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The Pantanal (Brazil) is a wetland region characterized by seasonal flooding. Hydrological cycles influence the water physicochemical parameters, causing seasonal variations in pH and nitrites. Thus, the objective of this study was to evaluate the effects of varying pH and nitrite concentrations on the toxicity of the cypermethrin-based pesticide Barrage®, considering both lethal (mortality) and sublethal endpoints (growth and development). Larvae of the endemic shrimp Macrobrachium pantanalense and of the estuarine Amazonian congener Macrobrachium amazonicum were exposed to cypermethrin (through Barrage®) under several pH levels (6.5, 7.5 and 8.5) or nitrite concentrations (0.1, 0.2 and 0.4 mg/L). The pH had direct effects on all the tested endpoints for both species. For M. pantanalense, the lethal effects of the cypermethrin formulation were more pronounced at low pH (96-h LC 50  = 0.004 μg/L at pH 6.5, and 0.146 μg/L at pH 8.5). For M. amazonicum, an opposite response was observed, with increased toxicity of the formulation at high pH (96-h LC 50  = 0.110 μg/L at pH 6.5 and 0.044 μg/L at pH 8.5). Variations in pH also seemed to modify the sublethal effects of the formulation on larval growth and development of M. pantanalense. Nitrite concentrations affected larval growth of both species, modifying also the effects of the cypermethrin formulation on the larval development of M. amazonicum. This work shows the importance of considering abiotic factors for risk assessment either due to possible direct effects on the physiology of organisms and/or due to interactions with other stressors, particularly in fragile biomes such as Pantanal., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Graphene nanocomposites are emerging carbon-based materials with interesting electrical, mechanical, optical and magnetic properties, relevant for applications in different fields. Despite this increased use, the impact of graphene nanocomposites residues in the environment has not been properly studied. Thus, the goal of this work was to assess the toxicity of two nickel/graphene nanocomposites (G/Ni1 and G/Ni2) differing in size and shape to Danio rerio embryos. Their toxicity was evaluated using apical (mortality, development and hatching), biochemical [cholinesterase (ChE), glutathione-S-transferase (GST), and catalase (CAT) activities] and behavioral (locomotor activity) endpoints. At the tested concentrations, neither of the nanocomposites presented lethal or developmental effects. Nevertheless, both nanocomposites induced behavioral effects, reducing swimming distances. This effect was, however detected at lower concentrations in the G/Ni1 nanocomposite. At biochemical level, only G/Ni1 nanocomposite showed to interfere with the measured parameters, increasing the activities of ChE, CAT and GST. Differences in the effects induced by the two nanocomposites seem to be related not only with their size, but also with the shape and the ability to continuously release nickel ions to aqueous medium. This work highlights the importance of studying graphene nanocomposites effects to aquatic organisms even when acute toxicity is not expected. The relevance of the effects found in this work need to be further analyzed in light of the consequences to the long-term fitness of the organisms and in light of the environmental concentrations expected for this type of compounds., (Copyright © 2019 Elsevier Inc. All rights reserved.)

In order to supply human demand for food, the aquaculture industry has been growing fast in the last years, being fish usually cultivated in overcrowded conditions. Hence, to prevent the rapidly disease spreading, antibiotics may be applied to both sick and healthy animals. Due to its broad spectrum, oxytetracycline (OTC) is one of the most used antibiotics in food-production. Yet, although useful to prevent infections, antibiotics may reshape aquatic animals' microbiome, disturbing hosts' welfare. However, the impact of this exposure to the organism microbiome and its surrounding environment is poorly understood. Then, the objective of this study was to analyze in detail the long-term effect of OTC in both zebrafish gut and water microbiomes. Zebrafish adults were exposed, via water, for two months to three concentrations of OTC (0, 10 and 10000 μg/L). Total DNA was extracted from gut and water samples and the V3-V4 region of the bacterial 16 S rRNA gene was sequenced using Illumina technology. Results of alpha and beta-diversity analyses revealed that long-term exposure to OTC impacted both zebrafish gut and water microbiomes. In water samples, effects were observed even at the lowest (10 μg/L) OTC concentration tested resulting in an increase in Deltaproteobacteria, namely the Myxococcales and Bdellovibrionales orders. On the other hand, effects on zebrafish gut were only observed at the highest concentration with the selection of Alphaproteobacteria and Actinobacteria classes. Although these classes are common in fish gut, the increase of Actinobacteria may represent a health problem since some genera like Gordonia are linked to some human infection disease. Nevertheless, in both gut and water, it was observed a decrease in Gamaproteobacteria, probably due to OTC mode of action. In silico functional metagenomic analysis revealed that OTC exposure selected general detoxification mechanisms. In addition, the abundance of functional genes involved in Quorum Sensing (QS) increased under OTC exposure suggesting that QS may help bacteria to survive OTC stress. Thus, future studies should consider post-exposure scenarios for a deeper analysis of the water and zebrafish gut resistome, since bacteria may react differently after exposure ceased., (Copyright © 2019 Elsevier Inc. All rights reserved.)

In recent years, the need for the development of alternative test methods for the conventional acute fish toxicity test (AFT) with adult fish has often been discussed. In addition, concerns have been raised on the potential risks related with environmentally realistic pesticide mixtures since risk evaluations have traditionally been based on individual pesticides. The insecticide/acaricide abamectin and the fungicide difenoconazole are the main pesticides that are intensively used in Brazilian strawberry crop and are hence likely to occur simultaneously in edge-of-field waterbodies. The aim of the present study was therefore to evaluate the lethal and sublethal toxicity of single and mixture exposures of these pesticides to zebrafish early life stages (embryos and juveniles). By comparing the derived toxicity data of the individual compounds with that previously determined for zebrafish adults, the order of life stage sensitivity was juvenile > adult > embryo. The pesticide mixture revealed a dose-level dependent deviation of the independent action model, with antagonism at low dose levels and synergism at high dose levels. Sublethal parameters (especially those related with locomotion) were considerably more sensitive than lethality. Subsequently, the inclusion of sublethal parameters may greatly improve the sensitivity of FET tests and hence its suitability as a substitution of adult fish testing in risk assessment evaluations., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

During the last century the increase in the mean global temperatures has been shown to impact on freshwater physicochemical parameters such as pH, dissolved oxygen, or ultraviolet (UV) light abundance. Changes in these parameters could modify the toxicity of environmental pollutants. Therefore, in the present study, the authors studied the tolerance (survival and sublethal endpoints) of zebrafish (Danio rerio) embryos to variations in pH (3-12), dissolved oxygen (3.9-237 μmol/L) and UV intensity (55-467 mW/m 2 ) using selected endpoints. Sublethal endpoint assessment included the quantification of hatching success, developmental delay, reduction of body length, frequency of edema, and morphological abnormalities. Median lethal concentrations (LC50s; 96-h) of 3.68 and 10.21 were determined for acid and alkaline pH, respectively. Embryo survival appeared to be relatively resistant to oxygen depletion with a 96-h LC50 of 0.42 mg/L. However, concentrations of 6 mg/L and below caused edema and developmental retardations. Continuous exposure to UV radiation affected zebrafish development by reducing survival and hatching rate and triggering a series of developmental abnormalities such as pericardial edema and deformities. A 72-h LC50 of 227 mW/m 2 was derived from intensity-response modeling. By generation of concentration-response parameters the authors' data provide a basis for the subsequent assessment of combined effect of environmental stress parameters and chemicals. Environ Toxicol Chem 2017;36:682-690. © 2016 SETAC., (© 2016 SETAC.)

Carbendazim is a widely used broad spectrum benzimidazole fungicide; however, its effects to non-target aquatic organisms are poorly studied. The aim of this study was to investigate the toxic effects of carbendazim to zebrafish early life stages at several levels of biological organization, including developmental, biochemical and behavioural levels. The embryo assay was done following the OECD guideline 236 and using a concentration range between 1.1 and 1.8mg/L. Lethal and developmental endpoints such as hatching, edemas, malformations, heart beat rate, body growth and delays were assessed in a 96h exposure. A sub-teratogenic range (from 0.16 to 500μg/L) was then used to assess effects at biochemical and behavioural levels. Biochemical markers included cholinesterase (ChE), glutathione-S-transferase (GST), lactate dehydrogenase (LDH) and catalase (CAT) and were assessed at 96h. The locomotor behaviour was assessed using an automated video tracking system at 120h. Carbendazim (96h-LC50 of 1.75mg/L) elicited several developmental anomalies in zebrafish embryos with EC50 values ranging from 0.85 to 1.6mg/L. ChE, GST and LDH activities were increased at concentrations equal or above 4μg/L. The locomotor assay showed to be extremely sensitive, detecting effects in time that larvae spent swimming at concentrations of 0.16μg/L and thus, being several orders of magnitude more sensitive that developmental parameters or lethality. These are ecological relevant concentrations and highlight the potential of behavioural endpoints as early warning signs for environmental stress. Further studies should focus on understanding how the behavioural disturbances measured in these types of studies translate into fitness impairment at the adult stage., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Pharmaceuticals are emerging contaminants as their worldwide consumption increases. Fibrates such as gemfibrozil (GEM) are used in human medicine to reduce blood concentrations of cholesterol and triacylglycerol and also are some of the most frequently reported pharmaceuticals in waste waters and surface waters. Despite some studies have already demonstrated the negative impact in physiological and/or reproductive endpoints in adult fish, data on survival and behavioral effects in fish larvae are lacking. This study aimed to assess the effects of GEM on zebrafish eleutheroembryo development and locomotor behavior. A fish embryo toxicity (FET) test was undertaken to evaluate GEM acute toxicity by exposing embryos to 0, 6.58, 9.87, 14.81, 22.22, 33.33 and 50mg/L. Developmental endpoints such as hatching success, edemas and malformations were recorded. A second test was undertaken by exposing embryos to 0, 1.5, 3 and 6mg/L in order to evaluate the effects of GEM on 120 and 144h post fertilization (hpf) larvae locomotor activity by video tracking, using a Zebrabox(®) (Viewpoint, France) device. From the data recorded, several parameters such as total swimming distance (TSD) and total swimming time (TST) in each 120s integration time were calculated. Data showed that this compound has a moderate toxic effect on fish embryo development, affecting both survival and hatching success with a calculated 96h LC50 of 11.01mg/L and no effects at the developmental level at 6mg/L. GEM seems to impair locomotor activity, even at concentrations where developmental abnormalities were unperceived, at concentrations as low as 1.5mg/L. Both TSD and TST were sensitive to GEM exposure. These effects do not seem to be independent of the developmental stage as 120hpf larvae seem to present a development bias with repercussions in locomotor behavior. This study highlights the need to include behavioral endpoints in ecotoxicological assays as this seems to be a more sensitive endpoint often disregarded., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This study aimed to assess the effect of ultraviolet radiation (UVR) and chemical stress (triclosan-TCS; potassium dichromate-PD; prochloraz-PCZ) on bacterial communities of zebrafish (Danio rerio) embryos (ZEBC). Embryos were exposed to two UVR intensities and two chemical concentrations not causing mortality or any developmental effect (equivalent to the No-Observed-Effect Concentration-NOEC; NOEC diluted by 10-NOEC/10). Effects on ZEBC were evaluated using denaturing gradient gel electrophoresis (DGGE) and interpreted considering structure, richness and diversity. ZEBC were affected by both stressors even at concentrations/doses not affecting the host-organism (survival/development). Yet, some stress-tolerant bacterial groups were revealed. The structure of the ZEBC was always affected, mainly due to xenobiotic presence. Richness and diversity decreased after exposure to NOEC of PD. Interactive effects occurred for TCS and UVR. Aquatic microbiota imbalance might have repercussions for the host/aquatic system, particularly in a realistic scenario/climate change perspective therefore, future ecotoxicological models should consider xenobiotics interactions with UVR., (Copyright © 2015 Elsevier Ltd. All rights reserved.)