mechanisms in both short- and long-term adaptation of species. Following this wrapping-up of current evidence, we particularly focused on bringing together the most promising future research avenues towards a better understanding of the effects of climate change on freshwater biodiversity, specifically highlighting potential molecular targets and the most suitable freshwater species for future epigenetic studies in this context.

Transgenerational Inheritance of DNA Hypomethylation in Daphnia magna in Response to Salinity Stress.

Epigenetic mechanisms have been found to play important roles in environmental stress response and regulation. These can, theoretically, be transmitted to future unexposed generations, yet few studies have shown persisting stress-induced transgenerational effects, particularly in invertebrates. Here, we focus on the aquatic microcrustacean Daphnia, a parthenogenetic model species, and its response to salinity stress. Salinity is a serious threat to freshwater ecosystems and a relevant form of environmental perturbation affecting freshwater ecosystems. We exposed one generation of D. magna to high levels of salinity (F0) and found that the exposure provoked specific methylation patterns that were transferred to the three consequent nonexposed generations (F1, F2, and F3). This was the case for the hypomethylation of six protein-coding genes with important roles in the organisms' response to environmental change: DNA damage repair, cytoskeleton organization, and protein synthesis. This suggests that epigenetic changes in Daphnia are particularly targeted to genes involved in coping with general cellular stress responses. Our results highlight that epigenetic marks are affected by environmental stressors and can be transferred to subsequent unexposed generations. Epigenetic marks could therefore prove to be useful indicators of past or historic pollution in this parthenogenetic model system. Furthermore, no life history costs seem to be associated with the maintenance of hypomethylation across unexposed generations in Daphnia following a single stress exposure.

The antagonist and synergist potential of cholinium-based deep eutectic solvents.

The design of environmentally friendlier solvents has gained increasing relevance in the last decade. Deep eutectic solvents (DES) have recently emerged, with advantages like low-cost and putative lower environmental impact. However, information about DES toxicity is still scarce. This work aims to contribute to profiling the ecotoxicity of DES based on cholinium chloride ([Chol]Cl). Six DES were addressed, combining [Chol]Cl (as hydrogen bond acceptor - HBA) with ethylene glycol, glycerol, 1,2-propanediol, propionic acid, 1-propanol, and urea as hydrogen bond donors (HBD), in different molar ratios. The Microtox® Acute Toxicity Test, was used for assessing their toxicity towards the marine bacteria Allivibrio fischeri . Because the dissociation of DES in water is expected, analysis appraising the mixtures toxicity theory should be considered, which is a step forward in this field. This analysis suggested that [Chol]Cl and all HBD with the exception of propionic acid:[Chol]Cl 1:2 and 4:1 behave antagonistically, which is contrary to what has been suggested previously. The most extreme cases are Urea:[Chol]Cl and 1-Propanol:[Chol]Cl, with EC50 values higher than their starting materials dosed singly, configuring very promising and biocompatible alternative solvents. Toxicity was found to be dependent on DES composition, as well as on molar proportions of the starting materials.

Ecotoxicological assessment of the herbicide Winner Top and its active substances-are the other formulants truly inert?

Formulants used in Plant Protection Products (PPPs) to promote their efficiency are normally undisclosed in the PPP documentation, unless they bear a human health or environmental hazardous potential per se. PPP regulation also demands the assessment of putative interactions among formulants within each product recipe and consequent effects, but these results are often unavailable. Such a case is that of the herbicide Winner Top (Selectis®, Portugal), which we selected as a model commercial formulation in the present study specifically aiming at (i) characterising its aquatic toxicity towards sensitive eco-receptors (Raphidocelis subcapitata, Chlorella vulgaris, Lemna minor and Lemna gibba), as well as that of its active substances (a.s.) nicosulfuron and terbuthylazine; (ii) comparing the ecotoxicity among the commercial formulation, the corresponding mixture of its a.s. and this a.s.'s mixture increasingly enriched with the formulants. Single chemical testing revealed that terbuthylazine was the strongest microalgae growth inhibitor and nicosulfuron was the strongest macrophyte growth inhibitor. On the other hand, the commercial formulation was consistently less toxic than the corresponding mixture of the a.s., suggesting that Winner Top formulants (72.9% of the commercial formulation) interact with the a.s., promoting less than additive effects in the selected non-target species. Importantly, this environmentally protective effect of the formulation can be apparent. Because macrophytes share most physiological features with the weeds targeted by the studied herbicide, it is likely that increased application doses are required to reach desired efficacy levels with the consequent detrimental increase of PPP residues load in edge-of-field freshwater ecosystems.

Toxicity of two pesticides in binary mixture on survival and enzymatic response of Cerastoderma edule - The warming influence.

Pesticide application increased by about 1 million tonnes in the last 3 decades. Pesticides' overuse, coupled with the need for several pesticides to control different pests in the same crop, and its application many times per year, results in dangerous chemical cocktails that enter in aquatic systems, with impacts to the ecosystems and its communities. Climatic changes are currently another great concern, is predicted by the end of the 21st century, the earth's surface temperature will increase by about 4 °C. Bivalve species are reported as essential to the ecosystems' balance. However, they are also indicated as the organisms that will suffer the most serious effects of the temperature increase. So, this work intends to: a) verify the harm of the sub-lethal concentrations of two worldwide used pesticides, oxyfluorfen and copper (Cu), when combined, to Cerastoderma edule at 15 °C and 20 °C; b) assess the changes in the antioxidant defence system, the activity of the neurological enzyme acetylcholinesterase and the nutritive value of C. edule, after exposure to sub-lethal concentrations of oxyfluorfen and Cu, single and in the mixture, at 15 °C and 20 °C; c) observe the interaction between Cu and oxyfluorfen, considering the different biomarkers. Bivalves were exposed to oxyfluorfen and Cu, single and combined, for 96 h, at 15 °C and 20 °C. Results showed lethal effects to the organisms exposed at 20 °C when exposed to the highest binary mixture concentrations. Biochemical effects were observed on the organisms exposed to 15 °C, despite not observing any lethal effects. Briefly, there was a reported increase in oxidative stress and a decrease in protein content, regardless of the increase in the activity of antioxidant enzymes. These results suggest the potentially dangerous effects of the chemicals' mixture combined with the temperature, on this species and its consumers, impacting the trophic chain, and consequently, the community structure and function.

Redox Homeostasis Disclosed in the Saltmarsh Plant Halimione portulacoides upon Short Waterborne Exposure to Inorganic Mercury.

The saltmarsh plant Halimione portulacoides was shortly exposed to realistic levels of inorganic mercury (iHg) with the aim of investigating the adaptative processes of the roots and leaves regarding redox homeostasis, physiology, and Hg accumulation. Plants were collected at a contaminated (CONT) and a reference (REF) site to address the interference of contamination backgrounds. The influence of major abiotic variables (i.e., temperature and light) was also examined. Total Hg levels, antioxidant enzymes, lipid peroxidation (LPO), and photosynthetic activity were analyzed after 2 and 4 h of exposure. A poor accumulation of Hg in the roots was noticed, and no translocation to the stems and leaves was found, but plants from the CONT site seemed more prone to iHg uptake (in winter). Despite this, antioxidant modulation in the roots and leaves was found, disclosing, in winter, higher thresholds for the induction of enzymatic antioxidants in CONT leaves compared to REF plants, denoting that the former are better prepared to cope with iHg redox pressure. Consistently, CONT leaves exposed to iHg had remarkably lower LPO levels. Exposure did not impair photosynthetic activity, pinpointing H. portulacoides' ability to cope with iHg toxicity under very-short-term exposure. Biochemical changes were noticed before enhancements in accumulation, reinforcing the relevance of these responses in precociously signaling iHg toxicity.

The Lethal and Sub-Lethal Effects of Fluorinated and Copper-Based Pesticides-A Review.

In recent decades, pollution levels have increased, mainly as a result of the intensive anthropogenic activities such industrial development, intensive agricultural practices, among others. The impact of metals and organic contaminants is, nowadays, a great concern to the scientific and political communities. Copper compounds are the main sold pesticides in Europe, as well as herbicides, including glyphosate. Diphenyl ethers are the second ones most sold. Glyphosate and copper compounds are intensively studied, but the opposite is seen in the case of diphenyl ethers, including fluorinated pesticides (e.g., oxyfluorfen). Some research has been performed to increase the knowledge about these contaminants, daily inputted on the aquatic systems and with dangerous effects at physical and biochemical levels on the organisms. A wide range of biomarkers (e.g., growth, survival, reproductive success, enzymatic activity, lipid metabolism) has been applied to determine the potential effects in many species. This review intends to: (a) perform a compilation of the knowledge in previous research about the action mode of organic (fluorinated-based herbicide) and inorganic (copper-based pesticides) contaminants; (b) carry out an information survey about the lethal and sub-lethal effects of the fluorinated-based pesticides, namely the oxyfluorfen and the copper-based pesticides, on aquatic species from different trophic levels, according to in vitro and in vivo studies; (c) understand the impact of oxyfluorfen and copper-based pesticides, considering their effects reported in in vitro studies and, simultaneously, the authorized concentrations by legal organizations and the effective concentrations of each pollutant found in the environment. The literature analyzed revealed noxious effects of Cu and oxyfluorfen to aquatic organisms, including freshwater and marine species, even when exposed to the reference as well as to environmental concentrations, thus highlighting the importance of more monitoring and ecotoxicological studies, to chemical pollutants and different species from different ecological niches, to sustain and improve the legislation.

Effects of Inorganic and Organic Pollutants on the Biomarkers' Response of Cerastoderma edule under Temperature Scenarios.

Currently, there is increased chemical pollution, and climate change is a major concern to scientific, political and social communities globally. Marine systems are very susceptible to changes, and considering the ecological and economic roles of bivalve species, like Cerastoderma edule, studies evaluating the effects of both stressors are of great importance. This study intends to (a) determine the toxicity of copper (Cu) and oxyfluorfen at the lethal level, considering the temperature; (b) assess the changes in the antioxidant defence enzymes as a consequence of the simultaneous exposure to chemical and warming pressures; and (c) determine if lipid peroxidation (LPO) and neurotoxic effects occur after the exposure to chemical and temperature stressors. C. edule was exposed to Cu and oxyfluorfen at different temperatures (15 °C, 20 °C and 25 °C) for 96 h. The ecotoxicological results reveal a higher tolerance of C. edule to oxyfluorfen than to Cu, regardless of the temperature. The antioxidant defence system revealed efficiency in fighting the chemicals' action, with no significant changes in the thiobarbituric reactive species (TBARS) levels to 15 °C and 20 °C. However, a significant inhibition of acetylcholinesterase (AChE) was observed on the organisms exposed to oxyfluorfen at 20 °C, as well as a decreasing trend on the ones exposed to Cu at this temperature. Moreover, the catalase (CAT) showed a significant increase in the organisms exposed to the two highest concentrations of Cu at 15 °C and in the ones exposed to the highest concentration of oxyfluorfen at 20 °C. Looking at the temperature as a single stressor, the organisms exposed to 25 °C revealed a significant increase in the TBARS level, suggesting potential LPO and explaining the great mortality at this condition.

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.

Venom of Viperidae: A Perspective of its Antibacterial and Antitumor Potential.

The emergence of multi-drug resistant bacteria and limitations on cancer treatment represent two important challenges in modern medicine. Biological compounds have been explored with a particular focus on venoms. Although they can be lethal or cause considerable damage to humans, venom is also a source rich in components with high therapeutic potential. Viperidae family is one of the most emblematic venomous snake families and several studies highlighted the antibacterial and antitumor potential of viper toxins. According to the literature, these activities are mainly associated to five protein families - svLAAO, Disintegrins, PLA2, SVMPs and C-type lectins- that act through different mechanisms leading to the inhibition of the growth of bacteria, as well as, cytotoxic effects and inhibition of metastasis process. In this review, we provide an overview of the venom toxins produced by species belonging to the Viperidae family, exploring their roles during the envenoming and their pharmacological properties, in order to demonstrate its antibacterial and antitumor potential.

Multigenerational DNA methylation responses to copper exposure in Daphnia: Potential targets for epigenetic biomarkers?

Epigenetic mechanisms are moving to the forefront of environmental sciences, as environmentally induced epigenetic changes shape biological responses to chemical contamination. This work focused on Daphnia as a representative of potentially threatened freshwater biota, aiming to gain an insight into the involvement of epigenetic mechanisms in their response and eventual adaptation to metal contamination. Copper-induced DNA methylation changes, their potential transgenerational inheritance, and life-history traits were assessed. Organisms with different histories of past exposure to copper were exposed to toxic levels of the element for one generation (F0) and then monitored for three subsequent unexposed generations (F1, F2, and F3). Overall, methylation changes targeted important genes for counteracting the effects of metals and oxidative stress, including dynein light chain, ribosomal kinase and nuclear fragile X mental retardation-interacting protein. Also, contrasting overall and gene-specific methylation responses were observed in organisms differing in their history of exposure to copper, with different transgenerational methylation responses being also identified among the two groups, without apparent life-history costs. Taken together, these results demonstrate the capacity of copper to promote epigenetic transgenerational inheritance in a manner related explicitly to history of exposure, thereby supporting the development and incorporation of epigenetic biomarkers in risk assessment frameworks.

Chemical characterization of riverine sediments affected by wastewater treatment plant effluent discharge.

This study was aimed at assessing the contribution of wastewater treatment effluents to the contamination profile of the sediments of receiving waterways. Three wastewater treatment plants (WWTP) were addressed, encompassing different population equivalent sizes, urbanization degrees and treatment methods translating differences in expected contamination patterns. Within each WWTP system, the assessment targeted the effluent and sediment samples collected upstream and downstream the effluent discharge point; contaminants belonging to several concerning chemical classes (metals and metalloids; pesticides; pharmaceuticals and personal care products, PPCPs; and polycyclic aromatic hydrocarbons, PAHs) were quantified both in effluent and sediment samples. Clear associations between contaminants present in the effluent and corresponding sediment samples were not always verified. In fact, a noticeable difference between the number or abundance of contaminants detected in effluents and in sediments, suggesting that effluents are not always the most likely source (e.g. PAHs). However, sediment contaminants that were likely sourced by the effluents were also identified (e.g. PPCPs). Sediment analysis offers an important historical view of contamination, especially in flowing recipient ecosystems where any characterization over the water matrix is ephemeral and linking exclusively to the moment of sampling. Hence, sediments should be considered for the establishment of WWTP operational benchmarks regulating the emission of contaminants, which is currently focused mostly on effluent composition thus potentially over/underestimating the longer-term impact of effluent discharge in the recipient waterways.

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.

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.