A nanogold sensor test for tire wear chemicals based on the plasmon ruler approach.

The release of tire wear substances in the environment is raising concerns about potential impacts on aquatic ecosystems. The purpose of this study was to develop a quick and inexpensive screening test for the following tire wear substances: 6-phenylphenyldiamine quinone (6-PPD quinone), hexamethoxymethylmelamine (HMMM), 1-3-diphenylguanidine (1,3-DPG), and melamine. A dual strategy consisting of nanogold (nAu) signal intensity and the plasmonic ruler principle was used based on the spectral shift from the unaggregated free-form nAu from 525 nm to aggregated nAu at higher wavelengths. The shift in resonance corresponded to the relative sizes of the tire wear substances at the surface of nAu: 6-PPD (560 nm), HMMM (590 nm), 1,3-DPG (620 nm), and melamine (660 nm) in a concentration-dependent manner. When present in mixtures, a large indiscriminate band between 550 and 660 nm with a maximum corresponding to the mean intermolecular distance of 0.43 nm from the tested individual substances suggests that all compounds indiscriminately interacted at the surface of nAu. An internal calibration methodology was developed for mixtures and biological extracts from mussels and biofilms and revealed a proportional increase in absorbance at the corresponding resonance line for each test compound. Application of this simple and quick methodology revealed the increased presence of melamine and HMMM compounds in mussels and biofilms collected at urban sites (downstream city, road runoffs), respectively. The data also showed that treated municipal effluent decreased somewhat melamine levels in mussels.

Toxicity of two heavy rare earth elements to freshwater mussels Dreissena polymorpha.

Rare earth elements (REE) are essential components of many electronic devices that could end-up in solid waste disposal sites and inadvertently released in the environment. The purpose of this study was to examine the toxicity of two heavy REEs, erbium (Er) and lutetium (Lu), in freshwater mussels Dreissena polymorpha. Mussels were exposed to 14 days to increasing concentration (10, 50, 250, and 1250 µg/L) of either Er and Lu at 15 °C and analyzed for gene expression in catalase (CAT), superoxide dismutase (SOD), metallothionein (MT), cytochrome c oxidase (CO1), and cyclin D for cell cycle. In addition, lipid peroxidation (LPO), DNA damage (DNAd), and arachidonate cyclooxygenase were also determined. The data revealed that mussels accumulated Er and Lu similarly and both REEs induced changes in mitochondrial COI activity. Er increased cell division, MT, and LPO, while Lu increased DNAd and decreased cell division. Tissue levels of Er were related to changes in MT (r = 0.7), LPO (r = 0.42), CO1 (r = 0.69), and CycD (r = 0.31). Lu tissue levels were related to changes in CO1 (r = 0.73), CycD (r = - 0.61), CAT (r = 0.31), DNAd (r = 0.43), and SOD (r = 0.34). Although the lethal threshold was similar between Er and Lu, the threshold response for LPO revealed that Er produced toxicity at concentrations 25 times lower than Lu suggesting that Er was more harmful than Lu in mussels. In conclusions, the data supports that the toxicity pattern differed between Er and Lu although they are accumulated in the same fashion.

Micro and Nanoplastic Contamination and Its Effects on Freshwater Mussels Caged in an Urban Area.

Plastic-based contamination has become a major cause of concern as it pervades many environments such as air, water, sediments, and soils. This study sought to examine the presence of microplastics (MPs) and nanoplastics (NPs) in freshwater mussels placed at rainfall/street runoff overflows, downstream (15 km) of the city centre of Montréal, and 8 km downstream of a municipal effluent dispersion plume. MPs and NPs were determined using flow cytometry and size exclusion chromatography using fluorescence detection. Following 3 months of exposure during the summer season, mussels contained elevated amounts of both MPs and NPs. The rainfall overflow and downstream of the city centre were the most contaminated sites. Lipid peroxidation, metallothioneins, and protein aggregates (amyloids) were significantly increased at the most contaminated sites and were significantly correlated with NPs in tissues. Based on the levels of MPs and NPs in mussels exposed to municipal effluent, wastewater treatment plants appear to mitigate plastic contamination albeit not completely. In conclusion, the data support the hypothesis that mussels placed in urbanized areas are more contaminated by plastics, which are associated with oxidative damage. The highest responses observed at the overflow site suggest that tire wear and/or asphalt (road) erosion MPs/NPs represent important sources of contamination for the aquatic biota.

The Comet Assay, a Sensitive Biomarker of Water Quality Improvement Following Adoption of Beneficial Agricultural Practices?

Numerous actions have been undertaken by farmers to attenuate the impact of agricultural activities on aquatic ecosystems. The identification of biomarkers that respond quickly to water quality improvement could facilitate the assessment of adopted alternative practices and help maintain mobilization among stakeholders. We evaluated the potential of the comet assay, a biomarker of genotoxic effects, using a freshwater mussel, Elliptio complanata, as a model animal. The frequency of DNA damage was assessed in hemocytes of mussels collected from a pristine habitat and caged for 8 weeks in the Pot au Beurre River, a tributary of the fluvial Lake St.-Pierre (Quebec, Canada) impacted by agricultural activities. We found that the level of DNA damage naturally induced in mussel hemocytes was low and showed very limited variations over time. Compared with these baseline levels and to laboratory controls, we observed a doubling in DNA alterations in mussels exposed to agricultural runoff in the third branch of the Pot au Beurre River. The genotoxic response was significantly lower in mussels caged in the first branch of the Pot au Beurre River, where longer stretches of shoreline have been restored as buffer strips. Glyphosate, mesotrione, imazethapyr, and metolachlor were the main discriminant pesticides between these two branches. Metolachlor was found in sufficient concentrations to induce DNA damage, but it is more likely that the observed genotoxicity was the result of a "cocktail effect," that is, the cumulative contribution of coexisting genotoxicants including the above-mentioned herbicides and ingredients in their formulation. Our findings suggest that the comet assay is a sensitive tool for the early detection of changes in water toxicity following the adoption of agricultural beneficial practices. Environ Toxicol Chem 2023;42:2201-2214. © 2023 Crown copyright and The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article is published with the permission of the Controller of HMSO and the King's Printer for Scotland.

Comparative toxicity of micro and nanopolystyrene particles in Mya arenaria clams.

The contamination of coastal marine environments by plastics of sizes ranging from mm down to the nanoscale (nm) could pose a threat to aquatic organisms. The purpose of this study was to examine the toxicity of polystyrene nanoparticles (PsNP) of various sizes (50, 100 and 1000 nm) to the marine clams Mya arenaria. Clams were exposed to concentrations of PsPP for 7 days at 15 °C and analyzed for uptake/transformation, changes in energy metabolism, oxidative stress, genotoxicity and circadian neural activity. The results revealed that PsNP accumulated in the digestive gland was 50 nm > 100 nm > 1000 nm. All sized increased oxidative stress as follows: 50 nm (peroxidase, antioxidant potential and LPO), 100 nm (LPO and antioxidant potential) and 1000 nm (LPO). Tissue damage was also size dependent by increasing genotoxicity. The 100 nm PsPP altered the levels of the circadian metabolite melatonin. We conclude that the toxicity of plastics is size dependent in clams.

In vitro assessment of the genotoxicity and immunotoxicity of treated and untreated municipal effluents and receiving waters in freshwater organisms.

Municipal wastewater effluent is one of the largest sources of pollution entering surface waters in the Laurentian Great Lakes. Exposure to wastewater effluent has been associated with impaired immune systems and induction of genotoxicity to aquatic animals. Due to habitat degradation and environmental pollution linked to industrial development and population growth, several regions of the Great Lakes have been designated Areas of Concern (AOCs). In this study, we assessed the effect of extracts of sewage influent, (treated) effluent and receiving surface waters from the Hamilton Harbour AOC and the Toronto and Region AOC (Ontario, Canada) on the phagocytic immune response of rainbow trout (Oncorhynchus mykiss) kidney leukocytes and the genotoxicity (DNA strand breaks) of these extracts on freshwater mussel (Eurynia dilatata) hemocytes. We identified and quantified numerous chemicals present in the various samples extracted for exposure. In freshwater mussels, extracts from Hamilton Harbour AOC induced DNA damage with the most frequency (12 out of 28 samples) regardless of sample type, reflecting past and present industrial activities. In contrast, extracts from Toronto and Region AOC induced DNA damage infrequently (2 out of 32 (summer) and 5 out of 32 (fall) samples, respectively) and from different WWTPs at different times. None of the extracts induced any significant effect on phagocytosis of rainbow trout kidney leukocytes. The present study indicates that despite overall improvements to effluent quality, treatment of influent by WWTPs may not result in a corresponding improvement of the genotoxicity of effluents. In vitro bioassays are useful and cost-effective rapid-screening tools for preliminary assessments of contamination of aquatic ecosystems.

Form-Dependent Toxicity of Silver Nanomaterials in Rainbow Trout Gills.

The toxicity of the form of nanoparticles is presently not well understood. The purpose of this study consists in comparing the toxicity of various forms of silver nanoparticles (nAg) in juvenile rainbow trout Oncorhynchus mykiss. Juveniles were exposed to various forms of polyvinyl-coated nAg of similar size for 96 h at 15 °C. After the exposure period, the gills were isolated and analyzed for Ag uptake/distribution, oxidative stress, glucose metabolism, and genotoxicity. Higher levels of Ag were detected in gills in fish exposed to dissolved Ag followed by spherical, cubic, and prismatic nAg. Size-exclusion chromatography of gill fractions revealed that the dissolution of nAg was observed for all forms of nAg where prismatic nAg released more important levels of Ag in the protein pool as in fish exposed to dissolved Ag as well. The aggregation of nAg was more important for cubic nAg in respect of the other forms of nAg. The data revealed that lipid peroxidation was closely associated with protein aggregation and viscosity. Biomarkers revealed changes in lipid/oxidative stress and genotoxicity, which were related to the loss of protein aggregation and inflammation (NO2 levels), respectively. In general, the observed effects were found for all forms of nAg where the effects from prismatic nAg were generally higher than for spherical and cubic nAg. The strong relationship between genotoxicity and inflammation response suggests the participation of the immune system in the observed responses of juvenile fish gills.

Investigation on the Toxicity of Nanoparticle Mixture in Rainbow Trout Juveniles.

The environmental impacts of nanoparticle mixtures in the aquatic environment is not well understood. The purpose of this study examined the sub-lethal toxicity of low concentrations (ug/L range) of selected nanoparticles alone and in mixtures in juvenile trout. Fish were exposed to to individual and two environmentally relevant mixtures of silver (nAg), copper oxide (nCuO) and cerium oxide (nCeO) nanoparticles for 96 h at 15 °C. After the exposure period, fish were depurated overnight and tissue levels in Ag, Ce, Cu and Zn were determined along with a suite of effects biomarkers such as oxidative stress/inflammation, denatured protein tagging (ubiquitin), DNA strand breaks (genotoxicity) and acetylcholinesterase (AChE) activity. The data showed that these nanoparticles behaved as suspended matter but were nevertheless bioavailable for fish with bioconcentration factors of 6, 8 and 2 for nAg, nCeO and nCuO respectively. Only nCuO alone increased malonaldehyde (lipid peroxidation) contents but all nanoparticles increased DNA damage, protein-ubiquitin labeling, and decreased AChE activity. Globally, the toxicity of nCeO and nCuO was generally stronger than nAg, and antagonist effects were found in the mixtures. The interactions involved in these antagonisms are not well understood but do not involve the liberation of free ions and labile zinc in tissues. In conclusion, the bioavailability and toxicity of these nanoparticles are influenced by mixtures of nanoparticles, which is likely to occur in contaminated environments.

Shape-Dependent Toxicity of Silver Nanoparticles on Freshwater Cnidarians.

Silver nanoparticles (AgNPs) are increasingly used in various consumer products, leading to their inadvertent release in aquatic ecosystems. The toxicity of AgNPs could be associated with the leaching of ionic Ag but also with the size, shape and surface properties. The purpose of this study was to test the null hypothesis that toxicity of AgNPs was independent of shape in the invertebrate Hydra vulgaris. The hydranths were exposed to increasing concentrations of ionic Ag and AgNPs of three different shapes (spherical, cubic and prismatic) with the same size and coating (polyvinylpyrrolidone). The data revealed that between 68% and 75% of total Ag remained in solution after the 96 h exposure period, while 85−90% of ionic Ag remained in solution. The 96 h lethal concentration (LC50) was lower with ionic (4 µg/L) and spherical AgNPs (56 µg/L), based on irreversible morphological changes such as loss of tentacles and body disintegration. Cubic and prismatic AgNPs were not toxic at a concentration of <100 µg/L. The sublethal toxicity was also determined at 96 h based on characteristic morphological changes (clubbed and/or shortened tentacles) and showed the following toxicity: ionic (2.6 µg/L), spherical (22 µg/L) and prismatic (32.5 µg/L) AgNPs. The nanocube was not toxic at this level. The data indicated that toxicity was shape-dependent where nanoparticles with a low aspect ratio in addition to high circularity and elongation properties were more toxic at both the lethal and sublethal levels. In conclusion, the shape of AgNPs could influence toxicity and warrants further research to better understand the mechanisms of action at play.

Isolation and Quantification of Polystyrene Nanoplastics in Tissues by Low Pressure Size Exclusion Chromatography.

Ecotoxicity investigations of plastic nanoparticles (NPs) should pay more attention to their ability to pass barriers, accumulate, and initiate toxicity in cells. The purpose of this study was to develop a simple size exclusion chromatography (SEC) methodology to measure plastic NPs in biological tissues. A SEC column was prepared using a high-resolution gel for large macromolecules to separate plastic NPs from the protein/lipid pools in tissues. It was necessary to prepare the samples in high salt and non-ionic detergent (0.5 M NaCl and 0.2% Tween-20) and apply 0.2% Tween-20 containing 14 mM NaCl for the elution buffer to limit proteins adsorption to NPs. This methodology was able to resolve 50 and 100 nm polystyrene NPs from the protein/lipid pools in tissue homogenates. The fluorescent dye neutral red (NR) was also used for transparent NPs. Moreover, a sample fractionation step was also proposed for plastic NPs concentration using a salting-out methodology with saturated NaCl (5 M) and acetonitrile. Polystyrene NPs partition in acetonitrile, which were further analyzed by SEC. This methodology was tested in two case studies with clams collected in a high boat traffic (harbor) area and with caged freshwater mussels downstream of a large urban area. Although the present methodology was developed with polystyrene NPs it should be amenable to other plastic polymers that react with the NR fluorescent probe.

Assessment of the toxicity of a mixture of five rare earth elements found in aquatic ecosystems in Hydra vulgaris.

Rare earth elements (REEs) are considered critical elements for technology and their extraction through mining activities is expected to increase in the future. Due to their chemical similarities, they often co-occur in minerals and thus their ecotoxicity should be assessed as a group/family. However, the available ecotoxicological studies focused mainly on the evaluation of the potential toxicological impacts of individual REEs rather than their mixtures. The aim of this study was therefore to evaluate the toxicity of a representative mixture of five REEs (La, Ce, Pr, Nd and Sm) spanning environmentally relevant concentrations ranging from 0.05X (29 µg REEs L-1) to 5X (2926 µg REEs L-1) to the test organism, Hydra vulgaris, at the morphological, reproductive and regenerative levels. The data showed that lethality occurred at concentrations near (2.5 fold) to those inducing sublethal effects after chronic exposure of 7 days. The mixture affected reproduction and head regeneration and even lethality at concentrations even below those reported at environmental concentration (0.5X = 293 µg REEs L-1) in lakes. This suggests that REEs concentrations found in lakes near mining activities could disrupt regeneration and impair embryonic development. Our data also revealed that combining the 5 REEs results in an antagonistic effect, suggesting that those elements share the same receptor and that low molecular weight and high radius elements (approaching iron) were less toxic. Taken together, hydra could be used as a sensitive model organism for the assessment of aquatic ecotoxicological risks of REE mixtures but further analyses of biochemical and gene expressions should improve our understanding of the long-term effects of REEs mixtures.

Lanthanides Release and Partitioning in Municipal Wastewater Effluents.

The use of lanthanides is increasing in our society, whether in communication technologies, transportation, electronics or medical imaging. Some lanthanides enter urban wastewater and flow through municipal wastewater treatment plants (WWTPs). However, little is known about the effectiveness of treatment processes to remove these elements and the concentrations released in effluents to receiving waters. The main objective of this study was to investigate the fate of lanthanides in various wastewater treatment processes. A secondary objective was to better understand the fate of medical gadolinium (Gd) complexes; anthropogenic inputs were differentiated from geological sources using an approach based on concentration normalization with respect to chondrite Post-Archean Australian Shale (PAAS). The hypothesis was that most lanthanides, especially of geological origin, are associated with the particulate phase and could be efficiently removed by WWTPs. To monitor these elements in different WWTPs, various urban influents and effluents from simple aerated lagoons to advanced treatments were sampled in Canada. The results showed that the rates of lanthanide removal by treatment processes decrease with their atomic number; from 95% for cerium (Ce) to 70% for lutetium (Lu), except for Gd, which was minimally removed. The normalization approach permitted the determination of the origin of Gd in these wastewaters, i.e., medical application versus the geological background. By distinguishing the geogenic Gd fraction from the anthropogenic one, the removal efficiency was evaluated according to the origin of the Gd; nearly 90% for geogenic Gd and a rate varying from 15% to 50% in the case of anthropogenic Gd. The processes using alum as the flocculating agent had the highest removal efficiency from wastewater.

Evaluation of uptake of the cytostatic methotrexate in Elliptio complanata mussels by LC-MS/MS.

Aquatic organisms are continuously exposed to emerging contaminants coming from urban effluents of wastewater treatment plants. The contamination of surface water by those effluents poses a number of environmental risks, and pharmaceuticals are part of this class of effluent contaminants. Various classes of pharmaceuticals are not treated by wastewater treatment plants and anticancer drugs are part of them. The chemotherapy drug methotrexate (MTX) is an emerging contaminant and its growing use with the increase in cancer cases worldwide raises potential risk to aquatic organisms exposed to effluent discharges. However, chemical analyses in exposed freshwater aquatic organisms for ecotoxicological studies are rarely available and no studies have been done yet to accompany ecotoxicological data of exposed filter-feeding organisms. The purpose of this study was to develop a specific and sensitive analytical LC-MS/MS method for the quantification of methotrexate uptake in mussels exposed at different concentrations of the drug. A solid/liquid extraction followed by solid phase extraction (SPE) using an MCX phase purification scheme was optimized. The optimal recovery of 65% and matrix effect of 38% allowed to achieve a limit of quantification of 0.25 ng g-1, with an accuracy of 99-106%, a precision of no more than 3% RSD, and linearity ranging from 0.25 to 25 ng g-1. This methodology was tested with mussels exposed for 96 h at different concentrations (4 to 100 µg L-1) of MTX. The data revealed tissue uptake at concentrations ranging from 0 to 2.53 ng g-1. This suggests that this drug has low uptake potential and this methodology could be used to examine tissue levels of this drug in organisms continuously exposed to urban pollution.

Occurrence and size distribution of silver nanoparticles in wastewater effluents from various treatment processes in Canada.

The occurrence of silver (Ag) in urban effluents is partly associated with the increasing use of silver nanoparticles (Ag NPs) as an antiseptic agent in various consumer products. Distinction among Ag forms must be taken into account in the assessment of exposure and toxicological effects to aquatic organisms. Wastewater treatment processes effectively remove Ag particles and colloids (mostly > 95%), but this still leaves notable concentrations (in order of ng/L) escaping to effluent-receiving waters. Total suspended Ag concentrations in various studied effluents ranged from 0.1 to 6 ng/L. The purpose of this study was then to measure and characterize Ag NPs in urban effluents for their concentrations and size distribution using the single particle ICP-MS technique (SP-ICP-MS). Wastewater influents and effluents from various treatment plants-from aerated lagoons to advanced treatment technology-were collected for three sampling days. Our results showed the presence of Ag NP in all samples with concentrations reaching 0.5 ng/L on a mass basis. However, on a particle number basis, Ag NP concentrations (expressed in particle/mL) in the 20-34-nm fraction (up to 3400 particles/mL) were much more abundant (> 700%) than in the > 35-nm larger fraction. The proportion of Ag at the nanoscale (1-100 nm) represents less than 8% of the total suspended Ag for all effluent samples, regardless of their origins. A significant correlation (linear regression: r2 > 0.7) was observed between Ag NP and total suspended Ag concentrations in investigated effluents. Because Ag nanotoxicity is size dependent, the determination of size distribution and exposure concentration on a particle number basis is urgently needed for risk assessment of this class of nanoparticles.

Occurrence and ecotoxicity of cytostatic drugs 5-fluorouracil and methotrexate in the freshwater unionid Elliptio complanata.

Municipal effluents continuously release cytostatic drugs with unknown consequences in aquatic organisms. The purpose of the study was to examine the sublethal toxicity of 2 commonly-found cytostatic drugs 5-fluouracile (5-FLU) and methotrexate (MTX) to endemic Elliptio complanata freshwater mussels. The mussels were exposed of each drugs at 0, 4, 20 and 100 µg/L for 96 h t 15 °C. After the exposure period, glutathione S-transferase (GST) and dehydrofolate reductase (DHFR) activities, DNA damage and lipid peroxidation (LPO) were determined. The drugs were detected in mussel tissues with no evidence of accumulation with either drugs. The drug 5-FLU gave a larger spectrum of effects than MTX such as increased DHFR, decreased LPO and DNA strand breaks (repair activity) suggesting that the mussels were metabolically hindered and reduced DNA repair activity. The drug MTX only increased DHFR activity in the gonad. Hence, the data suggest that these drugs are biologically active in freshwater mussels and based on the reported maximum levels of these drugs in municipal effluents, the observed effects are likely in sessile freshwater mussel species downstream urban sources of pollution.

Toxicity of representative mixture of five rare earth elements in juvenile rainbow trout (Oncorhynchus mykiss) juveniles.

Rare earth elements (REEs) are contaminants of increasing interest due to intense mining activities for commercial purposes and ultimately released in the environment. We exposed juvenile rainbow trout (Oncorhynchus mykiss) to a representative mixture of the five most abundant REEs for 96 h at concentrations similar found in lakes contaminated by mining activities at 0.1, 1, 10, and 100X whereas the 1x mixture contained cerium (Ce, 280 µg/L), lanthanum (La, 140 µg/L), neodymium (Nd, 120 µg/L), praseodymium (Pr, 28 µg/L), and samarium (Sm, 23 µg/L). We investigated the expression of 14 genes involved in oxidative stress, DNA repair, tissue growth/proliferation, protein chaperoning, xenobiotic biotransformation, and ammonia metabolism in the liver. In addition, DNA damage, oxidative stress (lipid peroxidation or LPO), inflammation (cyclooxygenase or COX activity), detoxification mechanisms (glutathione-S-transferase activity or GST), and labile zinc were determined in gills. The data revealed that genes involved in oxidative stress-catalase (cat), heat shock proteins 70 (hsp70), and glutamate dehydrogenase (glud) were upregulated while glutathione S-transferase (gst) and metallothionein (mt) gene expressions were downregulated. The mixture was genotoxic and increased labile Zn in gills of exposed trout. These changes occurred at concentrations 600 times lower than the LC50 for this mixture indicating effects below the 1X concentration. Based on principal component analysis and concentration-dependent reponses, the following sublethal effects were considered the most important/significant: DNA strand breaks (genotoxicity), labile Zn, cat, gst, hsp70, sparc, mt, and glud. These effects of fish juveniles are likely to occur in environments under the influence of mining activities.

Evaluation of general stress, detoxification pathways, and genotoxicity in rainbow trout exposed to rare earth elements dysprosium and lutetium.

Rare earth elements (REEs) have been recently identified as emergent contaminants because of their numerous and increasing applications in technology. The impact of REEs on downstream ecosystems, notably aquatic organisms, is of particular concern, but has to date been largely overlooked. The purpose of this study was thus to evaluate the toxicity of lanthanide metals, lutetium (Lu) and dysprosium (Dy) in rainbow trout after 96 h of exposure. The lethal concentration (LC50) was determined and the expression of 14 genes involved in different pathways such as oxidative stress, xenobiotic detoxification, mitochondrial respiration, DNA repair, protein folding and turnover, inflammation, calcium binding and ammonia metabolism were quantified in surviving fish. In parallel, lipid peroxidation (LPO), DNA damage (DSB), metallothionein level (MT) and cyclooxygenase activity (COX) were examined. The acute 96 h-LC50 data revealed that Lu was more toxic than Dy (1.9 and 11.0 mg/L, respectively) and was able to affect all investigated pathways by changing the expression of the studied genes, to the exception of superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST). It also induced a decrease in DNA repair at concentrations 29 times below the LC50. This suggests that Lu could trigger a general stress to disrupt the cell homeostasis leading to genotoxicity without promoting oxidative stress. However, Dy induced modulation in the expression of genes involved in the protection against oxidative stress, detoxification, mitochondrial respiration, immunomodulation, protein turnover and an increase in the DNA strand breaks at concentrations 170 times lower than LC50. Changes in mRNA level transcripts could represent an early signal to prevent against toxicity of Dy, which exhibited inflammatory and genotoxic effects. This study thus provides useful knowledge enhancing our understanding of survival strategies developed by rainbow trout to cope with the presence of lanthanides in the environment.

Crowding Effects of Polystyrene Nanoparticles on Lactate Dehydrogenase Activity in Hydra Attenuata.

Plastics pervade our environment and potentially release important quantities of plastic nanoparticles (NPs) from degradation in the environment. The purpose of this study was to examine the crowding effects of polystyrene NPs on lactate dehydrogenase (LDH) in vitro and following exposure to Hydra attenuata. First, LDH activity was measured in vitro in the presence of filamentous (F-)actin and NPs (50 and 100 nm diameter) to determine changes in viscosity and the fractal kinetics of LDH. The fractal dimension (fD) was also determined using the rescaled range analysis procedure. Secondly, these changes were examined in hydra exposed to NPs for 96h to concentrations of NPs. The data revealed that the addition of F-actin increased the rate of LDH at low substrate (pyruvate) concentrations compared to LDH alone with a gradual decrease in the rate with the addition of pyruvate, which is characteristic of the fractal behavior of enzymes in crowded environments. The addition of 50 and 100 nm NPs also produced these changes, which suggest that NPs could change the space properties of the LDH reaction. The fD was reduced to 0.85 and 0.91 with 50 and 100 nm NPs compared to 1.093 with LDH alone. Decrease in the fD was related with increased amplitudes and frequency in viscosity waves in the reaction media. Exposure of hydra to NPs confirmed the increase in LDH activity and the fD was significantly correlated with LDH activity (r = -0.5). Correction of LDH activity (residuals) still revealed an increase in LDH activity in hydra suggesting increased anaerobic metabolism by NPs. In conclusion, the presence of NPs in the intracellular space decreased the fD, which could influence LDH activity in organisms exposed to NPs.

Biophysical effects of polystyrene nanoparticles on Elliptio complanata mussels.

The presence of nanoplastics (NPs) in various  products and from the weathering of released plastic materials are of concern for the environment's safety. The purpose of this study was to examine the biophysical effects of polystyrene NPs on freshwater mussels. Mussels were exposed to a range of concentrations of NPs (0.1, 0.5, 1, and 5 mg/L) for 24 h and allowed to depurate for 12 h in clean aquarium water. The digestive gland was isolated and analyzed for NPs, lipids, viscosity, protein aggregation, anisotropic changes (liquid crystals: LCs), and the oscillatory modulation in viscosity during the formation of self-organizing enzyme complex of fumarase, malate dehydrogenase, and citrate synthase. The results revealed that mussels accumulated NPs in the digestive gland and their levels were significantly correlated with lipids levels, LCs, the increase in the malate dehydrogenase/citrate synthase activity ratio, and oscillations in viscosity. Protein aggregation was also found to be correlated with lipid levels. The data suggests that the presence of NPs in the digestive gland involves changes in lipid content and LC formation and perturbs the normal oscillations in viscosity during sequential enzyme reactions of the above enzymes. It is concluded that the uptake of NPs in cells could disrupt the internal organization of cells which can interfere with the normal association of enzymes involved in energy metabolism.