Arunava Pradhan

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.

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.

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.

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 <LC30. To better understand the contribution of ionic form in nanoCuO-induced stress, Cu2+ released from nanoCuO was quantified and the enzymatic responses to Cu2+ exposure at similar effective concentrations were compared. The highest activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GR) were observed at concentrations <LC5. The enzymatic activities decreased at effective concentrations between LC10 and LC30. GR activity remained higher than in control at all concentrations. The activity of glutathione S-transferase (GST) increased whereas that of catalase (CAT) decreased at concentrations between LC10 and LC30. The response patterns suggested that antioxidant enzymes could prevent oxidative stress at low concentrations (<LC10) of nanoCuO, thereby contributing to the survival of A. ligonifer. At concentrations between LC10 and LC30, effects of nanoparticulate or released ionic copper on enzyme activities were concentration-dependent, and led to oxidative stress and even to animal death. The activity of acetylcholinesterase (AChE) was strongly inhibited even at concentrations <LC10, suggesting neuronal stress in A. ligonifer.

Polyhydroxyfullerene Binds Cadmium Ions and Alleviates Metal-Induced Oxidative Stress in Saccharomyces cerevisiae

ABSTRACT 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.