Exploring the Effects of Graphene-Based Nanoparticles on Early Salmonids Cardiorespiratory Responses, Swimming and Nesting Behavior.

Graphene-based nanomaterials are exceptionally attractive for a wide range of applications, raising the likelihood of the release of graphene-containing nanoparticles into aquatic environments. The growing use of these carbon nanomaterials in different industries highlights the crucial need to investigate their environmental impact and evaluate potential risks to living organisms. The current investigation evaluated the nanotoxicity of graphene (nanoflakes) and graphene oxide (GO) nanoparticles on the cardiorespiratory responses (heart rate, gill ventilation frequency), as well as the swimming and nesting behavioral parameters of early stage larvae and juvenile salmonids. Both short-term (96 h) and long-term (23 days) exposure experiments were conducted using two common species: brown trout (Salmo trutta) and rainbow trout (Oncorhynchus mykiss). The findings demonstrated notable alterations in fish nesting behavior, swimming performance, and cardiorespiratory functions, indicating the potential toxicity of nanoparticles. This impact was observed at both physiological and whole-organismal levels in salmonids at early stages. Future investigations should explore different types of nanocarbons and their potential enduring effects on fish population structure, considering not only individual survival but also broader aspects of development, including feeding, reproductive, and other social dynamics.

Interactions of semiconductor Cd-based quantum dots and Cd2+ with gut bacteria isolated from wild Salmo trutta fry.

Background: With the rapid development of nanotechnology, more and more nanoproducts are being released into the environment where they may both pose ecological risks and be toxic to living organisms. The ecotoxicological impact of quantum dots (QDs), a class of nanoparticles (NPs), on aquatic organisms is becoming an emerging issue, this due to their nano-specific properties, to the physico-chemical transformation in the environment and to the possible release of toxic metals from their structure such as Cd. Methods: In this work, (i) spectroscopic measurements of commercially available Cd-based QDs (CdSe/ZnS-COOH) were made at various pH values (5.0 and 7.0) to study their interactions (at a concentration of 4 nm) with various strains of Gram-positive and Gram-negative gut bacteria after short-term exposure and (ii) the antibacterial efficacy of QDs and Cd2+ (at a concentration 0.09-3.56 mM) against gut bacteria isolated from wild freshwater Salmo trutta fry was studied at different temperatures (15 °C and 25 °C) and pH values (5.0 and 7.0) by applying a well-established disc diffusion assay. Results: Twenty-six gut bacterial isolates from wild Salmo trutta fry were identified as Aeromonas spp., A. popoffii, A. salmonicida, A. sobria, Carnobacterium maltaromaticum, Buttiauxella sp., Listeria sp., Microbacterium sp., Shewanella putrefaciens and Serratia sp. Cd-based (CdSe/ZnS-COOH) QDs at a concentration of 4 nm were found to be stable in aqueous media (with pH 7.0) or starting to form aggregates (at pH 5.0), thus, apparently, did not release heavy metals (HMs) into the media over 48 h in conditions of light or dark and did not show antibacterial efficacy on the gut bacteria isolated from wild Salmo trutta fry after short-term (9 h and 48 h) incubations. Cd2+ was found to produce significant dose-dependent toxic effects on bacterial growth, and the size of the inhibition zones on some of the tested strains significantly correlated with temperature. The most sensitive and the most resistant to Cd2+ were the Gram-positive bacteria, for which the minimum inhibitory concentration (MIC) values of Cd2+ were 0.09-0.27 mM and 3.11-3.29 mM respectively and varied significantly between the tested temperatures (15 °C and 25 °C). The MIC values of Cd2+ for the Gram-negative bacteria (18 out of 22 strains) ranged from 0.44 to 0.71 mM and did not differ significantly between the tested temperatures. Among the selected Gram-positive and Gram-negative strains, those with the higher sensitivity towards Cd2+ also revealed relatively stronger signals of QDs photoluminescence (PL) when transferred after incubation into fresh medium without QDs. In addition, the formation of endogenous metalloporphyrins observed spectroscopically in some bacterial strains indicates certain differences in metabolic activity that may play a protective role against potential oxidative damage.

The impact of co-treatment with graphene oxide and metal mixture on Salmo trutta at early development stages: The sorption capacity and potential toxicity.

Graphene oxide (GO) are novel nanomaterials with a wide range of applications due to their high absorption capacity. This study was undertaken with a view to assess the bioaccumulation and acute toxicity of GO used in combination with the heavy metal mixture (Cr, Cu, Ni and Zn) to fish embryos and larvae. For this purpose, Salmo trutta embryos and larvae were subjected to the 4-day long treatment with three different concentrations of GO, the metal mixture, which was prepared of four metals at the concentrations corresponding to the maximum-permissible-concentrations for EU inland waters (Cr-0.01, Cu-0.01, Ni-0.034, and Zn-0.1 mg/L), and with GO in combination with MIX (GO+MIX). When used in combination with the metal mixture, GO exhibited a high metal sorption capacity. The obtained confocal fluorescence microscopy results showed that GO located in the embryo chorion causing its damage; in larvae, however, GO were found only in the gill region. Results of these experiments confirmed the hypothesis that GO affects the accumulation of metals and mitigates their toxic effects on organism. In embryos, the acute toxicity of exposure to GO and co-exposure to MIX+GO was found to manifest itself through the decreased heart rate (HR) and malondialdehyde (MDA) level and through the increased metallothionein (MT) concentration. Meanwhile, in larvae, GO and MIX+GO were found to induce genotoxicity effects. However, changes in HR, MDA, MT, gill ventilation frequency, yolk sack absorption and cytotoxicity compared with those of the control group were not recorded in larvae. The obtained results confirmed our hypothesis: the combined effect of MIX and GO was less toxic to larvae (especially survival) than individual effects of MIX components. However, our results emphasize that fish exposure to GO alone and in combination with heavy metal contaminants (MIX+GO) even at environmentally relevant concentrations causes health risks that cannot be ignored.

Impact evaluation of marking Salmo trutta with Alizarin Red S produced by different manufacturers.

Fish otolith marking with the alizarin dye is a commonly used tool in sustainable fishery management. However, the reported effects of this dye on fish health are rather controversial and are possibly linked to differences in the composition of different brands of Alizarin red S (ARS). Laboratory experiments designed to elucidate effects of different concentrations of theoretically the same ARS as indicated by the CAS (Chemical Abstracts Service) number on fish at different development stages were carried out. The acute toxicity of ARS to Salmo trutta was found to be concentration- and fish developmental stage-dependant. Our study results showed that S. trutta sensitivity to ARS varies depending on its developmental stages as follows: fry (50-days after hatching) > alevins (30-days after hatching) > alevins (1-day after hatching). One of the tested ARS brands (purchased from VWR International LLC (Matsonford Road, USA)) was found to be several times more toxic to fish than another (purchased from Sigma-Aldrich (St. Louis, USA)), although according to the certificates of analysis, the tested substances were identical. Survival and growth of the S. trutta fry, which was marked with different ARS brands and stocked in the same natural stream, was investigated for two consecutive years. The results obtained indicate remarkable differences (p < 0.05) in the effects produced by the tested ARS brands, thus confirming our laboratory findings. The performed elemental analysis of the tested ARS dyes revealed significant differences in chemical impurities that these dyes contain. This study has, for the first time, expressed concern about the probable long-term impact of some ARS brands on the marked fish and their potential to bias the results of the studies dealing with ARS-marked fish.

Biological effects of multimetal (Ni, Cd, Pb, Cu, Cr, Zn) mixture in rainbow trout Oncorhynchus mykiss: Laboratory exposure and recovery study.

The present study tested the biological consequences of exposure to a multimetal mixture as a multiple chemical stressor on Oncorhynchus mykiss at molecular, cellular, physiological and whole-organism levels and on biomarker responses of this fish during the depuration period. To represent environmentally relevant multiple chemical stressors, in our study, we used the mixture of Zn, Cu, Ni, Cr, Pb and Cd at the concentrations corresponding to Maximum-Permissible-Concentrations (MPCs) acceptable for the EU inland waters. This study was undertaken with a view to elucidate if changes in the MPC of the test mixture components (Ni, Pb, Cd) could cause significantly different biomarker responses in O. mykiss from those previously determined in the carnivorous and omnivorous fishes exposed to the mixture of the same metals but at different MPCs of Ni, Pb and Cd. This study has revealed that exposure to mixtures of metals at MPC produces genotoxic effects in fish blood erythrocytes and a lethargic effect on O. mykiss behaviour, and, also, significantly increases the levels of Cd, Cr and Ni accumulated in the gills tissue. O. mykiss successfully depurated Cr and Ni in less than 28 days, however, the level of Cd decreased by only approximately 40% over the same period. A significant capacity of O. mykiss to restore its DNA integrity (Comet assay) after exposure to metal mixtures was revealed. However, the 28-day recovery period proved to be insufficiently long for erythrocytes with nuclear abnormalities to recover to the unexposed level. In conclusion, changes in the MPCs of Ni, Pb and Cd in the test mixture produce biological effects similar to those previously determined in S. salar, R. rutilus and P. fluviatilis exposed to the mixture of the same metals but at lower MPCs of Ni and Pb and at higher MPC of Cd.

Toxicological Potential of Cadmium Impact on Rainbow Trout (Oncorhynchus mykiss) in Early Development.

Cadmium (Cd) is a toxic element widely distributed in the aquatic environment and producing a wide variety of harmful effects. In this study, the acute toxicity (96 h LC50) of Cd to rainbow trout Oncorhynchus mykiss embryos and larvae was determined. The obtained results showed that hatched larvae were the most sensitive to Cd exposure. After 4 days of exposure, embryos were found to have accumulated greater concentrations of Cd than larvae. Exposure to Cd at sublethal concentrations produced deleterious, exposure duration-related effects on biological parameters (mortality, heart rate and gill ventilation frequency) of larvae. Cd induced a significant elevation of all the studied geno- and cytotoxicity endpoints in larval erythroblasts.

Embryotoxicity of Quantum Dots in Rainbow Trout Oncorhynchus mykiss During the Hatching Period.

Due to the active development and application of nanotechnology, nanoparticles have emerged as a new class of environmental pollutants. The aim of the study was to investigate quantum dots (QDs) access routes and distribution in embryos and larvae of rainbow trout Oncorhynchus mykiss and to determine the toxicity of QDs to rainbow trout larvae depending on the duration of exposure. CdSe/ZnS-COOH QDs at sublethal concentration was used during the toxicity test (1, 4 and 14 days). The results showed that QDs could get from the solutions into the larvae after hatching. QDs induced a significant increase in mortality, gill ventilation frequency and behavioral responses and a decrease in relative body mass in larvae at the end of the test. Larvae exposed to QDs were found to possess developmental malformations (blood clots). It was found that biological responses of larvae significantly depended on the duration of exposure to QDs.

Interaction of carboxylated CdSe/ZnS quantum dots with fish embryos: Towards understanding of nanoparticles toxicity.

Due to colloidal instability even with protective coatings, nanoparticles tend to aggregate in complex environments and possibly interact with biota. In this study, visualization of quantum dots (QDs) interaction with rainbow trout (Oncorhynchus mykiss) embryos was performed. Studies on zebrafish (Danio rerio) and pearl gourami (Trichogaster leerii) embryos have shown that QDs interact with embryos in a general manner and their affects are independent on the type of the embryo. It was demonstrated that carboxylated CdSe/ZnS QDs (4 nM) were aggregating in accumulation media and formed agglomerates on the surface of fish embryos under 1-12 days incubation in deep-well water. Detailed analysis of QDs distribution on fish embryos surface and investigation of the penetration of QDs through embryo's membrane showed that the chorion protects embryos from the penetration through the chorion and the accumulation of nanoparticles inside the embryos. Confocal microscopy and spectroscopy studies on rainbow trout embryos demonstrated that QDs cause chorion damage, due to QDs aggregation on the surface of chorion, even the formation of the agglomerates at the outer part of the embryos and/or with the mucus were detected. Aggregation of QDs and formation of agglomerates on the outer part of the embryo's membrane caused the intervention of the aggregates to the chorion and even partially destroyed the embryo's chorion. The incorporation of QDs in chorion was confirmed by two methods: in living embryos from a 3D reconstruction view, and in slices of embryos from a histology view. The damage of chorion integrity might have adverse effects on embryonic development. Moreover, for the first time the toxic effect of QDs was separated from the heavy metal toxicity, which is most commonly discussed in the literature to the toxicity of the QDs.