Toxicity of different-sized cobalt ferrite (CoFe2O4) nanoparticles to Oncorhynchus mykiss at early development stages.

As innovative and versatile agents with potential applications in a wide range of fields including medicine, electronics, wastewater treatment, cosmetics, and energy storage devices, magnetic nanoparticles (NPs) are significant attention. However, our knowledge of the harmful effects of different-sized NPs, particularly of their effects on aquatic animals, is limited. In this study, we evaluated the impact of different-sized (sub-2, 5, and 15 nm) cobalt ferrite (CoFe2O4) NPs on the biological parameters of rainbow trout (Oncorhynchus mykiss) embryos and larvae. The NPs were characterized using techniques such as high-resolution transmission electron microscopy (HRTEM) for imaging, X-ray diffraction (XRD) for crystallographic analysis, and energy-dispersive X-ray spectroscopy (EDX) for elemental analysis, and were tested for impact through a series of toxicity, genotoxicity, and biochemical assays at a concentration of 100 mg/L. The obtained results showed that toxicity of CoFe2O4 NPs depended on the size of NPs and the developmental stage of the fish. Our results, which revealed significant changes in biological parameters of O. mykiss under exposure to CoFe2O4 NPs, imply that these NPs may be not environmentally safe. The hierarchical cluster analysis showed that embryos of the control group were clearly separated from those exposed to NPs of various sizes. However, in the exposed larvae, the effects of control and the smallest-sized NPs (sub-2 nm) differed from those induced by larger NPs (5 nm and 15 nm). Additional research is necessary to comprehend the mechanisms underlying the observed variations, which would be advantageous for both managing the risk of NPs to humans and advancing the field of aquatic nanotoxicology.

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.

Juvenile fish responses to sublethal leachate concentrations: comparison of sensitivity of different behavioral endpoints.

The aim of the present study was to investigate changes in the locomotor activity of rainbow trout (Oncorhynchus mykiss) juveniles under the impact (2 h) of landfill leachate (as a multicomponent mixture) based on different endpoints such as average, maximum and angular velocities, movement duration, body mobility, and blood glucose level. Fish were exposed to five different sublethal leachate concentrations (0, 0.0625, 0.125, 0.25, and 0.5%). The locomotor activity of the leachate-exposed fish significantly decreased at 0.25 and 0.5% concentrations. Significant changes in fish behavior in response to sublethal leachate concentrations were determined during the first minutes of exposure. Angular velocity proved to be the most sensitive of all the endpoints tested. A positive correlation was observed among behavioral responses, but no correlation was established between the blood glucose level and behavioral endpoints. The blood glucose endpoint was found to be insensitive, and we suggest that it should be used only in combination with other endpoints to complement toxicity data. To enhance the understanding of rainbow trout behavioral characteristics in relation to time, and relations among behavioral endpoints of the fish under short-term exposure to a multicomponent mixture, in the current study, we investigated dynamics of the selected behavioral endpoints over time, relations among these endpoints and compared behavioral response rapidness and efficacy.

Rapidness- and Sensitivity-Based Comparison of Behavioral and Respiratory Responses of European Perch and Rainbow Trout to Metal Mixture Exposure.

The aim of the study was to investigate acute (96-h LC50) toxicity of the complex metal (Zn, Pb, Cd, Cu, Ni and Cr) mixture (MIX) to European perch (Perca fluviatilis) and rainbow trout (Oncorhynchus mykiss), and to examine differences between locomotor and respiratory endpoints based on rapidness and sensitivity of fish responses to short-term (2 h) metal mixture exposure. MIX was prepared taking into consideration the maximum-permissible-concentrations (MPC) set for these metals in EU inland waters (Directive 2008/105/EC). The studied behavioral and respiratory responses of O. mykiss, in contrast to those of P. fluviatilis, were found to be significantly affected by the relationship between exposure duration and treatment. In O. mykiss, gill ventilation frequency (GVF) was found to be the most rapid and sensitive endpoint of all the investigated ones. However, the investigated behavioral and respiratory endpoints of P. fluviatilis showed its behavioral insensitivity to MIX exposure.

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.

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.

Rapid Detection of Sublethal Toxicity Using Locomotor Activity of Rainbow Trout Juveniles.

This study examined changes in locomotor activity of the rainbow trout (Oncorhynchus mykiss) juveniles exposed to sublethal concentrations of hexavalent chromium (Cr6+) (as a single pollutant) and landfill leachate (as an complex mixture of mainly organic compounds, including trace amounts of metals). Fish were first examined for baseline behavior patterns for 3 h to determine the appropriate duration of acclimation and the control level. The average velocity of fish was found to be the most informative among other endpoints throughout the 3 h long acclimation. Under the effect of both test substances, fish locomotor activity significantly increased after 5 min reaching maximum values after 10 min of exposure. The juveniles exposed to leachate were more responsive than those exposed to Cr6+. However, in this study we did not succeed in identifying pollutants from the elicited fish behavioral response patterns.

The profound effect of harmful cyanobacterial blooms: From food-web and management perspectives.

Sustainable and effective water management plans must have a reliable risk assessment strategies for harmful cyanobacterial blooms (HABs) that would enable timely decisions to be made, thus avoiding the trespassing of ecological thresholds, leading to the collapse of ecosystem structure and function. Such strategies are usually based on cyanobacterial biomass and/or on the monitoring of known toxins, which may, however, in many cases, under- or over-represent the actual toxicity of the HAB. Therefore, in this study, by the application of growth-inhibition assays using different bacteria, algae, zooplankton and fish species, we assessed the toxicological potential of two cyanobacterial blooms that differed in total cyanobacterial biomass, species composition and cyanopeptide profiles. We demonstrated that neither cyanobacterial community composition nor its relative abundance, nor indeed concentrations of known toxins reflected the potential risk of HAB based on growth-inhibition assays. We discuss our findings in the context of food-web dynamics and ecosystem management, and suggest that toxicological tests should constitute a key element in the routine monitoring of water bodies so as to prevent under-/over-estimation of potential HAB risk for both ecosystem and public health.

Evaluation of toxic and genotoxic effects of low-level 137Cs ionising radiation on plants.

The aim of this study was to evaluate the impact of low internal exposure to 137Cs on L. sativum meristem cells and Tradescantia stamen hair cells. It also compared the impact of 137Cs internal and external irradiation of similar level on the plant seed germination and root growth. Compared to control, the tested internal (0.0007 mGy to 0.7 mGy) and external (0.04 mGy to 5.5 mGy) 137Cs ionising radiation doses stimulated the elongation of L. sativum roots by 11% to 12% and 24% to 33%, respectively. Internal 137Cs exposure (0.0003 mGy to 0.5 mGy) for 14 days caused 1.2% to 1.6% of somatic mutations and 19% to 87% of non-viable stamen hair in Tradescantia.

Toxicity to Tradescantia of technogenic radionuclides and their mixture with heavy metals.

The genotoxic effects on Tradescantia of (137)Cs, (90)Sr, and (236, 242)Pu, a heavy metal mixture [Cd, Cr(VI), Cu, Mn(II), Ni, Pb, Zn] and of a complex mixture of these toxicants were determined. The impact of radionuclides on plants subjected to ionizing radiation exposure was estimated. The number of somatic mutations and the quantity of nonviable stamen hairs were used as end points in the testing. An increase in the quantity of nonviable stamen hairs was observed with increasing internal exposure to (137)Cs, (90)Sr, and (236, 242)Pu; however, the number of somatic mutations was not observed to be dependent on ionizing radiation. The internal dose of individual radionuclides necessary to decrease the quantity of viable stamen hairs in Tradescantia by 50% can be arranged in the following sequence: (236, 242)Pu > (137)Cs > (90)Sr. Tradescantia died in the mixture of the radionuclides (90)Sr, (137)Cs, and (236)Pu (5 x 10(-2), 7 x 10(-5), and 4 x 10(-10) Gy, respectively) after 14 days, whereas the heavy metal mixture caused somatic mutations in 3% of the Tradescantia and nonviable stamen hairs in 7% but no mortality. However, the Tradescantia died in a combined mixture of these heavy metals and the radionuclides after 14 days. On the basis of all these observations, it can be concluded that the toxic effect of radionuclides was more significant than that of heavy metals.

Assessment of toxic interactions of heavy metals in a multicomponent mixture using Lepidium sativum and Spirodela polyrrhiza.

The toxicities of copper, chromium, cadmium, nickel, manganese, zinc, and lead ions and various concentrations of mixtures of them were studied using the aquatic plant Spirodela polyrrhiza and the terrestrial plant Lepidium sativum. The composition of the model mixture was based on average analytical data of the annual amounts of representative heavy metals (HM) in wastewater discharged from the Ignalina Nuclear Power Plant (Lithuania) during 1996. The observed and predicted effects of the HM mixture on tested plants were evaluated and compared with the prediction models used in describing the toxic interactions of heavy metals in the mixture. The type of toxic interaction at each tested concentration of the mixture was assessed by a statistical approach that tested the null hypothesis of additive toxicity (Ince et al., 1999) and the mixture toxicity index (MTI; Könemann, 1981). For both plant organisms the effect of the HM mixture calculated using the MTI was synergistic. However, assessment of the HM interaction type at 50% effect concentrations using the hypothesis of additive toxicity showed a synergistic effect for Spirodela polyrrhiza and an additive effect for Lepidium sativum. Though the results obtained using both prediction models for assessing the HM mixture's toxicity were similar, in our opinion, the additive toxicity model is more suitable than the MTI model because the former allows evaluation of the impact of various mixture concentrations, not only those with a 50% effect.