Nanomaterials and biochar mediated remediation of emerging contaminants.

The unrestricted release of various toxic substances into the environment is a critical global issue, gaining increased attention in modern society. Many of these substances are pristine to various environmental compartments known as contaminants/emerging contaminants (ECs). Nanoparticles and emerging sorbents enhanced remediation is a compelling methodology exhibiting great potential in addressing EC-related issues and facilitating their elimination from the environment, particularly those compounds that demonstrate eco-toxicity and pose considerable challenges in terms of removal. It provides a novel technique enabling the secure and sustainable removal of various ECs, including persistent organic compounds, microplastics, phthalate, etc. This extensive review presents a critical perspective on the current advancements and potential outcomes of nano-enhanced remediation techniques such as photocatalysis, nano-sensing, nano-enhanced sorbents, bio/phyto-remediation, which are applied to clean-up the natural environment. In addition, when dealing with residual contaminants, special attention is paid to both health and environmental implications; therefore, an evaluation of the long-term sustainability of nano-enhanced remediation methods has been considered. The integrated mechanical approaches were thoroughly discussed and presented in graphical forms. Thus, the critical evaluation of the integrated use of most emerging remediation technologies will open a new dimension in environmental safety and clean-up program.

Plastic Interactions with Pollutants and Consequences to Aquatic Ecosystems: What We Know and What We Do Not Know.

Plastics are a group of synthetic materials made of organic polymers and some additives with special characteristics. Plastics have become part of our daily life due to their many applications and uses. However, inappropriately managed plastic waste has raised concern regarding their ecotoxicological and human health risks in the long term. Due to the non-biodegradable nature of plastics, their waste may take several thousands of years to partially degrade in natural environments. Plastic fragments/particles can be very minute in size and are mistaken easily for prey or food by aquatic organisms (e.g., invertebrates, fishes). The surface properties of plastic particles, including large surface area, functional groups, surface topography, point zero charge, influence the sorption of various contaminants, including heavy metals, oil spills, PAHs, PCBs and DDT. Despite the fact that the number of studies on the biological effects of plastic particles on biota and humans has been increasing in recent years, studies on mixtures of plastics and other chemical contaminants in the aquatic environment are still limited. This review aims to gather information about the main characteristics of plastic particles that allow different types of contaminants to adsorb on their surfaces, the consequences of this adsorption, and the interactions of plastic particles with aquatic biota. Additionally, some missing links and potential solutions are presented to boost more research on this topic and achieve a holistic view on the effects of micro- and nanoplastics to biological systems in aquatic environments. It is urgent to implement measures to deal with plastic pollution that include improving waste management, monitoring key plastic particles, their hotspots, and developing their assessment techniques, using alternative products, determining concentrations of micro- and nanoplastics and the contaminants in freshwater and marine food-species consumed by humans, applying clean-up and remediation strategies, and biodegradation strategies.

Evidence of micro and macroplastic toxicity along a stream detrital food-chain.

Freshwater ecosystems are subjected to plastic extensive pollution because they are the direct link between plastic wastes and marine ecosystems. The aim of this study was to assess the impacts of different sizes of polyethylene plastics (micro: µPs and macroplastics: PBs) on freshwater decomposers of plant litter. We exposed leaf associated microbial assemblages to µPs (0.5 or 1.5 g L-1) and discs of PBs as follows: green plastic bags (PB-G) alone or in mixtures with transparent plastic bags (PB-Mix). Then, we conducted a feeding preference experiment with the invertebrate shredder Limnephilus sp. to assess their capacity to distinguish leaf discs from PB discs of the same size (12 mm). Leaf decomposition, activities of fungal enzymes and sporulation were inhibited by µPs and PB-Mix, and shifts in fungal community composition were observed. The invertebrate shredders preferred to feed on leaves treated with µPs avoiding those exposed to PB-G/PB-Mix. Our results demonstrated that plastics can have a direct effect on stream-dwelling microbial decomposers and an indirect effect on higher trophic levels (shredders), highlighting that trophic transfer is a route of plastic exposure. The plastic properties (size, concentration, colour) appear to influence plastic toxicity to microbes and shredders, indicating the importance of considering physicochemical properties when assessing their risks to freshwater ecosystems.

Can microplastics from personal care products affect stream microbial decomposers in the presence of silver nanoparticles?

Microplastics (MPs) are emerging contaminants of great concern due to their abundance and persistence over time in aquatic environments. However, studies on their impacts on freshwater organisms are scarce. In resemblance, silver nanoparticles (Ag-NPs) are incorporated into textiles and personal care products and are also classified as emerging contaminants. We used the leaf litter decomposition model system to investigate the effects of MPs from a commercially used personal care product, alone or in mixture with Ag-NPs, on the diversity and activities of freshwater microbial decomposers. We exposed stream microbial communities associated with leaf litter to increasing concentrations of MPs (polyethylene extracted from a personal care product; 100 µg L-1 up to 1 g L-1 5 concentrations plus 1 control) for 27 days in the absence or presence of Ag-NPs (0.1 mg L-1 and 1 mg L-1). The exposure to MPs, alone or in mixture with Ag-NPs, negatively affected fungal diversity and sporulation, with a reduction in leaf litter decomposition (Cohen's d > 1.5; r> 0.8; Bonferroni, P < 0.01). Shifts in community structure of sporulating fungi were observed, and effects were more pronounced in mixtures with Ag-NPs at the highest concentration. Mixtures of MPs with Ag-NPs (at the higher concentration) had the strongest impacts on extracellular enzymatic (ß-glucosidase, Cohen's d > 1; r > 0.5; phenol oxidase, Cohen's d > 1; r > 0.4) activities (ANOVAs, P < 0.05). Apart from sporulation rates, observed toxicity in mixtures was lower than that expected based on individual toxicity effects, mainly for higher concentrations (Bonferroni, P < 0.05). Our study provided evidence of the potential harmful effects of MPs, alone or in mixtures with Ag-NPs, on the activities of aquatic fungi and on a key ecosystem process, determinant to organic matter turnover in streams.

Elevated temperature may reduce functional but not taxonomic diversity of fungal assemblages on decomposing leaf litter in streams.

Mounting evidence points to a linkage between biodiversity and ecosystem functioning (B-EF). Global drivers, such as warming and nutrient enrichment, can alter species richness and composition of aquatic fungal assemblages associated with leaf-litter decomposition, a key ecosystem process in headwater streams. However, effects of biodiversity changes on ecosystem functions might be countered by the presumed high functional redundancy of fungal species. Here, we examined how environmental variables and leaf-litter traits (based on leaf chemistry) affect taxonomic and functional α- and ß-diversity of fungal decomposers. We analysed taxonomic diversity (DNA-fingerprinting profiles) and functional diversity (community-level physiological profiles) of fungal communities in four leaf-litter species from four subregions differing in stream-water characteristics and riparian vegetation. We hypothesized that increasing stream-water temperature and nutrients would alter taxonomic diversity more than functional diversity due to the functional redundancy among aquatic fungi. Contrary to our expectations, fungal taxonomic diversity varied little with stream-water characteristics across subregions, and instead taxon replacement occurred. Overall taxonomic ß-diversity was fourfold higher than functional diversity, suggesting a high degree of functional redundancy among aquatic fungi. Elevated temperature appeared to boost assemblage uniqueness by increasing ß-diversity while the increase in nutrient concentrations appeared to homogenize fungal assemblages. Functional richness showed a negative relationship with temperature. Nonetheless, a positive relationship between leaf-litter decomposition and functional richness suggests higher carbon use efficiency of fungal communities in cold waters.

Individual and mixed effects of anticancer drugs on freshwater rotifers: A multigenerational approach.

Human population growth has led to an increased release of chemical contaminants into aquatic environments. Emerging chemical contaminants (ECCs) are of increasing concern because they can affect non-target organisms in aquatic ecosystems. The application of anticancer drugs is increasing because of enhanced cancer rates and use of chemotherapy. We assessed the impacts of two widely used anticancer drugs known for their distinct modes of action, namely 5-fluorouracil (5-FU) and doxorubicin (DOX), on the freshwater rotifer Brachionus calyciflorus across generations. Rotifer mortality (24 h) and population growth (48 h) were assessed to determine initial lethal and sub-lethal effects. Exposure of rotifers to 5-FU (up to 200 mg L-1) did not cause mortality, while DOX caused mortality at high concentrations (EC50 = 15.6 mg L-1). Effects of 5-FU on population growth rate was higher than DOX (5-FU EC50 =10.49 µg L-1, DOX EC50 = 8.78 mg L-1). The effects of the drugs in binary mixture on population growth rates were dose dependent; significant antagonistic effects were found when 5-FU was present in the mixture at high concentrations. Finally, a transgenerational assay for five generations revealed that rotifers were able to recover their population growth rate after fourth generation when exposed to 5-FU; however, population became non-viable after the second generation of exposure to DOX. At the cellular level, accumulation of reactive oxygen species and plasma membrane damage were observed at EC10 and increased at EC50 for both drugs. After exposure of rotifers to 5-FU across generations, there were signs of oxidative stress recovery, as shown by a decrease in ROS accumulation and plasma membrane damage. Our results showed for the first time that the adverse effects of anticancer drugs on freshwater rotifer populations are drug and dose dependent and can persist or be attenuated along generations.

Importance of exposure route in determining nanosilver impacts on a stream detrital processing chain.

The commercial use and spread of silver nanoparticles (AgNPs) in freshwaters have greatly increased over the last decade. Both AgNPs and ionic silver (Ag+) released from nanoparticles are toxic to organisms and compromise ecosystem processes such as leaf litter decomposition. Yet little is known about how AgNPs affect multitrophic systems of interacting species. Furthermore, past work has focused on waterborne exposure with scarce attention given to effects mediated by the consumption of contaminated food. We assessed the importance of direct (via water) and indirect (via diet) AgNP exposure to a processing chain comprising leaf litter, fungi, a shredder (Gammarus pulex) and a collector (Habroleptoides confusa) in microcosms. Direct exposure to contaminated water for 15 days impaired microbial leaf decomposition by ∼50% and leaf-associated fungal biomass by ∼10%. Leaf consumption was reduced by ∼20% but only when G. pulex was exposed to silver via contaminated leaves. There was no effect on FPOM production. Ag + could impose oxidative stress on the shredders and collectors independent of exposure route, as indicated by increased catalase and glutathione S-transferase activities and decreased superoxide dismutase activity. The activity of a neuronal enzyme (cholinesterase) in collectors, but not shredders, also decreased by almost 50% when the animals were indirectly exposed to AgNP. Our results show that AgNPs and Ag+ may disrupt detrital processing chains through direct and indirect exposure routes, even at low concentrations. This highlights the importance of AgNP exposure pathways to interconnected stream biota and ecosystem processes for realistic assessments of risks to freshwater ecosystems.

Can photocatalytic and magnetic nanoparticles be a threat to aquatic detrital food webs?

Freshwaters are likely to serve as reservoirs for engineered nanomaterials (ENMs) due to their accelerated unintentional release, increasing the relevance of assessing their impacts on aquatic biota and the ecosystem processes they drive. Stream-dwelling microbes, particularly fungi, and invertebrate shredders play an essential role in the decomposition of organic matter and transfer of energy to higher trophic levels. We assessed the impacts of two photocatalytic (nano-TiO2 and erbium doped nano-TiO2) and one magnetic (nano-CoFe2O4) ENMs on detrital-based food webs in freshwaters by exposing chestnut leaves, colonized by stream-dwelling microbes, to a series of concentrations (0.25-150 mg L-1) of these ENMs. Microbial decomposition and biomass of fungal communities, associated with leaves, were not affected by the ENMs. However, the activities of antioxidant enzymes of microbial decomposers were significantly (P < 0.05) stimulated by ENMs in a concentration-dependent way, suggesting oxidative stress in stream microbial communities. The stronger responses of these stress biomarkers against nano-TiO2 (increase upto 837.5% for catalase, 1546.8% for glutathione peroxidase and 1154.6% for glutathione S-transferase) suggest a higher toxicity of this ENM comparing to the others. To determine whether the effects could be transferred across trophic levels, the invertebrate shredder Sericostoma sp. was exposed to ENMs (1 and 50 mg L-1) for 5 days either via contaminated water or contaminated food (leaf litter). Leaf consumption rate by shredders decreased significantly (P < 0.05) with increasing concentrations of ENMs via food or water; the effects were more pronounced when exposure occurred via contaminated food (up to 99.3%, 90.7% and 90.3% inhibition by nano-Er:TiO2, nano-CoFe2O4 and nano-TiO2, respectively). Overall, the tested photocatalytic and magnetic ENMs can be harmful to microbial decomposers and invertebrate shredders further compromising detrital-based food webs in streams.

Transcriptomics reveals the action mechanisms and cellular targets of citrate-coated silver nanoparticles in a ubiquitous aquatic fungus.

Silver nanoparticles (AgNPs) are among the major groups of contaminants of emerging concern for aquatic ecosystems. The massive application of AgNPs relies on the antimicrobial properties of Ag, raising concerns about their potential risk to ecologically important freshwater microbes and the processes they drive. Moreover, it is still uncertain whether the effects of AgNPs are driven by the same mechanisms underlying those of Ag ions (Ag+). We employed transcriptomics to better understand AgNP toxicity and disentangle the role of Ag+ in the overall toxicity towards aquatic fungi. To that end, the worldwide-distributed aquatic fungus Articulospora tetracladia, that plays a central role in organic matter turnover in freshwaters, was selected and exposed for 3 days to citrate-coated AgNPs (∼20 nm) and Ag+ at concentrations inhibiting 20% of growth (EC20). Responses revealed 258 up- and 162 down-regulated genes upon exposure to AgNPs and 448 up- and 84 down-regulated genes under exposure to Ag+. Different gene expression patterns were found after exposure to each silver form, suggesting distinct mechanisms of action. Gene ontology (GO) analyses showed that the major cellular targets likely affected by both silver forms were the biological membranes. GO-based biological processes indicated that AgNPs up-regulated the genes involved in transport, nucleobase metabolism and energy production, but down-regulated those associated with redox and carbohydrate metabolism. Ag+ up-regulated the genes involved in carbohydrate and steroid metabolism, whereas genes involved in localization and transport were down-regulated. Our results showed, for the first time, distinct profiles of gene expression in aquatic fungi exposed to AgNPs and Ag+, supporting different modes of toxicity of each silver form. Also, our results suggest that Ag+ had a negligible role in the toxicity induced by AgNPs. Finally, our study highlights the power of transcriptomics in portraying the stress induced by different silver forms in organisms.

Effects of metal nanoparticles on freshwater rotifers may persist across generations.

Nanotechnology has become one of the fastest growing industries in the current century because nanomaterials (NMs) are present in an ever-expanding range of consumer products increasing the chance of their release into natural environments. In this study, the impacts of two metal nanoparticles (Ag-NPs and CuO-NPs) and their equivalent ionic forms (Ag+ and Cu2+) were assessed on the lentic freshwater rotifer Brachionus calyciflorus and on its ability to adapt and recover through generations. In our study, Ag-NPs and CuO-NPs inhibited the rotifer population growth rate and caused mortality at low concentrations (< 100 µg L-1). Ag-NPs and CuO-NPs decreased in the medium when organisms were present (48 h exposure: 51.1 % and 66.9 %, respectively), similarly Ag+ and Cu2+ also decreased from medium in presence of the organisms (48 h: 35.2 % and 47.3 %, respectively); although the metal concentrations removed from the medium were higher for nanoparticles than metal ions, metal ions showed higher effects then their respective nanoparticle forms. Rotifer populations exposed for 4 generations to the toxicants were able to recover the population growth rate, but some rotifers showed developmental delay and inability to reproduce even after the removal of the toxicants. Intracellular accumulation of reactive oxygen species as well as plasma membrane damage were found in the rotifers at concentrations corresponding to EC10 (Ag-NPs = 1.7 µg L-1, Ag+ = 4.5 µg L-1, CuO-NPs = 46.9 µg L-1, Cu2+ = 35 µg L-1) of the population growth rate. Our results showed, for the first time, that effects of metal nanoparticles and metal ions on rotifer populations may persist along several generations. This should be taken into account when assessing risks of metal nanoparticles in freshwaters.

Biochemical and functional responses of stream invertebrate shredders to post-wildfire contamination.

Forests in Mediterranean Europe including Portugal are highly susceptible to wildfires. Freshwaters are often exposed to post-wildfire contamination that contains several toxic substances, which may impose risk to freshwater organisms and ecosystem functions. However, knowledge on the impacts of post-wildfire runoffs from different origins on freshwater biota is scarce. In forest streams, invertebrate shredders have a major contribution to aquatic detrital-based food webs, by translocating energy and nutrients from plant-litter to higher trophic levels. We investigated the leaf consumption behaviour and the responses of oxidative and neuronal stress enzymatic biomarkers in the freshwater invertebrate shredder Allogamus ligonifer after short-term exposure (96 h) to post-wildfire runoff samples from Pinus and Eucalyptus plantation forests and stream water from a burnt catchment in Portugal. Chemical analyses indicated the presence of various metals and PAHs at considerable concentrations in all samples, although the levels were higher in the runoff samples from forests than in the stream water. The shredding activity was severely inhibited by exposure to increased concentrations of post-wildfire runoff samples from both forests. The dose-response patterns of enzymatic biomarkers suggest oxidative and neuronal stress in the shredders upon exposure to increasing concentrations of post-wildfire runoffs. The impacts were more pronounced for the runoffs from the burnt forests. Moreover, the response patterns suggest that the energy from the feeding activity of shredders might have contributed to alleviate the stress in A. ligonifer. Overall, the outcomes suggest that the post-wildfire contamination can induce sublethal effects on invertebrate shredders with impacts on key ecological processes in streams.

Proteomic responses to silver nanoparticles vary with the fungal ecotype.

Enhanced commercial application of silver nanoparticles (AgNPs) is increasing the chance of their release into aquatic environments, potentially putting non-target microorganisms at risk. Impacts of AgNPs and Ag+ on two fungal ecotypes of Articulospora tetracladia, collected from a metal-polluted (At61) and a non-polluted (At72) stream, were assessed based on antioxidant enzymatic and proteomic responses. At61 showed more tolerance to AgNPs than At72 (EC20, 158.9 vs 7.5 µg L-1, respectively). Antioxidant enzyme activities were induced by AgNPs or Ag+ in both fungal ecotypes. Proteomic responses to AgNPs or Ag+ revealed that 41.3% of the total altered proteins were common in At72, while 27.3% were common in At61. In At72, gene ontology enrichment analyses indicated that Ag+ increased mainly the content of proteins involved in proteostasis and decreased the content of those related to vesicle-mediated transport; whereas the key group of proteins induced by AgNPs had functions in DNA repair and energy production. In At61, AgNPs induced proteins involved in energy production and protein biosynthesis, while both Ag forms induced proteins related to cell-redox and protein homeostasis, ascospore formation, fatty acid biosynthesis and nucleic acids metabolism. Both Ag forms induced stress-responsive proteins, and this was consistent with the responses of antioxidant enzymes. The negligible quantity of Ag+ released from AgNPs (<0.2 µg L-1) supported a minor role of dissolved ionic form in AgNP-induced toxicity to both fungal ecotypes. Overall, results unraveled distinct mechanisms of toxicity and cellular targets of nanoparticulate and ionic silver in aquatic fungi with different environmental background, and constitutes a proof of concept that toxicants induce adaptive responses in microbes to face emergent contaminants.

Wildfire impacts on freshwater detrital food webs depend on runoff load, exposure time and burnt forest type.

In the last decades, land-use changes have made Mediterranean forests highly susceptible to wildfires, which can cause several impacts not only on burnt areas, but also on adjacent aquatic ecosystems. Post-fire runoff from burnt areas may transport toxic substances to streams by surface runoff, including polycyclic aromatic hydrocarbons (PAHs) and metals, which can be noxious to aquatic organisms. Impacts on aquatic ecosystems can be related to fire severity, forest type and the exposure period; however, these factors have not been investigated in tandem. Here, we used the stream detrital system to determine the impacts of post-fire runoffs and stream water from a burnt catchment on trophic interactions between stream microbial communities and invertebrate shredders involved in leaf litter decomposition. Three distinct types of samples were collected from a burnt catchment: post-fire runoffs from high severity wildfires in Pinus and Eucalyptus forests, and stream water. Microbial decomposer communities (fungi and bacteria) and the invertebrate shredder Allogamus ligonifer were exposed for 10 and 20 days to increasing concentrations (0, 50, 75 and 100%) of runoff extracts. Our results showed that post-fire runoffs from high severity wildfires reduced microbially-driven leaf litter decomposition (up to 79%), invertebrate feeding (up to 75%), fungal biomass (up to 39%) and altered community composition; effects were more severe at the longer exposure time. The impacts varied with the runoff source and were related to the chemical composition in metals and total PAHs. This study emphasizes the importance of assessing the indirect effects of wildfires taking into account the effects of the runoff source, load and exposure time on freshwater biota and their ecological functions. Therefore, best forest management practices should be applied to minimize post-fire runoffs reaching aquatic ecosystems and to reduce the effects of these extreme events on freshwater biodiversity and ecosystem functioning.

Enzymatic biomarkers can portray nanoCuO-induced oxidative and neuronal stress in freshwater shredders.

Commercial applications of nanometal oxides have increased concern about their release into natural waters and consequent risks to aquatic biota and the processes they drive. In forest streams, the invertebrate shredder Allogamus ligonifer plays a key role in detritus food webs by transferring carbon and energy from plant litter to higher trophic levels. We assessed the response profiles of oxidative and neuronal stress enzymatic biomarkers in A. ligonifer after 96h exposure to nanoCuO at concentration ranges

Natural organic matter alters size-dependent effects of nanoCuO on the feeding behaviour of freshwater invertebrate shredders.

Nanoparticle size and the presence of natural organic matter (NOM) may influence the toxicity of nanoCuO to aquatic biota, but their interactive effects have been poorly investigated. We examined the feeding behaviour of the invertebrate shredder Allogamus ligonifer when exposed to sublethal concentrations of nanoCuO (50 and 100 mg L(-1)) with three particle sizes (12, 50 and 80 nm) in the absence or presence of humic acid (HA, 100 mg L(-1)) as a proxy of NOM. We further examined the ability of invertebrates to recover from the stressors. In the absence of nanoCuO and HA, the feeding rate of shredders was 0.416 mg leaf DM mg(-1 )animal DM day(-1). The exposure to increased nanoCuO concentrations inhibited the feeding rate and effects were stronger as nanoparticle size decreased (up to 83.3% inhibition for 12 nm particles). The exposure to HA alone inhibited the feeding activity by 52.7%. However, the co-exposure to nanoCuO and HA alleviated the inhibitory effects promoted by smaller and medium sized nanoCuO (up to 29.5%). The recovery of feeding activity by the shredders after stress removal was very low; maximum recovery (16.7%) was found for invertebrates rescued from pre-exposure to lower concentration of nanoCuO with larger size.

Fungi from metal-polluted streams may have high ability to cope with the oxidative stress induced by copper oxide nanoparticles.

Increased commercialization of products based on metal oxide nanoparticles increases the likelihood that these nanoparticles will be released into aquatic environments, thus making relevant the assessment of their potential impacts on aquatic biota. Aquatic fungi are distributed worldwide and play a key role in organic matter turnover in freshwater ecosystems. The present study investigated the impacts of copper oxide spherical nanoparticles (CuO-NPs; <50 nm powder, 5 levels ≤200 mg/L) on cellular targets and antioxidant defenses in 5 fungal isolates collected from metal-polluted or nonpolluted streams. The CuO-NPs induced oxidative stress in aquatic fungi, as evidenced by intracellular accumulation of reactive oxygen species, and led to plasma membrane damage and DNA strand breaks in a concentration-dependent manner. Effects were more pronounced with a longer exposure time (3 d vs 10 d). Under CuO-NP exposure, mycelia of fungi collected from metal-polluted streams showed less oxidative stress and higher activities of superoxide dismutase and glutathione reductase compared with fungi from nonpolluted streams. The latter fungi responded to CuO-NPs with a stronger stimulation of glutathione peroxidase activity. These findings may indicate that fungi isolated from metal-polluted streams had a greater ability to maintain the pool of reduced glutathione than those from nonpolluted streams. Overall, results suggest that populations adapted to metals may develop mechanisms to cope with the oxidative stress induced by metal nanoparticles.

Polyhydroxyfullerene binds cadmium ions and alleviates metal-induced oxidative stress in Saccharomyces cerevisiae.

The water-soluble polyhydroxyfullerene (PHF) is a functionalized carbon nanomaterial with several industrial and commercial applications. There have been controversial reports on the toxicity and/or antioxidant properties of fullerenes and their derivatives. Conversely, metals have been recognized as toxic mainly due to their ability to induce oxidative stress in living organisms. We investigated the interactive effects of PHF and cadmium ions (Cd) on the model yeast Saccharomyces cerevisiae by exposing cells to Cd (≤5 mg liter(-1)) in the absence or presence of PHF (≤500 mg liter(-1)) at different pHs (5.8 to 6.8). In the absence of Cd, PHF stimulated yeast growth up to 10.4%. Cd inhibited growth up to 79.7%, induced intracellular accumulation of reactive oxygen species (ROS), and promoted plasma membrane disruption in a dose- and pH-dependent manner. The negative effects of Cd on growth were attenuated by the presence of PHF, and maximum growth recovery (53.8%) was obtained at the highest PHF concentration and pH. The coexposure to Cd and PHF decreased ROS accumulation up to 36.7% and membrane disruption up to 30.7% in a dose- and pH-dependent manner. Two mechanisms helped to explain the role of PHF in alleviating Cd toxicity to yeasts: PHF decreased Cd-induced oxidative stress and bound significant amounts of Cd in the extracellular medium, reducing its bioavailability to the cells.

Physiological responses to nanoCuO in fungi from non-polluted and metal-polluted streams.

Nanocopper oxide (nanoCuO) is among the most widely used metal oxide nanoparticles which increases their chance of being released into freshwaters. Fungi are the major microbial decomposers of plant litter in streams. Fungal laccases are multicopper oxidase enzymes that are involved in the degradation of lignin and various xenobiotic compounds. We investigated the effects of nanoCuO (5 levels, ≤ 200 mg L(-1)) on four fungal isolates collected from metal-polluted and non-polluted streams by analyzing biomass production, changes in mycelial morphology, laccase activity, and quantifying copper adsorbed to mycelia, and ionic and nanoparticulate copper in the growth media. The exposure to nanoCuO decreased the biomass produced by all fungi in a concentration- and time-dependent manner. Inhibition of biomass production was stronger in fungi from non-polluted (EC50(10 days) ≤ 31 mg L(-1)) than from metal-polluted streams (EC50(10 days) ≥ 65.2 mg L(-1)). NanoCuO exposure led to cell shrinkage and mycelial degeneration, particularly in fungi collected from non-polluted streams. Adsorption of nanoCuO to fungal mycelia increased with the concentration of nanoCuO in the medium and was higher in fungi from non-polluted streams. Extracellular laccase activity was induced by nanoCuO in two fungal isolates in a concentration-dependent manner, and was highly correlated with adsorbed Cu and/or ionic Cu released by dissolution from nanoCuO. Putative laccase gene fragments were also detected in these fungi. Lack of substantial laccase activity in the other fungal isolates was corroborated by the absence of laccase-like gene fragments.

Copper oxide nanoparticles can induce toxicity to the freshwater shredder Allogamus ligonifer.

Increased commercialisation of nanometal-based products augments the possibility of their deposition into aquatic ecosystems; this, in turn, may pose risks to aquatic biota and associated ecological functions. Freshwater invertebrate shredders mostly use microbially-colonized plant litter as food resource and play an important role in aquatic detritus food webs. We assessed lethal effects of nanoCuO on the shredder Allogamus ligonifer (Trichoptera, Limnephilidae) by determining the concentration that induced 50% of death (LC(50)), and sublethal effects of nanoCuO on the feeding behaviour and growth of the shredder by exposing the animals to: (i) stream water supplemented with nanoCuO and microbially-colonized leaves, and (ii) stream water (without nanoCuO) and microbially-colonized leaves pre-exposed to nanoCuO. Results from acute lethal tests showed that the 96 h LC(50) of nanoCuO was very high (569 mg L(-1)). In the absence of nanoparticles, leaf consumption rate was 0.27 mg leaf DM mg(-1) animal DM d(-1) and the shredder growth rate was 56 µg animal DM mg(-1) animal DM d(-1). A significant inhibition in leaf consumption rate (up to 47%) and invertebrate growth rate (up to 46%) was observed when shredders were exposed to the higher tested sublethal concentration of nanoCuO (75 mg L(-1)) through either contaminated stream water or pre-contaminated food. The exposure to increased nanoCuO concentration via water or pre-contaminated food led to higher accumulation of copper in the larval body. Leached water-soluble ionic copper from the nanoCuO adsorbed or accumulated in the shredder (up to 10.2% of total Cu) seemed to influence the feeding behaviour and growth of the shredder.