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The eco-risk of roxarsone (ROX) was evaluated using multiple responses of earthworm biomarkers under different ROX concentrations for 28 d. With the increasing total arsenic accumulation (TAs-E), biological responses in earthworm generally showed a two-stage changes of homeostasis dysregulation and dose-dependent alterations. At the early periods, ROX stress increased the reactive oxygen species (ROS) and lipid peroxidation (LPO) in a similar manner, and apparently disrupted mitochondrial calcium ([Ca2+]m). But earthworms regulated their mitochondrial and redox homeostasis through stable mitochondrial membrane potential (MMP) and increase of ATP level, superoxide dismutase (SOD) and catalase (CAT). After 14 d, the positively correlated mitochondrial effects of ROS, [Ca2+]m, MMP and ATP were related to the behavioral inhibition of burrow length, depth and reuse rate as well as antioxidant up-regulation of Nrf2, HO-1, sod1 and cat. These results contributed possible biomarkers from the dose-dependent relationship between mitochondrial, antioxidant and behavioral responses. Multiple biological detection in earthworms can better reflect the sub-chronic ecotoxicity of phenylarsenic pollutants in soil.

Numerous byproducts resulting from chlorinated disinfection are constantly being generated during water treatment processes. The potential risks of these new emerging pollutions remain largely unknown. Here, we determined the risks of chlorinated disinfection byproducts of diazepam (DZP) in the cellular and zebrafish exposure experiments. The cytotoxicity of disinfection byproducts (MACB and MBCC) was greater than DZP in macrophage raw 264.7 cells at 10 mg/L. We further found that the effects of MBCC on the metabolism of glycine, serine, threonine and riboflavin were far greater than DZP by the targeted metabolomics methods. Moreover, MBCC significantly decreased the peak amplitude of neuronal action potential in primary embryonic rat (Spragu-Dawley SD) hippocampal neurons. We finally determined behavioral toxicity of DZP and byproducts in zebrafish larvae. MBCC significantly decreased the maximal swim-activity and peak duration of zebrafish after 72 h exposure. Altogether, these findings indicate the MBCC pose serious pressures on public health.

Aquatic animals are exposed to various cyanobacterial products released concomitantly to the environment by decaying blooms. Although there exist results on the toxicity of cyanobacterial extracts little is known on the influence of pure oligopeptides or their mixtures and elucidated mechanisms of behavioral toxicity in zooplanktonic organisms. Therefore, the aim of the present study was to assess the effects of single and mixed pure cyanobacterial oligopeptides: microginin FR-1 (MG-FR1), anabaenopeptin-A (ANA-A) and microcystin-LR (MC-LR) at various concentrations on the swimming behavior and catecholamine neurotransmitter activity, muscular F-actin structure, DNA nuclear content and cell viability of a model rotifer Brachionus calyciflorus. Swimming behavior was analyzed with the use of video digital analysis. Fluorescent microscopy imaging was used to analyze neuromotoric biomarkers in the whole organisms: neuromediator release (by staining with EC517 probe), muscle F-actin filaments (by staining with blue phalloidin dye). DNA content and cytotoxicity was also determined by Hoechst 34580 and propidium iodide double staining, respectively. The results showed that single oligopeptides inhibited all the tested endpoints. The binary mixtures induced synergistic interaction on swimming speed except for MG-FR1 +MC-LR which was nearly additive. Both binary and ternary mixtures also synergistically degraded F-actin and triggered cytotoxic effects visible in the whole organisms. Antagonistic inhibitory effects of all the binary mixtures were found on catecholamine neurotransmitter activity, however the ternary mixture induced additive toxicity. Antagonistic effects of both binary and ternary mixtures were also noted on nuclear DNA content. The results of the study suggest that both depression of neurotransmission and impairment of muscle F-actin structure in muscles may contribute to mechanisms of Brachionus swimming speed inhibition by the tested single cyanobacterial oligopeptides and their mixtures. The study also showed that natural exposure of rotifers to mixtures of these cyanobacterial metabolites may result in different level of interactive toxicity with antagonistic, additive synergistic effects depending on the variants and concentrations present in the environment., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The typical tire manufacturing additive 6PPD, its metabolites 6PPDQ and 4-Hydroxy should be monitored because of their ubiquitous presence in the environment and the high toxicity of 6PPDQ to coho salmon. The toxic effect of 6PPD and its metabolites have been revealed superficially, especially on behavioral characteristics. However, the behavioral indicators explored so far are relatively simple and the toxic causes are poorly understood. With this in mind, our work investigated the toxic effects of 6PPD, 6PPDQ and 4-Hydroxy on adult zebrafish penetratingly through machine vision, and the meandering, body angle, top time and 3D trajectory are used for the first time to show the abnormal behaviors induced by 6PPD and its metabolites. Moreover, neurotransmitter changes in the zebrafish brain were measured to explore the causes of abnormal behavior. The results showed that high-dose treatment of 6PPD reduced the velocity by 42.4% and decreased the time at the top of the tank by 91.0%, suggesting significant activity inhibition and anxiety. In addition, γ-aminobutyric acid and acetylcholine were significantly impacted by 6PPD, while dopamine exhibited a slight variation, which can explain the bradykinesia, unbalance and anxiety of zebrafish and presented similar symptoms as Huntingdon's disease. Our study explored new abnormal behaviors of zebrafish induced by 6PPD and its metabolites in detail, and the toxic causes were revealed for the first time by studying the changes of neurotransmitters, thus providing an important reference for further studies of the biological toxicity of 6PPD and its metabolites., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interest or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The eco-risk of roxarsone (ROX) was evaluated using multiple responses of earthworm biomarkers under different ROX concentrations for 28 d. With the increasing total arsenic accumulation (TAs-E), biological responses in earthworm generally showed a two-stage changes of homeostasis dysregulation and dose-dependent alterations. At the early periods, ROX stress increased the reactive oxygen species (ROS) and lipid peroxidation (LPO) in a similar manner, and apparently disrupted mitochondrial calcium ([Ca 2+ ] m ). But earthworms regulated their mitochondrial and redox homeostasis through stable mitochondrial membrane potential (MMP) and increase of ATP level, superoxide dismutase (SOD) and catalase (CAT). After 14 d, the positively correlated mitochondrial effects of ROS, [Ca 2+ ] m , MMP and ATP were related to the behavioral inhibition of burrow length, depth and reuse rate as well as antioxidant up-regulation of Nrf2, HO-1, sod1 and cat. These results contributed possible biomarkers from the dose-dependent relationship between mitochondrial, antioxidant and behavioral responses. Multiple biological detection in earthworms can better reflect the sub-chronic ecotoxicity of phenylarsenic pollutants in soil., (Copyright © 2021. Published by Elsevier Inc.)

Numerous byproducts resulting from chlorinated disinfection are constantly being generated during water treatment processes. The potential risks of these new emerging pollutions remain largely unknown. Here, we determined the risks of chlorinated disinfection byproducts of diazepam (DZP) in the cellular and zebrafish exposure experiments. The cytotoxicity of disinfection byproducts (MACB and MBCC) was greater than DZP in macrophage raw 264.7 cells at 10 mg/L. We further found that the effects of MBCC on the metabolism of glycine, serine, threonine and riboflavin were far greater than DZP by the targeted metabolomics methods. Moreover, MBCC significantly decreased the peak amplitude of neuronal action potential in primary embryonic rat (Spragu-Dawley SD) hippocampal neurons. We finally determined behavioral toxicity of DZP and byproducts in zebrafish larvae. MBCC significantly decreased the maximal swim-activity and peak duration of zebrafish after 72 h exposure. Altogether, these findings indicate the MBCC pose serious pressures on public health., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Carbamate pesticides (CMs) and organophosphorus pesticides (OPs) have been widely used in agriculture and toxicologically affect non-target organisms. Although there are many reports about their toxicities, the combined behavioral toxicities of CM/OP mixtures on Caenorhabditis elegans have rarely been studied. In this study, body bend inhibition (BBI), head thrash inhibition (HTI), and swimming speed inhibition (SSI) by CMs and OPs were chosen as the toxicity endpoints. The locomotion behavioral toxicities of individual pesticides (carbofuran (CAR), methomyl (MET), chlorpyrifos (CPF), and triazophos (TAP)) and their binary mixtures on C. elegans were determined systematically and the toxicological interaction profiles of various CM/OP mixture rays constructed using the combination index. It was shown that four pesticides and their binary mixture rays have significant inhibitory effects on the locomotion behavior of C. elegans; that is, they produce locomotion behavioral toxicities and the toxicity of two OPs is higher than those of two CMs. The toxicological interactions in the binary CM and OP mixtures are different from each other. For example, one mixture ray (CAR-MET-R1) in the CM system on the SSI endpoint exhibits synergism at all concentration levels, another ray (CAR-MET-R3) displays low-dose synergism and high-dose additive action on BBI and HTI endpoints, and weak synergism at high-dose on SSI, and other rays perform additive action. Two rays (CPF-TAP-R1 and CPF-TAP-R2) in the OP mixture system display low-dose additive action and high-dose antagonism on the three endpoints. Another ray (CPF-TAP-R3) shows the additive action at all concentration levels. It can be concluded that it is not sufficient to evaluate the combined toxicity of binary CM/OP mixtures using only one concentration ratio ray and that it is necessary to examine multiple concentration ratios., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The literature has largely shown the toxicity of petroleum-based PLA biomicroplastics (PLABioMPs) and encouraged the production of alternative materials to replace their use, such as biopolymers. However, knowledge concerning the effects of biopolymers on aquatic organisms remains under development. The hypothesis that the acute exposure (five days) to polylactic acid (PLA) biopolymers may lead to behavioral and biochemical changes and to their accumulation in Danio rerio larvae was tested. Based on the results, PLA biomicroplastics (PLA BioMPs) at concentration of 3 and 9 mg/L decreased swimming distance and speed of larvae in the open field test. This outcome suggests effects on animals' locomotor and exploration activities. Larvae's longer immobility time and greater permanence in the peripheral zone of the apparatus is indicative of anxiety-like behavior caused by the exposure to PLA BioMPs. Zebrafish larvae accumulated PLA BioMPs and their acetylcholinesterase activity was inhibited by their presence, which reinforces the accumulative potential of biopolymers and their direct or indirect role as anxiogenic agents, even at sublethal concentrations. The decreased activity of acetylcholinesterase reinforces the neurotoxic action in groups exposed to PLA BioMPs. The current study has confirmed the initial hypothesis and is an insight about the toxicity of these biopolymers in D. rerio larvae, since it deepens the discussion about the environmental risk of these substances in freshwater ecosystems., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Mercury, and particularly its organic form, methylmercury (MeHg), is a ubiquitous environmental contaminant with documented dosage-dependent adverse effects on endpoints spanning many levels of biological organization. However, relatively little is known about the sublethal impacts of environmentally-relevant exposures on behavioral characteristics that may impact predator-prey relationships, and thus the potential for Hg bioaccumulation within food webs due to behavioral impairments. This study investigated the potential for dietary mercury exposure to impair two behavioral outcomes in the highly invasive rusty crayfish, Faxonius rusticus, which are expected to influence interactions with their fish predators: the tail-flip escape response and chelae pinch strength. Field-caught animals were randomly assigned to four dietary treatments with mean (±1 SE) dry-weight total Hg (THg) concentrations of 3.52 ± 0.57, 114.01 ± 4.05, 274.10 ± 0.38, and 565.79 ± 1.33 ng/g in the control, low, moderate and high exposure treatments, respectively, for 16 weeks. After initial observations, exposures began and mass and behavior were measured every two weeks. At the end of the experiment, THg concentrations in tail muscle tissue were significantly higher in the high exposure treatment than in the control and low exposure treatments (Tukey's HSD, family-wise α = 0.05). Exposure-dependent declines in survival, mass, pinch strength, or tail-flip escape response velocity were not detected within the 12- to 16-week experimental exposure period, which represents one season of the crayfish's 3-4 year lifespan. This suggests that crayfish may be relatively tolerant of dietary exposure to sublethal concentrations of mercury within a single season. Further investigation of the physiological underpinnings of this tolerance is warranted., (Copyright © 2019 Elsevier Inc. All rights reserved.)